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WELCOME LETTER<br />
Dear Colleague,<br />
As Presidents of the European Association of Tissue Banks (EATB) and the Asociación Española de Bancos de Tejidos (AEBT), Chairs<br />
of the Organizing Committee of the conference, it is our great pleasure to welcome you to the 6th World Congress on Tissue Banking<br />
jointly with the 20th EATB and the 12th AEBT Congresses.<br />
With the collaboration of the American Association of Tissue Banks (AATB), the Asociación Latinoamericana de Bancos de Tejidos<br />
(ALABAT), the Asia-Pacific Association of Surgical Tissue Banking (APASTB) and the Australasian Tissue and Biotherapeutics Forum (ATBF),<br />
this edition of the congress joins all the associations involved in the field of Tissue Banking, making this meeting a very unique event.<br />
The scientific topics will cover updated aspects on Tissue Banking, being specially focused on the new challenges of Paediatric Tissue<br />
Banking. The Congress will give to the world renowned experts in the field of tissue banking, the opportunity to share difficulties,<br />
experiences and specific particularities of children s tissues and cells transplantation and has a primary goal of establishing new and<br />
permanent contacts to promote worldwide collaboration.<br />
Barcelona is a modern cosmopolitan city, which has managed to preserve an important cultural, architectural and monumental historic<br />
legacy. Barcelona is almost as lively at night as it is during the day and it has a unique, exciting atmosphere, which provides the visitor<br />
with a truly unforgettable experience.<br />
We are looking forward to sharing with you all your knowledge and thoughts in order to give to this edition a special flavor both<br />
scientifically and personally.<br />
Esteve Trias<br />
President EATB<br />
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Josep M. Segur<br />
President AEBT<br />
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PRESIDENT OF THE HONOUR COMMITTEE<br />
S.M. la Reina de España<br />
MEMBERS<br />
Molt Hble. Sr. Artur Mas i Gavarró<br />
President of the Generalitat of Catalonia<br />
Excm. Sr. Xavier Trias i Vidal de Llobetera<br />
Mayor of Barcelona<br />
Hble. Sr. Boi Ruiz i Garcia<br />
Health Minister of the Goverment of the Generalitat of Catalonia<br />
Dr. D. Rafael Matesanz Acedos<br />
President of the Organización Nacional de Trasplantes (ONT)<br />
Dr. Pere Montserrat<br />
Director of the Organització Catalana de Trasplantaments (OCATT)<br />
Dra. Rosa Deulofeu<br />
Past Director of the Organització Catalana de Trasplantaments (OCATT)<br />
Sr. Antonio Esteve i Cruellas<br />
President of the Banc de Sang i Teixits (BST)<br />
Dr. Josep M. Piqué i Badia<br />
General Director of Hospital Clinic of Barcelona
COMMITTEES<br />
Organising Committee<br />
Presidents<br />
Esteve Trias Clinic-TSF / EATB<br />
Josep M. Segur Clinic-TSF / AEBT<br />
Members<br />
María Jesus Felix OCATT<br />
Alfredo Adan Clinic-TSF<br />
Elba Agustí Clinic-TSF<br />
Ricardo Cassaroli Clinic-TSF<br />
José Luis Poma Clinic-TSF<br />
Santiago Suso Clinic-TSF<br />
Maria Zardoya Clinic-TSF<br />
Aurora Navarro Banc de Sang i Teixits<br />
Ramon Pau Pla Banc de Sang i Teixits<br />
Scientific Committee<br />
President<br />
Deirdre Fehily Ireland<br />
Members<br />
Mercé Alsina Spain<br />
Juan Álvarez de Toled Spain<br />
Martha W. Anderson USA<br />
John Armitage UK<br />
Arlinke Bokhorst The Netherlands<br />
Anne Cathrine Bollerup Denmark<br />
Scott Brubaker USA<br />
Miguel Casares Spain<br />
Ricardo Casaroli Spain<br />
Akila Chandrasekar UK<br />
Natividad Cuende Spain<br />
Iva Dekaris Croatia<br />
Ana Fernandez Spain<br />
Gregorio Garrido Spain<br />
Luca Gianaroli Italy<br />
Ramón Gomis Spain<br />
Marisa Roma Herson Brazil<br />
Ramón Huguet Spain<br />
Ramadan Jashari Belgium<br />
Artur Kaminsky Poland<br />
John Kearney UK<br />
Johann Kurz Austria<br />
Jose Luis Pomar Spain<br />
Stefan Poniatowski UK<br />
Diego Ponzin Italy<br />
Axel Pruss Germany<br />
Lluis Puig Spain<br />
Nelleke Richters The Netherlands<br />
Jacinto Sanchez Spain<br />
Santiago Suso Spain<br />
Anna Veiga Spain
The 6th World Congress on Tissue Banking, the 20th Congress of European Association of Tissue Banks and<br />
the 12º Congreso de la Asociación Española de Banco de Tejidos gratefully acknowledges the following<br />
companies for their support<br />
ALCHIMIA<br />
AL.CHI.MI.A. S.r.l. is a leading European manufacturer of medical devices. The newest line of products, dedicated to the processing of<br />
human tissues and cells intended for transplantation, includes solutions for tissue and cell decontamination, rinsing and cryopreservation.<br />
All products meet the requirements of the European Directives on tissue and cells.<br />
AL.CHI.MI.A. s products (Human Tissue Processing):<br />
BASE.128, 250 ml — 500 ml<br />
BASE.128 RED, 250 ml — 500 ml<br />
BASE, 500 ml<br />
BASE RED, 500 ml<br />
CRYO.ON, 10 ml — 100 ml<br />
GLYO.ON, 250 ml — 500 ml<br />
www.alchimiasrl.com<br />
DR. FRANZ KOEKLER CHEMIE GMBH<br />
Dr. F. K hler Chemie is an independent, family owned business with representations on all five continents. Since 1959, we provide products<br />
in the fields of electrolytes, contrast media, antidotes, anaesthetics, as well as organ protective and cardioplegic solutions. We are a<br />
committed partner of cardiac and transplant surgery worldwide.<br />
www.koehler-chemie.de<br />
GRIFOLS<br />
Serving people s health<br />
Grifols is a Spanish holding company specialized in the pharmaceutical-hospital sector. We have three primary divisions -- Bioscience,<br />
Diagnostic and Hospital — which develop, produce and market our innovative products and services to medical professionals in more<br />
than 90 countries around the world.<br />
In the range of Hospital division products, the company develops complete systems for preparing intravenous solutions and advanced<br />
therapy drugs under sterile conditions inside hospital pharmacies, including the Misterium“ Modular Clean-Room which adapts and grows<br />
to meet your needs.<br />
ICCBBA<br />
ISBT 128 is the global information standard for coding and labeling of human tissue, blood, and cellular therapy products. ICCBBA is a<br />
not-for-profit organization that manages the ISBT 128 Standard.<br />
Information about ISBT 128 may be found at www.iccbba.org<br />
IGL GROUP — STEL ALPHA<br />
IGL GROUP (France, Spain, America Latina, USA) was created to facilitate research relationships, and develop products used in the fields<br />
or organ recovery, preservation, and transplantation. Leader in Organ Preservation, we are proud to introduce STEM ALPHA, a French<br />
innovative company, specialized in tissue preservation. IGL Group and Stem Alpha continuously explore new research opportunities, with<br />
the goal of enhancing organs & tissues quality for transplantation. We welcome you to our booth #3<br />
www.igl-group.com. www.stemalpha.com<br />
LABS, INC.<br />
LABS Inc., is a global, full-service testing laboratory with over 30 years of expertise in regulated testing for human organs, cells, tissues<br />
and implantable biologic products and devices. LABS Inc. provides an intimate customer service experience, which results in personalized<br />
laboratory solutions for clients in any stage of the product development lifecycle: Donor eligibility, R&D, Processing/Validation and Final<br />
lot release.<br />
www.labs-inc.org
LIFENET HEALTH<br />
LifeNet Health, a non-profit global leader in regenerative medicine, is the world s largest provider of bio-implants for transplantation whose<br />
mission is saving lives and restoring health by advancing the field of tissue engineering. The LifeNet Health Bio-Implants Division is a<br />
leader in the engineering and processing of dental, cardiovascular, spinal and orthopedic bio-implants and distributes more than 400,000<br />
bio-implants every year to restore health to patients around the world.<br />
www.lifenethealth.org<br />
MACOPHARMA<br />
www.macopharma.com<br />
MAK SYSTEM<br />
With more than 30 years of success in 60 countries worldwide, MAK-SYSTEM is the leading software solution provider in Web Technology<br />
for Tissue Bank, Cord Blood Bank, Cell Therapy Lab, Blood Bank and transfusion services. Our team will welcome you at our booth for<br />
a presentation of the WEB Software solution T.C.S. dedicated for Tissue, Cord Blood and Stem Cells.<br />
www.mak-system.net<br />
MASTERCONTROL, INC<br />
Despite being primarily focused on helping people and diagnosing disease, Blood, Biologic, and Tissue organizations still have pressing<br />
business needs. Reducing overall costs and increasing efficiency is critical to such organisations because they must be cost sensitive, cope<br />
with limited resources, and manage thin operating margins. MasterControl produces software solutions that meet these needs by enabling<br />
organisations around the world and streamlining businesses processes<br />
For more information visit: www.mastercontrol.com<br />
PLANER/TELSTAR<br />
Planer are a leader in cell care, manufacturing Precision Benchtop Incubators, Controlled Rate Freezers, Temperature/Level/Gas Monitoring<br />
and Alarm systems. They also supply Liquid Nitrogen Storage/Supply Systems, cryogenic and safety accessories. They are accredited<br />
manufacturers of medical devices meeting international standards.<br />
www.planer.com<br />
SPIERINGS ORTHOPAEDICS BV.<br />
Spierings Medische Techniek BV is a medical technical engineering company focusing on researching, designing, developing and testing<br />
medical devices for the orthopaedic, plastic and general surgery market. One of the areas of expertise is the design and development<br />
of bone processing systems.<br />
Spierings Orthopaedics BV is a medical company focusing on production, regulatory affairs, outsourcing, quality control and CE marking<br />
of orthopaedic implants and instruments. Two main products are the Noviomagus Bone Mill and Noviomagus Femoral Head Reamer<br />
Set. www.spierings.biz<br />
TELOS GMBH<br />
The Marburg Bonebank System was designed in 1992 due to the growing problems with graft material including: viral and bacterial<br />
transmissions, quality concerns over gamma irradiated bone, costs of purchasing bone from tissue banks, non-standardization of tissue<br />
bank processing procedures among other concerns. Since 1992, there are 650 units in use and over 180,000 bone processed worldwide.<br />
The Marburg Bonebank System is a thermal processing method that allows a hospital to retrieve the femoral head from a surgical living<br />
donor during a total hip arthroplasty.<br />
The Telos GmbH is a German family business, which was founded 1977. It is specialized in Orthopedic and Traumatology. Our products<br />
are sold in 22 countries by distribution partners.<br />
www.telos-marburg.de<br />
TRANSPLANT ENTERPRISE SYSTEM<br />
Transplant Connect is the global leader in Electronic Medical Records Systems and Enterprise Management Software for Tissue Banks,<br />
Eye Banks and Organ Procurement Organizations.˚ We also provide Online Donor Registries, Research and Analytics Systems and custom<br />
solutions.˚ A certified Public Benefit Company, our mission is to help improve quality, increase efficiency and reduce errors — all to help<br />
maximize the donor gift.˚ Our web-based iTransplant Systems are comprehensive, intuitive and very cost-effective.˚ iTransplant is the most<br />
widely-adopted software of its kind in the world. ˚Visit our booth for a demonstration and visit our website for additional information -<br />
www.transplantconnect.com.
DGFG<br />
The German Society for Tissue Transplantation (DGFG) organizes tissue donation, processing and allocation of tissue transplants. As a<br />
non-profit network, consisting of 16 tissue banks and numerous hospitals and transplantation centers, we are open for further constructive<br />
partnerships in order to optimize patient care.<br />
www.gewebenetzwerk.de<br />
GUBENER PLASTINATE GMBH<br />
The Gubener Plastinate GmbH is a global acting company in Plastination; it manufactures high-quality human anatomical specimens for<br />
medical education, research and the BODY WORLDS exhibitions. Our established body donation program comprises over 12.000 living<br />
donors, from which we receive 2-4 deceased donors per week. For further information please visit our websites:<br />
www.vonhagens-products.com and www.koerperspende.de/en.html<br />
TISSUEREGENIX<br />
Tissue Regenix Group Plc is a regenerative medical products company founded on its patented dCELL process. The technology allows<br />
tissues to be de-cellularised without incurring protein cross linking which can be seen when harsh chemical agents are employed to remove<br />
the cells within tissue. The remaining matrix retains its stereo chemical structure, encouraging rapid re-cellularisation by the patients own<br />
cells, while resisting the onset of re-calcification when the surface is in contact with circulating blood. The technology is available for<br />
licensing and commercialised through the companies dCELL Vascular Patch.<br />
www.tissueregenix.com<br />
TRANSPLANT CONNECT<br />
Transplant Connect is the global leader in Electronic Medical Records Systems and Enterprise Management Software for Tissue Banks,<br />
Eye Banks and Organ Procurement Organizations. We also provide Online Donor Registries, Research and Analytics Systems and custom<br />
solutions. A certified Public Benefit Company, our mission is to help improve quality, increase efficiency and reduce errors — all to help<br />
maximize the donor gift. Our web-based iTransplant Systems are comprehensive, intuitive and very cost-effective. iTransplant is the most<br />
widely-adopted software of its kind in the world. Visit our booth for a demonstration and visit our website for additional information -<br />
www.transplantconnect.com
GENERAL INFORMATION<br />
Websites<br />
www.eatb2011.com<br />
www.eatb.org<br />
www.aebt.org<br />
Meeting venue<br />
AXA Auditori<br />
Avda. Diagonal, 547<br />
08029 Barcelona - Spain<br />
www.axa.es<br />
Slide Preview Rooms<br />
For presentations in Auditori please go to 2nd floor using the<br />
elevator.<br />
For presentations in Room 1 and Room 3 please go to -1 floor<br />
(Deu I Mata entrance or using the elevator).<br />
Your presentation has to be loaded at least two hours in advance.<br />
If you will use your own laptop (not allowed for free papers and<br />
invited short papers presentations) kindly inform as well to the<br />
technicians in the slide preview room.<br />
Credits<br />
20th Congress of the European Association of Tissue Banks<br />
(Barcelona, Spain, 9.—11.11.2011)<br />
Event code:˚6611<br />
was granted˚18˚European CME credits (ECMEC) by the European<br />
Accreditation Council for Continuing Medical Education<br />
(EACCME).<br />
Badges<br />
Please wear your conference badge at all times during the<br />
conference. Wearing your badge is mandatory during breakfast,<br />
lunch and welcome reception.<br />
Social events<br />
Wednesday, 9th November - 07.00pm<br />
Welcome Reception at the Barcelona City Hall<br />
Coach Service - Departure from Auditori AXA at 18.30h<br />
Thursday, 10th November — 08.30pm<br />
Gala Dinner at Can Cortada<br />
Coach Service - Departure from Auditori AXA at 20.00h<br />
Technical Secretariat<br />
Torres Pardo<br />
N pols, 187 2… - 08013 Barcelona - Spain<br />
Phone: +34 93 246 35 66<br />
Fax +34 93 231 79 72<br />
E-mail: m.velazquez@torrespardo.com<br />
Registration fees<br />
ON-SITE<br />
Members* 525 Eur.<br />
Non members 575 Eur.<br />
8% VAT included<br />
The registration fee for participant includes participation in scientific<br />
session, congress bag with program and abstract book, Welcome<br />
Cocktail, working lunch, coffee breaks and access to industrial<br />
exhibition and participation in the Jornada Iberoamericana.<br />
ON-SITE<br />
Day ticket for members* Non available<br />
Day ticket for non members Non available<br />
8% VAT included<br />
The registration fee for one day includes participation in scientific session,<br />
congress bag with program and abstract book, Welcome Cocktail<br />
(November 9th), working lunch, coffee breaks and access to industrial<br />
exhibition.<br />
* Members: of the EATB - AEBT - AATB - ALABAT - APASTB - ATBF-<br />
EATB sister associations<br />
Social Events<br />
GALA DINNER: 70 Eur.<br />
Certificate of attendance<br />
Confirmations of attendance will be issued at the registration desk.<br />
Changes<br />
The organizers reserve the right to adjust or change the program<br />
as necessary.<br />
Insurance and liability<br />
In registering for the <strong>EATB2011</strong> participants agree that neither the<br />
congress nor the European Association of Tissue Banks assume<br />
any liability whatsoever. Participants are requested to make their<br />
own arrangements in respect of health and travel insurance.<br />
Official language<br />
The official congress language is English.<br />
No simultaneous translation will be available.<br />
Official Travel Agency<br />
Viajes El Corte Ingl s<br />
Dpto. Sociedades Cient fico-M dicas<br />
Casado del Alisal, 14 Æ 28014 Madrid,Spain<br />
Tel. 91 330 07 55 — fax 91 420 39 52<br />
e-mail: secretariaturistica@viajeseci.es
PROGRAM AT A GLANCE<br />
07.30<br />
08.30 09.30<br />
09.30 10.30<br />
10.30 11.00<br />
11.00 12.00<br />
12.00 12.30<br />
12.30 13.30<br />
13.30 14.30<br />
14.30 16.00<br />
16.00 16.30<br />
16.30 17.15<br />
17.15 18.00<br />
18.00 19.00<br />
19.00 20.30<br />
AUDITORI ROOM 1 ROOM 3<br />
Registration desk opens<br />
Welcome addresses<br />
Presentations by Collaborating Associations<br />
Session: Ethical themes of common interest<br />
Session: Ethical themes of common interest - II<br />
Free papers: Ethical themes of common interest<br />
Session:Biobanking and banking for research:<br />
interaction with Tissue Banking<br />
Session:<br />
Specific challenges of donation, processing<br />
and clinical application in paediatrics<br />
Session:<br />
Specific challenges of donation, processing<br />
and clinical application in paediatrics - II<br />
Invited short papers:<br />
Specific challenges of donation, processing<br />
and clinical application in paediatrics<br />
WEDNESDAY, NOVEMBER 9TH<br />
Coffee break - Visit to the exhibition area<br />
11.30 h<br />
Free papers:<br />
International Experience on Tissue Banking<br />
Lunch<br />
Coffee break - Visit to the exhibition area<br />
Welcome Reception at the Barcelona City Hall<br />
WEDNESDAY, NOVEMBER 9TH
PROGRAM AT A GLANCE<br />
07.30<br />
08.00 09.15<br />
09.15 09.30<br />
09.30 10.30<br />
10.30 11.00<br />
11.00 12.30<br />
12.30 13.00<br />
13.00 15.00<br />
15.00 15.45<br />
15.45 16.30<br />
16.30 17.00<br />
17.00 17.30<br />
17.30 18.30<br />
18.30 20.30<br />
20.30<br />
AUDITORI ROOM 1 ROOM 3<br />
Registration desk opens<br />
Session:<br />
Donation promotion and selection to ensure<br />
safety of recipients<br />
Invited short paper: Donation<br />
Free papers: Donation<br />
Session: Cardiovascular I<br />
EATB ASSEMBLY<br />
Session: Cardiovascular II<br />
Free papers: Cardiovascular I<br />
Session: Cardiovascular III<br />
Free papers: Cardiovascular II<br />
THURSDAY, NOVEMBER 10TH<br />
Coffee break - Visit to the exhibition area<br />
Session: Reproductive<br />
Session: Musculoskeletal<br />
Free papers: Musculoskeletal I<br />
Coffee break - Visit to the exhibition area<br />
Free papers: Musculoskeletal II<br />
Gala Dinner at Can Cortada<br />
LUNCH<br />
Session: Skin & Amniotic membrane<br />
Free papers - Skin & Amniotic Membrane I<br />
Invited short paper & Free papers:<br />
Skin & Amniotic Membrane<br />
Session: Cord Blood<br />
THURSDAY, NOVEMBER 10TH
PROGRAM AT A GLANCE<br />
08.00<br />
08.30 09.45<br />
09.45 09.55<br />
09.55 10.15<br />
10.15 10.45<br />
10.45 11.15<br />
11.15 13.00<br />
13.00 14.00<br />
14.00 15.45<br />
15.45 16.45<br />
16.45 17.15<br />
17.15 18.10<br />
18.10 18.25<br />
AUDITORI ROOM 1 ROOM 3<br />
Registration desk opens<br />
Session: Learning from Biovigilance<br />
Invited short paper: Learning from Biovigilance<br />
Free papers: Learning from Biovigilance<br />
CODING in the EU;<br />
Update from the European Commission<br />
to the scientific associations<br />
Session:<br />
Glimpse into the future; advanced therapies<br />
Invited short paper & Free papers:<br />
Glimpse into the future; advanced therapies<br />
Session: European good practice standards<br />
Panel discussion: Association interaction<br />
Presidential Closing remarks.<br />
Awards & Announcement of Meeting 2012<br />
FRIDAY, NOVEMBER 11TH<br />
Coffee break - Visit to the exhibition area<br />
LUNCH<br />
Session: Cornea<br />
15.15 - 16.55 h<br />
Free papers: Cornea<br />
Coffee break - Visit to the exhibition area<br />
FRIDAY, NOVEMBER 11TH
SCIENTIFIC PROGRAM - 9th November 2011<br />
AUDITORI<br />
08.30-08.40 h WELCOME<br />
Objectives of the Congress<br />
08.40-09.00 h WELCOME ADDRESSES BY KEY INSTITUTIONAL REPRESENTATIVES:<br />
Presentation of their mission and activities in the field of tissue banking<br />
European Commission<br />
Council of Europe<br />
World Health Organisation<br />
International Atomic Energy Agency<br />
09.00-09.30 h PRESENTATIONS BY COLLABORATING ASSOCIATIONS: description of the association and its key activities<br />
World Union of Tissue Banking Associations:<br />
European Association of Tissue Banks (EATB)<br />
American Association of Tissue Banks (AATB)<br />
Asociación Latinoamericana de Bancos de Tejidos (ALABAT)<br />
Asia-Pacific Association of Surgical Tissue Banking (APASTB)<br />
Australasian Tissue and Biotherapeutics Forum (ATBF)<br />
Other Collaborating Associations<br />
European Eye Banking Association (EEBA)<br />
Eye Banking Association of America (EBAA)<br />
Asociación Pan Americana de Bancos de Ojos (APABO)<br />
European Society for Human Reproduction and Embryology (ESHRE)<br />
Asociación Española de Bancos de Tejidos (AEBT)<br />
09.30-10.30 h ETHICAL THEMES OF COMMON INTEREST (I)<br />
Moderators: M. Lopez and N. Terribas<br />
09.30-09.45 h Protecting Donors and Patients - monitoring unethical practices in tissue and cell donation<br />
M. Carmona, Switzerland<br />
09.45-10.00 h The Ethics of Consent for Different Tissue and Cell Uses (transplant, research, biobanking)<br />
A. Goldenberg, US<br />
10.00-10.15 h Detecting and Preventing Illegal and fraudulent activities (IFA)˚<br />
D. Fehily, Ireland<br />
10.15-10.30 h Discussion<br />
10.30-11.00 h COFFEE BREAK<br />
AUDITORI<br />
11.00-12.30 h ETHICAL THEMES OF COMMON INTEREST (II)<br />
Moderators: M. Lopez and N. Terribas<br />
11.00-11.15 h Updating AATB consent and authorisation terminology<br />
C. Strong, US<br />
11.15-11.30 h Gamete donation and the movement of donors and patients<br />
A. Veiga, Spain<br />
11.30- 12.00 h Discussion<br />
12.00-12.30 h FREE PAPERS - ETHICAL THEMES OF COMMON INTEREST<br />
Moderators: M. Lopez and N. Terribas<br />
12.00-12.10h Model of coexistence for public and family cord blood banks<br />
Alvarez Ramos , A.; Losada Pescador A.<br />
VidaCord, Spain
SCIENTIFIC PROGRAM - 9th November 2011<br />
12.10-12.20 h A fundamental right to decide whether and how our body will be used in research<br />
Zardoya , M.; Vilarrodona A.; Rodriguez C.; Ruiz A.; Paredes D.; Trias E.<br />
Transplant Services Foundation. Hospital Clinic of Barcelona. Spain.<br />
12.20-12.30h Cord Blood: Public and Private<br />
Golubic, B.<br />
Department for Blood, Tissues and Cells Inspection and Monitoring Inspection Service Ministry of health<br />
and social welfare, Zagreb, Croatia<br />
ROOM 1<br />
11.30-13.30 h FREE PAPERS - INTERNATIONAL EXPERIENCE ON TISSUE BANKING<br />
Moderators: M. Manyalich and I. Uhrynowska-Tyszkiewicz<br />
11.30-11.40 h TPM as international educational model in tissue banking<br />
Navarro , A.; Duque E.; Paez G.; Manyalich M.<br />
Banc de Sang Teixits. Barcelona. Spain<br />
11.40-11.50 h Basic conditions for a network in tissue medicine: Quality, Transparency and Efficiency<br />
B rgel , M.<br />
DGFG - German Society for Tissue Transplantation<br />
11.50-12.00 h Transforming tissue donation and transplantation in Canada into a single integrated inter-provincial system.<br />
Haun , M.; Mohr J.; Derksen P.; Parsons C.; Sher G.<br />
Canadian Blood Services<br />
12.00-12.10 h Minutes from the first edition experience of the University of Barcelona Master Degree in Donation & Transplantation<br />
of Organs, Tissues and Cells.<br />
Ballest , C.; Segur JM.; Casaroli R.; Pomar JL.; Alsina M.; Suso S.; Manyalich M.<br />
Facultat de Medicina, Universitat de Barcelona. Spain<br />
12.10-12.20 h Speed is Life. Aplying Lean Healthcare to Tissue Banks.<br />
Tornos J., I.; Serigo X.; Giralt E.<br />
AUREN. Spain<br />
12.20-12.30 h Tissue recovery team: 8 years of experience<br />
Fari as , O.; Vilarrodona A.; Savio Manuel A.; Luque S.; Oliva R.; P rez ˚ML.; Trias E.<br />
Transplant Services Foundation - Hospital Clinic. Barcelona. Spain<br />
12.30-12.40 h The Donor Tissue Bank Replacement Facility — Meeting Future Demands in Tissue / Cell banking<br />
Herson Roma, M.; Poniatowski S.; Adamas F.; Cordner S.<br />
Donor Tissue Bank of Victoria / Victorian Institute of Forensic Medicine. Australia<br />
12.40-12.50 h Current status of tissue banking in Korea<br />
Kang , Y.; Chung Y.; Lim J.; Kim Y.<br />
St. Vincent’s Hospital, Catholic University of Korea<br />
12.50-13.00 h Optimizing the process of tissue banking in a blood bank network<br />
Rodr guez , L.; S ez M.; Genis X.; Bosch A.; Salinas R.; Castella D.; Navarro A.<br />
Banc de Sang i Teixits. Barcelona. Spain<br />
13.00-13.10 h Retrospective analysis of 26 years of homograft heart valve banking in central South Africa<br />
Van Den Heever Jacobus, J.; Neethling Morris Leonard W.; Smit Edwin F.; Botes L.<br />
Dept of Cardiothoracic Surgery, University of the Free State, Bloemfontein, South Africa.<br />
13.10-13.20 h Clean Rooms and Tissue Banking: How Happy I Could be with Either GMP or GTP ?<br />
Klykens , J.; Pirnay J.; Verbeken G.; Giet O.; Baudoux E.; Jashari R.<br />
University Hospital Leuven. Belgium<br />
13.20-13.30 h Banking of Cryopreserved Vascular Allografts in Europe: 20 years of activity in European Homograft Bank (EHB)<br />
in Brussels<br />
Jashari , R.; Van Hoeck B.; Goffin Y.; Fan Y.; Rousse N.<br />
European Homograft Bank (EHB), International Association
SCIENTIFIC PROGRAM - 9th November 2011<br />
AUDITORI<br />
12.30-13.30 h BIOBANKING AND BANKING FOR RESEARCH: INTERACTION WITH TISSUE BANKING˚˚ ˚<br />
Moderators: J. S nchez and A. Bosch˚<br />
12.30-12.45 h Ethics of Donation for Research<br />
M. Anderson, US<br />
12.45-13.00 h Review of interactions between banks of tissues for clinical and non-clinical uses<br />
A. Bokhorst, The Netherlands<br />
13.00-13.20 h ˚ Legal issues surrounding consent and anonymisation of clinical samples for bio-banking<br />
A. Emaldi, Spain<br />
13.20-13.30 h Discussion<br />
13.30-14.30 h LUNCH<br />
14.30-16.00 h SPECIFIC CHALLENGES OF DONATION, PROCESSING AND CLINICAL APPLICATION IN PAEDIATRICS<br />
Moderators: R. Matesanz and E. Trias<br />
14.30-14.45 h Specific challenges of donation, processing and clinical application in paediatrics<br />
M. López Santamar a / F. Hernández Spain<br />
14.45-15.00 h Launch of a Spanish Network for paediatric tissue transplantation<br />
J.L. Pomar, Spain<br />
15.00-15-15 h 20 years of experience on bone allografting˚ in children and adolescents<br />
R.Huguet, F. Torner and A. Muset, Spain<br />
15.10-15-30 h The Challenges of Musculoskeletal Transplantation in children<br />
M. San Julian, Spain<br />
15.30-15-45 h Assisted Reproduction Technology (ART): preserving fertility in children with cancer<br />
C. Andersen, Denmark<br />
15.45-16.00 h Discussion<br />
16.00-16.30 h COFFEE BREAK<br />
16.30-18.00 h SPECIFIC CHALLENGES OF DONATION, PROCESSING AND CLINICAL APPLICATION IN PAEDIATRICS (II)<br />
Moderators: O. Schwint and Mº J. F lix<br />
16.30-16.45 h Cardiovascular: the benefits of decellularised valves for children<br />
F. Da Costa, Brazil<br />
16.45-17.00 h Cornea: the Challenges of Cornea Transplant in Children<br />
J. Alvarez Toledo, Spain<br />
17.00-17.15 h Skin grafting for burns - the particular challenges of growing children<br />
M. Herson, Australia<br />
Invited Short Papers<br />
17.15-17.25 h vCJD risk reduction measures for children<br />
A. Chandrasekar, UK<br />
17.25-17.35 h The need for a Paediatric Tissue Bank: a surgeon s perspective<br />
X. Soldado, Spain<br />
17.35-17.45 h Case studies in Paediatric Cornea Transplantation<br />
N. Planas, Spain<br />
17.45-17.55 h Treatment of Paediatric Bone Tumors with Bone Banking<br />
J. de Las Heras, Spain<br />
19.00-20.30 h WELCOME RECEPTION at the Barcelona City Hall
SCIENTIFIC PROGRAM - 10th November 2011<br />
AUDITORI<br />
08.00-10.30 h DONATION PROMOTION AND SELECTION TO ENSURE SAFETY OF RECIPIENTS<br />
Moderators: A. Lobo and M. Anderson<br />
08.00-08.15 h Donation Promotion in the USA<br />
D. Fleming, US<br />
08.15-08.30 h Donation Promotion in Italy; the Mestre Eye Donation Program<br />
D. Ponzin, Italy<br />
08.30-08.45 h A Risk Assessment Model for Donor Testing<br />
W. Oei, The Netherlands<br />
08.45-09.15 h Open discussion<br />
09.15-09.25 h Invited Short Paper<br />
Collection programs; Donor eligibility, obtention and quality parameters<br />
M. Torrabadella, Spain<br />
09.30-10.30 h FREE PAPERS — DONATION<br />
Moderators: A. Lobo and M. Anderson<br />
09.30-09.40 h Developing Tissue Donation in Northern Germany - actual status, strategies and challenges<br />
Wulff , B.; Heinemann A.; Montenero M.; Pueschel K.<br />
Institute of Legal Medicine. Germany<br />
09.40-09.50 h Postmortem Blood Assays for HIV, HBV and HCV in tissue donors. Systematic literature review<br />
Mieth , K.; Muñoz O.; Soto C.; Navas J.; Gonzalez J.<br />
Fundacion Cosme y Damian. Colombia<br />
09.50-10.00 h Tissue Donation in Emergency Medicine Departments- The Scottish Experience<br />
Galea G.; Donalldson S.<br />
SNBTS Tissues and Cells Directorate. Scotland<br />
10.00-10.10 h The role of forensic institutes in tissue donation in Germany — the Munich example<br />
Braun , C.; Wulff B.; Graw M.<br />
Institute for Legal Medicine Munich. Germany<br />
10.10-10.20 h Q fever in tissue donors in the Netherlands<br />
Van Wijk J, M.; Maas E W.; Hogema B.; Koot M.; Renders C N.; Hermans H M.; Bokhorst G A.<br />
BISLIFE Foundation. Netherlands<br />
10.20-10.30 h Tissue donation and organ donation: a competitive or a cooperative System - Experiences after the tissue act<br />
from 2007 in Germany<br />
Nitschke , F.; Manecke A.; Wille D.<br />
DGFG - German Society for Tissue Transplantation. Germany<br />
10.30-11.00 h COFFEE BREAK<br />
AUDITORI<br />
11.00-12.40 h CARDIOVASCULAR (I)<br />
Moderators: S. Brubaker and JL Pomar<br />
11.00-11.55 h Report from the Consensus Meetings of European Cardiovascular Tissue Banks<br />
T. de By, The Netherlands; R. Hetzer, Germany; R. Parker, UK; C. McDonald, UK<br />
11.55-12.07 h Report Report BATB Microbiology Survey 2010<br />
R. Lomas, UK<br />
12.07-12.10 h AATB CV Survey Results 2007<br />
S. Brubaker, US<br />
12.10-12.25 h CV Tissue Banking: Methods Used in Qu bec<br />
J. Tremblay, Canada<br />
12.25-12.40 h CV Tissue Banking: Variations among Technician Grading<br />
O. Schwint, Argentina
SCIENTIFIC PROGRAM - 10th November 2011<br />
ROOM 1<br />
11.00-13.00 h REPRODUCTIVE<br />
Moderators: D. Fehily and A. Veiga<br />
11.00-11-20 h The power of Demography: how demographic dynamics affect society, politics and economy<br />
J. de Mouzon, France<br />
11.20-11.40 h Viral testing of partner gamete donors — time for a change to EU requirements?<br />
AC. Bollerup, Denmark<br />
11.40-12.00 h The importance of viral screening<br />
E. Mocanu, Ireland<br />
12.00-12.20 h The role of cryopreservation of gametes and embryos<br />
G. Calderon, Spain<br />
12.20-12.40 h ESHRE; The role of the scientific society.<br />
L. Gianaroli, Italy<br />
12.40-13.00 h Open discussion<br />
13.00-15.00 h LUNCH<br />
EATB ASSEMBLY<br />
AUDITORI<br />
15.00-16.30 h CARDIOVASCULAR (II)<br />
Moderators: A. Cowie and R. Parker<br />
CV Tissue Decellularization Protocols<br />
15.00-15.15 h M Brenner da Costa, Brazil and UK<br />
15.15-15.30 h N. Lyon, US<br />
15.30-15.45 h F. Peycelon, US<br />
15.45-16.25 h Free papers — Cardiovascular I<br />
Moderators: A. Cowie and R. Parker<br />
15.45-15.55 h Antibiotic Decontamination of Heart Valve Allografts: Historical Trends Before and After the Implementation of a Higher<br />
Temperature of Incubation.<br />
Tremblay , J.; Paquet I.; B liveau L.; Germain M.<br />
H ma-Québec<br />
15.55-16.05 h Comparison of the effects of ischaemic times of harvested homografts on histological appearance and tissue strength<br />
Van Den Heever Jacobus, J.; Bester D.; Smit Edwin F.; Botes L.<br />
Dept of Cardiothoracic Surgery, University of the Free State, Bloemfontein, South Africa<br />
16.05-16.15 h Pulmonary homografts for right ventricular outflow tract reconstruction during Ross procedure: nineteen years results<br />
Juthier , F.; Jashari R.; Rousse N.; Van Hoeck B.; Banfi C.; Vincentelli A.; Prat A.<br />
Chru Lille. France<br />
16.15-16.25 h Decontamination of cryopreserved cardio-vascular allografts: detection and eradication of the slow growing skin germs<br />
Jashari , R.; Fan Y.; Van Hoeck B.; Le Mercier N.; De Gelas S.<br />
European Homograft Bank (EHB), International Association. Belgium<br />
ROOM 1<br />
15.00-16.30 h MUSCULOSKELETAL (I)<br />
Moderators: S. Poniatowski, JM. Segur<br />
15.00-15.15 h Meniscus Recovery and Processing<br />
O.Fariñas, Spain<br />
15.15-15.30 h Meniscus Transplantation<br />
R. Cugat, Spain<br />
15.30-15.45 h Viable meniscal allograft transplantation<br />
A. Dhollander, Belgium<br />
15.45-16-00 h Open discussion
SCIENTIFIC PROGRAM - 10th November 2011<br />
16.00-16.30 h FREE PAPERS — MUSCULOSKELETAL I<br />
Moderators: S. Poniatowski, JM. Segur<br />
16.00-16.10 h The Inactivation Effect of Standard and Fractionated Electron Beam Irradiation on Enveloped and Non-enveloped Viruses<br />
Schmidt , T.; Gohs U.; Hoburg A.; Schumann W.; Scheffler S.; Nitsche A.; Pruss A.<br />
Charit University Medicine Berlin, Julius Wolff Institut, Germany<br />
16.10-16.20 h Effect of gamma irradiation on mechanical properties of human cortical bone: influence of different processing methods<br />
Jastrzebska , A.; Grazka E.; Gut G.; Uhrynowska-tyszkiewicz I.; Marowska J.; Kaminski A.<br />
National Centre for Tissue and Cell Banking, Warsaw, Poland<br />
16.20-16.30 h Effect of electron beam irradiation on mechanical properties of human cortical bone: influence of different<br />
processing methods<br />
Grazka , E.; Jastrzebska A.; Gut G.; Uhrynowska-tyszkiewicz I.; Marowska J.; Kaminski A.<br />
National Centre for Tissue and Cell Banking, Warsaw, Poland<br />
ROOM 3<br />
15.00-16.30 h SKIN AND AMNIOTIC MEMBRANE (I)<br />
Moderators: N. Richters and M. Herson<br />
15.00-15.15 h State of the art in skin processing<br />
E. Pianigiani, Italy<br />
15.15-15.30 h State of the art in amniotic membrane processing<br />
J.Koller, Slovakia<br />
15.30-16.30 h Free papers - Skin & Amniotic Membrane I<br />
Moderators: N. Richters and M. Herson<br />
15.30-15.40 h Skin decontamination and impact of antibiotic residues on microbiological tests.<br />
Vassanelli , A.; Bosinelli A.; Rizzi S.; Lucchini E.; Gatto C.; Giurgola L.; Tothova J.<br />
Tissue Bank of Verona. Italy<br />
15.40-15.50 h Inactivation of Borrelia Burgdorferi by glycerol<br />
Richters , C.; Oei A.; De Wever B.; Hovius J.<br />
Euro Skin Bank. Netherlands<br />
15.50-16.00 h Establishment of a rinsing procedure to eliminate antibiotic and glycerol residues from cryopreserved skin decontaminated<br />
with BASE.128<br />
Pianigiani , E.; Gatto C.; Giurgola L.; Ierardi F.; D’amato Tothova J.<br />
Siena Skin Bank. Italy<br />
16.00-16.10 h The influence of radiation sterilization and preservation on cell morphology of Human Amniotic Membrane under Scanning<br />
Electron Microscope<br />
Suzina Sheikh Ab, H.; Suzina Sheikh Ab H.; Kamalia Z.; Yusof N.; Asnah H.<br />
Tissue Bank, Universiti Sains Malaysia<br />
16.10-16.20 h Effect of incubation temperature on viable skin decontamination.<br />
Rodriguez , L.; Tarragona E.; Gomariz E.; Ortega I.; Rodriguez V.; Genis X.; Navarro A.<br />
Banc de Sang i Teixits. Barcelona. Spain<br />
16.20-16.30 h Gamma Radiation Sterilized Amnios: Recent Clinical Applications in Mexico.<br />
Martínez-pardo , M.; Valencia-fonseca Elena L.; León-téllez Y.; Vázquez-maya L.<br />
Instituto Nacional de Investigaciones Nucleares. Mexico<br />
16.30-17.00 h COFFEE BREAK
SCIENTIFIC PROGRAM - 10th November 2011<br />
AUDITORI<br />
17.00-18.30 h CARDIOVASCULAR (III)<br />
Moderators: R. Jashari and A. Navarro<br />
17.00-17.15 h Development of Universal Cardiovascular Nomenclature<br />
I. Tyszkiewicz, Poland and R. Jashari, Belgium<br />
17.15-17.30 h CV Tissue Banking: Methods Used in Barcelona<br />
E. Agusti, Spain<br />
17.30-18.20 h FREE PAPERS - CARDIOVASCULAR II — IMPROVING SAFETY<br />
Moderators: R. Jashari and A. Navarro<br />
17.30-17.40 h Optimization of cardiovascular tissue decontamination and rinsing with BASE.128 and BASE.<br />
Terzi , A.; Buzzi M.; Guarino A.; Dainese L.; Vasuri F.; Gatto C.; Tothova J.<br />
banca dei tessuti cardiovascolari regione emilia romagna. Italy<br />
17.40-17.50 h Impact of antibiotic residues on microbiological analysis: implication of tissue banking<br />
Gatto , C.; Giurgola L.; Beccaro M.; Lipartiti M.; D’amato Tothova J.<br />
R&D ALCHIMIA SRL. Italy<br />
17.50-18.00 h New insights of antibiotic solution: antibacterial susceptibility in the 4 degree medium condition.<br />
Motomura , N.; Saito A.; Tamura K.; Noguchi N.; Hattori O.; Oda N.; Seki M.<br />
University of Tokyo. Japan<br />
18.00-18.10 h Quantification of residual antibiotics in cardiovascular tissue preparations<br />
Broecker , S.; Hartwig S.; Meyer R.<br />
Charit — University Hospital Berlin. Germany<br />
18.10-18.20 h Use of antibiotics/antimycotics in the new tissue preservation solution TiProtec ®<br />
Rauen , U.; Ebner A.; Deussen A.<br />
Institut f r Physiologische Chemie, Universit tsklinikum Essen. Germany<br />
18.20-18.30 h Assessment of Bioburden on Samples of Human and Animal Tissues: Part 1- Results of Method Development<br />
and Validation Studies<br />
Osborne , J.; Kowalski J.; Mosley G.; Merritt K.<br />
MTF<br />
ROOM 1<br />
17.00-18.40 h FREE PAPERS - MUSCULOSKELETAL (II)<br />
Moderators: M. San Julián and A. Pruss<br />
17.00-17.10 h Benefits of using a decontamination method in bone processing<br />
Fari as , O.; Vilarrodona A.; Vito S.; Tabera J.; Hinojosa Angel M.; Segur JM.; Trias E.<br />
Transplant Services Foundation - Hospital Clinic. Barcelona. Spain<br />
17.10-17.20 h Evidence based bone and tissue bank processes standardization<br />
Navas , J.; Mieth K.; Soto C.; Gonzalez J.<br />
Fundacion Cosme y Damian. Colombia<br />
17.20-17.30 h Comparison of frozen and freeze-dried cancellous bone graft in cavitary defects<br />
Camacho, P.; Segur JM.; García E.; García R.; Torner P.; Fariñas Ó.; Suso S.<br />
Hospital Clínic. Barcelona. Spain<br />
17.30-17.40 h Validity of an automatic measure protocol in distal femur for allograft selection from a threedimensional virtual bone<br />
bank system.<br />
Aponte-tinao Alberto, L.; Milano Edgardo F.; Farfalli Luis G.; Ritacco Eduardo L.; Schwint O.; Seiler C.; Reyes M.<br />
Italian Hospital of Buenos Aires. Argentina<br />
17.40-17.50 h Whole acetabulum allografts : a twenty years experience<br />
Ribas , M.; Vilarrubias J.; Ginebreda I.; De La Torre B.; Bellotti V.; De Meo F.; Cavaliere P.<br />
University Hospital Dexeus. Barcelona. Spain<br />
17.50-18.00 h Onlay Cortical Strut Allografting in Total Hip Arthroplasty (THA) Revision<br />
Medrano , C.; Bori G.; Gallart X.; Segur JM.; Fernandez-valencia J.; Riba J.; Garcia S.<br />
Hospital Clínic. Barcelona. Spain<br />
18.00-18.10 h Treatment of benign bone tumors with demineralized bone matrix after curetagge<br />
Soto , C.; Soto C.; Navas J.; Mieth K.; Gonzalez J.<br />
Fundacion Cosme y Damián. Colombia
SCIENTIFIC PROGRAM - 10th November 2011<br />
18.10-18.20 h An overview of the operations, traceability and safety aspects of human bone allografts at the Centre for Tissue Engineering-<br />
Bone Bank in South Africa<br />
Karakatsanis , E.<br />
Centre for Tissue Engineering - Bone Bank (Tshwane University of Technology). South Africa<br />
18.20-18.30 h Test system for osteoinductive activity of materials containing recombinant morphogenetic protein rhBMP-2<br />
Ryabov , A.; Akatov V.; Chekanov A.; Lekishvili M.; Fadeeva I.; Sklyanchuk E.; Guriev V.<br />
"Interdentos", Korolev. Russian Federation<br />
18.30-18.40 h Freeze-dried human serum albumin improves the adherence and proliferation of mesenchymal stem cells on mineralized<br />
human bone allografts<br />
Cs nge , L.; Skaliczky G.; Klara T.; Weszl M.; Schandl K.; Szendroi M.; Lacza Z.<br />
West Hungarian Regional Tissue Bank. Hungary<br />
ROOM 3<br />
17.00-17.10 h INVITED SHORT PAPER: SKIN & AMNIOTIC MEMBRANE<br />
Moderators:N. Richters and M. Herson<br />
Allografts and critical burned patients: experience and future<br />
J L. Fernández Cañamaque<br />
Plastic Surgery Department - Hospital Universitario de Getafe, Madrid. Spain<br />
17.10-17.30 h FREE PAPERS - SKIN & AMNIOTIC MEMBRANE II<br />
Moderators: N. Richters and M. Herson<br />
17.10-17.20 h Microbiological controls in skin tissue bank<br />
Pérez ML.; Vilarrodona A.; Savio A.; Otero N.; Martinez-Conesa E.; Nuñez V.<br />
Transplant Services Foundation-Hospital Clinic. Barcelona. Spain<br />
17.20-17.30 h Amniotic membrane in use for various preclinical and clinical indications<br />
Hennerbichler , S.; Peterbauer-Scherb A.; Petter-Puchner A.; Kesting M.; Redl H.; Gabriel C.<br />
Red Cross Blood Transfusion Service of Upper Austria, Linz/ Austrian Cluster for Tissue Regeneration. Austria<br />
17.30-18.30 h CORD BLOOD<br />
Moderators: L. Ponce and B. Golubic<br />
17.30-17.45 h Obstetric and perinatal issues in cord blood collection<br />
F. Figueras, Spain<br />
17.45-18.00 h Cord blood processing and storage<br />
S. Querol, Spain<br />
18.00-18.15 h Cord blood selection, release and transplantation<br />
G. Sanz, Spain<br />
18.15-18.30 h Discussion
SCIENTIFIC PROGRAM - 11th November 2011<br />
AUDITORI<br />
08.30-10.15 h LEARNING FROM BIOVIGILANCE<br />
Moderators: R. Marazuela and Y.-K. Kang<br />
˚<br />
08.30-08.45 h Project NOTIFY<br />
A. Nanni Costa, Italy<br />
08.45-09.00 h SOHO V&S<br />
D. Fehily, Italy<br />
09.00-09.15 h Eye Banking experience in the USA<br />
M. Macsai, US<br />
09.15-09-45 h Case discussion<br />
09.45-09.55 h Invited Short Paper<br />
Traceability and biovigilance: the role of the EUROCET registry<br />
A. Ghirardini, Italy<br />
09.55-10.15 h FREE PAPERS —LEARNING FROM BIOVIGILANCE<br />
Moderators: R. Marazuela and Y.-K. Kang<br />
09.55-10.05 h Catalonian Surveillance Register<br />
Barrio , R.; Felix MJ.; Aran B.; Deulofeu R.<br />
OCATT. Barcelona. Spain<br />
10.05-10.15 h National Histovigilance Education System Improves Professional Understanding of Severe Adverse Reactions and Events<br />
Cebulc , G.; Avsec D.<br />
Slovenija-Transplant. Slovenia<br />
10.15-10.45 h COFFEE BREAK<br />
AUDITORI<br />
10.45-11.15 h CODING IN THE EU; UPDATE FROM THE EUROPEAN COMMISSION TO THE SCIENTIFIC ASSOCIATIONS<br />
Moderators: R. Hitchev and L. Gianaroli<br />
EU Commission<br />
AUDITORI<br />
11.15-13.00 h GLIMPSE INTO THE FUTURE; ADVANCED THERAPIES (I)<br />
Moderators: J. Kurz and N. Calvente<br />
˚<br />
11.15-11.30 h Tissue engineering<br />
J. Kearney, UK<br />
11.30-11.45 h Cell therapy<br />
N. Cuende, Spain<br />
11.45-12.00 h Tissues and ATMP: A marriage of convenience. Aiming for a single quality standard<br />
J. Tabera, Spain<br />
12.00.12-15 h Artificial corneal tissue: A new challenge<br />
M. González, Spain<br />
12.15-12.30 h XCELIA Advanced Therapies. Towards the development of cellular medicines<br />
J. García, Spain<br />
12.30-13.00 h Open Discussion<br />
13.00-14.00h LUNCH
SCIENTIFIC PROGRAM - 11th November 2011<br />
AUDITORI<br />
14.00-15.40 h GLIMPSE INTO THE FUTURE; ADVANCED THERAPIES (II)<br />
Moderators: J. Kearney and N. Cuende<br />
14.00-14.10 h Invited short paper<br />
Tolerogenic Dendritic Cell Therapy in Refractory Crohn s Disease<br />
D.Benitez, Spain<br />
14.10-15.40 h FREE PAPERS - GLIMPSE INTO THE FUTURE; ADVANCED THERAPIES<br />
Moderators: J. Kearney and N. Cuende<br />
14.10-14.20 h Microbiological contamination monitoring of cleanroom facilities in a tissue and cell establishment<br />
Vilarrodona , A.; Tabera J.; Nuñez V.; Gonzalez V.; Fariñas O.; Trias E.<br />
Transplant Services Foundation-Hospital Clinic. Barcelona, Spain<br />
14.20-14.30 h Presentation of the Project of the Support Office to the Clinical Research in Advanced Therapies.<br />
Aran , B.; Felix M.; Barrio R.; Deulofeu R.<br />
Organització Catalana de Trasplantaments. OCATT. Barcelona. Spain<br />
14.30-14.50 h Biomechanical properties camparation between freeze dried flexor tendon and composite of freeze dried flexor tendonbone<br />
marrow mesenchymal stem cell in rabbit model<br />
Suroto , H.; Ferdiansyah F.; Rantam Abdul F.; Roeshadi D.<br />
Biomaterial Center and Tissue Bank - Dr Soetomo General Hospital. Indonesia<br />
14.40-14.50 h Improvement of limbal stem cells and mucosal epithelial cells isolation and cultivation method.<br />
Dragunova , J.<br />
Central Tissue Bank, University Hospital Bratislava. Slovenia<br />
14.50-15.00 h Testing on cell cultures of biogenic and synthetic materials applied in dentistry<br />
Volova , L.<br />
Institute of experimental medicine and biotechnologies. Russian Federation<br />
15.00-15.10 h BMP-2 and BMP-7 content in DBM putty produced by tissuelab in a gmp pharmaceutical plant<br />
Ramelli , M.; Della Valle E.; Mariniello R.<br />
Tissuelab S.p.a. Italy<br />
15.10-15.20 h Differentiation of porcine dermal cells inside autologous fibrin scaffolds<br />
Puente , P.; Lude a D.; López M.; Ramos J.; Aranda Luis J.; Varela G.; Iglesias J.<br />
Tissue Bank San Francisco Clinic Foundation. US<br />
15.20-15.30 h Comparision of different seeding strategies to enhance fibroblast penetration within a human acellular dermis for soft<br />
tissue augmentation<br />
Vitacolonna , M.; Smith M.; Hohenberger P.; Roessner E.<br />
Universit tsmedizin Mannheim ; Chirurgische Klinik. Germany<br />
˚<br />
ROOM 1<br />
14.00-15.15 h CORNEA (I)<br />
Moderators: D. Ponzin and R. Casaroli<br />
14.00-14.15 h The Eye Bank Association of America, 1961 - 2011: Celebrating Fifty Years of the EBAA<br />
M. Macsai, US<br />
14.15-14.30 h Eye Banking in Australia and New Zealand: Current trends and future challenges<br />
N. Nuttall, Australia<br />
14.30-14.45 h The 2008 EBAA; Strategic Plan: Moving Forward to Serving the World Community- Key Steps<br />
M. Macsai, US<br />
14.45-15.00 h The impact of the Pan-American Association of Eye Banks’ mission on the Latin-American Eye Banks<br />
L. Barbosa de Sousa, Brazil<br />
15.00-15.15 h The European Eye Bank Association — History, Present and Future<br />
I. Dekaris, Croatia
SCIENTIFIC PROGRAM - 11th November 2011<br />
15.15-16.55 h Free papers - CORNEA (II)<br />
Moderators: I. Dekaris and A. Fernández<br />
15.15-15.25 h Corneal assessment using light microscopy: the difference between healthy and pathological tissue<br />
Jirsova, K.<br />
Ocular Tissue Bank and Laboratory of the Biology and Pathology of the Eye, Institute of Inherited Metabolic Disorders,<br />
General and Teaching Hospital and 1st Faculty of Medicine, Charles University in Prague, Czech Republic<br />
15.25-15.35 h Corneal procurement, banking and transplantation in France<br />
Martinache, I.<br />
France<br />
15.35-15.45 h Parameters influencing microbiological safety of corneas<br />
Van Geyt, C.<br />
Belgium<br />
15.45-15.55 h Quality management within the eye bank<br />
Andrea Gareiss-Lok<br />
CEBT CEO, Hornhautbank Muenchen gGmbH, Munich. Germany<br />
15.55-16.05 h Pre-cutting of grafts for posterior lamellar keratoplasty<br />
Hjortdal, J.<br />
The Danish Eye Bank, Department of Opthalmology, Aarhus University Hospital. Danemark<br />
16.05-16.15 h Serology and microbiological testing in eye banks: legal aspects in Europe<br />
Maier, Ph.<br />
University Eye Hospital Freiburg. Germany<br />
16.15-16.25 h Implementation of European legislation in eye banking: the Czech experience.<br />
Krabcova I., Zlacka D., Jirsova K.<br />
Laboratory of the Biology and Pathology of the Eye, Institute of Inherited Metabolic Disorders and Ocular Tissue Bank,<br />
General Teaching Hospital and 1st Faculty of Medicine, Charles University in Prague, Czech Republic<br />
16.25-16.35 h Prospective development of Eye Banking in the Russian Federation<br />
Borzenok, S. A., Komakh<br />
Russian Federation<br />
16.35-16.45 h Organotipic culture preservation as strategy to increase the number of corneas for transplants<br />
Otero N.; Vilarrodona A.; Martinez-Conesa Mar ía E.; Nuñez V.; Pérez Luisa M.; Agustí E.<br />
Transplant Services Foundation-Hospital Clínic. Barcelona. Spain<br />
16.45-6.55 h Microbial Risk Assessment in Eye Banking<br />
Majore , I.; Scheider U.; Wittmershaus I.; B rgel M.; Blomberg L.; Knipper A.<br />
German Society of Tissue Transplantation, Germany<br />
AUDITORI<br />
15.45-16.45 h EUROPEAN GOOD PRACTICE STANDARDS<br />
Moderators: P. Van Grevestein and M. Amil<br />
15.30 -16.00 h EuroGTPs PROJECT<br />
Project Partners<br />
16.00-16.15 h Keeping the Council of Europe Guidance up-to-date<br />
M. López, France<br />
16.15-16.30 h The role of EATB in good practice standarization<br />
EATB<br />
16.45-17.15 h COFFEE BREAK<br />
AUDITORI<br />
17.15-18.10 h Panel Discussion ASSOCIATION INTERACTION<br />
Moderators: M. Macsai and D. Fehily<br />
18.10-18.25 h PRESIDENTIAL CLOSING REMARKS<br />
AWARDS, ANNOUNCEMENT OF MEETING 2012
LIST OF <strong>POSTERS</strong><br />
INTERNATIONAL EXPERIENCE ON TISSUE BANKING<br />
P-01 ACTIVITY REPORT OF THE MULTI-TISSUE BANK OF THE RED CROSS BLOOD TRANSFUSION SERVICE OF UPPER AUSTRIA<br />
Hennerbichler , S.; Peterbauer-Scherb A.; Plöderl K.; Gabriel C.<br />
Red Cross Blood Transfusion Service of Upper Austria, Linz/ Austrian Cluster for Tissue Regeneration. Austria.<br />
P-03 ISO 9001:2008 AUDITS AND NONCONFORMITY REPORTING IN A TISSUE BANK<br />
Kalaydjieva, V.; Popova P.<br />
Tissue Bank "Osteocentre Bulgaria". Sofia. Bulgaria.<br />
P-04 A 6 YEAR RETROSPECTIVE REVIEW OF THE KALEIDOSCOPE OF CHALLENGES ENCOUNTERED BY A SOUTH AFRICAN TISSUE<br />
BANK AND THEIR INFLUENCE ON DONOR STATISTICS AND ALLOGRAFT AVAILABILITY.<br />
Karakatsanis , E.<br />
Centre for Tissue Engineering - Bone Bank (Tshwane University of Technology). South Africa.<br />
P-05 COMMUNICATION & COSTUMER SERVICE IN A TISSUE BANK.<br />
Sobrevilla , E.; Merayo N.; Gallardo G.; Redondo M.; Miranda I.; Jardí F.; Trias E.<br />
Transplant Services Foundation. Hospital Clínic. Barcelona. Spain.<br />
P-06 TISSUE PROCESSING RESULTS IN 2010 IN THE BANK TISSUE AREA OF BLOOD AND TISSUE BANK OF ARAGON<br />
Martínez-Lorenzo , M.; Piñero Pimpinela E.; Ibañez Solla L.; Vicente Borruel O.; Fernández-pacheco J.; Callén Sevilla L.<br />
Banco de Sangre y Tejidos de Aragón. Spain<br />
SPECIFIC CHALLENGES OF DONATION; PROCESSING AND CLINICAL APPLICATION IN PAEDIATRICS<br />
P-07 EFFECTS OF NEW EU DIRECTIVE ON POSTMORTEM DONATION ACTIVITY IN SWEDEN<br />
Druid , H.; Wallberg L.; Odö B.; Josephsson S.; Dahlberg A.; Alkass K.<br />
Karolinska Institutet. Sweden.<br />
P-08 LEGAL MEDICINE VERSUS HEART VALVE DONATION<br />
Braun , C.; Wittmann G.; Jeckle J.; Schramm W.; Matthias G.<br />
Institute for Legal Medicine Munich. Germany.<br />
P-09 DONATION FOR RESEARCH USING A PROSPECTIVE STRATEGY<br />
Druid , H.; Alkass K.; Bäcklin S.<br />
Karolinska Institutet. Sweden.<br />
P-10 DESIGN OF AN INTEGRATED CAMPAIGN FOR DONATION AND PROCUREMENT OF SKIN AND TISSUES THROUGH SOCIAL<br />
NETWORKING AND MICROBLOGGING SERVICES: THE MEXICAN EXPERIENCE OF THE NATIONAL RESEARCH CENTER AND BURN<br />
CARE (CENIAQ)<br />
Martínez-f , F.; Sandoval-z H.<br />
Skin and Tissue Bank of the National Institute of Rehabilitation. Ministry of Health, Mexico.<br />
P-11 THE CHALLENGE TO COORDINATE DEATH IN THE NAME OF LIFE<br />
Popova , P.; Kalaydjieva V.<br />
Tissue bank "Osteocenre Bulgaria". Sofia. Bulgaria.<br />
LIST OF<br />
<strong>POSTERS</strong><br />
P-12 IMPACT ASSESSMENT OF SKIN DONATION PROGRAM IN COSTUMERS OF A EMERGENCY ROOM AND UNIT OF INTENSIVE<br />
CARE.<br />
Martínez-f , F.; Guzmán-m K.; Lomeli-r C.; Madinaveitia-v J.; Elizondo B.; Carrillo E.<br />
Skin and Tissue Bank of CENIAQ. National Institute of Rehabilitation. Mexico.
P-13 REVIEW OF THE TISSUE DONATION PROCESS AT THE TATA MEMORIAL HOSPITAL<br />
Samant Chandrashekhar, U.; Gajiwala Lobo A.; Lima C.; Mayekar V.<br />
Tata Memorial Hospital. India.<br />
P-14 CT SCAN IMAGING AS A TOOL IN TISSUE DONOR SCREENING PROCESS<br />
Herson Roma, M.; Morgan N.; Fitzsimmons A.; Poniatowski S.; O'donnell C.; Woodford N.; Cordner S.<br />
Donor Tissue Bank of Victoria / Victorian Institute of Forensic Medicine. Australia.<br />
CARDIOVASCULAR<br />
P-15 IN VITRO SUSCEPTIBILITY OF HIGH VIRULENCE MICROORGANISMS ISOLATED IN HEART VALVE BANKING<br />
Villalba , R.; Solis F.; Fornés G.; Jiménez A.; Eisman M.; González A.; Gómez Villagrán J.<br />
Centro Regional de Transfusión Sanguínea. Spain.<br />
P-16 SECURING DONOR SAFETY AND OPTIMIZING RECIPIENT OUTCOMES IN LIVING DONOR LIVER TRANSPLANTATION:<br />
SYSTEMATIC SELECTION OF GRAFT TYPES AND USE OF CRYO-PRESERVED VEIN GRAFT<br />
Tamura , S.; Motomura N.; Saito A.; Ono M.; Sugawara Y.; Hasegawa K.; Kokudo N.<br />
Tokyo University. Japan.<br />
P-17 THE EFFECT OF FREEZING RATES ON MOISTURE CONTENT AND SURFACE TEXTURE OF FREEZE-DRIED BOVINE PERICARDIUM<br />
Rusidah , M.; Adam Z.; Chee Wei T.; Suhaili Y.; Suzina S.<br />
Tissue Bank, School of Medical Sciences, Health Campus, Universiti Sains Malaysia.<br />
P-18 ANATOMICAL FRECUENT AANOMALIES IN CARDIAC VALVES. 15 YEARS OF EXPERIENCE<br />
Alvarez Viviana M.; Alvarez Veronica P.; Favaloro Rene R.; Bertolotti Mario A.<br />
Favaloro´s Foudation, Buenos Aires. Argentina.<br />
P-19 MECHANICAL PROPERTIES OF MITRAL ALLOGRAFTS ARE NOT REASONABLY INFLUENCED BY CRYOPRESERVATION<br />
Spatenka J, J.; Hlubocky J J.; Novacek V.; Spatenka J J.; Mokracek A.; Kochova P.; Habrmanova A.<br />
University Hospital Motol, Prague. Czech Republic.<br />
P-20 FACTORS AFFECTING THE EFFECTIVENESS OF A CARDIOVASCULAR RECOVERY TEAM.<br />
Vito , S.; Oliva R.; Luque S.; Hinojosa M.; Miranda M.; Calamardo M.; Pérez M.<br />
Transplant Services Foundation (TSF). Hospital Clínic. Barcelona. Spain.<br />
P-21 MINIMISING INFECTION & MAXIMISING POTENTIAL DURING CARDIECTOMY OF OXFORD DONORS<br />
Davies, J.; Auckburally F.; Charlesworth K.; Thomas Y.; Ratntunga C.<br />
Oxford Heart Valve Bank. UK.<br />
P-22 RECIPIENT OUTCOME STUDIES IN OXFORD<br />
Davies , J.; Charlesworth K.; Foster R.; Thomas Y.; Auckburally F.; Ratntunga C.<br />
Oxford Heart Valve Bank. UK.<br />
REPRODUCTIVE<br />
P-23 OVARIAN TISSUE BANKING - THE UPPER AUSTRIAN NETWORK FERTISAVE<br />
Hennerbichler , S.<br />
Red Cross Blood Transfusion Service of Upper Austria, Linz/ Austrian Cluster for Tissue Regeneration. Austria.<br />
LIST OF<br />
<strong>POSTERS</strong><br />
P-24 FREEZING OF SEMEN PEARL SAMPLES AND SPERMATOZOIDS FROM TESTICLE BIOPSIES<br />
Rendal Vázquez , M.; García García M.; Rendal Vazquez M.; López Piñón M.; Carballal Rodríguez M.; Rodríguez Cabarcos M.; Sola<br />
Rodríguez A.<br />
Complejo Hospitalario Universitario A Coruña. Spain.
P-25 OPTIMUM CRYOPRESERVATION OF SEMEN AND SPERMATOZOIDES FROM TESTICLE BIOPSIES<br />
Rendal Vázquez , M.; García García M.; Rendal Vázquez M.; Carballal Rodríguez M.; López Piñón M.; Rodríguez Cabarcos M.; Sola<br />
Rodríguez A.<br />
Complejo Hospitalario Universitario A Coruña. Spain.<br />
P-26 SYSTEM FOR THE STORAGE AT ULTRALOW TEMPERATURES OF BIOLOGIC MATERIAL FOR IVF USE<br />
Rendal Vazquez , M.; Andion Núñez C.; Rendal Vázquez M.; Rodríguez Cabarcos M.; Fernández Lago C.<br />
Tissue Bank Complejo Hospitalario Universitario A Coruña. Spain.<br />
P-27 CASE REPORT: THE USE OF PLATELET-RICH PLASMA IN ORTHOTOPIC CRYOPRESERVED OVARIAN TISSUE TRANSPLANTATION<br />
Murcia, N.; Tauste C.; Rodríguez L.; González S.; Almeida L.; Salvador C.; Callejo J.<br />
Hospital Sant Joan de Déu. Spain.<br />
MUSCULOSKELETAL<br />
P-28 APOPTOSIS MEDIATED CELL LOSS AFTER HUMAN MENISCI CRYOPRESERVATION.<br />
Villalba , R.; Peña J.; Navarro P.; Luque E.; Jimena I.; Romero A.; Gómez Villagrán J.<br />
Centro Regional de Transfusión Sanguínea. Spain.<br />
P-29 DEVELOPMENT AND EVALUATION OF THE NEW METHOD OF GLUCOCORTICOID OSTEOPOROSIS TREATMENT<br />
Volova , L.<br />
Institute of experimental medicine and biotechnologies. Russian Federation.<br />
P-30 DOES THE MENISCUS TRANSPLANT PREVENT OSTEOARTHRITIS? FUNCTIONAL OUTCOME AND RADIOGRAPHIC 5-YEAR<br />
FOLLOW-UP<br />
Valencia-García , H.; Fahandezh-Saddi H.; López-Hualda Á.; Martínez-Marín J.; Torrejón-de La Cal M.; Marín-Aguado M.; Vaquero-<br />
Mateo J.<br />
Hospital Universitario Fundación Alcorcón. Spain.<br />
P-31 ALLOGRAFT SELECTION FOR TRANSEPIPHYSEAL TUMOR RESECTION AROUND THE KNEE USING THREE-DIMENSIONAL<br />
SURFACE REGISTRATION<br />
Aponte-Tinao Alberto, L.; Schwint O.; Ritacco Eduardo L.; Farfalli Luis G.; Milano Edgardo F.; Bou-Sleiman H.; Reyes M.<br />
Italian Hospital of Buenos Aires. Argentina.<br />
P-32 THREE-DIMENSIONAL MORPHOMETRIC ANALYSIS OF THE DISTAL FEMUR: A VALIDITY METHOD FOR ALLOGRAFT SELECTION<br />
USING A VIRTUAL BONE BANK<br />
Aponte-Tinao Alberto, L.; Farfalli Luis G.; Ritacco Eduardo L.; Milano Edgardo F.; Schwint O.<br />
Italian Hospital of Buenos Aires. Argentina.<br />
P-33 DEEP-FROZEN FEMORAL HEADS: THE HIGHER THE WEIGHT, THE BETTER THE VIABILITY<br />
Mirabet , V.; Solves P.; Larrea L.; Ródenas T.; Pamplona T.; Alavés F.; Roig R.<br />
Banco de Céulas y Tejidos de la Comunidad Valenciana. Spain.<br />
P-34 ALLOGENIC CARTILAGE APPLICATION IN RHINO- AND GENIOPLASTY<br />
Pavluk-pavluchenko L.; Lekishvili M.; Dubinin S.; Ryabov , A.<br />
Peoples’ Friendship University of Russian. Russian Federation.<br />
P-35 INFLUENCE OF BISPHOSPHONATES AS A PART OF A BIOCOMPOSITE MATERIAL ON AN OSTEOGENESIS.<br />
Lekishvili M.; Rodionova S.; Yurasova Y.; Torgashin A.; Ryabov , A.<br />
N.N. Priorov Central Institute of Traumatology and Orthopaedics (CITO) Tissue bank, Moscow. Russian Federation.<br />
LIST OF<br />
<strong>POSTERS</strong>
P-36 USING DEMINERALIZED BONE GRAFTS AT RECONSTRUCTION OF NASAL SEPTUM.<br />
Khamidov, A., Lekishvili, M., Ahmedov, S., Vasiliev, M., Ryabov, A.<br />
Scientific Research Centre Otorhinolaryngology. Moscow. Russian Federation.<br />
P-37 APPLICATION OF DEMINERALIZED XENOGENIC IMPLANTS IN ORAL SURGERY.<br />
Lekishvili, M., Yurasova, Y., Reznikova, N., Ryabov, A.<br />
N.N. Priorov Central Institute of Traumatology and Orthopaedics (CITO) Tissue bank, Moscow. Russian Federation.<br />
P-38 OSTEOGENIC EFFECT OF TETRACYCLIN IN COMPOUND XENOGENEIC BONE IMPLANTS.<br />
Zunino, J. H. 1 , Filomeno, A. 2 , Cigliutti, G.2, Semiglia, G. 2<br />
1 - INDT, Tissue Bank, Hospital de Clínicas, 2 - Cátedra de Pequeños Animales, Orientación Quirúrgica, Facultad de Veterinaria.<br />
Uruguay.<br />
P-39 COMPARATIVE STUDIES ON BIOMECHANICAL PROPERTIES OF IRRADIATED PORCINE TENDON.<br />
Spinosa, M.1, Santoro, N. C. 1 , Blangino, A. E. 2 , Kairiyama, E. 1<br />
1 - National Commission of Atomic Energy, 2 - Faculty of Engineering, University of Buenos Aires. Argentina.<br />
P-40 OSTEOINDUCTIVE AND OSTEOGENIC PROPERTIES OF TWO XENOGENEIC BONE MATRICES.<br />
Zunino Hamlet, J.; Semiglia G.; De Pro C.; Carzoglio J.; Filomeno A.<br />
INDT, Tissue Bank, Hospital de Clinicas. Uruguay.<br />
P-41 COMPUTER ASSISTED NAVIGATION FOR TUMOR RESECTION AND ALLOGRAFT TRANSPLANTATION<br />
Aponte-tinao Alberto, L.; Farfalli Luis G.; Ritacco Eduardo L.; Milano F.; Ayerza Angel M.; Muscolo Luis D.<br />
Italian Hospital of Buenos Aires. Argentina.<br />
P-42 ACHILLES ALLOGRAFT AS A NEW EXTENSOR SYSTEM OF THE KNEE<br />
Méndez, A.; Ares O.; Lozano L.; Segur JM.; Martínez- J.; Popescu D.; Garcia E.<br />
Hospital Clínic. Barcelona. Spain.<br />
P-43 IMPROVEMENT PLAN FOR USE OF GRAFT FEMORAL HEAD OF LIVING DONOR.<br />
Valencia-García , H.; Marín-Aguado M.; Torrejón-de La Cal M.; López-Hualda A.; Martínez-Martín J.; Vaquero-Mateo J.; Fahandezh-<br />
Saddi H.<br />
Hospital Universitario Fundación Alcorcón. Spain.<br />
P-44 AUDIT OF THE FIRST 13 YEARS OF EXISTENCE OF BONE AND TISSUE BANK OF HOSPITAL UNIVERSITARIO FUNDACIÓN<br />
ALCORCÓN<br />
Valencia-García , H.; Torrejón-de La Cal M.; Marín-Aguado M.; Fahandezh-Saddi H.; Martínez-Martín J.; Vaquero-mateo J.; López-<br />
Hualda A.<br />
Hospital Universitario Fundación Alcorcón. Spain.<br />
P-45 THE INFLUENCE OF SINTERING TEMPERATURE ON COMPOSITION AND SURFACE MORPHOLOGY OF BOVINE BONE<br />
Firdaus , A.; Adam Z.; Hawani A.; Chee Wei T.; Hidayu C.; Suzina S.<br />
Tissue Bank, School of Medical Sciences Health Campus Universiti Sains Malaysia.<br />
P-46 SEARCH FOR METHODS TO INCREASE THE NUMBER OF PROCUREMENTS IN A TEMPORAL BONE BANK<br />
Caremans , J.; Muylle L.; Hamans E.; Van De Heyning P.<br />
University Hospital Antwerp. Belgium.<br />
LIST OF<br />
<strong>POSTERS</strong><br />
P-47 THE USE OF BONE ALLOGRAFT IN REVISION TOTAL KNEE ARTHROPLASTY<br />
López-Hualda , A.; Valencia-García H.; Matínez-Martín J.; Fahandezh-Saddi H.; Marín-Aguado M.; Torrejón-de La Cal M.; Vaqueromateo<br />
J.<br />
Hospital Universitario Fundación Alcorcón. Spain.
P-48 INFLUENCE OF DONOR TYPE IN BONE AND TISSUE GRAFTS OUTCOMES IN THE QUARANTINE PERIOD<br />
Mieth , K.; González J.; Navas J.; Soto C.<br />
Fundacion Cosme y Damian. Colombia.<br />
P-49 FUNDACIÓN COSME Y DAMIAN: A MODEL OF A PRIVATE, NON FOR PROFIT SINGLE TISSUE BANK<br />
Soto , C.; Navas J.; Mieth K.; González J.<br />
Fundacion Cosme y Damián. Colombia.<br />
P-50 OSSICULAR PROSTHESES FROM BANKED BONE VIA NUMERICALLY CONTROLLED MICROMILLING<br />
Ramelli , M.; Liscio M.; Carriaggio F.; Berrettini S.; Danti S.; Mancini I.<br />
Tissuelab S.p.a. Italy.<br />
P-51 NEW METHODS FOR CARTILAGE GRAFT EVALUATION: OPTICAL COHERENCE TOMOGRAPHY, POLARIZATION SENSITIVE<br />
OPTICAL COHERENCE TOMOGRAPHY AND THERMOGRAVIMETRIC ANALYSIS<br />
Martínho Junior Carlos, A.; Freitas Zanardi A.; Brocardo Diva Machado L.; Santin Plumeri S.; Soares Augusto Neves F.; Mathor Beatriz M.<br />
Nuclear and Energy Research Institute. Brazil.<br />
P-52 THE USE OF THE OF SPONGY BONE IMPLANT IN THE CORRECTION OF THE IDIOPATIC SCOLIOSIS<br />
Jacas Torne F, M.; Garcia Mesa N.; Alvarez Cambra R.<br />
Complejo Cientifico Ortopedico Internacional "Frank Pais". Cuba.<br />
P-53 BONE ALLOGRAFT USE FOR SALVAGE OF FAILED TREATMENT OF INTERTROCHANTERIC HIP FRACTURES<br />
Garcia, E.; Zumbado Alonso J.; Camacho P.; Méndez A.; López I.; Tornero E.; Segur JM<br />
Hospital Clínic. Barcelona. Spain.<br />
P-54 BIOMECHANICAL CHARACTERIZATION OF A PROCESSED, ANTIGEN-FREE BONE XENOIMPLANT FOR SKELETAL<br />
RECONSTRUCTION.<br />
Zunino Hamlet, J.; Lantero Carlos J.; Aguiar S.; Deri E.<br />
INDT, Tissue Bank, Hospital de Clínicas. Uruguay.<br />
P-55 PROCESSING METHODS VALIDATION FOR PATELLAR TENDON ALLOGRAFT USING X-RAY DIFFRACTION<br />
Pérez Campos , H.; Wodowoz O.; Saldias M.; Faccio R.; Mombru A.; Alvarez I.<br />
Instituto Nacional de Donación y Trasplante de Células, Tejidos y Órganos (INDT) - Laboratorio de Cristalografía Facultad de Química<br />
Udelar. Uruguay.<br />
P-56 EVOLUTION OF THE ACL ALLOGRAFT IN THE ACL RECONSTRUCTION SURGERY. A 15 YEAR FOLLOW UP<br />
López Zabala , I.; Claret García G.; Pereira E.; Granados N.; Garcia Oltra E.; Sastre Solsona S.<br />
Hospital Clínic. Barcelona. Spain<br />
P-57 TRANEXAMIC ACID AS A SUBSTITUTE OF APROTININ FOR CHONDROCYTE GRAFT PREPARATION<br />
Bursig , H.; Sitek P.; Wysocka A.; Kurzak A.; Król D.; Dylag S.; Nozynski J.<br />
Tissue Bank. Poland.<br />
P-58 AUTOLOGOUS CHONDROCYTE CULTURE – 5 YEARS EXPERIENCE<br />
Bursig , H.; Wysocka A.; Sitek S.; Kurzak A.; Kepski F.; Dylag S.; Gazdzik T.<br />
Tissue Bank. Poland.<br />
P-59 DEVELOPMENT OF AN EFFICIENT METHOD TO DEMINERALISE BONE<br />
Calamardo Alba, M.; Vito S.; Fariñas O.; Luque S.; Vilarrodona A.; Segur JM.; Trias E.<br />
Transplant Services Foundation - Hospital Clínic. Barcelona. Spain.<br />
LIST OF<br />
<strong>POSTERS</strong>
P-60 DISTAL TIBIA FAILURE IS THE MOST COMMON COMPLICATION IN MASSIVE INTERCALARY MASSIVE BONE ALLOGRAFT<br />
RECONSTRUCTION IN PEDIATRIC TUMOR<br />
Martanto , T.; Martanto T.; Edward M.<br />
Dept. of Orthopaedic Dr. Soetomo General Hospital/Airlangga University. Indonesia.<br />
P-61 INFECTION AND LOOSENING PROBLEM IN BONE ALLOGRAFT APPLICATIONS IN TUMOR CASES<br />
Martanto , T.; Edward M.<br />
Dept. of Orthopaedic Dr. Soetomo General Hospital/Airlangga University. Indonesia.<br />
P-62 TENDON ALLOGRAFT FOR MULTILIGAMENT KNEE INSTABILITY<br />
Morales, J.; Popescu D.; Segur JM.; Suso S.<br />
Hospital Clínic. Barcelona. Spain.<br />
P-63 EFFECTS OF CHONDROPROTECTIVE AGENTS: GLUCOSAMINE, CHONDROITIN SULFATE, AND HYALURONIC ACID ON<br />
CHONDROCYTES CULTURED FOR TRANSPLANTATION<br />
Olender , E.; Uhrynowska-tyszkiewicz I.; Kaminski A.<br />
Medical University of Warsaw; National Centre for Tissue and Cell Banking. Poland.<br />
P-64 STRUCTURAL BONE ALLOGRAFT FOR RECONSTRUCTION OF THE ACETABULAR COMPONENT IN SEVERE HIP ARTHROPLASTY<br />
LOOSENING.<br />
Fernández-valencia , J.; Medrano C.; Tornero E.; Tomás X.; Bori G.; Segur JM.; Gallart X.<br />
Hospital Clínic. University of Barcelona. Spain.<br />
P-65 HOW OPTICAL COHERENCE TOMOGRAPHY CAN BE USED TO EVALUATE QUANTITATIVELY GRAFTS BEFORE ITS USE.<br />
Mathor Beatriz, M.; Martínho Junior Carlos A.; Santin Plumeri S.; Soares Augusto Neves F.; Freitas Zanardi A.<br />
Nuclear and Energy Research Insitute. Brazil.<br />
P-66 ENDOGENOUS AND EXOGENOUS RESISTANCE FACTORS OF CONNECTIVE TISSUE TRANSPLANTS TO THE EXPOSURE OF THE<br />
RADIATIVE EMISSION<br />
Shangina , O.; Shangina O.<br />
Russian Eye and Plastic Syrgery Center. Russian Federation.<br />
P-66 bis LABRAL TRANSPLANTATION IN HIP INSTABILITY<br />
Ribas M, Tey M, Bellotti V, Erquicia J<br />
ICATME – Institut Català de Traumatologìa i Medicina de l’Esport<br />
University Hospital Dexeus – USP, Barcelona. Spain<br />
P-66 bis 2 INVETERATE SERIOUS DEFECTS OF ACHILLES TENDON. RECONSTRUCTION PROCEDURES<br />
Núñez-Samper, M.<br />
Clínica Virgen del Mar. Madrid. Spain.<br />
SKIN & AMNIOTIC MEMBRANE<br />
P-67 IN VITRO RECONSTRUCTION OF AN ARTIFICIAL BILAMINAR MULTICELLULAR SKIN<br />
Sánchez-Muñoz , I.; Granados R.; Sánchez-Toribio M.; Casares M.<br />
Hospital Universitario de Getafe. Spain.<br />
LIST OF<br />
<strong>POSTERS</strong><br />
P-68 HUMAN AMNIOTIC MEMBRANE AS SCAFFOLD FOR KERATINOCYTES AND FIBROBLAST CULTURE<br />
San Luis Verdes , A.; Rendal Vázquez M.; Domenech García N.; Yebra-pimentel Vilar M.; García Barreiro J.; Andión Núñez C.<br />
Unit of Criobiology Complejo Hospitalario Universitario A Coruña. Spain.
P-69 PRODUCTION OF RADIOSTERILIZED PIG SKIN AS BIOLOGICAL DRESSING<br />
Reyes Frías , M.; Reboyo Barrios D.<br />
Instituto Nacional de Investigaciones Nucleares. Mexico<br />
P-70 MORPHOMETRICS ANALYSIS OF IRRADIATED SKIN FROM TRANSMISSION ELECTRON MICROSCOPE IMAGES<br />
Spinosa , M.; Pachado Alberto J.; Horak Inés C.; Kairiyama E.<br />
National Commission of Atomic Energy. Argentina.<br />
P-71 DONOR PARAMETERS AFFECTING SKIN TISSUE RECOVERY<br />
Savío Manuel, A.; Pérez Luisa M.; Fariñas O.; Alemany X.; Vito S.; Oliva R.; Luque S.<br />
Transplant Services Foundation-Hospital Clínic. Barcelona. Spain.<br />
P-72 TIME-KILL STUDIES OF BASE.128 AGAINST S. EPIDERMIDIS II, S. CAPITIS AND P. ACNES ISOLATED FROM DONOR SKIN.<br />
Gatto , C.; Pianigiani E.; Ierardi F.; Giurgola L.; D'Amato Tothova J.<br />
R&D Alchimia SRL. Italy.<br />
P-73 UV LIGHT STIMULATES CELL PROLIFERATION BY EGR-1 ACTIVATION IN HUMAN PRIMARY DERMAL FIBROBLASTS OBTAINED<br />
FROM MULTIORGAN DONORS.<br />
Martínez-f , F.; Barrera-l A.; Lomeli-r C.; Madinaveitia-v J.; Sandoval-z H.; Machuca-r C.; Villegas-c H.<br />
Skin and Tissue Bank of CENIAQ, National Institute of Rehabilitation. Mexico.<br />
P-74 MICROBIOLOGICAL ANALYSIS OF SKINGRAFTS PROCESSED BY THE SKIN & TISSUE BANK OF THE INR IN MÉXICO.<br />
Martínez-f , F.; Lomeli-r C.; Barrera-l A.; Vizcaino-d A.; Guzmán-m K.; Sandoval-z H.; Madinaveitia-v J.<br />
Skin and Tissue Bank of CENIAQ. National Institute of Rehabilitation. Mexico<br />
P-75 THE EXPERIENCE OF THE OPERATIVE MODEL THE SKIN AND TISSUE BANK OF INR IN MÉXICO<br />
Martínez-f , F.; Vizcaino-d A.; Lomeli-r C.; Barrera-l A.; Estrada-v E.; Villegas-c H.; Madinaveitia-v J.<br />
Skin and Tissue Bank of CENIAQ. National Institute of Rehabilitation. Mexico<br />
P-76 ACTIVITY IN THE RECENTLY ESTABLISH SKIN BANK OF TSF.<br />
Savío , A.; Pérez M.; Fariñas O.; Hinojosa M.; Miranda M.; Manotas C.; Zardoya M.<br />
Tissue Bank, Transplant Services Foundation (TSF) - Hospital Clínic de Barcelona. Spain.<br />
P-77 USE OF AMNION IN THIRD DEGREE BURNS AND NON-HEALING WOUNDS<br />
Lobo Gajiwala , A.; Gajiwala K.<br />
Tata Memorial Hospital. India.<br />
CORD BLOOD<br />
P-78 EVOLUTION FROM 2007-2010 IN MALAGA CORD BLOOD BANK<br />
Rizo Alfaro , P.; Ponce Verdugo L.; Gómez Maldonado P.; Galeote Padilla A.; Hernández Lamas M.; Prat Arrojo I.<br />
Malaga Cord Blood Bank. Spain.<br />
LIST OF<br />
<strong>POSTERS</strong><br />
P-79 VIABILITY AND FORMATION OF ERYTHROID, GRANULOCYTE/MACROPHAGE AND MEGAKARYOCYTIC COLONIES IN BLOOD<br />
HEMATOPOIETIC PROGENITORS AFTER 20 YEARS STORED IN LÍQUID NITROGEN<br />
Romero Vega, E.; Romero Vega E.; Meca Rovayo M.; Rivero Arias G.; Salat Martí A.<br />
Banco Sectorial de Tejidos de Cádiz. Spain.<br />
P-80 NEW GMP METHOD FOR CORD BLOOD CRYOPROCESSING WITH CLOSED TUBING SYSTEM AND PRE-COOLED GEL SLEEVES<br />
Kubesova , B.; Karkoskova L.; Matejkova E.<br />
Tissue Bank University Hospital Brno. Czech Republic.
P-81 INVENTORIES: MALAGA CORD BLOOD BANKING<br />
Rizo-Alfaro , P.; Ponce-Verdugo L.; Gómez-Maldonado P.; Falcón-Morales F.; Mayorga-Pastrana C.; Hernández-Lamas Carmen M.; Prat-<br />
Arrojo I.<br />
Malaga Cord Blood Bank. Spain.<br />
P-82 DONOR SELECTION OF UMBILICAL CORD BLOOD<br />
Ponce-Verdugo , L.; Bellido-Estevez I.; Hernández-Lamas Carmen M.; Jaen-García J.; Pérez-Salazar Carmen M.; González-Conejo Maria A.;<br />
Prat-Arrojo I.<br />
Malaga Cord Blood Bank. Spain.<br />
P-83 INFLUENCE ON TIME TO CRYOPRESERVATION<br />
Ponce-Verdugo , L.; Gómez-Maldonado P.; Rizo-Alfaro P.; Galeote-Padilla A.; Martín-Bejar Teresa M.; Hernández-Lamas Carmen M.;<br />
Prat-Arrojo I.<br />
Malaga Cord Blood Bank. Spain.<br />
P-84 UMBILICAL CORD BLOOD: ECONOMIC STUDY<br />
Ponce-Verdugo , L.; Hernández-Lamas Carmen M.; Rizo-Alfaro P.; Pérez-Parrado S.; España-Ramos T.; Arenas-Aguilar P.; Prat-Arrojo I.<br />
Malaga Cord Blood Bank. Spain.<br />
P-85 MALAGA CORD BLOOD BANK: EVALUATION THE UNITS SENT FOR TRANSPLANT<br />
Ponce-Verdugo , L.; Hernández-Lamas Carmen M.; Rodríguez-Ruiz Y.; Nieto-Chups V.; Roman-Morillas C.; Tornay-Gómez Azucena A.;<br />
Prat-Arrojo I.<br />
Malaga Cord Blood Bank. Spain.<br />
P-86 CONTINUING EDUCATION MALAGA CORD BLOOD BANK<br />
Gómez-Maldonado , P.; Ponce-Verdugo L.; Hernández-Lamas Carmen M.; Rizo-Alfaro P.; Prat-Arrojo I.<br />
Malaga Cord Blood Bank. Spain.<br />
P-87 THE CORD BLOOD BANK OF THE RED CROSS BLOOD TRANSFUSION SERVICE OF UPPER AUSTRIA<br />
Hennerbichler , S.; Peterbauer-Scherb A.; Plöderl K.; Havlicek-Pötscher M.; Süssner S.; Gabriel C.<br />
Red Cross Blood Transfusion Service Of Upper Austria, Linz/ Austrian Cluster For Tissue Regeneration. Austria.<br />
P-88 SEMI-AUTOMATED HIGH-THROUGHPUT NEXT GENERATION SEQUENCING BASED HLA-TYPING FOR CORD BLOOD SAMPLES<br />
Hennerbichler , S.; Stabentheiner S.; Niklas N.; Hofer K.; Pröll J.; Danzer M.; Gabriel C.<br />
Red Cross Blood Transfusion Service of Upper Austria, Linz/ Austrian Cluster for Tissue Regeneration. Austria.<br />
GLIMPSE INTO THE FUTURE; ADVANCED THERAPIES<br />
P-89 HEPATIC CELLULAR THERAPY UNIT, IIS-HOSPITAL LA FE: ESTABLISHMENT OF HEPATIC CELL BANK FOR CELLULAR<br />
TRANSPLANTATION.<br />
Bonora-Centelles , A.; Pareja E.; Cortés M.; Mirabet V.; Mir J.; Castell J.; Gómez-Lechón M.<br />
Instituto de Investigación Sanitaria - Fundación para la Investigación Hospital La Fe. Spain.<br />
P-90 CLINICAL TRIAL COMPARING PLATELET-RICH PLASMA TO BETAMETASONA AND BUPIVACAIN SUBACROMIAL INJECTION IN<br />
ROTATOR CUFF TENDYNOSIS. PRELIMINARY RESULTS<br />
Pacha Vicente D.; Llusá M.; Nardi J.; Navarro A.; Rodríguez L.; Genis X.<br />
Vall d’Hebron Hospital. Spain.<br />
P-91 ESTABLISHING THE SNBTS ISLET ISOLATION LABORATORY<br />
Mitchell Leigh, D.; Mcquillan T.; Drain J.; Casey J.; Galea G.; Mcgowan N.<br />
SNBTS Tissues & Cells Directorate. UK.<br />
LIST OF<br />
<strong>POSTERS</strong>
P-92 HUMAN AMNIOTIC MEMBRANE AS A SOURCE OF PROGENITOR CELLS FOR HUMAN ARTICULAR CARTILAGE REPAIR<br />
Rendal Vazquez , M.; Muiños López E.; Diaz Prado S.; Hermida Gómez T.; Sanluis Verdes A.; Fuentes Boquete I.; Blanco Garcia F.<br />
Cryobiology Unit Complejo Hospitalario Universitario A Coruña. Spain.<br />
P-93 COMPARISON OF DIFFERENT METHODS OF GENOMIC DNA EXTRACTION AND PURIFICATION FROM TISSUE.<br />
Isidro-Marrón , P.; Astudillo-González A.; Martínez-Camblor P.; Vallina-Álvarez A.; Fernández-Juárez L.<br />
BioBanco Hospital Universitario Central de Asturias-Oficina de Investigación Biosanitaria del Principado de Asturias (OIB). Spain.<br />
P-94 HUMAN MESENCHYMAL CELLS OF ADIPOSE TISSUE INHIBITS THE PRODUCTION OF PGE2 IN HUMAN MONOCYTES<br />
Platas , J.; Mirabet V.; Villamón E.; Olivar T.; Alcaraz José M.; Guillén lM.<br />
Dpt. Pharmacology, University of Valencia and IDM. Spain.<br />
P-95 HEPATOCYTE ISOLATION FROM LIVERS NOT SUITABLE FOR WHOLE ORGAN TRANSPLANTION. LIVER CELL THERAPY<br />
Gómez Gómez M.; Manyalich M.<br />
DTI FOUNDTION. Spain.<br />
P-96 DIFFERENCES IN SURFACE MARKERS EXPRESSION AND CHONDROGENIC POTENTIAL AMONG DIFFERENT TISSUE-DERIVED<br />
MESENCHYMAL CELLS FROM ELDERLY PATIENTS WITH OSTEOARTHRITIS<br />
Sanz-Moreno , J.; Desportes Bielsa P.; Alegre-Aguarón E.; García-Álvarez F.; Martínez-Lorenzo M.<br />
Servicio de Inmunología del Hospital Clínico Universitario Lozano Blesa. Spain.<br />
P-97 ADIPOSE TISSUE BANKING FUTURE PROSPECTS: FIRST STEP, SET UP OF CRYOPRESERVATION METHODOLOGY.<br />
Agusti , E.; Brugada P.; Vilarrodona A.; Martínez L.; Pérez Luisa M.; Martínez-Conesa E.; Trias E.<br />
Transplant Services Foundation-Hospital Clínic. Barcelona Spain.<br />
P-98 INTRA-ARTICULAR KNEE INJECTION OF PLATELETS RICH PLASMA TO TREAT OSTEOARTHRITIS.<br />
Rodríguez , L.; Genis X.; Tarragona E.; Ortega I.; Gomariz E.; Navarro A.; Joshi N.<br />
Banc de Sang i Teixits. Spain.<br />
P-99 COLD STORAGE OF PRIMARY HEPATOCYTES IN TIPROTEC VARIANTS<br />
Rauen , U.; Pless-petig G.; Sauer Maximilian I.<br />
Institut für Physiologische Chemie, Universitätsklinikum Essen. Germany.<br />
P-100 BIOLOGICAL PROPERTIES OF POROUS BIOMATERIALS OF THE CONNECTIVE TISSUE ORIGIN AND POTENTIALS OF THEIR<br />
USE IN SURGERY<br />
Muslimov S.; Khasanov R.; Shangina O.; Musina L.; Kornilaeva G.; Khasanova Y. - Russian Eye and Plastic Surgery Center. Russian<br />
Federation.<br />
P-101 A DECELLULARIZATION PROCESS IN TISSUE BANKING<br />
Ramos, J.; Iglesias F.; López M.; De La Puente Pilar M.; Vázquez José J.; Fernández A.<br />
Tissue Establishment San Francisco Clinic Foundation. Spain.<br />
CORNEA<br />
P-102 ASSESSMENT OF SUBJECTIVE PARAMETERS OF PATIENTS WITH AUTOLOGOUS Vs HETEROLOGOUS EYE DROPS TREATMENT<br />
IN OCULAR PATHOLOGY<br />
Guijarro , L.; Guijarro-Hernández L.; Benítez Del Castillo J.; Ponce-Verdugo L.; Gómez-Maldonado P.; Hernández-Lamas M.; Prat-Arrojo I.<br />
Hospital de Jerez De La Frontera. Spain.<br />
P-103 PROSPECTIVE DEVELOPMENT OF EYE BANKING IN THE RUSSIAN FEDERATION<br />
Komakh Alekseevich, Y.; Borzenok Anatolievich S.<br />
The S. Fyodorov Eye Microsurgery Federal State. Russian Federation.<br />
LIST OF<br />
<strong>POSTERS</strong>
P-104 MARKERS TO IDENTIFY LIMBAL STEM CELLS ON HUMAN AMNIOTIC MEMBRANE<br />
Rendal, M.; Sanluis A.; Yebra- Vilar M.; López I.; Domenech García N.; Andión C.; Blanco F.<br />
Unit of Criobiology Complejo Hospitalario Universitario A Coruña. Spain.<br />
P-105 CIRCUIT FOR IMPLEMENTING A DONATION OF EYE TISSUE<br />
Cabrejas Ayuso, A.; Cabrejas Ayuso A.; Cano Puig M. - Hospital de Bellvitge. Spain.<br />
P-106 EVOLUTION OF OCULAR TISSUE COLLECTION IN AN ACUTE CARE HOSPITAL FROM 1995 TO PRESENT DAY<br />
Marin, I.; Vallejo P.; Tobaruela P.; Rusiñol J.; Azagra M.; Quintana S.<br />
Hospital Universitari Mútua Terrassa, Social Work. Spain.<br />
P-107 DONOR AGE AND CORNEA ENDOTHELIAL CELL DENSITY, INFLUENCE ON TRANSPLANT OUTCOME.<br />
Santos , S.; Santamaria A.; Echevarria J.; Rodríguez P.; Azcarate M.; Pérez-Vaquero M.; Vesga M.<br />
Centro Vasco De Transfusión y Tejidos Humanos. Spain.<br />
P-108 ANALYSIS OF FACTORS AFFECTING THE VIABILITY OF CONEAL TISSUE.<br />
Rodríguez , L.; Genis X.; Rodríguez L.; Tarragona E.; Ortega I.; Gomariz E.; Navarro A. - Banc de Sang i Teixits. Spain.<br />
LIST OF<br />
<strong>POSTERS</strong><br />
P-109 COMMUNICATION BETWEEN TISSUE BANK AND IMPLANTING SURGEON: SHORT-TERM FOLLOW-UP OF TISSUES FOR<br />
OCULAR SURGERIES.<br />
Nuñez , V.; Agustí E.; Martínez E.; Otero N.; Pérez M.; Casaroli-Marano R.; Vilarrodona A.<br />
Transplant Services Foundation-Hospital Clínic. Barcelona. Spain.<br />
P-110 A COMPARISON OF DIFFERENT CULTURE MEDIA FOR THE STORAGE OF HUMAN DONOR CORNEAS FOR GRAFTING<br />
Zlacka D., Krabcova I., Kortusova J. and Jirsova K.<br />
Laboratory of the Biology and Pathology of the Eye, Institute of Inherited Metabolic Disorders and Ocular Tissue Bank, General Teaching<br />
Hospital and 1st Faculty of Medicine, Charles University in Prague, Czech Republic.
INVITED<br />
SPEAKER<br />
LECTURES
BIOBANKING AND BANKING FOR RESEARCH: INTERACTION WITH<br />
TISSUE BANKING<br />
THE ETHICS OF RESEARCH CONSENT<br />
Martha Anderson, Musculoskeletal Transplant Foundation.<br />
Introduction:<br />
The use of donated human tissues, organs and other body parts is critical to the development of new pharmaceuticals,<br />
medical devices and products, as well as to the on-going training of medical professionals. Unlike donation for<br />
transplantation, donation for research is far less regulated and less-well organized. In 2011, the AATB published standards<br />
for Non-Transplantable Anatomic Donation Organizations (NADO) that, for the first time, established specific guidelines for<br />
handling non-transplantable tissues (NTT). Many individuals who wish to donate for transplant upon their death are<br />
precluded from doing so, due to advanced age or disease processes; research donation provides them an opportunity to<br />
contribute to science and mankind.<br />
In the U.S, tissue banks and recovery agencies (organ, eye and tissue) provide NT organs and tissues to researchers. A<br />
variety of uses and users include:<br />
Internal Use: Tissue Banks require NTT to conduct new product development and processing validation.<br />
Academic Use: Academic centers use NTT in research projects such as evaluating the impact of processing techniques<br />
on tissue quality, development of new surgical techniques and training medical professionals.<br />
Medical Industry Use: Bio-banks, Bio-skills labs and pharmaceutical and medical device companies use human specimens<br />
to train surgeons in new surgical techniques and to test new medical devices and pharmaceuticals.<br />
Non-medical Use: Search and Rescue teams use NTT for training cadaver dogs; the military and automotive industries<br />
use specimens for testing safety equipment.<br />
Ethical issues associated with research tissues abound. They include:<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
Obtaining adequate, legal authorization for research from a donor or the next-of-kin of a recently deceased person<br />
Using donated tissues and organs for research rather than transplant<br />
Use of human tissues for non-medical use<br />
Approval process for non-medical and medical use<br />
Use of human specimens by for-profit entities<br />
Eliminating the cost of funeral expenses in exchange for research donation.<br />
Background:<br />
For purposes of this abstract and presentation, I will discuss how the Musculoskeletal Transplant Foundation (MTF) and our<br />
subsidiary the International Institute for the Advancement of Medicine (IIAM) address these ethical issues.<br />
Specimen Type Use Number of Specimens Provided<br />
Organ<br />
Pharmaceutical Research, Other Medical Device Research<br />
& Development<br />
Anatomic Tissue BioSkills labs, BioBanks, Medical Device R&D 1450<br />
Anatomic Tissue Internal (MTF) Research & Quality Assurance 4500<br />
Anatomic Tissue Academic Research 50<br />
830
INVITED<br />
SPEAKER<br />
LECTURES<br />
MTF and IIAM provide NT tissues and organs to a variety of entities. On an annual basis, we provide approximately the<br />
following number of research specimens:<br />
Donor authorization process<br />
On average, 89% of MTF donors give consent or authorization for “transplant, medical education and research”. When<br />
tissue is rejected for transplant (18% of recovered donors), the donor chart is reviewed and if medical education and/or<br />
research authorization has been given, we will endeavor to place that tissue for research, either internally or externally.<br />
Approximately 800 donors per year are potential research donors.<br />
IIAM handles NT organs, tissues and whole body donors (WBD) that are not suitable for transplantation. WB donation is<br />
often coordinated with a dying patient and/or his/her family during the final phase of life. Nursing Homes, Hospices,<br />
Funeral Homes etc. are the primary referral sources. Many NADOs in the US allow patients to pre-register their desire to<br />
be a WBD; in these instances, the patient and/or family fills out a consent form, medical/social history form and is preauthorized<br />
for donation prior to death. In other instances, the NADO is contacted at or near the time of death and evaluates<br />
the potential donor’s suitability. Research donor criteria is much less restrictive than transplant criteria. Donor authorization<br />
is obtained by tissue bank staff who also conduct a medical/social history interview. Authorization for non-medical uses<br />
(e.g., automotive safety testing, search and rescue training) is specifically documented.<br />
Determining apropiate use of NT Tissues and organs<br />
Although there is no mandatory allocation system for NTT in the U.S., it is MTF and IIAM’s philosophy that the priority for<br />
placement of donated organs and tissues should be:<br />
1. Transplant<br />
2. Internal research for MTF’s R&D and QA activities<br />
3. Academic or non-profit medical research and surgical training<br />
4. For-profit medical research and/or surgical training<br />
All research entities must execute an application that specifies exactly what types and amounts of specimens are requested<br />
and a Biomaterials Transfer Agreement (BMTA). The BMTA includes a wide range of responsibilities of parties including:<br />
providing a clear explanation of the proposed uses of the donated specimens<br />
the requirement for IIAM to de-identify the specimens and donor-related documentation<br />
requirement that all specimens be tested for infectious diseases prior to use<br />
prohibition against re-selling any specimens to a third party without prior written permission, and<br />
appropriate disposal of specimens following completion of the project.<br />
Each request for tissue is reviewed by senior staff at MTF, a medical director and other relevant departments such as<br />
Processing and QA. MTF’s Ethics Committee reviews all non-medical requests such as cadaver dog search and rescue<br />
training or the use of specimens in the development of military and automotive safety equipment, and requires supplemental<br />
consents from donor families for these types of research. Additionally, the MTF Donor Family Council is available to review<br />
research protocols and specific research requests. Because these specimens come from deceased donors, Internal Review<br />
Board (IRB) review and approval is not necessary, which can dramatically speed the review process.<br />
Financial questions<br />
Tissues from MTF are provided to academic researchers gratis; researchers who receive specimens from IIAM are charged<br />
a service fee that covers the costs associated with operating that business unit, with for-profit researchers typically being<br />
charged a higher service fee than those from non-profit agencies.<br />
One controversial area of WBD is the practice of advertising free cremation to anyone who becomes a WBD. With WBD,<br />
there is no need for funeral services such as embalming, but it is the practice of WBD tissue banks to return ‘cremains’ to<br />
the family following donation. NADOs often cremate a small portion of a body, and return those remains to a family for<br />
their memorial service. This practice could be seen as an incentive for a family member to donate their elderly loved one’s<br />
body to avoid funeral expenses. Advertising by some WBD tissue banks underscores this concern:
INVITED<br />
SPEAKER<br />
LECTURES<br />
“With cremation prices being so high and the need for body donors to support medical sciences so critical, the<br />
lasting impact of whole body donation makes so much sense. So please do look at all of your options. There is<br />
high cost cremation, low cost cremation and then there is free cremation. We are sure you will agree that while<br />
low cost is pretty good, a free service is hard to beat. Cremation prices are always covered by XYZ when<br />
donating your body to science.”<br />
The AATB’s new standards for Non-Transplant Anatomic Donation Organizations (NADOs) specifically preclude any<br />
monetary inducement:<br />
NT-D3.000 monetary compensation<br />
Monetary inducement or other valuable consideration, including goods or services, shall not be offered to a donor,<br />
Authorizing Person, the donor’s estate, or any other third party, except that the NADO may reimburse responsible third<br />
parties for costs directly associated with a donation and may reimburse Living Donors for costs associated with an acceptable<br />
donation, including compensation for restoration of lost earnings when directly attributable to donation, if and as authorized<br />
by law. Donors or their families should not be required to pay for any expenses related to the Acquisition of NAM for<br />
education and/or research.<br />
Conclusion:<br />
Donation for research offers potential donors and their families an additional opportunity to contribute to science and<br />
mankind. Tissue Banks, medical teaching institutions, pharmaceutical and medical device companies depend on human<br />
specimens for new medical education and training, product development and quality assurance validation studies. Ethical<br />
issues in the field of Research Donation are similar to those faced by transplant tissue banks including: Donor Authorization,<br />
allocation and financial incentives. A Non-Transplant Anatomic Donation Organization should have systems in place to<br />
assure that ethical policies are implemented and followed.
SPECIFIC CHALLENGES OF DONATION, PROCESSING AND CLINICAL<br />
APPLICATION IN PAEDIATRICS<br />
20 YEARS OF EXPERIENCE ON BONE ALLOGRAFTING IN CHILDREN AND<br />
ADOLESCENTS<br />
Dr. R. Huguet Carol; Dr. F. Torner Rubies; Dr. Alex Muset Lara, Hospital Sant Joan de Déu. Barcelona, Spain<br />
We can consider that in children and adolescents, there are currently 2 main groups of indications for the use of bone<br />
allografts in orthopedic surgery and traumatology.<br />
– The reconstruction of bone defects in limb salvage for cancer surgery.<br />
– The performance of spinal arthrodesis<br />
Limb salvage surgery in the treatment of malignant bone tumors has now become the therapeutic option in most cases. This<br />
surgery requires several reconstructive techniques and the use of bone allografts on numerous occasions.<br />
Allografts in orthopedic cancer surgery can be non-structural allograft used for filling cavities, or structural used to replace<br />
large bone defects secondary to bone resections.<br />
We may use in 4 ways structural bone allografts in children and adolescents:<br />
– Intercalary allografts<br />
– Osteoarticular Allografts<br />
– Composite allograft-prosthesis<br />
– Composite allograft-vascularized bones.<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
We can find a high rate of complications by the use of structural allografts in some series published. Among the complications<br />
of allografts we can find nonunion, infection or allograft failure, complications that in some cases could be increased by the<br />
use of radio and chemotherapy.<br />
In recent times, we have experimented with the composite use of vascularized bone and allograft, joint to a stable<br />
osteosynthesis that could significantly reduce these complications.<br />
We consider that the use of structural allografts in children remains a valid option of treatment that may be permanent in<br />
some cases and in others allows growth and preserve bone stock during skeletal maturation.
THE CHALLENGES OF MUSCULOSKELETAL TISSUE TRANSPLANT IN CHILDREN<br />
Mikel San-Julian, MD, PhD, Department of Orthopaedic Surgery. University of Navarra, Pamplona, Spain.<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
From 1987, 4661 musculoskeletal allografts have been managed in our bone bank. Cortical allografts are mainly used for<br />
surgical treatment of paediatric bone sarcomas. Increase in survival combined with the demanding functional requirements<br />
of these young patients challenge the surgeon with limb salvage and reconstruction procedures. Since the early ’80s, it has<br />
been well known that conservative limb surgery for bone sarcomas does not reduce the possibilities of survival. Thus, in most<br />
centres with experience, conservative limb surgery is the first treatment option. This approach, however, is much more<br />
disputed in the case of child patients. Conservative surgery in children is a challenge for the orthopaedic surgeon because<br />
of the small size of the patients and, the possibility of secondary limb length discrepancies and functional requirements.<br />
Surgery is carried out in two steps. First, there is en bloc resection. The second step is reconstruction. The different types of<br />
limb salvage surgery can be classified by location of the tumour: Joint resections or diaphysis resections. According to the<br />
implanted material, reconstructions can be: Biological (allografts or autografts), non-biological (megaprosthesis or<br />
expandable prosthesis) or combination of prosthesis and allograft. Growth plate represents a temporary barrier to the<br />
tumour spread. When the tumour does not affect the articular end of the bone, it is possible to conserve the joint. This has<br />
numerous advantages for the patient. In the case of paediatric patients, the advantages are even greater, because the longterm<br />
problems of prosthetic reconstruction in children can be avoided and because conservation of the growth cartilage<br />
makes it possible to avoid many subsequent problems. Loss of the joint (as occurs, by definition, under any articular resection)<br />
can later lead to a significant degree of functional deficit. It was for this reason, to avoid loss of the joint, that Professor<br />
Cañadell developed, in 1984, a technique to allow conservation of the articulation and with it conservation of a great part<br />
of growth potential: epiphysiolysis before excision of the tumour. When the joint surface needs to be resected in children,<br />
an osteoarticular allograft can be used for reconstruction. This reconstruction method can be used as a temporary solution,<br />
while the patient is still growing.<br />
Chemotherapy and radiation therapy have a negative influence on the healing of the allograft. The average consolidation<br />
time for a metaphyseal osteotomy is 6.5 months. In contrast, for a diaphyseal osteotomy, consolidation takes on average<br />
16 months. Cortical bone allografts are associated with a higher rate of complications than those of cancellous bone.<br />
Stabilisation of the graft must be sufficiently secure to allow consolidation. Depending on the location, there are different<br />
techniques: Kirschner wires, osteosynthesis plates, intramedullary nails, etc. At the metaphyseal level, it has been<br />
demonstrated that consolidation takes place quite easily whichever method is used, and so complex and laborious syntheses<br />
are not necessary; crossed Kirschner wires are sufficient in most cases. The younger the patient, the faster is the consolidation<br />
of the allograft.<br />
When possible, preservation of the growth cartilage will avert the problem of later dissymmetry. Note that graft osteosynthesis<br />
(if this method of reconstruction is used) must respect the physis if we wish growth to continue. For tumours that are near to<br />
or even in contact with the physis, eipiphysiolysis prior to resection can conserve the greater part of potential for growth.<br />
Finally, if resection implies loss of the growth cartilage, we can employ a reconstruction method with an implant somewhat<br />
longer (2 or 3 cm at the most) than the resected piece, thereby diminishing the final dissymmetry. Sometimes, a complication<br />
can be made into an opportunity for elongation, for example, when a graft or insert has to be removed due to infection or<br />
fracture. Elongation can be through the patient’s healthy bone, or the reconstruction material can be substituted for a larger<br />
version and elongation applied through the soft parts.<br />
These grafts are usually extracted such that a part of each tendon is left, for example, the insertion of the gluteal tendons,<br />
rotator cuffs or patellar tendons, for soft tissue reconstruction.<br />
In special locations, such as foot bones, allografts can also be used for reconstruction after resection of tumours.<br />
Conclusion:<br />
In the last few years it has become clear that in most paediatric cases of bone tumour it is possible to use reconstructive<br />
surgery with limb preservation and thereby improve these patients’ functionality and quality of life.
SKIN GRAFTING FOR BURNS – THE PARTICULAR CHALLENGES OF GROWING<br />
CHILDREN<br />
HERSON, M. Australia.<br />
Quite familiar is the saying: “children are like small adults”. Therefore, it should come of no surprise that children, similar<br />
to any other age group, become highly vulnerable to infection and prone to intense fluid depletion at the loss of continuity<br />
of the skin envelope. The difference lies in the intensity – that is when age and ‘size’ matters and responses are more<br />
dramatic.<br />
Unfortunately, children remain as the largest age group prone to burns, quite often a result of a minute of parental distraction<br />
in the home setting. Emotions become high, challenges in treatment are multiple, and end functional or esthetical results quite<br />
often concerning, if not disappointing.<br />
Burns scar for life – and children who survive severe burns, become otherwise healthy adults who have the largest part of<br />
their lives ahead of them. To compound the problem, different from the adult population children are, inevitably emotionally<br />
immature and ...must grow. Whilst within time, some scarring may subside into acceptable aesthetic levels, hypertrophic<br />
scarring and keloids particularly around articular surfaces may conform into true armours which prevent normal development<br />
and function. Progressive restriction of movement occurs, leading to severe functional deformity if not growth impairment.<br />
The ultimate consequence is social isolation.<br />
The medical experience in the 50’s Korean War increased the understanding around fluid replacement in the hypovolemic<br />
shock phase – this was a milestone in improved survival rates. Additional gains derived from care in specialised Units and<br />
the advent of topical anti microbials. However, the next important step in the survival rate increase was the acknowledgment<br />
of the benefits of early removal of damaged tissue. This has changed the way burns are managed today, in particular large<br />
ones. However, aggressive removal of tissues bring the need to cover the newly opened wounds; as alternatives were looked<br />
into, banked human skin became the gold standard as temporary wound cover.<br />
Skin allografts can be applied immediately after in depth wound cleaning or surgical debridement and become accepted<br />
as “own skin”. Once applied to the wound, the ensuing renewed barrier promotes modulation of the healing response,<br />
reduction in fluid loss and in bioburden. By protecting the exposed nervous terminals, pain is alleviated. Children, of all<br />
patient populations, can highly benefit from this almost collateral effect, as has been described by the Dutch in the use of<br />
glycerol processed allografts as coverage for deep, and painful, second degree burns.<br />
Unfortunately, within a couple of weeks of transplantation, allogeneic tissues are recognized by the immunocompetent<br />
recipient as ‘non-self’, and rejection is triggered manifested as sloughing of the epidermal component and absorption of<br />
the dermal component. This apparent tissue dissolution has made surgeons to consider skin allografts to be, albeit the best<br />
possible, only temporary wound dressing alternative.<br />
As burn victims survive the initial trauma, wounds start to heal. Wound closure can become complete either through<br />
spontaneous re-epidermisation of more superficial (1 st degree) burns, or by permanent replacement of damaged skin by auto<br />
grafting. Left to Nature, deep wounds will heal through a combination of re-epidermisation from the healthy borders and<br />
most important, through wound contraction; the end result will involve various degrees of hypertrophic scarring and wound<br />
bed contracture.<br />
Is allograft skin truly anything else but a ‘temporary wound cover’?<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
Increasingly, there is the recognition that this may not be all the truth. Despite the apparent liquefaction of the allograft as<br />
the manifestation of the rejection phenomena, there seems to be a degree of allo dermal collagen integration into the wound.
INVITED<br />
SPEAKER<br />
LECTURES<br />
Areas treated with allografts have lower tendency to hypertrophic granulation tissue and following auto grafting seem to<br />
evolve in the long term, as more pliable and contour improved areas. Potentially, the increased elasticity leads to less<br />
contractures. Therefore, improved outcomes of healed burn wounds closure seem to have a direct correlation between the<br />
presence, or absence, of dermal components.<br />
This being the case and in the premise that dermal elements of the allografted skin can become permanently incorporated<br />
to the wound bed, allogeneic skin grafts can be processed and its use expanded from the concept of a just very sophisticated<br />
‘like for like’ temporary wound dressing into a regenerative matrix of the so welcome neo dermal component.<br />
Biotechnology enhanced products will eventually supersede traditional banked split-thickness human skin allografts. However,<br />
for the moment being, allogeneic skin tissue remain as a most versatile alternative where expanding it use from mere wound<br />
dressings into providers of essential elements for dermal regeneration may be of particular value in growing children.
CARDIOVASCULAR (I)<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
REPORT FROM THE CONSENSUS MEETINGS OF EUROPEAN CARDIOVASCULAR<br />
TISSUE BANKS<br />
Theo de By 1 , Roland Hetzer 1 , Robert Parker 2 and Carl MacDonald 3<br />
1 Foundation of European Tissue Banks, Berlin, Germany. 2 Royal Brompton Cardiovascular Tissue Bank, London, UK.<br />
3 National Bacteriology Laboratory NHS Blood and Transplant, Colindale, London, UK.<br />
The report with respect to the Consensus Meetings of European Cardiovascular Tissue Banks comes in four sections:<br />
1. General purpose of the consensus meetings<br />
2. The surgeon’s evaluation of the importance of the cardiovascular tissue banking methods<br />
3. Directory of European Cardiovascular Tissue Banks and overview of addresses world wide<br />
4. Decontamination methods in cardiovascular banks, validation, science and consensus<br />
1. General purpose of the consensus meetings<br />
Consensus meetings of cardiovascular tissue banks have been held since the 1990’s. Its purpose is for cardiovascular tissue<br />
banks to compare and discuss the many different methods that are used for standardization. The methods discussed cover<br />
the entire range of the tissue bank process, from donor recruitment and selection criteria to thawing and implantation.<br />
After an interval of some years, The Foundation of European Tissue Banks decided to revive these useful meetings. As a result,<br />
2 meetings took place in 2011; one of these was entirely focused on prevention of contamination, microbiology detection<br />
methods and different decontamination processes. Thirty-six professional representatives of 22 cardiovascular banks<br />
contributed to the programs and discussions of those consensus meetings.<br />
The result was a “cross fertilization” of insights, knowledge, evidence, experience and learning, and a certain degree of<br />
consensus has been achieved. Summaries of the meetings were made available to the participants and ensured progress<br />
in the consensus, aimed at continuous improvement of cardiovascular tissue banking methods.<br />
The Foundation of European Tissue Banks intends to continue the meetings in 2012 and subsequent years.<br />
2. The surgeon’s evaluation of the importance of the cardiovascular tissue bank methods<br />
Unfortunately, some surgeons may take it for granted that cardiovascular tissue grafts of good quality are available for<br />
those patients for whom a homograft is the implant of choice.<br />
Demonstrating the experience with the treatment of endocarditis patients and right ventricular outflow tract reconstruction<br />
in infants and children at the German Heart Institute in Berlin, it becomes clear that a good communication between surgeons<br />
and the cardiovascular tissue bankers contributes to a better understanding of the surgical needs and a continuous<br />
improvement of methods and specifications issued by the tissue bank. For the treatment of active endocarditis, infecting the<br />
patient’s heart valve(s), a combination of a treatment with antibiotics, combined with surgery is the effective method of<br />
choice. The specification and characteristics of the homografts used by the surgeon must be standardized, and predictable<br />
in terms of short- and long term outcome. The cases of endocarditis at the German Heart Institute show a great variety in<br />
morbidity. Most homografts, implanted in those patients, were tailored according to the patients’ needs. As the homograft<br />
itself is not produced as an industrial product, both qualitatively and quantitatively, communication between tissue bankers<br />
and surgeons is of high importance. Besides standardizing information from tissue bank to surgeons, the feedback of the<br />
surgeons to the tissue banks is highly necessary for tissue bankers to improve methods where possible and to anticipate the<br />
surgeon’s future needs. The Foundation of European Tissue Banks brings both together to achieve a consensus about the<br />
best methods for the benefit of our patients.<br />
3. Directory of European Cardiovascular Tissue Banks.<br />
and overview of addresses world wide.<br />
Bringing together of cardiovascular tissue bankers requires an overview of the organisations involved. Prior to and during<br />
the consensus meetings the need was felt to create a directory in which we could easily identify the different tissue bank<br />
organisations as well as colleagues in Europe and other parts of the world. For the first time we have been able to compose<br />
such a directory for publication.
INVITED<br />
SPEAKER<br />
LECTURES<br />
The template for our Directory was generously provided by the EEBA, thus it follows more or less the same format. There is<br />
an overview per European country, including the tissue banks which have been identified.. In addition, the Directory contains<br />
specific statistics provided by 19 cardiovascular tissue banks in 10 countries. The numbers provided by the tissue banks cover<br />
the use of about 1700 hearts during the period 2007-2010. The responding banks issued 1486 homografts in 2010. Of<br />
these grafts 34% were aortic and 66% were pulmonary valves.<br />
Statistics about donor age, microbiology and serology do give an insight in the dynamics influencing the results of the<br />
contributing tissue banks. The addresses of colleagues world wide have been listed, as far as we could trace them.<br />
The Directory is clearly a first edition. We had to work with the data that were graciously made available by our colleagues.<br />
We realize that this edition is not yet optimal. The Foundation of European Tissue Banks strives to issue a second edition in<br />
2012.<br />
4. Decontamination methods in cardiovascular banks, validation, science and consensus<br />
Comparing the methods of detection, decontamination and validation used by 17 European Cardiovascular Tissue Banks,<br />
it was learned that there is an existing wide spectrum of methods. At the Consensus Meetings, organized by the Foundation<br />
of European Tissue Banks, it became clear that there is a large variation both in use of antibiotics as well as in concentrations<br />
of these antibiotics and the time period over which these antibiotics are used. Some of these methods are supported with<br />
scientific studies, others with extensive validation methods as used in the pharmaceutical industry. Those cardiovascular<br />
banks that had not yet documented the rationale of their methods have certainly been able to benefit from the work done<br />
by their colleagues. It has become clear that even if methods are different, they may lead to the same desired result, namely<br />
an allograft without micro-organisms which could infect the recipient. The exchange of methods and results will be continued,<br />
a.o.? by setting up an exchange of samples? between cardiovascular banks and their microbiology laboratories, to further<br />
prove the effectiveness of the different decontamination methods.
BRITISH ASSOCIATION FOR TISSUE BANKING CARDIOVASCULAR MICROBIOLOGY<br />
SURVEY 2010<br />
R.J. Lomas, Affiliation: National Health Service Blood and Transplant Tissue Services, 14 Estuary Banks, Liverpool L24 8RB,<br />
United Kingdom.<br />
Abstract:<br />
In 2010, the Cardiovascular Special Interest Group of the British Association for Tissue Banking (BATB) surveyed the<br />
microbiology testing, decontamination and evaluation practices of its members. This survey was subsequently extended to<br />
heart valve banks in Europe, North America and Australia. It addressed different aspects of microbiology protocols in detail:<br />
How banks tested the microbiological status of allografts both pre and post decontamination<br />
How banks decontaminated allografts<br />
How banks determined if allografts were suitable for clinical use based on microbiological data<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
The survey was circulated using an online survey tool, although written responses were also facilitated. It was carried out<br />
under the promise of anonymity to encourage participation.<br />
Initially, fourteen responses were received. These were reviewed and summarised, and the outcome report was circulated<br />
to all participants in May 2011. Two further responses were received after this date, and the report updated accordingly in<br />
October 2011. Copies of the report have been circulated to all banks that participated in the survey, and may be obtained<br />
from the author on request.<br />
The objective of the survey was to collate information on microbiology protocols used in different banks, so that banks could<br />
review the protocols used by their peers and determine where improvements in their own practises might be implemented.<br />
The collated data revealed considerable differences in decontamination, testing and acceptance protocols, even between<br />
banks operating in the same country. This highlights the need for close liaison between heart valve bankers, leading ideally<br />
to international harmonization of microbiology protocols to agreed best practice.
DEVELOPMENT OF UNIVERSAL CARDIOVASCULAR NOMENCLATURE<br />
Izabela Uhrynowska-Tyszkiewicz (1), Ramadan Jashari (2)<br />
(1) Medical University of Warsaw / National Centre for Tissue and Cell Banking, Warsaw, Poland<br />
(2) European Homograft Bank, Brussels, Belgium<br />
Aim:<br />
The development of universal cardiovascular nomenclature is one of the current aims of ICCBBA.<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
Background:<br />
ICCBBA is an international, non-governmental, non-profit, information standards organization which manages the ISBT<br />
128 standard - a global standard for the identification, labeling, and information transfer of human blood, cell, tissue, and<br />
organ grafts across international borders and disparate health care systems. The organization is staffed by eight, has a volunteer<br />
Board of Directors from around the world, and includes various technical advisory groups (TAGs) made up of 270+<br />
volunteer experts. There are 3 TAGs which deal with tissues: North America Tissue TAG (NATTAG), European Tissue TAG<br />
(ETTAG) and Eye Bank TAG (EBTAG).<br />
Methods:<br />
At the beginning of 2011 two Joint ETTAG-NATTAG Terminology Working Groups have been established to ensure a universal<br />
approach during revision of existing ISBT128 tissue graft terminology (Joint ETTAG-NATTAG Terminology Work<br />
Group for Cardiovascular Tissues and Joint ETTAG-NATTAG Terminology Work Group for Tendons). All the terminologies<br />
developed through Joint ETTAG-NATTAG Terminology Working Groups will be public and published on the ICCBBA website<br />
in a standards terminology document. The developed terminologies can be used for any coding system including ISBT<br />
128. Therefore in order to ensure as global approach as possible the experts not only from Europe and North America but<br />
from other parts of the world (South America, Asia-Pacific region, Australia) were invited to participate in both groups. The<br />
groups work by consensus. Meetings are held through conference call and e-mail discussions. Anticipated time to complete<br />
each terminology is twelve months.<br />
Results:<br />
To date, 14 individuals form 8 organizations were interested to offer their time and knowledge by taking part in cardiovascular<br />
project. They met 2 times by teleconference calls and stared developing a consensus for class and attribute definitions<br />
of cardiovascular grafts.<br />
Summary:<br />
The Joint ETTAG-NATTAG Terminology Work Group for Cardiovascular Tissues encourages everyone who has an interest<br />
in their work to participate in calls and meetings. To learn more, contact the ICCBBA office.
REPRODUCTIVE<br />
THE POWER OF DEMOGRAPHY. HOW DEMOGRAPHY DYNAMICS AFFECTS<br />
SOCIETY, POLITICS AND ECONOMY. THE PARTICULAR PROBLEM OF ART<br />
J. de Mouzon, MD, MPH, EIM past chairman<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
Introduction:<br />
Access to assisted reproductive technology (ART) shows huge variations in the World, from a few cycles to several thousands<br />
per million in other countries. These differences can be viewed as inequity for couples suffering of infertility in those countries<br />
where access is low, and would need to be considered by health authorities and national policies. Many factors can play<br />
a role on the access, in particular, demography. But demography also interacts with economic and other societal aspects.<br />
METHODS: ART data were obtained from World and EIM-ESHRE registries on ART, in which data are collected from<br />
established national or regional registers of various origin. Demographic data were obtained from the U.S. Census Bureau,<br />
Population Division, and the<br />
CIA World fact book, in population section, through their website portal. Economic data were obtained in World<br />
development indicators (World bank). The analysis concern 2007 data from 52 countries participating in the ART registries.<br />
They belong to Europe, North and Latin America, Asia, Australia / New Zealand. Analyzed demographic indicators were<br />
the total fertility rate (total number of children per woman at the end of reproductive age), the mean women’s age, the<br />
proportion of women of reproductive age (15-45 years) the women expectancy of life, infant mortality. The economic<br />
variables considered were the gross domestic product, total and per capita, the funding of ART cycles and the distribution<br />
of Public/Private Health Expenditure.<br />
Results:<br />
Human reproduction has to be considered in a general context of decreasing fertility rates, not only in developed countries,<br />
but also in developing ones, this being probably pat of the demographic transition. However, the fertility rate is generally<br />
higher in countries with a low mean women’s age, high proportion of women aged 15-45, a low life expectation, high infant<br />
mortality, and low GDP. On the opposite, access to ART is high in countries with high women’s age, low proportion of<br />
women aged 15-45, a high life expectation, low infant mortality, and high GDP. ART access is also much higher when<br />
access is free or reimbursement important and in countries where the general balance between public and private health<br />
expenditure is in favour or public.<br />
Conclusions:<br />
Fertility decreases in all countries, and particularly in developed countries (
INVITED<br />
SPEAKER<br />
LECTURES<br />
VIRAL TESTING OF PARTNER GAMETE DONORS – TIME FOR A CHANGE TO EU<br />
REQUIREMENTS?<br />
Anne Cathrine Bollerup, MD, Danish Medicines Agency, Denmark, Peter Saugmann-Jensen, MD, National Board of<br />
Health, Denmark<br />
Introduction:<br />
The European Tissues and Cells Directive (EUTCD) 2006/17/EC, Annex III defines the requirements for partner donation<br />
(not direct use). The text states that donors (both members of the treated couple) of reproductive cells, which are processed<br />
and/or stored and reproductive cells that will result in the cryopreservation of embryos must be tested for HIV, Hepatitis B<br />
and C at the time of donation to assess the risk of cross-contamination.<br />
Background:<br />
Many couples treated with medically Assisted Reproductive Technology (ART) receive more than a single transferral of<br />
gametes and embryos. In 2009 the European practices were discussed at a meeting of Competent Authorities for Tissues<br />
and Cells, with a view to establishing the level of compliance with the requirements of Annex III. The collated information<br />
from various Member States (MS’s) of the EU revealed different interpretations and national practices with regard to the time<br />
frame and frequency of testing. Only a few MS’s had implemented testing literally at each transferral event (i.e. ‘each<br />
donation’). The majority reported testing every 6, 12 or 24 months e.g., Denmark interprets “donation” as comprising of a<br />
sequence of procurements events. A consecutive series of such events within the same clinic, and within the same healthy<br />
and tested couple, is regarded as part of one treatment regime and donation; so long as the treatment regime does not<br />
exceed a twenty-four month period.<br />
The MS’s also noted that testing at each time of donation is costly. Since a typical fertility treatment may extend over a 1 –<br />
2 years period, with treatment intervals rarely exceeding 4 months, it is projected that each partner in the fertility treatment<br />
would have to be tested between 3 to 6 times. In addition, the high frequency of testing in presumed healthy couples, where<br />
the “risk behaviour” has been excluded by health professional interrogation, were seen as disproportionate compared with<br />
the risk reduction; where the latter being assessed as marginal or nil, therefore not adding appreciably to overall safety.<br />
However, the minutes of the Commission meeting reflect the Commission’s view the Directive was clear, and that MS’s not<br />
testing at ‘each donation’ were in infringement of the regulations.<br />
On the above background, a Working Group in which participated three Competent Authorities (Denmark, Ireland and<br />
Belgium) and ESHRE (European Society of Human Reproduction and Embryology) was set up by the Commission in 2009.<br />
Its defined scope was to collect and analyse the available evidence-based data to formulate a modified proposal of a testing<br />
protocol – for partner donations - based on current scientific data. The working group was tasked to provide a comprehensive<br />
overview and a quantitative assessment of the potential risks related to quality and safety, if the testing frequency was<br />
reduced from once per donation, to once per year or once per 2 years. Areas of identified interest were the probability of<br />
infection of embryos, the potential or known cases for transmitting HIV, Hepatitis B and C during processing, preservation<br />
and cryo-storage, as well as the potential mix-up of gametes. In connection with this work, ECDC (European Centre for<br />
Disease Prevention and Control) was commissioned to perform a study and make a report, including risk assessments of<br />
various regimes of testing requirements for reproductive cells in partner donation, from ‘each donation’ to ‘periodic testing’<br />
(e.g. every 12 or 24 months).<br />
Methodology:<br />
The ECDC risk assessment was based on a model for the estimation of residual risk (RR) using available estimates of<br />
prevalence and incidence of the infections concerned. The residual risk is typically expressed as the probability of an infected<br />
donation being used or as the number of donations that need to be screened before one is missed. The residual risks under<br />
the different testing procedures were compared to estimate the change in risk.
A literature search of published studies on viral infections among MAR service patients and the risk of cross-contamination.<br />
Thereafter estimating the risk was based on the experience of the infection of the partner, when there is a test error or seroconversion<br />
in the patient; infection of the non- partner in case of sero-conversion and mix up of gametes; infection of the<br />
non- partner in case of sero-conversion and transmission of virus during cryo-storage of embryos, as well as infection of<br />
the non- partner in case of sero-conversion and embryos infecting embryos.<br />
Results:<br />
The residual risk of missing a case with the current testing requirements<br />
■ HIV: 17.83 undetected cases / million person years i.e. one case missed for every 56,101 tests<br />
■ HBV: 32.08 undetected cases / million person years i.e. one case missed for every 31,172 tests<br />
■ HCV: 267 undetected cases / million person years i.e. one case missed for every 3,745 tests<br />
There was only a modest increase in the residual risk, when comparing the testing every 6 months, 12 months or 24 months<br />
with the base case of testing at each donation (which was assumed to take place in 4 month intervals)<br />
The absolute numbers of additional cases missed when applying 24 month schedule were found to be:<br />
■ HIV: 0.46 more cases /million person years<br />
■ HBV: 1.93 more cases / million person years<br />
■ HCV: 8.6 more cases / million person years<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
The major contribution of risk comes from the prevalence, and this is practically removed in all of the potential scenarios by<br />
performing the first test itself.<br />
Among ~14000 MAR service patients, including some patients found to be HBV-, HCV- or HIV –positive, the occurrence of<br />
cross-contamination in the ART facility, or the transmission of virus, and cross-contamination in storage, as well as horizontal<br />
or vertical transmission to a partner or neonate has never been documented. (Wingfield and Cottell, 2010).<br />
Additional information in published journals have shown the risk of cross-contamination during processing, cryo-preservation<br />
and storage of reproductive cells to be negligible or non-existing.<br />
Sero-conversion after tests of ~14.000 patients in Ireland were not seen in the study reported by Wingfield and Cottell. Risk<br />
of infecting the partner is much higher at home in an intimate relationship<br />
We have never seen a sero-conversion in a viral negative ART patient. In 2010, 3 cases of gamete mix up were reported<br />
out of 42.000 cycles in the UK. Transmission of a virus through an embryo has never been seen. (Risk= never x 1 / 14000<br />
x never)<br />
We have never seen a sero-conversion in a viral negative ART patient. We have never seen transmission of virus between<br />
embryos & gametes in cryo storage. We have never seen transmission of a virus through an embryo. (Risk = Never x Never<br />
X Never)<br />
We have never seen a sero-conversion in a viral negative ART patient. We have never seen airborne transmission of virus.<br />
We have never seen transmission of a virus through an embryo. (Risk = Never x Never X Never)<br />
What is the problem?<br />
■ Fertility clinics work with patients screened for infection<br />
■ For each couple it is known whether they are infectious or not<br />
■ More than 99 % of the patients seeking fertility treatment are free from HIV and Hepatitis virus<br />
■ All patients are tested e.g. ~100 % of patients in the normal ART program are free from HIV and Hepatitis virus<br />
Conclusion:<br />
Based on the results presented at the meeting for national competent authorities (NCA’s) for tissues and cells in June 2011<br />
the NCA’s group concluded that it was not needed to maintain the current testing requirements for partner donation as laid<br />
down in Annex III of Directive 2006/17/EC, seeing a testing regimen of up to 24 months would not add significantly to risk<br />
of transmission of infection. Taking this formally into account would require an amendment of the Directive, through a<br />
regulatory procedure. It is likely the proposed amendment to Directive 2006/17/EC may be formalised at the end of this<br />
year, with its transposition into national regulations taking a little longer.<br />
From a financial perspective there are considerable cost savings in relation to testing for HIV, Hepatitis B and C for partner<br />
donors of reproductive cells in fertility treatment in Denmark and in the whole Europe.
INVITED<br />
SPEAKER<br />
LECTURES<br />
REFERENCES<br />
1. Directive 2006/17/EC of 8 February 2006 implementing Directive 2004/23/EC of the European Parliament and of the<br />
Council as regards certain technical requirements for the donation, procurement and testing of human tissues and cells.<br />
2. ECDC, Risk assessment on change of testing requirements for reproductive cells in partner donation in Technical report,<br />
M. Salminen et al., 2011 European Centre for Disease Prevention and Control: Stockholm<br />
3. M. Wingfield and E. Cottell. Viral screening of couples undergoing partner donation in assisted reproduction with regard<br />
to EU Directives 2004/23/EC, 2006/17/EC and 2006/86/EC: what is the evidence for repeated screening? Hum<br />
Reprod, 2010
SKIN AND AMNIOTIC MEMBRANE (I)<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
CHALLENGES ON AMNIOTIC MEMBRANE - PROCESSING AND CLINICAL USE.<br />
Koller J., Comenius University School of Medicine and University Hospital Bratislava, Burn Department and Central Tissue<br />
Bank, Slovakia<br />
Introduction:<br />
Human foetal membranes (HFM), from an anatomical point of view, represent two loosely connected membranes engulfing<br />
the human foetus during embryogenesis and organogenesis until delivery. The inner, thinner, strong and shiny membrane<br />
is the amnion (HAM), whereas the outer, thicker, less homogenous and dull membrane is the chorion (HCHM). Processing<br />
of HFM for use as a biological skin substitute/dressing was first described back in the 19th Century by Davis (1910) and<br />
Sabella (1913). Currently HFM, particularly HAM. are used as biological dressings or carriers for different purposes.<br />
Material and Methods:<br />
This presentation is aiming to review the relation between anatomical structure, biomechanical properties and content of<br />
various active substances included in HAM to functionality of HAM used in various clinical and therapeutic indications. In<br />
addition to approved methods of clinical applications of HAM, progress in research, particularly development of more<br />
sophisticated both somatic and embryonic in vitro cell culture methods, new horizons for enlarged use of HAM in research<br />
as well as clinical practices are going to be opened.<br />
Current processing and clinical use:<br />
Availability of HAM is good, as they are procured from living donors at childbirth. Donor screening and testing are provided<br />
according to current European regulations. Procurement is done either in clean environment such as delivery rooms, or<br />
under aseptic conditions during Caesarean sections at operating theatres. The procured material is transported in sterile<br />
containers to tissue establishments for processing. Processing is provided according to standard operative procedures applied<br />
in particular establishments. Preservation can be performed by air drying, freeze drying, deep freezing, or glycerolization.<br />
In case where clean non-sterile processing is provided, terminal sterilisation by irradiation, or other validated method is<br />
mandatory.<br />
Current clinical use of HAM include treatment of burns, skin donor sites, different both acute and chronic wounds, ocular<br />
diseases and injuries and as a separating membrane to prevent adhesions in various surgical situations.<br />
Challenges and perspectives:<br />
In the recent few years a lot of research is going on in the use of HAM as a source of pluripotent stem cells, as a source of<br />
growth factors and cytokines for wound healing purposes, and as a carrier of in vitro cultured cells for clinical applications<br />
in both ophtalmology and plastic surgery.<br />
Summary:<br />
HAM have been used clinically for already more than one century in various clinical situations with good results. Their<br />
beneficial effects include barrier function, wound protection from outer environment and infection, wound preparation for<br />
optimal take of skin grafts, pain reduction, enhancement of angiogenesis, epithelialisation and reduction of scarring. Their<br />
advantages include availability, simple procurement, preparation and preservation methods, low antigenicity, and, last, but<br />
not least, relatively low costs.
LEARNING FROM BIOVIGILANCE<br />
THE NOTIFY PROJECT<br />
Alessandro Nanni Costa, Italian National Transplant Centre<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
From September 2010 to February 2011, the World Health Organization (WHO), the Italian National Transplant Centre<br />
(CNT) and the EU-funded Project ‘Vigilance and Surveillance of Substances of Human Origin’ (SOHO V&S) joined forces<br />
to organise a global initiative aimed at raising the profile of vigilance and surveillance of substances of human origin; the<br />
initiative was called Project NOTIFY. The scope of the project included organs, tissues and cells for transplantation and for<br />
assisted reproduction. Ten international expert groups were tasked by WHO with gathering documented cases of reactions<br />
and events across the scope of the substances under consideration, using published articles and vigilance system reports as<br />
their information sources. The work was conducted on a Google site where over 100 participants (regulators, clinicians,<br />
professional society representatives, scientific experts) inserted more that 1,500 published references and added comment<br />
regarding alerting signals and the methodology applied for confirmation of imputability. The cases were used as the basis<br />
for developing draft guidance on detection and confirmation of reactions and events, with an emphasis on the key role of<br />
the treating physician.<br />
The NOTIFY project culminated in a meeting that took place in Bologna from February 7 th to 9 th 2011. The 113 invited<br />
experts from 36 countries represented regulatory and non-regulatory government agencies, professional societies and<br />
scientific and clinical specialities from all WHO regions. The meeting was made possible with funds raised by CNT together<br />
with those allocated within the SOHO V&S project for an international meeting on vigilance reporting and investigation.<br />
The meeting explored the work already carried out on-line and agreed on priorities for the future development of global<br />
V&S for organs, tissues and cells.<br />
From the meeting, the Bologna Initiative for Global Vigilance and Surveillance (BIG V&S) was established and a number of<br />
outcomes will result. A detailed report of the meeting is in the process of publication, together with 5 didactic guidance<br />
documents, each one addressing one category of adverse reaction or event. The SOHO V&0S project is using the data and<br />
commentary in the development of its guidance for tissue and cell V&S in the EU. WHO will publish a booklet for clinicians<br />
that will summarise the guidance on detection and investigation of adverse reactions and events that was developed by<br />
project Notify.<br />
A new dedicated website is being developed by CNT, as part of a sustained collaboration with WHO, for the promotion of<br />
V&S. The site will host the Notify compendium of adverse incident cases in a database that will be publicly accessible. These<br />
cases, and new cases as they arise, will be posted on the site using key words and a minimum data set which will enable<br />
searching by, for instance, type of human substance, type of infectious disease transmission agent, type of logistical error.<br />
The tool will be a source of information for clinicians, potential donors and patients who wish to understand better the risks<br />
associated with particular types of donation or human application; for professionals who need information when deciding<br />
on the suitability of a potential donor and for regulators who need information on previous experiences of specific types of<br />
reported events and reactions.<br />
An international Editorial Board with regulatory and professional representatives from the fields of organs, tissues and cells,<br />
has been established to oversee the work of the new website and to take forward the other outputs of the Bologna Initiative<br />
including the development of correspondence tables for terminology and agreement on common definitions, where possible.<br />
It is intended that this initiative will facilitate global sharing of V&S information and guidance for the enhancement of donor<br />
and recipient safety and for greater public transparency in transplantation and assisted reproduction.
VIGILANCE AND SURVEILLANCE OF SUBSTANCES OF HUMAN ORIGIN - THE SOHO<br />
V&S PROJECT<br />
Deirdre Fehily PhD, Italian National Transplant Centre.<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
The project ‘Vigilance and Surveillance of Substances of Human Origin’ (SOHO V&S) was launched in March 2010 and<br />
will run for 3 years, co-funded by the European Union Public Health programme. The key objective is to develop a shared<br />
view of how serious adverse events and reactions associated with tissue and cell donation or human application should be<br />
reported, evaluated and investigated within the EU. A series of work packages address specific areas of concern: vigilance<br />
in assisted reproduction, vigilance for living donors, how investigation of adverse incidents should be performed, the<br />
promotion of vigilance and surveillance among clinicians and the detection and prevention of illegal and fraudulent activity.<br />
The project aims to facilitate harmonisation of terminology and documentation and consensus on how information should<br />
be exchanged between EU Member States, the European Commission and third countries.<br />
The project is co-ordinated by the Italian National Transplant Centre (CNT). It has a Steering Committee and a large number<br />
of collaborating partner organisations, including all of the major European professional societies in the field and a number<br />
of professional. The involvement of the World Health Organisation and many collaborating partners, both professional<br />
societies and regulators, from outside the EU ensures that the guidance developed in this project reflects international needs<br />
and realities, in the context of global movement of human tissues and cells for human application.<br />
Although the EU Directives include gametes and embryos in their scope, it emerged during the EUSTITE project that the<br />
Directive definitions and the EUSTITE vigilance tools were not fully adapted to the field of assisted reproduction and the<br />
reporting requirements are interpreted in different ways in Member States. A SOHO V&S work-package led by the French<br />
Biomedicine Agency has explored these issues with the active participation of the European Society for Human Reproduction<br />
and Embryology, ESHRE. The group has amended the EUSTITE vigilance tools to make them more relevant to the field and<br />
has developed specific guidance for EU vigilance and surveillance in assisted reproduction. The document has been submitted<br />
to the European Commission and the tissue and cell Competent Authorities of the EU.<br />
Up to now, most EU Competent Authorities for tissues and cells have focused their efforts on putting in place systems and<br />
procedures to implement the regulatory functions that are required by the tissues and cells Directives, notably inspection,<br />
authorisation and vigilance. Many, however, lack experience and training, as well as procedures to be followed, for the<br />
investigation of cases where illegal or fraudulent activity is suspected. A SOHO V&S work-package led by the French<br />
Agency for the Safety of Health Products has gathered information from Member States and non-EU countries on cases that<br />
have been investigated and concluded, in some cases with enforcement action. The work-package has drawn on this<br />
experience to developed guidance, including tools and recommendations, to support EU Member States in this particular<br />
area of work.<br />
Guidelines on the investigation of adverse reactions and events will be delivered by two work-packages in SOHO V&S. CNT<br />
is leading a work-package developing guidance for authorities and the Polish partner is leading a work-package developing<br />
guidance for hospitals and clinics where tissues and cells are applied to patients. Both groups will draw on the outcomes<br />
of the NOTIFY project, an initiative led by the World Health Organization in which this project participated, where experts<br />
from across the globe gathered information on documented cases of adverse reactions and events for organs, tissues, cells,<br />
gametes and embryos, drawing on the data to develop didactic guidance. The database created by the project will be<br />
publicly available as an interactive searchable tool on a dedicated website, hosted by CNT on behalf of WHO, and will<br />
provide data on how different types of reactions and events have been detected and confirmed. The new website with the<br />
database will be publicly available early in 2012 (www.notifylibrary.org).<br />
In the later stages of this three-year project, the Irish Medicines Board will lead a work-package delivering training for EU<br />
Vigilance officeres, based on the various principles of good practice identified and documented by all of the work-packages.<br />
The courses will be delivered following the successful model developed in the EUSTITE project, with a combination of<br />
e-learning followed by a residential module.
An effective vigilance and surveillance system plays a pivotal role in enhancing the safety of tissue and cells for human<br />
application. In some cases it facilitates rapid intervention by professionals or regulators to prevent further harm to patients.<br />
In general, it ensures the sharing of invaluable information to support improvements in systems and procedures for the<br />
benefit of donors and patients. The SOHO V&S project aims to maximise this learning opportunity through international<br />
collaboration between regulators and professionals.<br />
Further information and updates can be found at www.sohovs.org<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
The SOHO V&S project is supported by co-funding from the Department of Health and Consumer Protection of the European<br />
Commission. Grant agreement n. 20091110
GLIMPSE INTO THE FUTURE; ADVANCED THERAPIES (I)<br />
TISSUES AND ATMPS; MARRIAGE OF CONVENIENCE. AIMING A UNIQUE QUALITY<br />
STANDARD<br />
Tabera J., Vilarrodona A., Trías E.<br />
Transplant Services Foundation-Hospital Clínic. Barcelona. Spain.<br />
Introduction:<br />
The Transplant Services Foundation (TSF) is a good example of organization that develops different activities under different<br />
regulations frameworks. Donation, procurement, evaluation, processing, preservation, storing, distribution and biovigilance<br />
of tissues and cells are regulated by the 2004/23/EC, 2006/17/EC and 2006/86/EC Directives, with the objective<br />
of guarantying their quality and safety, whereas the production of ATMPs (Advanced Therapy Medicinal Products) must fulfil<br />
the EU Good Manufacturing Practices (GMP) and are regulated by the European legislation that covers the Advanced Therapies<br />
and by the European rules for medicinal products.<br />
Aim:<br />
The purpose of this challenge is to assess the benefits and inconveniences on the integration of the tissues quality standards<br />
and the ATMPs ones.<br />
Methods:<br />
The starting point of the GMP requirements implementation was an ISO 9001 certification and a TE authorisation. Such implementation<br />
was planned through a risk assessment basis in order to assess the positive, negative or neutral impact of the<br />
diverse adaptations on the quality system in place. The requirements related to the Quality Management (Product Quality<br />
Review, Risk Management), organizational requirements related to the personnel roles (independency between Quality Control<br />
and Production), requirements on the design and use of facilities (A/B grade for aseptic processes, clean rooms and<br />
equipment qualification, microbes and airborne particles monitoring), requirements related to the documentation used (production<br />
and packaging batch records, archive), requirements related to production (processes validation), requirements related<br />
to quality control activities (good laboratory practices, stability studies), requirements related to outsourced phases and<br />
products and services suppliers (suppliers management, quality agreements), requirements derived from out of specifications<br />
products (recall and return systematic) and requirements related to the self-inspections (integrated checklist), were all of<br />
them analysed to determine if the implementation could be performed immediately, with some adaptations or were not suitable<br />
to implement.<br />
Hence, the study to assess the suitability of the Good Manufacturing Practices requirements implementation is based in a<br />
risk assessment methodology, assuming in all cases that the subsequent residual risk was acceptable. This analysis was combined<br />
with a change control exercise to assess the actions that should be taken and their impact.<br />
Results:<br />
In this practical approach, 23 requirements such as PQR, facilities monitoring during the process, media-fill validation, air<br />
quality, cleaning systematic, training requirements for the Responsible Person, relationship between production and quality<br />
control affairs, retention samples, on going stability studies, product specifications file/registration dossier, extensive clinical<br />
studies, batch definition, parametric release, investigational products, validation of processes, primary materials suppliers<br />
evaluation, archive retention period, traceability and biovigilance, have been assessed with the following summarized<br />
results:<br />
7 requirements were implemented without adaptations<br />
14 requirements were implemented with adaptations<br />
2 requirements were not implemented<br />
INVITED<br />
SPEAKER<br />
LECTURES
INVITED<br />
SPEAKER<br />
LECTURES<br />
Conclusions:<br />
The Transplant Services Foundation decided to extend the scope of the strict requirements to all the processes, understanding<br />
that it was the problem and the solution at the same time. Fulfil the GMP requirements in all the processes means a significant<br />
effort in terms of personnel and materials, but at the same time is understood as the solution to the coexistence of<br />
different quality standards.<br />
The fact of providing the Hospital Clínic de Barcelona (HCB) with facilities, personnel and a whole quality system that respect<br />
the GMP requirements allowing to develop and obtain the Advanced Therapy Medicinal Products, supposes the mandatory<br />
satisfaction of the needs arising from the scientific advance that houses and promotes the HCB. Furthermore, it allows<br />
merging in an optimal manner the steps of detection of the therapeutic need, development of the solution and the assessment<br />
of its efficacy.<br />
On the other hand, the bet to extend the scope of the certification, not only to the obtention of ATMPs but also to the tissue<br />
processing, answers to a realistic and regulatory need of the scenario where both therapeutic solutions do coexist.<br />
The need of adapting and completing the current regulation, to make it cover the process running from the donation till the<br />
tissular and/or cellular products obtention, is been identified by the experience of the TSF. To have an unique process covered<br />
by different regulations, that at the same time do not respond to the reality of both of them, is an unresolved matter<br />
within the European Union that arises as a immediate challenge for the TSF in line with its compromise to stand as a reference<br />
in the tissular and cellular field.<br />
The production of ATMPs is currently regulated within the European regulation of medicinal products and it is with no doubt<br />
and excellent reference for them to achieve an adequate grade of quality and safety. However, the coincidence on the initial<br />
steps of donation, with regard to the processing of tissues for transplant, the similarity between tissues and ATMPs concerning<br />
the specifications and quality controls needed, the unclear borders between the different types of tissues and ATMPs<br />
and the fact that both therapeutic solutions share actors and scenario in the moment of transplant and application, leads to<br />
believe in the necessity of a specific, adapted and integrated regulation to guarantee the c
ARTIFICIAL CORNEAL TISSUE: A NEW CHALLENGE<br />
Miguel González-Andrades. Spain.<br />
Corneal diseases are one of the most important causes of visual impairment in the world. Thus, many patients affected by<br />
this pathology end up needing a corneal transplant. This treatment implies two major problems: the lack of donors and the<br />
possibility of graft rejection. Therefore, this necessity has encouraged the generation of a corneal substitute in the laboratory.<br />
In order to accomplish this challenge, tissue engineering appears as promising science, whose aim is to generate artificial<br />
tissues and organs that can replace the damaged one within the human body. Regarding tissue engineering of the cornea,<br />
different approaches have been attempted.<br />
We present two models of artificial corneas based on fibrin-agarose scaffolds and acellular xenografts, developed by our<br />
research group. Both models are based on the combination of corneal cells and scaffolds. Corneal epithelial cells and<br />
keratocytes were obtained from sclerocorneal limbus of human cadavers. On the one hand, the fibrin-agarose model was<br />
developed seeding these cells into an artificial matrix, which was constructed with a mixture of agarose VII and fibrin from<br />
human donors. On the other hand, the acellular xenograft model was created applying a decellularization process to pig<br />
corneas, based on NaCl. After obtaining an acellular corneal stroma from pig corneas, the human keratocytes were seeded<br />
over it. Once we developed both artificial corneal models, histological, genetic expression profile and optical analyses were<br />
carried out.<br />
Both models showed a well-developed stroma based on the presence of collagen and proteoglycans. The keratocytes<br />
proliferated and spread, migrating across the corneal matrix. Some immunohistochemical assays were performed, showing<br />
the differentiation and characteristic expression of corneal proteins. Optical analyses revealed the high transparency level<br />
that both models presented, observing that UV-light was mostly absorbed by the corneal substitutes. All these results suggest<br />
that corneal substitutes made by tissue engineering show similar characteristics to human corneas. Thus, artificial corneas<br />
could represent a promising treatment for many corneal diseases that do not currently have an adequate and established<br />
therapeutic procedure.<br />
Supported by grant P10-CTS-6060 by Junta de Andalucia<br />
INVITED<br />
SPEAKER<br />
LECTURES
CORNEA (I)<br />
DEKARIS IVA<br />
European Eye Bank Association – EEBA<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
The European Eye Bank Association (EEBA) is a tehnical-scientific organization comprising individual members from 84 eyebanks<br />
from 24 European countries and 4 international colleague eye-banks. EEBA is today the leading pan-national<br />
association in Europe dedicated to the advancement of eye-banking and an authoritative reference point for eye banks<br />
wishing to work according to quality standards.<br />
Aims:<br />
The Association is formed: to contribute to the development of standards for eye banking in Europe, to establish an agreed<br />
set of EEBA Standards, to promote data collection on graft outcome, to facilitate the information interchange between eye<br />
banks, to provide opportunities of eye banking practice, to encourage relevant research and development, to provide<br />
informed comment to external agencies, to foster education in eye banking, to maintain national and international links with<br />
corneal transplant communities, to make knowledge of eye banking available to any person for the general good of society.<br />
Activities:<br />
In the period from 2005-2009 the number of corneas received in EEBA member eye-banks was 31587 (±3034) corneas.<br />
The overall percentage of corneas issued for transplanation in 2009 was 49% (lower as compared to previous 5 years); one<br />
hypothesis for this decrease may be the implementation of the „24h-regulation“ on taking blood samples in several EU<br />
countries, which causes a high loss of potential donor material. Most of the corneas were transplanted in the hospital housing<br />
the eye-bank or in the own country. According to their activities, expressed as number of corneas issued per year, 36<br />
European eye banks are issuing 100-500 corneas, 7 are issuing 50-100 corneas and 7 eye banks >1000 corneas. Of the<br />
received corneas, 47 % was retrieved by enucleation and 53 % by corneoscleral disc excision. Organ-culture remains the<br />
preferred storage methods in EEBA (71% of preserved corneas). In recent years EEBA Techinical guidelines have been<br />
developed and published describing selection criteria deemed acceptable for cornea evaluation. Of all corneas that were<br />
not selected for grafting, 16.2% were excluded because of abnormal morphology and other reasons were serology (7.9%),<br />
medical history (2.7%), microbiology (3.3%) and others. All banks test all donors for HIV Abs, Hepatitis B and C, either<br />
antigen, antibodies or both and most eye banks additionally test for syphilis. Mean frequency of positive serology ranges<br />
from 5.0-8.5% in the period from 2005-2009. In 2009, 16% of corneas was selected and issued for lamellar grafting, and<br />
sometimes lamellar grafts were pretreated in the bank. Only a small proportion of the transplanted corneas were tissue-typed;<br />
typing for HLA I was done for 10.4 % of all donors and for HLA II for 8.7 %. Adverse reactions (such as primary failures<br />
and endophthalmitis) occured at 0.2 and 0.05 % risk. Of other tissues used for ocular surgery EEBA banks were issuing<br />
sclera, limbal graft, stem cells and amniotic membranes.
EUROPEAN GOOD PRACTICE STANDARD<br />
EURO-GTP PROJECT<br />
Trias E., Zardoya M., Vilarrodona A., Tabera J.<br />
Transplant Services Foundation-Hospital Clinic. Barcelona, Spain<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
Introduction<br />
In Europe, there are currently neither common procedures for how the procurement, the processing and the preservation of<br />
tissues for transplant should be carried out, nor for how the donor selection and screening of tissue donors according to the<br />
type of tissue retrieved should be done.<br />
It is important that the methodology of tissue banking activities is harmonized among all tissue establishments in Europe so<br />
that high quality and the safety of transplanted tissues can be guaranteed. There are regulations which provide the basis<br />
for performing tissue banking activities in a professional and coherent manner. However, this project aims to go one step<br />
further, and provide the specific tissue practices (GTPs) to be followed in European tissue banks, in order to harmonize these<br />
decisive procedures. This will also contribute to a higher confidence in the exchange of tissues for transplant among the<br />
Member States. To guarantee the highest level of quality and safety of the tissue grafts for recipients it is very important to<br />
carry out the tissue banking procedures in adequate environmental conditions and facilities. The GTPs will be very helpful<br />
tools for all kinds of TEs and TEs that are in different phases as concerns their development and evolution.<br />
Objectives<br />
The main objective of this project is to develop common Good Tissue Practices (GTPs) for European tissue establishments (TE),<br />
as well as a Training Model for TE personnel concerning the activities that are carried out in TE, including donor selection<br />
and recovery, and processing, preservation and storage of tissues for transplant. The aim is to apply these practices<br />
European-wide to increase the know-how and the level of performance of tissue banking staff, and to harmonize the<br />
techniques used in order to provide tissues for transplant of high quality and safety, hence reducing the risk of disease<br />
transmission to recipients.<br />
Methods<br />
– To develop European Good Tissue Practices (Euro-GTPs Guide) for Tissue Establishments activities (such as donor<br />
selection process, recovery, processing, preservation and storage of tissues).<br />
– To develop a training model for TE personnel.<br />
– The development of generic Euro-GTPs for TEs personnel<br />
– The development of tissue-specific Euro-GTPs<br />
– Approval of the Euro-GTPs<br />
– Design of a Training Model for TE personnel<br />
Results<br />
Before starting the elaboration of the GTPs, the following preliminary activities help us to know the current situation of the<br />
TEs both at regulatory and practical level;<br />
To have a better understating of the above mentioned issues, a questionnaire has been developed and addressed to TEs for<br />
recollecting data on the methods used for tissue banking processes. The objective of this questionnaire is to find out: (i)<br />
information on the quality processes that are being used and (ii) whether TEs carry out activities related to tissue recovery<br />
and processing.<br />
The questionnaire has been distributed to 147 TEs and completed by 22 TEs from several Member States through the EU<br />
(collaboration asked from some national authorities in order to have a representative analysis of the EU). Additionally, it will<br />
be uploaded in the project website and TEs from the countries of the consortium that are not direct participants in the project
(and other EU countries if possible) will be invited to complete it so that the project can have a better global idea of the current<br />
situation in Europe.<br />
The project coordinator elaborated a survey to be distributed among the Eustite Final Conference participants (25 EU<br />
inspectors). The current or future inspectors were asked to answer some questions regarding their audit findings, the<br />
difficulties they find to apply the current regulation and their opinion about the worth of a more detailed and realistic<br />
guidance such as GTPs at European Level.<br />
The following documents and regulation have been analyzed:<br />
– The different European and national existing legal regulations<br />
– Quality systems applied in TEs<br />
– Procedures followed by TEs<br />
– Standards followed in TEs<br />
– EQSTB project results<br />
– Good Manufacturing Practices<br />
As a result of the analysis of the above mentioned questionnaires and documents and more specifically the GMP, we have<br />
identified the processes to be included in the GTPs guide.<br />
As regards the generic GTP, these include requirements related to:<br />
– Personnel<br />
– Facilities<br />
– Documentation<br />
– Donor selection and evaluation<br />
– Recovery<br />
– Processing<br />
– Storing and distribution<br />
– Quality management<br />
– Risk assessment<br />
– Validation methodologies<br />
– Documentation<br />
As regards the specific GTPs, they contain the following:<br />
– Donor selection<br />
– Recovery methods<br />
– Minimum quality criteria<br />
– Preservation methods<br />
– Storage and distribution<br />
The key issues and the issues not covered or covered in a weaker way by standards or regulation have been identified and<br />
highlighted as “HOT-TOPIC”. The web-based HOT-TOPICS forum supplements the guidelines by highlights, practical<br />
examples and special tools for developing areas /conversation pieces of tissue banking practices. These HOT-TOPICS issues<br />
are under regular revision and tissue bankers are urged to participate in the HOT-TOPICS discussion on the web page.<br />
This document covers the following:<br />
– Donor selection<br />
– Recovery and processing environments<br />
– Tissue specific quality criteria<br />
– Risk management<br />
– Traceability and vigilance<br />
– Validation<br />
– Critical third party agreements<br />
INVITED<br />
SPEAKER<br />
LECTURES
SPECIFIC CHALLENGES OF DONATION, PROCESSING AND CLINICAL<br />
APPLICATION IN PAEDIATRICS (II)<br />
vCJD RISK REDUCTION MEASURES FOR CHILDREN<br />
Dr Akila Chandrasekar FRCPath., NHS Blood and Transplant, Liverpool, United Kingdom<br />
INVITED<br />
SHORT<br />
PAPER<br />
Abstract:<br />
Variant Creutzfeldt-Jakob disease (vCJD) is a rare and fatal human neurodegenerative condition caused by abnormal prions.<br />
The incidence of vCJD in the UK is much higher than elsewhere in the world. The most likely route of transmission of vCJD<br />
is by exposure to prion from contaminated food infected with bovine spongiform encephalopathy (BSE). Various control<br />
measures were introduced in the UK to minimise the risk of passing BSE from cattle to humans.<br />
Transmission of prions has occurred (Iatrogenic CJD) during medical care through neurosurgical instruments, transplantation<br />
of corneal and dura mater grafts and administration of cadaveric-derived pituitary growth hormone and gonadotrophins.<br />
Transmission of vCJD by blood transfusions has been reported in the UK. The number of current or past blood or tissue donors<br />
incubating the disease at the time of donation is not certain.<br />
To minimise the potential risk of secondary transmission from human to human by tissue transplantation, the tissue services<br />
associated with UK blood services have introduced a number of risk reduction measures. These include donor exclusion of<br />
transfusion and transplant recipients, additional vCJD testing for deceased tissue donors where possible and improved<br />
processing steps to remove as much blood and marrow content from tissue grafts as possible. In addition, for children under<br />
the age of sixteen, who would not have had exposure to vCJD through the dietary route, imported skin allografts from<br />
countries with low risk for vCJD were made available.<br />
This presentation gives overview of challenges facing the tissue establishments due to emerging infections across the world.
– Import and export<br />
– Continuity plans<br />
– Tissue establishment dossier<br />
INVITED<br />
SPEAKER<br />
LECTURES<br />
Conclusions<br />
The European Good Tissue Practice (GTP) guidelines and the adjacent training model have been established as an outcome<br />
of the EU-funded project - Euro-GTPs - to provide a complete and detailed tissue banking information package for tissue<br />
bankers as well as for tissue establishment (TE) inspectors in Europe. These guidelines bring together the current minimum<br />
regulatory requirements of the European tissue and cells Directives and go one step further - incorporating useful good<br />
manufacturing practice (GMP) principles and utilizing the expertise of tissue bank experts to provide a set of practical<br />
recommendations for good practice in European TEs. The GTPs are developed to be a helpful tool for all kinds of TEs in<br />
different phases of their development and evolution as well as for competent authorities (CA) when performing TE inspections.<br />
The web-based HOT TOPICS forum is an important supplement to the guidelines. It highlights areas where it is generally<br />
acknowledged that greater harmonization is needed, where consensus is lacking on the best practice to be applied, or<br />
where it is commonly thought that tools are needed to support improvements in practice. Practical examples and some<br />
particular tools for developing areas and where conversations can proceed on specific tissue banking topics are proposed.<br />
These HOT TOPICS issues are under regular revision and tissue bankers are urged to participate in the HOT TOPICS<br />
discussion on the web page. As consensus is reached and good practice on these topics is more clearly defined, these texts<br />
will move into the full guidance document. New hotspot topics will be
PRIMARY PEDIATRIC KERATOPLASTY: DONOR AGE AND GRAFT SURVIVAL<br />
Planas Domènech, N. Fernández Guardiola, A. Physician of the Department of Pediatric Ophthalmology.<br />
Hospital St. Joan de Déu. Barcelona. Spain.<br />
Blackground:<br />
The ECD declines with age in the normal cornea. This process of cell loss is greatly accelerated after penetrating keratoplasty<br />
and persists for years after transplantation. Corneal clarity after penetrating keratoplasty can be affected by endothelial cell<br />
loss over time. The exact causa of postoperative cell loss is unknown but may be a result of donor or preservation factors,<br />
surgical stress, cellular interactions between the donor and recipient, immune reaction, normal or accelerated cellular aging,<br />
or glaucoma. Past studies evaluating endothelial cell loss after corneal transplantation have produced conflicting results with<br />
regard to the effect of donor age. Penetratin keratoplasty in children has been documented to have a higher rate of graft<br />
failure than adult keratoplasty.<br />
Purpose:<br />
To report our experience of primary penetrating keratoplasty in children focusing on the donor age.<br />
INVITED<br />
SHORT<br />
PAPER<br />
Methods:<br />
We undertook a retrospective review of penetrating keraroplasty performed in children 16 years and younger between<br />
1995 and 2010 at the Department of Ophtalmology, Hospital Sant Joan de Déu. Barcelona, Spain.<br />
Results:<br />
A total of 37 primary penetrating keratoplasties were perfomed in 34 patitnets during the study interval. The surgical<br />
indications were congenital opacities in 12 eyes (32,4%), acquired nontraumatic opacities in 21 eyes (56,8%), and acquired<br />
traumatic opacities in 4 eyes (10.8%). The mean recipient age was 7,8 years (range 5 months - 16 years). The mean donor<br />
age was 31,5 years (range 7 hours - 68 years). The mean donor endothelial cell density was 2968 (range 2157 - 4700).<br />
Overall graft survival at 1 year was 70%. Graft survival at 1 year was different among the surgical indications categories<br />
(congenital opacities 42%, acquired nontraumatic opacities 86%, acquired traumatic opacities 75%). Graft survival at 1 year<br />
was different among the different age groups (50% for patients younger than 6 months, 44% for patients 6 months to 5 years,<br />
83% for patients older than 5 years). Graft survival at 1 years was different among the different donor age groups (73%<br />
for donors younger than 16 years, 50% for donors 16 to 45 years, 88% for donors older than 45 years). Graft survival at<br />
1 year was similar among the different methods of corneal preservation (Optisol 68%, tissue culture 67%).<br />
Conclusions:<br />
Primary pediatric keratoplasty has an overall prognosis for graft survival of 70% at 1 year. Patients who received a cornea<br />
form a donor 16 to 45 years had worse 1-year graft survival comapred with patients who received a cornea form a donor<br />
younger than 16 years or older than 45 years. however, it is difficult to compare graft survival among the different donor<br />
age groups because there is considerable variation in the indications for transplantation and in the recipient age, in each<br />
group.
SKIN AND AMNIOTIC MEMBRANE (I)<br />
ALLOGRAFTS AND CRITICAL BURNED PATIENTS: EXPERIENCE AND FUTURE<br />
Fernández Cañamaque, J.L. Plastic Surgery Department - Hospital Universitario de Getafe, Madrid. Spain<br />
INVITED<br />
SHORT<br />
PAPER<br />
Nowadays skin allografts or homografts represent a base of the huge surgical stock for treatment of critical burned patient.<br />
Cadaver skin, started to work as allograft at the beginning of 50´s decade; allografts used as a temporary dressing have<br />
an important function in preventing infection and reducing fluid loss trough these burn wounds. Moreover, it carries out a<br />
basic function in order to prepare the surgical bed after debriding the burn wound, so autologous skin grafts could be inset,<br />
and as protection for this skin grafts when they are placed in the same surgical time, as it is described in the Alexander<br />
technique, also called Sandwich technique.<br />
Extraction conditions must be the same than in other organ extracted before transplantation, but they are not used<br />
immediately so it is mandatory to process them in order to preserve and store the allograft.<br />
There are 2 basic techniques in processing, cryopreservation and Glycerol preservation. Both have advantages and<br />
disadvantages. Because we have great experience in Glycerol preservation, and the bigger capacity in bacterial and viral<br />
inactivation of this technique is the reason we prefer this one.<br />
In this short paper we present the full process, starting with the extraction in the donor cadaver, the preservation and the<br />
surgical utilization in the critical burned patient surgery, going on with the postoperatory evolution up to the elimination of<br />
the graft.<br />
We present too how must, or at least how we would like the future of allografts be in the wide context of critical burned<br />
surgical treatment.
GLIMSE INTO THE FUTURE; ADVANCED THERAPIES (I)<br />
TOLEROGENIC DENDRITIC CELL THERAPY IN REFRACTORY CROHN’S DISEASE<br />
Elena Ricart 1,2 , Julián Panés 1,2 , Jordi Rimola 3 , Raquel Cabezon 1 , Carolina España 1 and Daniel Benítez-Ribas 1 .<br />
1 Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (Ciberehd) and<br />
2 Department of Gastroenterology and 3 Radiology. IDIBAPS, Hospital Clínic. Barcelona. Spain.<br />
INVITED<br />
SHORT<br />
PAPER<br />
Introduction:<br />
Cronh’s disease (CD) is a chronic transmural inflammation that can involve any part of the intestinal tract, predominantly<br />
the small and/or large intestine. Since CD is a consequence of loss of tolerance against the normal gut flora, a novel<br />
approach based on the modulation of dendritic cell (DC) function is especially attractive. DCs are the most important of<br />
professional antigen presenting cells in the intestine. It is known that DCs are involved in inducing tolerance against<br />
self/harmless antigens. Despite the promise of cellular therapy with immunogenic DCs in cancer patients, no clinical studies<br />
have taken advantage of their specific immunosuppressive properties so far.<br />
Methods:<br />
Tolerogenic dendritic cells (Tol-DCs) were generated from isolated monocytes using clinical grade reagents (IL-4 and GM-<br />
CSF) and synthetic culture media. DCs were cultured for 6 days and the cells activated by adding a cocktail of cytokines<br />
(TNF-α, IL-6, IL-1β and PGE2). Dexamethasone (1*10 -6 M) and vitamin A (10 -9 M) were added at day 3 of differentiation<br />
as immunosuppressive agents. The stability of DCs was evaluated by adding second stimuli after washing Dex away from<br />
the culture.<br />
Results:<br />
Tol-DCs had limited expression of costimulatory molecules and impaired alloresponse induction. Tol-DCs depicted a semimature<br />
phenotype. Production of the anti-inflammatory cytokine IL-10 was significantly increased on tol-DCs whereas IL-12<br />
was completely absent. Furthermore, tol-DCs were poorer stimulators of antigen-specific T lymphocyte response (PBLs or naïve<br />
T cells) than control DCs, showing the role of tol-DCs in dampening the specific T lymphocyte activation. Moreover, tol-DCs<br />
were stable to second stimuli preserving the phenotype and cytokine production pattern; keeping high levels of IL-10 secretion<br />
without IL-12. Interestingly, we were able to generate tol-DCs from Crohn’s disease patients with the same tolerogenic profile<br />
than can be applied in future clinical trials.<br />
Conclusions:<br />
Our findings highlight that the combination of dexamethasone together with a cytokine cocktail renders clinical grade<br />
tolerogenic DCs, displaying a semi-mature phenotype, a shifted balance towards anti-inflammatory cytokines, low T cell<br />
stimulatory properties. The ability of these cells to induce tolerance represents a potential and novel therapeutic approach<br />
to treat autoimmune diseases to silence unwanted immune reactions.<br />
This work was supported by grant SAF 2009-07272 from the Ministerio de Ciencia e Innovacion and Ciberehd. Ciberehd<br />
is funded by the Instituto de Salud Carlos III.
ORAL<br />
PRESENTATIONS
ORAL - NOVEMBER 9TH - ETHICAL THEMES OF COMMON INTEREST<br />
AUDITORI - 12.00-12.30<br />
O-1<br />
MODEL OF COEXISTENCE FOR PUBLIC AND FAMILY CORD BLOOD BANKS<br />
ÁLVAREZ RAMOS, A.; LOSADA PESCADOR A. VidaCord. Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
Cord blood from HLA-matched sibling is the best option to save the life of a child suffering from a subsidiary of transplant<br />
disease. Despite this clinical evidence, there is much controversy about family UCB banks: It is argued that storing UCB in<br />
a private bank for autologous use is useless (for UCB steam cells already carry the same genetic error that caused the<br />
disease) and that family UCB banks are a menace to key social values such as solidarity, altruism and fairness. To address<br />
these issues Spain published the Royal Decree 1301/2006, which places special emphasis on the risk of potential misleading<br />
information family CBB could spread about the real clinical applications UCB may have on what is defined as "potential<br />
autologous use," as well as the fact that any UCB unit stored in a family bank in Spain must be necessarily made universally<br />
available. There is no consideration in this RD about the potential allogeneic use of UCB within the family.<br />
Material and Methods:<br />
We analyzed the RD 1301/2006, the Spanish Umbilical Cord Blood National Plan (PNSCU), public statements made by<br />
the director of the Spanish National Transplant Organization, VidaCord (first UCBB licensed in Spain) Mission Statement,<br />
as well as studies on CBT done by Eliane Gluckman and the Spanish Constitution. DISCUSSION UCB of an HLA-matched<br />
sibling is the best option for hematopoietic transplant. Therefore, it is legitimate for parents to decide to store in private<br />
banks their children UCB . VidaCord objective, in its third point ("the consented transfer...") argues that individual and<br />
families’ rights can be reconciled with the overall population’s interests. The PNSCU states that the whole Spanish population<br />
CB transplant needs are met with 60,000 units stored in public banks, which increase their inventories from altruistic<br />
donations from part of the 450,000 annual births in Spain.<br />
Conclusions:<br />
1. Having stored the cord blood in a family bank is the best medical alternative in the event of illness subsidiary of allogeneic<br />
sibling transplant.<br />
2. The right to health protection is prior to the likely incidence of a disease.<br />
3. Public health authorities must balance the individual right to give your children the best clinical option with the whole<br />
population care.<br />
4. It is possible to reconcile the two positions, favoring this way society interests as a whole. To achieve this, the value of<br />
solidarity must be subordinated to individual freedom and the principle of autonomy.
O-2<br />
A FUNDAMENTAL RIGHT TO DECIDE WHETHER AND HOW OUR BODY WILL BE<br />
USED IN RESEARCH<br />
ZARDOYA, M.; VILARRODONA A.; RODRÍGUEZ C.; RUÍZ A.; PAREDES D.; TRÍAS E.<br />
Transplant Services Foundation. Hospital Clinic of Barcelona. Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract / Introduction:<br />
Human organs, tissues and cells can be used not only for clinical applications but also for teaching and research with or<br />
without commercial purposes. The supply of human tissue with this aim is increasingly demanded and sometimes<br />
indispensable. The main ethical and legal issues to be considered are related to confidentiality, data protection, secondary<br />
use of samples, return of results and informed consent. Taken into account the movement of tissues at European level and<br />
the International exchange we also face the problem of lack of harmonization and a common regulatory framework to<br />
protect autonomy. This study will mainly focus on consent requirements to guarantee and protect autonomy both from living<br />
and deceased donors. Aim The main objective is to analyze if current medical standards and legal regulations guarantee<br />
the autonomy of the will. A secondary aim is to examine the opinions and attitude of donors and donor families towards<br />
donation for teaching and research.<br />
Materials and Methods:<br />
A study of legal regulations and ethical requirements in different European countries has been done to find out differences<br />
in consent regulations and clinical practices to protect the donor. Not only legal regulations have been studied but also the<br />
kind of information given and methods used prior to request consent, as informed consent encourages difficulties when a<br />
broader consent is needed for a secondary use of the samples or for the involvement of private companies in the projects.<br />
It has also been analyzed the clinical practices used in different hospitals in our country to guarantee donor’s autonomy when<br />
legislation does not differentiate between specific consent when the tissues, organs and cells are going to be used for<br />
research, for teaching or for research with commercial purposes. This information has been obtained from regional and<br />
hospitals transplant coordination units. The willingness to donate for teaching or investigation has been also studied,<br />
considering the organ and tissue donors from July 2009 to 30 June 2011 in our hospital.<br />
Results:<br />
Ethical, legal and social issues have not been regulated in a precise and uniform manner in the analyzed countries. Most<br />
of them are developing a binding legal document that regulates the distribution of samples for research. However, specific<br />
consent is not always required for using tissues and cells for research. A recent study conducted in our hospital has concluded<br />
that most of the families of potential donors are in favour of the use of their organs, cells and tissues in research (14.5%<br />
refused donation for other purposes than clinical)<br />
Conclusions:<br />
The recovery and distribution of tissue for research is a need in growing demand. It is a must to organize the activities and<br />
design the distribution procedure according to the current legal framework and respecting the ethical requirements. It will<br />
protect donor’s autonomy concerning the use of the body or body parts for non clinical aims and it will contribute to creating<br />
a more favourable public opinion towards the donation of organs, tissues and cells for research and teaching aims. The<br />
following should be guarantee: - Donor must understand what the sample is to be used for - Donors are informed that their<br />
sample might be used in commercial research - Patients know that material left over following diagnosis might be used for<br />
research By protecting autonomy through the process of obtaining consent we will allow individuals to exercise the<br />
fundamental right to decide how their tissues, organs and cells will be used in research.
O-2 BIS<br />
CORD BLOOD: PUBLIC AND PRIVATE<br />
BRANKA GOLIVICH<br />
ORAL - NOVEMBER 9TH - INTERNATIONAL EXPERIENCE ON TB<br />
ROOM 1 - 11.30-13.30 H.<br />
O-3<br />
TPM AS INTERNATIONAL EDUCATIONAL MODEL IN TISSUE BANKING<br />
NAVARRO, A.; DUQUE E.; PAEZ G.; MANYALICH M. Banc de Sang Teixits. Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract / Background:<br />
The tissue and cell donation have enabled the development of a health care field which has improved the quality of life of<br />
numerous patients. As its success relies on the staff professionalism, all personnel involved in the tissue and cell donation<br />
process should be appropriately qualified and able to access continuing knowledge. The Transplant Procurement<br />
Management (TPM) has been providing for nine years academic training courses on tissue banking in compliance with the<br />
agreed professional requirements, current practices and recognized standards of proficiency in the field. OBJECTIVE: To<br />
provide academic training courses on tissue banking in compliance with the agreed professional requirements, good current<br />
tissue practices and recognized standards of proficiency in the field.<br />
Method:<br />
The Tissue Banking Courses provided by TPM are meant to develop and reinforce the core skills and competences of<br />
professionals involved in the tissue and cell donation, recovery, processing and transplantation. They consist in a blended<br />
learning program (Face-to-Face and On-Line) and are structured in modules for a better assimilation of concepts. Participants<br />
perform practical activities throughout the whole course. General debates allow them to share opinions and draw on their<br />
practical knowledge and experience. Continuous and final assessment is performed.<br />
Results:<br />
A total of 376 professionals have been trained within the TPM courses on tissue banking as following: 265 participants from<br />
41 countries (172 Europe, 51 America, 24 Oceania, 15 Asia and 3 Africa) attended the on-line courses and 111<br />
participants (80 Europe, 12 Asia, 10 America, 6 Oceania and 3 Africa) the face-to-face ones. The overall assessment<br />
shows that the courses’ objectives have been successfully accomplished with an average of: 4.0 in the on-line and 4.4 in<br />
the face-to-face course. The applicability to the daily job has also been evaluated positively with an average of 4.0 in the<br />
on-line and 4.1 in the face-to-face course (scoring performed on a 1-poor to 5-excellent scale).<br />
Conclusion:<br />
By providing specialized training with practical applicability in the tissue banking field, TPM allows practitioners to improve<br />
their core skills and competences, establish optimal policies and practices for human tissue and cell donation. Due to its<br />
international approach, specialists may become part of a wide network of colleagues who manage the same challenges<br />
worldwide.
O-4<br />
BASIC CONDITIONS FOR A NETWORK IN TISSUE MEDICINE: QUALITY,<br />
TRANSPARENCY AND EFFICIENCY<br />
BÖRGEL, M. DGFG - German Society for Tissue Transplantation. Germany.<br />
Abstract:<br />
The embedding of tissue medicine in the non-profit sector, primarily established at major university hospitals, is one goal of<br />
the German Society for Tissue Transplantation (DGFG). To meet the increasing regulatory requirements of the Human Tissue<br />
Act, the factors of success for a non-profit network in tissue medicine are quality, transparency and efficiency in many ways.<br />
Tissue donation and tissue processing in accordance with high uniform standards ensure high quality in all processes and<br />
tissue graft quality at the best. Uniform standards enable efficient and cost-saving banking processes and exploit synergies,<br />
as well. This continuously improves the supply of high-quality tissue grafts through DGFG. Transparency in all activities is<br />
necessary for an authentic and altruistic commitment in the sensitive issue of tissue donation. The annual report of DGFG<br />
unfolds transparently the non-profit character of all activities of the network.<br />
O-5<br />
TRANSFORMING TISSUE DONATION AND TRANSPLANTATION IN CANADA INTO A<br />
SINGLE INTEGRATED INTER-PROVINCIAL SYSTEM<br />
HAUN, M.; MOHR J.; DERKSEN P.; PARSONS C.; SHER G. Canadian Blood Services. Canada.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Tissue banking in Canada consists of more than 20 independent tissue banks that recover, process, store and distribute<br />
tissue (In Quebec the provincial blood agency Hema-Quebec manages the tissue supply). Delivery of healthcare services is<br />
a provincial jurisdiction in Canada, which can present challenges with respect to standardization, equity of access for<br />
patients, and co-operation between programs. In August 2008, Canadian Blood Services was asked by the federal and<br />
provincial governments to leverage its unique role and expertise as the operator of the Canadian blood system (outside<br />
Quebec) to design an inter-provincial system for tissue (and organ) donation and transplantation. Canadian Blood Services<br />
has undertaken an exhaustive consultation with tissue donation and transplantation stakeholders, end users and experts<br />
across the country and internationally, documenting their concerns, advice and proposed solutions. A committee of tissue<br />
experts has developed a series of recommendations characterized by three key themes: safety and quality, equitable and<br />
timely access to tissue products, and efficiency. The plan recommends the existing independent and un-coordinated tissue<br />
banks transition to a system that will double tissue donation and recovery activity, consolidate tissue processing, standardize<br />
quality programs, maintain a single shared inter-provincial inventory, and be managed by one organization. The<br />
recommendations, a proposed implementation plan and cost estimates were delivered to governments in April 2011.
ORAL<br />
PRESENTATIONS<br />
O-6<br />
MINUTES FROM THE FIRST EDITION EXPERIENCE OF THE UNIVERSITY OF<br />
BARCELONA MASTER DEGREE IN DONATION & TRANSPLANTATION OF ORGANS,<br />
TISSUES AND CELLS<br />
BALLESTÉ, C.; SEGUR J.M.; CASAROLI R.; POMAR LUIS J.; ALSINA M.; SUSO S.; MANYALICH M. Facultat de Medicina,<br />
Universitat de Barcelona. Spain.<br />
Abstract/Introduction:<br />
The first Master´s degree in the field of Donation & Transplantation was offered by the University of Barcelona, 2010-2011.<br />
This official program, integrated within the EHEA, was designed to provide participants with the necessary knowledge,<br />
skills and competencies for their future clinical care activity as well as helping them to carry out any scientific research. The<br />
Master’s degree was structured in 3 common modules: Research in donation, Research in organ transplantation, Research<br />
in tissues, cells & hematopoietic progenitors transplants and 2 specialized: Research & Professional Path.<br />
Objectives:<br />
It is with lot of interest to analyse and to define the feedback from this first edition. The results will allow knowing the strong<br />
and the week points in several aspects.<br />
Methodology:<br />
The experience and the motivation of the candidates to continue their career in this field were evaluated. An evaluation<br />
questionnaire for the organizational aspects was used to know the participant´s opinion for the following aspects: Content<br />
of the materials; the way of presenting; the way of running the question/answer minutes; personal benefits from the<br />
participation in the class. The percentage of the participation was also an indicator showing the interest of the students.<br />
Results:<br />
Participant’s profile The total number of the participants was 25, with a heterogeneous cultural and professional profile<br />
background; 17 Medical Doctors, 4 Nurses, 2 Biologists and 1 Psychologist. 22 of 25 were graduated; 3 left the programme<br />
for personal or professional reasons. Internal subjects evaluations The evaluation showed the following scores: Content of<br />
the materials (4,29±0.26); presentation (4,22±0.32); Questions/answers minutes (4,32±0.29);Personal benefits<br />
(4,06±0.31) On- line results Online learning system was used. The participants were able to express their opinion, asking<br />
questions or raising discussions in the Open Forums. The evaluation was a summary of the student activities and the tasks<br />
fulfilled by them. Research projects Divided in 2 different fields: Research & Investigation (77% of the participants) and<br />
Clinical and/or Laboratory Practice (23% of the participants). 72.6% of the participants manage to fulfil the study in the<br />
scheduled time. All but 2 were considered as adequate by the ad-hoc tribunal and passed it.<br />
Conclusions:<br />
The great and heterogeneous participation showed the interest of the participants. This Master offers the possibility to enlarge<br />
the knowledge and skills of a large number of foreign professionals providing them with an official degree, known in the<br />
scientific and academic community. Analyzing the results, this programme achieved the expected results giving a great<br />
support to the following edition.
O-7<br />
SPEED IS LIFE. APLYING LEAN HEALTHCARE TO TISSUE BANKS<br />
TORNOS JUAN, I.; SERIGO X.; GIRALT E. Auren. Spain.<br />
Abstract:<br />
Combat pilots have since the very beginning of military actions believed that “Speed is Life”. In a Sense this principle is also<br />
applicable to Tissue Banks. - Processing tissues after extraction needs to be done at maximum speed to assure quality of the stored<br />
“product”. - Service to Hospitals needs to be at maximum speed to assure reaching patients on time. Of course, speed is always<br />
subject to the Quality of the Processes involved. What is really needed is to perform the right steps, in the right sequence, with<br />
the right quality at the maximum speed possible. Being capable to achieve this goal is nothing else than getting as close to<br />
perfection as possible. Up till now, a high amount of effort has been placed on assuring that the correct steps are performed,<br />
and robust and strict Quality Systems have been developed and implemented in many Tissue Banks all over the world. This is in<br />
most cases not enough to guarantee the fastest process possible. Lean Healthcare is the correct tool to achieve this goal applying<br />
different methods and tools that start on a deep and solid analysis of the processes to look for anything not adding value to it.<br />
This is frequently called MUDA using the Japanese word. - Any time a tissue is waiting to be collected or queuing to be tested,<br />
MUDA is being generated. - Wrong extractions of stem cells generate in many cases storage of unusable tissues. This is also<br />
MUDA. - Errors in decisions about volumes of blood to be stored at hospitals generate stock breakdowns or excesses which are<br />
also MUDA. - Returns of unused blood from hospitals to Tissue Banks consume time and resources adding no value and are again<br />
a source of MUDA. - Etc. Systematic and exhaustive elimination of waste (MUDA) is the path to get closer to perfection and thus<br />
to be capable to perform processes at the “speed of light”. In this talk we will propose a number of methods and tools inherited<br />
from the manufacturing world that will allow Tissue Banks to go a step forward in developing an Excellent process, providing<br />
exceptional service to hospitals and thus to patients. All of these methods are grouped today within the Lean Healthcare approach<br />
that is growing fast in its application in most of Health Services in the USA and Europe.<br />
O-8<br />
TISSUE RECOVERY TEAM: 8 YEARS OF EXPERIENCE<br />
ORAL<br />
PRESENTATIONS<br />
FARIÑAS, O.; VILARRODONA A.; SAVIO MANUEL A.; LUQUE S.; OLIVA R.; PÉREZ LUISA M.; TRÍAS E. Transplant Services<br />
Foundation - Hospital Clinic. Spain.<br />
Abstract/Introduction:<br />
Traditionally in Europe each tissue type from cadaveric donors is recovered by a different recovery team (orthopaedic<br />
surgeons, plastic surgeons, cardiovascular surgeons, ophtalmologists). In 2002 we developed a new model of tissue recovery<br />
team with the objective of improving our recovery system. This new model of tissue recovery team, that started its activity in<br />
2003, was conceived from a combination of the European model where it is almost always present a medical doctor in the<br />
retrieval, and the American model where their members recover all the tissues (skin, cornias, cardiovascular and<br />
musculoskeletal tissue). Our new concept of tissue recovery team is composed by three members (medical doctor, nurse and<br />
technician) that are in charge of recover all the tissues. It is always mandatory that a medical doctor (team leader) is present<br />
in the team due to its medical knowledge, and the other two members can be combined in different ways (nurse-technician,<br />
nurse-nurse, technician-technician). Aim To evaluate the results and effectiveness of this new tissue recovery team model<br />
during the period between 2003 and 2010.<br />
Materials and Methods:<br />
This retrospective study evaluates the skin, cardiovascular and musculoskeletal retrievals performed by the new tissue recovery<br />
team during the period from 2003 to 2010. The analysis focused on different variables depending on the tissue recovered.
ORAL<br />
PRESENTATIONS<br />
From skin tissue the total number of retrievals, total amount of skin obtained and the average skin surface obtained per donor<br />
were analyzed. From musculoskeletal tissue the total number of retrievals, % of recovery errors and the contamination rate<br />
were analyzed. From cardiovascular tissue the total number of heart-valves and arteries recovered were analyzed. Results<br />
The skin tissue recovery started in 2008. The number of skin retrievals increased from 91 (2008) to 139 (2010). It was<br />
observed the importance of the personnel learning curve because the average skin surface obtained per donor raised from<br />
2489.67 cm 2 (2008) to 3257.01 cm 2 (2010). Regarding the musculoskeletal tissue recovery two main variables were<br />
analyzed: recovery error and contamination rate. The recovery error rate decreased drastically since the establishment of<br />
the new tissue recovery team (average from 2003 to 2010 was 1.04) comparing with the results obtained by the traditional<br />
team (average from 2000 to 2002 was 2.54%). The contamination rate was influenced by the incorporation of new<br />
personnel and the development of a new donor cleaning methodology (23.01% from 2003 to 2007 and 17.5% from 2008<br />
to 2010).<br />
Conclusions:<br />
The new model of tissue recovery team provides many benefits to our tissue establishment. A more accurate control of the<br />
recovery procedures can be performed increasing its achievement as well as the possibility of personnel training. To conclude<br />
we obtained a more professional recovery team decreasing the recovery errors and the contamination rates during tissue<br />
retrieval.<br />
O-9<br />
THE DONOR TISSUE BANK REPLACEMENT FACILITY – MEETING FUTURE DEMANDS<br />
IN TISSUE / CELL BANKING<br />
HERSON ROMA, M.; PONIATOWSKI S.; ADAMAS F.; CORDNER S. Donor Tissue Bank of Victoria / Victorian Institute of<br />
Forensic Medicine. Australian.<br />
Abstract:<br />
This presentation shares the considerations given to the design of a replacement facility for the Donor Tissue Bank of Victoria<br />
(DTBV), Melbourne, Australia The DTBV was established in 1989, by the Victorian Institute of Forensic Medicine in awareness<br />
of the privileged access to human tissue for transplantation. The DTBV is the only multi-tissue tissue bank in Australia with<br />
all the relevant services under the one roof which include a team of transplant coordinators, processing scientists and<br />
technicians and a fully NATA/ TGA accredited microbiology and serology laboratory. As a multi-tissue banking facility<br />
(skin, cardiac and muscle-skeletal), the DTBV has provided increasing number of grafts in Victoria and Australia. DTBV’s<br />
original purpose built 440m 2 tissue banking facility opened in 1992; by 2009, although still fully compliant with the<br />
requirements of the Therapeutic Goods Administration (TGA) weaknesses in the architectural design and structure of the<br />
processing core suite were becoming a liability. The facility could become non-compliant to the stricter incoming codes of<br />
manufacturing practice and inadequate to absorb growth of the DTBV’s banking activity. Plans for future also acknowledged<br />
that current tissue grafts will be surpassed by biotechnology enhanced products incorporating cells and scaffolds. Tissue<br />
banks such as the DTBV are uniquely positioned to become translation platforms to convert successful bench-level results into<br />
quality compliant and cost effective transplantable products. In 2009, DTBV was successful in its submission to<br />
Commonwealth for a AUD$13 million grant to to build a state of the art replacement tissue and cell banking facility, at the<br />
Victorian Institute of Forensic Medicine (Coronial Services Centre) site. The new facility is currently under construction, with<br />
a design aimed to incorporate the elements of incoming increased tissue and cell manufacturing regulatory requirements,<br />
as well as attain the capacity to manufacture biotechnology enhanced tissue and cell products. The envisioned 1,600 m2 gross<br />
built area, will be been split into: ground level (Tissue Retrieval, Stores, Despatch); first floor (Offices, Micro Lab, R&D Lab)<br />
and second floor (processing core and CSSD). There have been quite a number of challenges in the design considering the<br />
need to ensure a versatile and long lasting plant, to allow for future changes in room use dictated by specific product<br />
requirements, and to incorporate the specific requirements of cell cultures within a GMP controlled environment. Whilst the<br />
new facility will ensure business continuity, only time and use will confirm if the goals have been met.
O-10<br />
CURRENT STATUS OF TISSUE BANKING IN KOREA<br />
KANG , Y.; CHUNG Y.; LIM J.; KIM Y. St. Vincent's Hospital, Catholic University of Korea. Korea.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
In 1971, first bone bank was established in Korea. The first clinical case of allograft transplantation was reported at the<br />
Journal of Korean Orthopaedic Association in 1973. After then, more than 60’s hospital based surgical bone banks were<br />
established throughout the country. Since the law on safety and control for human tissues had been established in 2005,<br />
tissue transplantation was tremendously grown in Korean medical society. Currently, 57 hospital based bone banks, 5<br />
processing tissue banks, three regional tissue banks and 70 tissue distributors are working in Korea. From 2005, every<br />
tissue banks have to report their activities to Korea Food & Drug Administration(KFDA) including number of donors, products<br />
and distribution of the tissues, and etc. KFDA with professional and academic associations annually has been performed<br />
quality assessment for the tissue banks within the country. Varieties of the quality failure were detected by KFDA inspection<br />
including inadequate facilities, equipments, manpower, documents and achieves etc. In every year, KFDA reported the<br />
activities of tissue banks. In 2005, 55,512 tissues were used in the country. At that time, only 10,158(18%) of tissues were<br />
produced by tissue banks in the country. Majority of the tissues (45,354 tissues; 82%) were imported from foreign countries<br />
due to lack of tissues produced by the country. The country had been paid to import tissues from abroad for approximately<br />
several million dollars every year. Those findings were due to shortage of tissue donors, lack of infrastructures of the tissue<br />
banks and shortage of tissue bank operators. In 2009, 223,158 tissues were used. However, 138,739 of tissues (62%)<br />
were produced in the country, and 84,419 (38%) tissues were imported from abroad. In 2003, Korea started the<br />
implementation of a national training program for tissue bank operators and the establishment of a National Training<br />
Centre(NTC) for tissue bank operators. Through past 7 years educational efforts a basic core of trained physicians and<br />
tissue bank operators has been established providing a professional training of tissue bank operators. These individuals and<br />
their respective banks have provided an increasing number of high quality grafts to the communities they serve at a cost far<br />
less than if they were acquired from abroad. The tissue bank operators trained so far in the NTC were 100s. These figures<br />
indicate the quality improvements of tissue banks and lots of cost saving of the country. Within 5 years, NTC will train<br />
another 100s tissue bank operators.<br />
O-11<br />
OPTIMIZING THE PROCESS OF TISSUE BANKING IN A BLOOD BANK NETWORK<br />
SAEZ, M., RODRÍGUEZ, L., GENIS, X., GRIFOLS, R., MASSUET, LL., PROFITOS, J., CASTELLA, D., CALLAO, V.,<br />
MOYA, F., PANADES, M., SALINAS, R., BOSCH, A., NAVARRO, A. Banc de Sang i Teixits. Spain.<br />
Abstract:<br />
Outcomes Banc de Sang i Teixits de Catalunya (BST) has developed a unique model which takes advantage of blood bank<br />
network to provide tissue banking services abroad in Catalonia. This approach is intended to bring tissue banking activities<br />
closer to donors, patients and specialists while reducing the structural cost of the services rendered. In our scheme, a central<br />
facility processes the tissues recovered by the local teams, stores and distributes them to the hospitals through the blood bank<br />
network. A close collaboration between BST staff and hospital specialists plays a central role in our model. The program of<br />
tissue services in our blood bank includes:<br />
– Blood bank services assisting living donors for autologous eye drops and platelet rich plasma treatments.<br />
– Specialized multitissue and cornea cadaveric recovery teams.<br />
– Centralized processing and preservation facility of living and cadaveric tissues.
– BST is in charge of tissue availability in hospitals as well as delivery and biovigilance. Methods and materials From<br />
January 2006 to December 2010, BST engaged tissue banking activities in 14 hospital blood banks in a regular basis,<br />
including the assistance of living donors and patients for eye drops production, fertility preservation and platelet rich plasma<br />
processing. Also two cadaveric recovery teams for cornea, musculoskeletal, skin and cardiovascular tissues recovery have<br />
been trained and 12 autonomous cornea recovery centres established. Additionally to facilitate tissues availability in the<br />
territory, tissues are sent to transfusion services allowing a supply 24 hours 365 days a year. Results In 2006 BST managed<br />
397 tissue donors (180 living and 217 cadaveric). After 5 years of implementing new tissue strategy activity increased 70%<br />
obtaining in 2010 a total of 674 donors (411 living and 263 cadaveric). Regarding tissue distribution we have observed<br />
an increase of 66 % of corneas transplanted (194 corneas transplanted in 2006 vs 322 in 2010), 61 % in musculoskeletal<br />
transplant (1152 tissues in 2006 vs 1858 in 2010) and 18% and 29% of skin and cardiovascular transplants respectively.<br />
Regarding living patients we provided in 2006 a total of 3762 eyedropper bottles compared to 6691 in 2010.<br />
Conclusions:<br />
Tissue services model in our blood bank has allowed:<br />
– Better and closer care of living tissue donors through BST network services resources.<br />
– Improving tissue quality before processing by our trained recovery teams.<br />
– Overall control from tissue donation to transplantation assuring complete traceability.<br />
– Responding specialist needs by increasing our human tissue portfolio.<br />
ORAL<br />
PRESENTATIONS<br />
O-12<br />
RETROSPECTIVE ANALYSIS OF 26 YEARS OF HOMOGRAFT HEART VALVE BANKING<br />
IN CENTRAL SOUTH AFRICA<br />
VAN DEN HEEVER JACOBUS, J.; NEETHLING MORRIS LEONARD W.; SMIT EDWIN F.; BOTES L. Dept of Cardiothoracic<br />
Surgery, University of the Free State, Bloemfontein, South Africa.<br />
Abstract/Introduction:<br />
The history of using homologous cardiac valves dates back more than 30 years. Through the years emphasis was placed<br />
on the optimization of graft retrieval, preservation techniques and clinical application. A cardiac homograft valve bank<br />
was established at the Department of Cardiothoracic Surgery, University of the Free State, Bloemfontein in 1982. Methods:<br />
A retrospective analysis was performed on all allograft data since 1984. Results: Since the first valve was successfully<br />
procured and transplanted in 1984, 2743 aortic and pulmonary homografts were harvested from 1927 donors, of which<br />
1667 [1067 (64%) aortic and 600 (36%) pulmonary] were released for clinical use. Road accidents (36%) and violent<br />
deaths (56%) make up the majority of unnatural causes of donor deaths. 1076 (39.23%) of valves were discarded for<br />
various reasons, the main reasons being Human Immunodeficiency Virus (32%), Structural abnormalities (22%), Hepatitis<br />
B (7.5%), Positive cultures (10.9%) and venereal diseases (8.9%). The mean donor age was 26.34 years with a male<br />
predominance of 1459 males versus 468 females. The average ischemic time was 33 hours mainly due to medico-legal<br />
autopsies exceeding the desired 24 hour time limit. The valves were disinfected in an antibiotic cocktail of Mefoxin,<br />
Piperacillin, Amikacin and Amphotericin B prior to cryopreservation. The surgical procedures utilizing the majority of<br />
homografts were aortic valve replacements (42.9%), aortic root replacements (19.3%) and right ventricular-pulmonary<br />
artery conduits (33.3%). The bank also supplied 23 other centers with homografts (426 aortic and 301 pulmonary).<br />
Conclusion:<br />
The Bloemfontein bank has established itself over the years as a leading cardiac homograft bank in South Africa. However,<br />
availability of suitable donors and homografts of optimal quality still remains a major concern, and further research in this<br />
field to help alleviate the problem is constantly required.
ORAL<br />
PRESENTATIONS<br />
O-13<br />
CLEAN ROOMS AND TISSUE BANKING: HOW HAPPY I COULD BE WITH EITHER<br />
GMP OR GTP?<br />
KLYKENS, J. 1 , PIRNAY, J. 2 , VERBEKEN, G. 2 , GIET, O. 3 , BAUDOUX, E. 3 , JASHARI, R. 4 , VANDERKELEN, A. 2 , ECTORS, N. 1<br />
1 - University Hospital Leuven, 2 - Queen Astrid Military Hospital, Brussels, 3 - University Hospital Liège, 4 - European Homograft<br />
Bank, Brussels. Belgium.<br />
Abstract:<br />
The regulatory framework of tissue banking introduces a number of requirements for monitoring clean rooms for processing<br />
tissue or cell grafts. Although a number of requirements were clearly defined, some requirements are open for interpretation.<br />
This study aims to contribute to the interpretation of GMP or GTP guidelines for tissue banking. A multi center study was carried<br />
out in 4 centers: Cell and tissue banks from University Hospitals Leuven, (Leuven, Belgium), Cell and tissue banks from the<br />
Queen Astrid Military Hospital (Brussels, Belgium), Cell and tissue banks from University Hospital Liège (Liège, Belgium),<br />
European Homograft Bank (Brussels, Belgium). These centers contain a total of 17 cell and tissue banks. All centers use<br />
monitoring programs compliant to ISO 14698 and United States Pharmacopeia 29 (2005). Based on the experience of the<br />
participating centers, the results of the monitoring program were evaluated to determine the feasibility of a clean room in tissue<br />
banking and the monitoring program. Controlled environments of Grade A in B, Grade A in C and Grade A in D were<br />
evaluated. Grade A is obtained with laminar air flow cabinets. Also the microbial efficacy of an incubator in a clean room<br />
environment was evaluated. This study indicated that a monitoring program of a clean room at rest in combination with (final)<br />
product testing is a feasible approach. Although a limited amount of out of spec situations was recorded, the main contributor<br />
were measurements of 2 CFU in a laminar air flow cabinet. Although no statistical significance (p between 0.90 and 0.95) was<br />
found compared with a Grade A in B, further evaluation shows a strong indication that a Grade D environment is not the ideal<br />
background environment for a Grade A obtained through a laminar airflow cabinet, The microbial contamination of an<br />
incubator in a clean room were evaluated using air samples and contact plates. Results indicate a limited microbiological load.<br />
A controlled environment is mandatory for tissue and cell processing. A monitoring program based on at rest measurements<br />
is feasible for all cell and tissue banks and gives sufficient assurance, in combination with adequate standard operating<br />
procedures and (final) product testing, that safe allografts are delivered to the patients. A GMP Grade D environment does not<br />
seem to be ideal background for a Grade A environment obtained through a laminar air flow cabinet. Further measurements<br />
are required to evaluate the efficacy of a GMP Grade C environment as a background for a Grade A laminar airflow cabinet.<br />
The contamination level in incubators is limited in standard operations, although closed containers seem to be necessary to<br />
protect products where background environments don’t qualify for open processing.<br />
O-14<br />
BANKING OF CRYOPRESERVED VASCULAR ALLOGRAFTS IN EUROPE: 20 YEARS OF<br />
ACTIVITY IN EUROPEAN HOMOGRAFT BANK (EHB) IN BRUSSELS<br />
JASHARI, R.; VAN HOECK B.; GOFFIN Y.; FAN Y.; ROUSSE N. European Homograft Bank (EHB), International Association.<br />
Belgium.<br />
Abstract:<br />
EHB has been banking the Cryopreserved Human Heart Valves and Arteries since 1989 and 1991, respectively. The heart<br />
beating (MOD) and the non-heart beating cadaver donors (NHBD) of age 12 to 55 years, fulfilling the donor acceptance<br />
criteria, were the donors of arteries. The main procurement centers were in Belgium, France and Switzerland. The surgical<br />
preparation and morphological evaluation are performed by the trained surgeons that are assisted by experienced technical
assistants, in the cleanroom class A with B/C background. Incubation in antibiotic cocktail for 20-48 hours and<br />
cryopreservation are carried out by technical staff. Storage is carried out in the vapors of liquid nitrogen (LN) at ≤-150°C<br />
and shipment in dry shipper in vapors of LN (≤-150°C) or dry ice (-76°C). 3533 arterial allografts are evaluated during<br />
last 20 years: 2421 (68.5%) are accepted and cryopreserved whereas 1112 (31.5%) discarded for morphology (46%),<br />
contamination (35%), serology (6%), stock surplus (4.5%), histology (3.5%) and surgical damages at procurement (3%).<br />
2297 arterial segments are implanted: 110 ascending and 396 descending thoracic aortas, 256 bifurcations, 138 iliac and<br />
1176 femoral arteries for infected native and prosthetic arteries (80%) or critical limb ischemia without available autologous<br />
venous material (15%). In 5% of cases the arteries are used in congenital cardiac surgery. Post-thawing evaluation and<br />
implantation information are required from implanting surgeon and are received in almost 100% of cases. However the FU<br />
information is received only in about 50% of cases. Not all requests for arterial allografts are fulfilled due to shortage of<br />
available allografts. A multicentric FU study might give clarification on the durability of cryopreserved arterial allografts since<br />
there are some reports on aneurysm formation on long term.<br />
ORAL - NOVEMBER 10TH - DONATION I<br />
AUDITORI - 09.30-10.30<br />
O-15<br />
DEVELOPING TISSUE DONATION IN NORTHERN GERMANY - ACTUAL STATUS,<br />
STRATEGIES AND CHALLENGES-<br />
WULFF, B.; HEINEMANN A.; MONTENERO M.; PUESCHEL K. Institute of Legal Medicine. Germany.<br />
Abstract:<br />
For a long time post mortem donation of the cornea had been the only possibility to donate in Hamburg, but during the last<br />
four years the Hamburg Institute of Legal Medicine started an additional program for the retrieval of musculoskeletal and<br />
cardiovascular tissues. New contacts were tied to tissue banks, the University Medical Center´s wards, hospitals in the city<br />
state of Hamburg and its surrounding, that means in the medical field. But tissue donation is nearly unknown by the<br />
inhabitants of our area, so we addressed ourselves to the task of "Spreading the News" in addition to the daily work of<br />
guiding the consenting process and improving surgical techniques. We present the different aspects, the actual status and<br />
also the problems of our ressources, capacities and networking arising from our felt obligation to contribute to the supply<br />
of tissue transplants for the patients not only in our region.<br />
O-16<br />
POSTMORTEM BLOOD ASSAYS FOR HIV, HBV AND HCV IN TISSUE DONORS.<br />
SYSTEMATIC LITERATURE REVIEW<br />
MIETH, K.; MUÑOZ O.; SOTO C.; NAVAS J.; GONZÁLEZ J. Fundación Cosme y Damián. Colombia.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
The sensitivity, specificity and predictive values of the serologic assays for HIV, HBV and HCV in screening of living donors<br />
are well defined. The operative characteristics of these serologic tests are not well defined for cadaveric donors. There is a<br />
potential risk of disease transmission in association to false negative results. The frequency of discharging donors can be<br />
affected by the false positive results.
Objective:<br />
Definition of the sensitivity, specificity and predictive values of the serological assays for HIV, HBV and HCV in the evaluation<br />
of cadaveric tissue donors. Evaluation of the quality of the blood samples in cadaveric donors an the association to the<br />
serological assay operative characteristics. Literature search A systematic review of the published literature in the last 25 years<br />
was done. The electronic data base Medline, Embase, Cochrane, LILACS and BIREME were used. Secondary manual search<br />
was added. Inclusion criteria Diagnostic test studies that evaluate serological assays for HIV, HBV and HCV in cadaveric tissue<br />
donors. Data extraction and analysis Two of the authors , in a standardized and independent way , revised the evidence,<br />
selected the papers for inclusion and evaluated the quality by using the QUADAS instrument. The findings were discussed<br />
and the final recommendations were produced using a consensus process.<br />
Results:<br />
The quality of the evidence is low. Most of the published evidence does not meet the standards to define sensitivity, specificity<br />
and predictive values for the serological assays in cadaveric tissue donors. There is a high variability in demographic<br />
characteristics, risk factors of the donors, time after dead and time elapsed for the process of the samples. These findings<br />
avoid definitive conclusions. The evidence suggests that the risk of having false negative results in these serological assays<br />
in cadaveric donors is low. The risk of false positive results is high, it is associated to serological assays for hepatitis B and<br />
C and when the time elapsed after dead is longer than 24 hours. More research is needed in this area.<br />
O-17<br />
TISSUE DONATION IN EMERGENCY MEDICINE DEPARTMENTS-<br />
THE SCOTTISH EXPERIENCE<br />
GALEA, G.; GALEA G.; DONALLDSON S. SNBTS Tissues and Cells Directorate. United Kingdom.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Since 2000 SNBTS Tissue and Cells Directorate (TCD)has been the preferred provider of tissues forn Scottish patients. In order<br />
to ensure sufficiency, it was necessary to identify the best sources of donors for tissue donation. To this end a study was<br />
conducted to assess the number of potential tissue donors across Scotland using ICD code profiles over a 5 year period. The<br />
study identified 3 main hospital sites of potential tissue donors - Emergency medicine departments (EMD), Intensive Care<br />
Units and Coronary Care units- the former providing the highest potential numbers. Moreover the EMDs in the more densely<br />
populated areas were shown to provide the highest yields. Therefore the TCD team has focussed its activities in EMDs in<br />
Central Scotland and has worked in colaboration with clinical EMD teams to develop a programme for clinical staff to<br />
become Designated Requestors. Since this collaboration started the approach rate to families has increased significantly, the<br />
number of authorisations (consents) has increased from 38 to 48%. This level of consenting has fluctuated over the years<br />
and the reasons for this are being examined on an ongoing basis. A Potential Donor Audit shows that at least 20 tissue<br />
donors and 38 cornea donors are obtained via this route. This is a significant proportion of our tissue donor pool.
O-18<br />
THE ROLE OF FORENSIC INSTITUTES IN TISSUE DONATION IN GERMANY –<br />
THE MUNICH EXAMPLE<br />
BRAUN, C.; WULFF B.; GRAW M. Institute for Legal Medicine Munich. Germany.<br />
Abstract:<br />
In 2010 the existing tissue donation program concerning cornea and heart valves at the Institute of Legal Medicine in Munich<br />
was reorganized to better meet the conditions of the new German Tissue Law and to expand the Institute's activity to<br />
musculoskeletal tissues in cooperation with the German Institute of Cell and Tissue Replacement (DIZG). Experiences gained<br />
at the Institute of Legal Medicine in Hamburg with its 5 year old tissue donation program proved valuable for establishing<br />
a similar program in Munich. However, due to differences between the city state of Hamburg and the 'area' state of Bavaria<br />
there are many organisational and procedural problems, making it necessary to find unique solutions. Considering the<br />
different settings we will present the underlying problems as well as the resulting organisational structure of the Munich<br />
tissue donation program to date. Furthermore, an outlook and comparison is given on the part different forensic institutes<br />
in Germany can play in tissue donation.<br />
O-19<br />
Q FEVER IN TISSUE DONORS IN THE NETHERLANDS<br />
VAN WIJK J, M.; MAAS E W.; HOGEMA B.; KOOT M.; RENDERS C N.; HERMANS H M.; BOKHORST G A.<br />
BISLIFE Foundation. The Netherlands.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
In the Netherlands, large outbreaks of Q fever occurred between 2007 and 2009. After implementation of various measures<br />
the outbreak is currently decreasing. Although no C. burnetii transmission through tissue transplantation has been described<br />
in literature, there is some evidence that suggests this is possible. This study aimed to determine the seroprevalence of C. burnetii<br />
in Dutch tissue donors and to determine whether C. burnetii DNA can be present in tissues used for transplantation (cornea,<br />
skin, heart valves, tendons and bone). Methods Starting October 2010, 1000 consecutive Dutch tissue donors, of whom at least<br />
one tissue was approved at initial assessment, were tested for previous infection with C. burnetii with a commercially available<br />
IgG Phase 2 ELISA. During the study period donors with increased occupational hazard, signs suspect for acute Q fever or<br />
known previous Q-fever were excluded from donation. Of all donors who tested IgG positive, donated tissues were tested by<br />
PCR for the presence of C. burnetii DNA. Results Between October 2010 and June 2011, 1018 donors were tested for phase<br />
2 IgG antibodies against C. burnetii. Of these donors 50 (4.9%) tested positive. Donated tissues were corneas (N=47), skin<br />
(N=9), cardiovascular tissues (N=7) and musculoskeletal tissues (N=8). Some tissues could not be tested by PCR, because there<br />
was no permission for research (N=3), corneas (N=4) or heart valves (N=1) were rejected for morphological reasons or<br />
because no bone marrow sample was available (N=1). In 39 of the 40 tested corneas (with rim) no C. burnetii DNA was<br />
detected. In one cornea the result of PCR was indeterminate. In 8 of the 9 tested skin donors, the only cardiovascular tissue donor<br />
tested thus far and in 6 of the 7 tested bone marrow samples of musculoskeletal tissue donors no C. burnetii DNA was detected.<br />
In 1 skin donor and 1 musculoskeletal tissue donor the PCR result was indeterminate.<br />
Conclusion:<br />
Almost 5% of the tissue donors in the Netherlands tested positive for anti-Coxiella IgG. In none of the donated tissues thus<br />
far tested C. burnetii DNA was detected. The results of this study could be used to optimize donor selection criteria.
O-20<br />
TISSUE DONATION AND ORGAN DONATION: A COMPETITIVE OR A COOPERATIVE<br />
SYSTEM - EXPERIENCES AFTER THE TISSUE ACT FROM 2007 IN GERMANY<br />
NITSCHKE, F.; MANECKE A.; WILLE D. DGFG - German Society for Tissue Transplantation. Germany.<br />
Abstract:<br />
The use of tissue transplants such as cornea, heart valves, blood vessels and musculoskeletal tissue is an important component<br />
in the treatment of tissue defects. The necessary tissues can be obtained according to standard procedures from organ<br />
donors also agreeing to tissue donation. In the German Society for Tissue Transplantation (DGFG) in the north east region<br />
(Mecklenburg-Vorpommern) such a donation program has been in place and under continuous development since the early<br />
nineties. By cooperation between transplant surgeons in hospitals, German Foundation for Organ Transplantation (DSO)<br />
transplant coordinators and local tissue banks, it was possible to obtain Cornea from non heart beeating donors 98 %<br />
(1893/1925) and musculoskeletal tissues from 21% (402/1925). Additional were 71% (215/304) Cornea and 60%<br />
(181/304) musculoskeletal Tissues harvested from organ donors in the years 2004 to 2010. Agreement to donation of<br />
heart valves and vessels from non-usable donated hearts and vessels was 33% (101/304). According to the new German<br />
Tissue Law, tissue donation and procurement is subject to German Transplantation Law and German Drug Law. Appropriate<br />
standards that protect the priority of organ donation, but also enable tissue donation must therefore be established and put<br />
into practise.<br />
ORAL - NOVEMBER 10TH - CARDIOVASCULAR I<br />
AUDITORI - 15.45-16.25<br />
O-21<br />
ANTIBIOTIC DECONTAMINATION OF HEART VALVE ALLOGRAFTS: HISTORICAL<br />
TRENDS BEFORE AND AFTER THE IMPLEMENTATION OF A HIGHER TEMPERATURE<br />
OF INCUBATION<br />
TREMBLAY, J.; PAQUET I.; BÉLIVEAU L.; GERMAIN M. Héma-Québec. Canada.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Background:<br />
We previously reported experimental data showing that antibiotic decontamination of heart valve allografts is inefficient at<br />
4 ªC and optimal at 37 ªC. We hereby compared the efficacy of our decontamination protocol before and after our tissue<br />
bank increased the temperature of incubation of the antibiotic soak. Methods: Hearts are rinsed and put into cold, nonantibiotic<br />
containing transport solution. Prior to dissection, 1 ml of the transport solution is placed into a thioglycollate broth,<br />
another 1 ml in TSB, both incubated for 14 days at 35 ªC and 22 ªC, respectively. Two small pieces of dissected tissues are<br />
cultured in a similar fashion. Dissected valves and accompanying pieces of residual tissues are soaked for 18 to 26 hours<br />
into a solution containing cefoxitin, gentamicin and vancomycin and then rinsed three times with Ringer’s lactate. Two pieces<br />
of control residual tissues and the filtered final rinse solution are cultured as above. The valves are then put into DMSO<br />
containing medium package and a 100 ml aliquot of this medium is filtered and cultured prior to sealing the package. Prior<br />
to the end of June 2010, heart valves were prªcessed at 4 ªC from procurement until final packaging, including the antibiotic<br />
decontamination soak. After that date, the procedure remained unchanged except for the 24 hour antibiotic decontamination<br />
step which is now done at 37 ªC. In order to qualify for transplantation, a graft must have negative cultures on samples taken<br />
post decontamination and no high pathogenicity microbes on any of the cultures.
Results:<br />
Prior to June 2010, 272 valves were prªcessed and decontaminated at 4 ªC, of which 153 (56.3%) had positive cultures on<br />
samples taken prior to the decontamination step. Of those 153 valves, 77 had negative cultures after antibiotic<br />
decontamination. Since June 2010, 66 valves were decontaminated at 37 ªC, of which 20 (30.3%) had positive cultures on<br />
samples taken prior to the decontamination step. Of those 20 valves, 17 had negative cultures after antibiotic<br />
decontamination. The efficacy of the decontamination prªcedure at 37 ªC is therefore 85.0% (17/20), compared to 50.3%<br />
(77/153) at 4 ªC (p=0.004, Fisher exact test). When taking into consideration the presence of high pathogenicity microbes,<br />
the proportion of valves that were acceptable for transplantation went from 61.0% (166/272) during the days of soaking<br />
at 4ªC, to 80.3% (53/66) after the temperature of incubation was raised to 37 ªC (p=0.003, Chi-square test). The rate of<br />
positive cultures pre-decontamination also decreased significantly between both periods (p=0.0002, Chi-square test).<br />
Conclusions:<br />
Compared with our historical results of placing heart valves in antibiotics at 4 ªC, our new procedure of antibiotic soaking at 37<br />
ªC achieves a significantly higher rate of successful decontamination. That, in combination with a secular decrease in our rates<br />
of pre-processing contamination, has led to a major increase in the proportion of tissues deemed suitable for transplantation.<br />
O-22<br />
COMPARISON OF THE EFFECTS OF ISCHAEMIC TIMES OF HARVESTED<br />
HOMOGRAFTS ON HISTOLOGICAL APPEARANCE AND TISSUE STRENGTH<br />
ORAL<br />
PRESENTATIONS<br />
VAN DEN HEEVER JACOBUS, J.; BESTER D.; SMIT EDWIN F.; BOTES L. Dept of Cardiothoracic Surgery, University of the<br />
Free State, Bloemfontein, South Africa.<br />
Abstract/ Introduction:<br />
Homografts in cardiac surgery are well established but availability remains the main limiting factor in their clinical<br />
application. Homografts are procured from beating heart donors and cadavers with a limited ischaemic time of 12-24 h<br />
post mortem. However, this time limitation has not been established scientifically. Several publications suggest that it would<br />
be acceptable to extend the harvesting times. Homograft availability is an international problem and the purpose of the study<br />
is to extend the harvesting time beyond 24 h, benefiting all cadaver donor based programs.<br />
Methods:<br />
The study focused on three groups: (a) Group A (n=5) consisted of homograft valves subjected to < 6 h ischaemic time and<br />
stored at 4⁰C prior to cryopreservation; (b) Group B (n=15), subjected to 24 h, 48 h and 72 h cold (4ªC) ischemic times<br />
prior to processing and cryopreservation; (c) Group C (n=15), subjected to 6 h room temperature warm (23ºC) ischaemic<br />
time, followed by 18 h, 42 h and 66 h cold (4ºC) ischaemia, after which the valves were processed and cryopreserved. Tissue<br />
strength was determined by thermal denaturation temperature (Td) and tensile strength. Tissue morphology was assessed<br />
by Scanning Electron Microscopy (SEM) and Haematoxylin and Eosin(H&E) stain.<br />
Results:<br />
No statistically significant difference (p>0.05) in tensile strength could be demonstrated between Group A, B and C with no<br />
differences between the three ischaemic time intervals. The results were confirmed by Td analysis. Tissue strength did not<br />
decrease as a result of prolonged ischaemic times or elevated temperatures. Autolysis were only observed in the 48 h (40%)<br />
and 72 h (100%) tissue of Group C but was not sufficient to affect tissue strength. The reduction of endothelial cells over time<br />
in both Group B and Group C did not influence tissue strength up to 72 h.<br />
Conclusion:<br />
Based on scientific evidence regarding tissue strength it seems acceptable to extend harvesting time to 48 h. However,<br />
animal studies need to be performed to substantiate these results, as the in vivo graft host interactions might produce<br />
calcification and/or host rejection results that might still affect the safe clinical use of such tissue.
O-23<br />
PULMONARY HOMOGRAFTS FOR RIGHT VENTRICULAR OUTFLOW TRACT<br />
RECONSTRUCTION DURING ROSS PROCEDURE: NINETEEN YEARS RESULTS<br />
ORAL<br />
PRESENTATIONS<br />
JUTHIER, F.; JASHARI R.; ROUSSE N.; VAN HOECK B.; BANFI C.; VINCENTELLI A.; PRAT A. CHRU LILLE. France.<br />
Abstract/Background:<br />
Replacement of the aortic valve or aortic root with a pulmonary autograft (Ross procedure) is widely used for aortic valve<br />
disease in growing patients and young adults. In most cases, a cryopreserved pulmonary homograft is used for reconstruction<br />
of the right ventricular outflow tract (RVOT). Main drawbacks of this procedure are progressive dilatation of the pulmonary<br />
autograft and RVOT failure, the predominant indication for reoperation of the pulmonary conduit being stenosis. The aim<br />
of this study was to evaluate the long-term hemodynamic behaviour of pulmonary homografts in pulmonary position after<br />
Ross procedure.<br />
Methods:<br />
Three hundred seventy-one patients had a Ross procedure in our institution between March 1992 and May 2011. Among<br />
them, 307 patients received a pulmonary homograft (supplied by the European Homograft Bank) in pulmonary position and<br />
they represent the study population. Mean age was 28 ± 10.7 years. Mean homograft diameter was 25.8 ± 2.2 mm. A<br />
comprehensive echocardiography was performed at discharge, at 6 months and then on an annual basis. Pulmonary<br />
stenosis was defined as a mean transvalvular gradient of more than 20 mm Hg across the homograft. Median follow-up<br />
was 5.5 years (range, 7 days-18.9 years).<br />
Results:<br />
Perioperative mortality was 2.6% (8 patients). Late mortality was 2.6% (8 patients). During follow-up, 10 (3.3%) patients had<br />
reoperation on the RVOT with a mean time-interval of 9.0 ± 4.0 years. Three of them received percutaneous implantation<br />
of transcatheter pulmonary valve prosthesis, 12.2 ± 1.7 years after the Ross procedure. Causes of reoperation were<br />
homograft failure in 5 (1.6 %) cases and pulmonary endocarditis in 5 (1.6%) cases. 5 (1.6%) other patients developed a<br />
pulmonary endocarditis medically treated. Mean transpulmonary gradients were respectively of 4.1± 2.5 mm Hg; 8.6 ± 6.3<br />
mm Hg; 11.9 ± 10.9 mm Hg and 10.8 ± 6.9 mm Hg post-operatively and at 5, 10 and 15 years. Freedom from pulmonary<br />
stenosis was 99.0% (IC 95%; 98.3-99.7%); 97.5% (IC 95%; 96.2- 98.8%) and 88.6% (IC 95%; 88.2- 92.4%) at 5 and 10<br />
and 15 years.<br />
Conclusion:<br />
Pulmonary homografts represent a safe valvular substitute for RVOT reconstruction during Ross procedure in our institution.<br />
This may be due to our policy of systematic oversizing of the allograft and the liberal use of anti-inflammatory drugs in the<br />
post-operative period. The gradual transvalvular gradient increase during follow-up however requires continued<br />
echocardiographic monitoring. Patients who meet criteria for isolated RVOT replacement can be successfully treated with<br />
catheter-based pulmonary valve implantation.
ORAL<br />
PRESENTATIONS<br />
O-24<br />
DECONTAMINATION OF CRYOPRESERVED CARDIO-VASCULAR ALLOGRAFTS:<br />
DETECTION AND ERADICATION OF THE SLOW GROWING SKIN GERMS<br />
JASHARI, R.; FAN Y.; VAN HOECK B.; LE MERCIER N.; DE GELAS S. European Homograft Bank (EHB), International<br />
Association. Belgium.<br />
Abstract/Introduction:<br />
We have previously shown that up to 30% of raw material, retrieved for heart valve and vascular allograft preparation is<br />
initially contaminated. Although the majority of the allografts are sterilized by decontamination procedure, about 8-10% of<br />
the tissues remain germ- positive in the final step of the processing, decreasing importantly the yield of the allografts at the<br />
end of the processing. About 50% of contaminant germs are the slow growing skin anaerobs such as Propionibacterium.<br />
The aim of this study was to examine whether those contaminated tissues remain germ- positive after a long term period of<br />
cryopreservation.<br />
Methods:<br />
Total of 28 allografts (7 arteries and 21 valves), which were discarded for contamination with Propionibacterium after being<br />
incubated in the antibiotic cocktail, were cryopreserved and stored in the vapor phase of liquid nitrogen for 33 to 81 months<br />
(median 61 months). They were thawed and diluted according to the standard procedure of EHB. The tissue samples are<br />
cultured for aerobe and anaerobe germs (in the thyoglycolate and rezasurine medium) at 30-35°C and for the fungi and<br />
yeasts (in the tryptic soy bouillon) at 20-25°C following the European pharmacopeia. The results are presented at 7, 14 and<br />
21 of incubation. Results. 32.1% of the allografts tested positive at 7 days and 64.3% at 14 day. No new samples tested<br />
positive at day 21. These preliminary data indicate that Propionibacterium is capable of resisting to a very low temperature<br />
of below -187°C as long as 81 months and remaining its “slow-grow” nature.<br />
Conclusion:<br />
Minimum 14 days’ culture is essential for the accurate detection of this germ. Extension of incubation to 21 day is not<br />
necessary for its detection. Success in decontaminating this common bacterium is important for the safety of cardio-vascular<br />
allograft recipients.
ORAL<br />
PRESENTATIONS<br />
ORAL - NOVEMBER 10TH - CARDIOVASCULAR II - IMPROVING SAFETY<br />
AUDITORI - 17.30-18.20<br />
O-25<br />
OPTIMIZATION OF CARDIOVASCULAR TISSUE DECONTAMINATION AND RINSING<br />
WITH BASE.128 AND BASE<br />
TERZI, A. 1 , BUZZI, M. 1 , GUARINO, A. 2 , DAINESE, L. 2 , VASURI, F. 3 , TOTHOVA, J. 4 , GATTO, C. 5<br />
1 - Banca Dei Tessuti Cardiovascolari Regione Emilia Romagna, 2 - Lombardia Cardiovascular Tissue Bank, Milano, Italy,<br />
3 - Pathology Unit, “f. Addarii” Institute Of Oncology And Pathology, S. Orsola-malpighi Hospital, Bologna, Italy, 4 - R&d<br />
Al.chi.mi.a Srl, Ponte San Nicolò (pd), Italy R&d Al.chi.mi.a Srl, Ponte San Nicolò (pd), Italy, 5 - R&d Al.chi.mi.a Srl, Ponte<br />
San Nicolò (pd), Italy R&d Al.chi.mi.a Srl, Ponte San Nicolò (pd), Italy.<br />
Abstract:<br />
Purpose To define the optimal time and temperature conditions for decontamination and rinsing of cardiovascular tissues<br />
using the BASE.128 and BASE medical devices in order to decontaminate efficiently the tissue and minimize the presence<br />
of antibiotic residues. Methods Ten cardiovascular tissues from heart beating and non heart beating donors, including valves<br />
and blood vessels, were retrieved by two different cardiovascular tissue banks. After transport to the bank, tissues were<br />
divided in three equivalent segments, which were decontaminated with BASE.128 (AL.CHI.MI.A., Italy) at 4°C for 24h,<br />
22°C for 8h and 37°C for 6h, respectively. All tissues were rinsed with BASE (AL.CHI.MI.A., Italy) at 4°C overnight before<br />
freezing at -80°C. Before and after tissue processing, bacteriological tests were performed on tissues and transport and<br />
rinsing liquids using BACT-ALERT and Thioglycollate and TSB mediums. After thawing, the presence of antibiotic residues<br />
was evaluated on tissue homogenates by disk diffusion method, and sterility test performed according to the European<br />
Pharmacopeia after removing potential interfering antibiotics with a specific resin mixture. Each tissue was sampled before<br />
and after processing and fixed in formalin for subsequent histological examination (Hematoxylin-Eosin and Weigert’s stain).<br />
Results Almost all investigated tissues were initially contaminated with one or more bacterial spp. (Staphylococcus spp.,<br />
Streptococcus spp., Propionibacterium spp.). Contamination was more often detected in the transport media rather than<br />
tissue. Post-processing microbiological tests of all investigated tissues and liquids were negative in all investigated time and<br />
temperature conditions. Despite the overnight rinsing, disk diffusion analysis showed that some antibiotic residues were<br />
present, mainly depending on the decontamination condition. The highest residue content was found after decontamination<br />
at 4° C for 24h. Residues were lower after decontamination at 22°C for 8h and almost absent after decontamination at 37°c<br />
for 6h. Inhibition zones were detected in Staphyloccoccus aureus seeded plates, indicating the possible presence of<br />
Vancomycin residues. Significantly smaller inhibition zones were found on Candida albicans seeded plates, indicating the<br />
presences of traces of antimycotic. No inhibition zones were found in Pseudomonas aeruginosa seeded plates, indicating<br />
the absence of any antibiotic active against Gram negative – bacteria (Gentamicin and Cefotaxime). Histological analysis<br />
showed no differences among tissues in terms of integrity. Conclusions Tissue decontamination with BASE.128 allowed to<br />
eliminate efficiently all contaminants from human cardiovascular tissues in all investigated conditions, without affecting the<br />
tissue integrity. Decontamination at 37°C for 6h followed by an overnight rinsing allowed to eliminate antibiotic residues,<br />
thus minimizing both antibiotic transmission to recipient and interference with microbiology tests.
O-26<br />
IMPACT OF ANTIBIOTIC RESIDUES ON MICROBIOLOGICAL ANALYSIS: IMPLICATION<br />
OF TISSUE BANKING<br />
GATTO, C.; GIURGOLA L.; BECCARO M.; LIPARTITI M.; D'AMATO TOTHOVA J. R&D Alchimia SRL. Italy.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Decontamination of tissue allografts in antibiotic cocktails can lead to the antibiotic carry-over effect, which in turn can<br />
result in the false negative microbiological analysis. Purpose Investigate impact of antibiotic residues induced by tissue<br />
decontamination on microbiological analysis. Methods Cryopreserved human cardiovascular tissues and skin or corneas<br />
were retrieved, processed and cryopreserved by four different tissue banks. Cardiovascular tissue and skin were<br />
decontaminated at 4°C for 24h/72h (cardiovascular) or at 22°C for 90 min. (skin) either with bank solution or BASE.128<br />
(AL.Chi.MI.A.SRL) and cryopreserved in RPMI 1640 with addition of 10% DMSO. Corneas were processed and stored in<br />
organ culture conditions. Microbiological analysis on tissue and processing liquids were performed according to bank<br />
standard procedures using BACT-ALERT, BACTEC and Thioglycollate/ TSB mediums. After thawing, the presence of antibiotic<br />
residues was evaluated on tissue homogenates by disk diffusion and sterility test performed according to the European<br />
Pharmacopeia after removing potential interfering antibiotics with a specific resin mixture. Contaminants were genetically<br />
identified by ribosomal RNA sequencing.<br />
Results:<br />
Bacteriological analysis reports on cardiovascular tissues, before processing, showed tissue contamination by one or more<br />
microorganisms. Except for one cardiovascular tissue, the bacteriological test reports after the decontamination showed<br />
negative results for all tissues and liquids. Bacteriological analysis reports on skin samples showed negative results before<br />
and after processing. Microbiological analysis performed on corneal storage liquids reported negative results. Disk diffusion<br />
test of thawed cardiovascular tissue and skin or cornea homogenates showed inhibition zones mainly on S. Aureus and on<br />
P. Aeruginosa seeded plates, indicating the presence of antibiotic residues active mainly against gram-positive rather than<br />
gram-negative bacteria. Inhibition zones were significantly reduced (S. Aureus plates) or completely eliminated (P.<br />
Aeruginosa plates) after the treatment of tissues and liquids with a resin mixture for elimination of antibiotic residues. Sterility<br />
test on the tissue and liquid samples after elimination of the residual antibiotics, showed significative positive (turbid) results<br />
for cardiovascular tissues treated with the bank solution. Turbidity was not detected on cardiovascular tissues decontaminated<br />
with BASE.128RED. Significative number of skin samples was found positive in sterility test, independently on used antibiotic<br />
cocktails, indicating inappropriate decontamination conditions. Sterility test on corneal storage liquids after removing<br />
antibiotic residues, showed significative number of positive (turbid) samples not always detected on tissue sample. Ribosomal<br />
DNA sequencing identification demonstrated the presence of clinical and environmental contaminants.<br />
Conclusions:<br />
The presence of the residual antibiotics in tissue and liquid samples submitted to bacteriological analysis can result in<br />
significant number of false negative results. Validation of decontamination processes and of microbiological methods are<br />
necessary in order to guarantee the safety of tissue allografts.
O-27<br />
NEW INSIGHTS OF ANTIBIOTIC SOLUTION: ANTIBACTERIAL SUSCEPTIBILITY IN THE<br />
4 DEGREE MEDIUM CONDITION<br />
MOTOMURA , N.; SAITO A.; TAMURA K.; NOGUCHI N.; HATTORI O.; ODA N.; SEKI M. University of Tokyo. Japan.<br />
Abstract/Background:<br />
In many tissue banks, disinfection of retrieved tissues by antibiotics cocktail has been done under the 4 degree condition in<br />
a refrigerator. Antibiotics have been designed to use at the body temperature per se, and some combination of the cocktail<br />
may not exert the best performance as an antibacterial agent. We examined the susceptibility of several commonly used<br />
antibiotics at the 4 degree condition in order to find out the better combination under the refrigerator atmosphere.<br />
Methods:<br />
Number of survival cells after treatment of antibiotic agents at 4 degree for 24 hours was compared. Following bacterias<br />
were used; P. aeruginosa, MRSA, En. faecalis, Propionebacterium acnes, E. coli, and S. epidermidis. Used antibiotic cocktails<br />
(concentration, microgram/ml) were as follows. Solution A<br />
We use this solution at present:<br />
cefmetazole (240), lincomycin (120), vancomycin (50), polymixin B (1000 u/ml). Solution B: gentamicin (50), sitafloxacin<br />
(50), vancomycin (50), polymixin B (1000 u/ml). Solution C: gentamicin (50), sitafloxacin (50), clindamycin (100), polymixin<br />
B (1000 u/ml). Solution D: gentamicin (50), sitafloxacin (50), vancomycin (50), clindamycin (100), polymixin B (1000<br />
u/ml).<br />
Results:<br />
In P. aeruginosa and E. coli, all the 4 solutions showed complete remission. In P. acnes and S. epidermidis, Solution A did<br />
not show any effectiveness. In MRSA and E. feacalis, Solution B, C, and D worked well when the amount of bacterias was<br />
less.<br />
Conclusion:<br />
Cefmetazole which is used in our Tissue Bank at present did not show better performance than other new generations under<br />
the 4 degree condition. This result may encourage revising the antibiotic cocktails even in other tissue banks.<br />
O-28<br />
QUANTIFICATION OF RESIDUAL ANTIBIOTICS IN CARDIOVASCULAR TISSUE<br />
PREPARATIONS<br />
BROECKER, S.; HARTWIG S.; MEYER R. Charité – University Hospital Berlin. Germany.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Keywords:<br />
Cardiovascular tissue preparations, liquid chromatography mass spectrometry (LC-MS), antibiotics, residual quantification<br />
Aims: In the preparation of cardiovascular tissues for transplantation purposes an antibiosis step with a mixture of different<br />
antibiotics is included. In order to determine the degree of possible effects of the antibiotics in the recipient organism by<br />
transplantation, the presence and concentrations of the antibiotic residues in the cardiovascular tissue preparations were<br />
examined. Furthermore the systemic effect on the organism by the detected antibiotics was estimated.
Methods:<br />
The investigated samples (vascular walls or myocardial tissue) were collected in the Deutsches Herzzentrum Berlin from post<br />
mortem cases during autopsy. The preparation of the samples was performed in the same way the homografts were treated.<br />
They were incubated in 100 ml of a “medium 199” and 2 ml of an antibiotic solution consisting of amikacin, ciprofloxacin,<br />
flucytosine, metronidazole and vancomycin at 5 ° C for 24 hours. After that, they were washed three times with 0.9% NaCl<br />
and blot dried. An aliquot of approximately 20 mg vascular wall or myocardial tissue were accurately weighed and 500<br />
µl acetonitrile + 1 % formic acid were added. Then the sample was homogenized using a vibrating ball mill. After<br />
centrifugation, 400 µl of the supernatant were removed and evaporated to dryness. The residue was dissolved in 100 µl<br />
water + 1 % formic acid. 1 µl was directly injected for analysis.<br />
Results:<br />
The determination of concentrations was performed by external calibration and by standard addition. The results were in<br />
good agreement. The limit of detections (LOD) were: amikacin (10 µg/g), ciprofloxacin (0.1 µg/g), flucytosine (0.1 µg/g),<br />
metronidazole (0.1 µg/g) and vancomycin (20 µg/g). The ten investigated samples from five homografts showed that the<br />
residues lead to concentrations in the recipient organism in concentrations that are three to four orders of magnitude below<br />
the therapeutic level of the individual antibiotics.<br />
Conclusions:<br />
The detected residual amounts of antibiotics have no specific risk for the transplantation recipients. Allergic reactions cannot<br />
be excluded. For this reason, the user has to be informed about the composition of the antibiotic mixture and the risk of<br />
possible allergic reactions. According to this study, a change of the composition or method of application of the antibiotic<br />
mixture in the procedure of cardiovascular tissue preparation is not necessary.<br />
References:<br />
[1] M. Schulz, A. Schmoldt, Therapeutic and toxic blood concentrations of more than 800 drugs and other xenobiotics,<br />
Pharmazie 58 (2003), 447–474.<br />
O-29<br />
USE OF ANTIBIOTICS/ANTIMYCOTICS IN THE NEW TISSUE PRESERVATION<br />
SOLUTION TIPROTEC®<br />
RAUEN, U.; EBNER A.; DEUSSEN A. Institut für Physiologische Chemie, Universitätsklinikum Essen. Germany.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
We have previously developed the tissue preservation solution TiProtec®, which has proved to provide largely superior<br />
protection of porcine aortic segments, rat mesenteric arteries, human internal mammary arteries and human saphenous veins<br />
than diverse current cold storage solutions, suggesting its use in the tissue banking of vascular grafts. However, in many<br />
countries antibiotic/antimycotic treatment of allografts is required by the regulatory authorities. Therefore, we here assessed<br />
whether antibiotics/antimycotics can safely be added to TiProtec® without compromising its protective potential. To this<br />
end, an antibiotic/antimycotic cocktail widely used in tissue banking in Germany, consisting of gentamicin (40 µg/mL),<br />
piperacillin (1 mg/mL), flucloxacillin (1 mg/mL), metronidazole (200 µg/mL) and amphotericin B (100 µg/mL), was added<br />
to TiProtec® solution and assessed in cold storage of cultured porcine aortic endothelial cells and of the A. saphena of the<br />
rat. During 7 days of cold storage, TiProtec® solution without the antibiotic/antimycotic cocktail strongly protected the<br />
endothelial cells against cold-induced injury (LDH release 8 ± 4%) compared to cold storage in Krebs-Henseleit buffer or<br />
HTK solution (LDH release > 70%). The addition of the antibiotic/antimycotic cocktail, however, eliminated this protective<br />
effect (LDH release 87 ± 6%). Further experiments revealed that the toxicity of the cocktail was due to its antimycotic<br />
component amphotericin B, while the four antibiotics did not have any effect on endothelial cell survival. The antimycotic´s<br />
toxicity proved to be due to both toxicity of the amphotericin B itself and toxicity of the additives in the galenic preparation
used. Reduction of the antimycotic´s concentration to 10 µg/mL did not avoid the toxicity. Functional studies in the rat A.<br />
saphena confirmed these results: A. saphena cold-stored in TiProtec® for 7 days showed norepinephrine-induced vessel tone<br />
development and endothelium-dependent relaxation equivalent to fresh controls. When the antibiotic/antimycotic cocktail<br />
was added during cold storage no vasoreactivity at all could be detected, whereas omission of amphotericin B from the<br />
cocktail gave almost identical results to TiProtec® without antibiotic/antimycotics. Taken together, these results show that –<br />
with regard to cell/tissue protection – the antibiotics gentamicin, piperacillin, flucloxacillin and metronidazole can safely be<br />
added to TiProtec® solution whereas the antimycotic amphotericin B should be avoided. Testing of alternative antimycotics<br />
and of other currently used antibiotic cocktails is under way and the preservation of the antibiotic/antimycotic activities in<br />
TiProtec® solution will be assessed thereafter.<br />
O-29BIS<br />
ASSESSMENT OF BIOBURDEN ON SAMPLES OF HUMAN AND ANIMAL TISSUES:<br />
PART 1- RESULTS OF METHOD DEVELOPMENT AND VALIDATION STUDIES<br />
OSBORNE, J.; KOWALSKI, J.; MOSLEY, G.; MERRITT, K., MTF. US.<br />
Abstract:<br />
Recovered human and animal tissues are used extensively in surgery for wound repair and reconstruction of various<br />
anatomical sites. In preparation for the validation of chemical disinfection and radiation sterilization processes, studies were<br />
performed on the development of bioburden recovery methods and validation of recovery efficiency factors for human bone<br />
and soft tissue and also for porcine dermis. The “inoculated product” approach was used in combination with four repetitive<br />
extractions. Although each of the three recovery methods tested appeared to reach “exhaustion”, less than 10% recovery<br />
when compared to the first rinse, sonication plus mechanical shaking gave the highest recovery efficiency in most cases when<br />
compared to the inoculation control. The highest recovery efficiency was generally observed with Fluid D as the rinse<br />
medium. The results demonstrated the importance of performing bioburden method development and validations studies.<br />
The method validation strategy described here, using a combination of tissue inoculation and repetitive treatment, showed<br />
the superiority of sonication plus mechanical shaking using Fluid D as the rinse medium. In addition, the use of only the<br />
exhaustive extraction approach could have resulted in the development of a methodology that consistently underestimated<br />
the bioburden present on/in recovered tissue.<br />
ORAL - NOVEMBER 10TH - MUSCULOSKELETAL I<br />
ROOM 1 - 16.00-16.30<br />
ORAL<br />
PRESENTATIONS<br />
O-30<br />
THE INACTIVATION EFFECT OF STANDARD AND FRACTIONATED ELECTRON BEAM<br />
IRRADIATION ON ENVELOPED AND NON-ENVELOPED VIRUSES<br />
SCHMIDT, T.; GOHS U.; HOBURG A.; SCHUMANN W.; SCHEFFLER S.; NITSCHE A.; PRUSS A. Charité University Medicine<br />
Berlin, Julius Wolff Institut, Germany.<br />
Abstract/Introduction:<br />
As a basic safety measure, donated tendon grafts, to be used in anterior cruciate ligament replacement, currently undergo<br />
serological screening for markers of virus infections to avoid the transmission of pathogens. However, current terminal<br />
sterilization methods which inactivate all pathogens also impair the biomechanical properties of the grafts. Gamma
ORAL<br />
PRESENTATIONS<br />
irradiation shows dose dependent detrimental effects. As an additional safety tool we investigated Electron beam (Ebeam)<br />
radiation in vitro and found favourable biomechanical results which could be further improved if the required absorbed dose<br />
of 34 kGy was applied in 10 fractions of 3.4 kGy. In this study, we aimed to investigate the virus inactivation kinetics of<br />
standard and fractionated Ebeam irradiation to evaluate its impact on the virus safety of transplants.<br />
Methods:<br />
We investigated the following viruses: the enveloped human immunodeficiency type 2 (HIV-2) and pseudorabies virus (PRV,<br />
a model for human herpesviruses) and the non-enveloped hepatitis A (HAV) and porcine parvovirus (PPV, a model for<br />
parvovirus B19). All virus stocks were prepared from the supernatant of cultured infected cells. The Ebeam treatment was<br />
performed in eppendorf plastic vials in CO2 gas atmosphere at −70 ± 5°C. For evaluation of standard Ebeam (SEbeam)<br />
treatment, virus stocks were irradiated with 3.4; 6.8; 13.6-34 kGy within one step. In the case of fractionated Ebeam<br />
(FEbeam) process, virus stocks were irradiated with 1 x 3.4 kGy; 2 x 3.4 kGy; 3 x 3.4 kGy;-10 x 3.4kGy. The log (10)<br />
reduction was measured by cytopathogenic effects after virus titration (TCID(50)/ml) and the D10 values (kGy) were<br />
calculated for the different viruses.<br />
Results:<br />
We determined the following D10 values: HIV-2: SEbeam 9.0±0.5 kGy; FEbeam: 8.0±0.5 kGy, PRV: SEbeam: 5.6±0.4 kGy;<br />
FEbeam: 5.8±0.4 kGy, HAV: SEbeam: 6.5±0.2 kGy; FEbeam: 5.9±0.2 kGy, PPV: SEbeam 8.6±0.6 kGy; FEbeam: 7.5±0.6<br />
kGy. A dose of at least 36.0 kGy at -70°C for SEbeam treatment and a dose of 32.0 kGy for FEbeam treatment were<br />
necessary to achieve a sufficient reduction of 4 log steps in the case of HIV-2, which was the most resistant of all viruses<br />
investigated in this study.<br />
Discussion:<br />
For both Ebeam processes comparable virus inactivation kinetics and D10 values were determined. The superior<br />
biomechanical in vitro results using the fractionated Ebeam process compared to standard Ebeam or gamma treatment<br />
suggest that this novel procedure is a safe and effective option for a terminal sterilization method which achieves full pathogen<br />
inactivation without impairing the biomechanical properties of the grafts. However, the biological effects must be confirmed<br />
in an animal model before it can be used for human graft sterilization.<br />
O-31<br />
EFFECT OF GAMMA IRRADIATION ON MECHANICAL PROPERTIES OF HUMAN<br />
CORTICAL BONE: INFLUENCE OF DIFFERENT PROCESSING METHODS<br />
JASTRZEBSKA, A.; GRAZKA E.; GUT G.; UHRYNOWSKA-TYSZKIEWICZ I.; MAROWSKA J.; KAMINSKI A. National<br />
Centre for Tissue and Cell Banking, Warsaw, Poland.<br />
Abstract/Objectives:<br />
Gamma irradiation of cortical bone grafts utilized in reconstructive medicine is used to prevent infective disease transmission.<br />
However, gamma irradiation usually performed at low dose rate in combination with a high exposure period, has been<br />
supposed to impair mechanical properties of bone tissue, essential for bone graft clinical functionality. To adress this issue,<br />
we evaluated mechanical effects in human cortical bone grafts, induced by irradiation with two doses of gamma rays (25<br />
and 35 kGy) in different processing conditions*.<br />
Material and methods:<br />
Left and right femoral shafts procured from six male cadaveric donors (mean age 51+/-3 yrs) were cleaned of soft tissues,<br />
transversely cut into slices of 10 mm height from which bone marrow was removed, and assigned to eight experimental<br />
groups of twelve specimens each, according to different processing methods (defatted or non-defatted), gamma irradiation<br />
dose (25 or 35 kGy, Co-60 Gamma Irradiator ISOGAMMA-LLCo, Hungary), as well as temperature conditions of irradiation<br />
(ambient temperature or dry ice). Specimens from control groups (defatted or non-defatted) were not irradiated. Prior to<br />
mechanical testing for compression, cross-sectional area of each bone slice was measured using computer tomography
(Toshiba Aquilion CT scan system TSX-101A). The compressive strength of bone rings was measured in the wet state with<br />
the speed of compression of 1mm/s, using Material Testing Machine Z250 (Zwick/Roell, Germany), and mechanical<br />
properties calculated from the load-deformation curve. Results We did not observe any statistically significant differences<br />
in strength at fracture and Young’s modulus of gamma-irradiated cortical bone rings as compared to control ones (nonirradiated),<br />
irrespective of gamma dose applied (25 or 35 kGy), temperature of irradiation (ambient temperature or dry<br />
ice), as well as processing procedure (defatting or non-defatting) prior to irradiation.<br />
Conclusion:<br />
Gamma irradiation of compact bone graft at the doses applied seems not to impair their mechanical competence under<br />
compression within elastic strain region, irrespective of irradiation temperature and processing method. * The study was<br />
partially supported by International programme „Safety and Optimisation of Radiation Sterilization in Tissue Banking:<br />
Studies on Functional Properties of Irradiated Tissue Grafts (CRP E3.10.06), IAEA Research Contract No.16114/R0.<br />
O-32<br />
EFFECT OF ELECTRON BEAM IRRADIATION ON MECHANICAL PROPERTIES OF<br />
HUMAN CORTICAL BONE: INFLUENCE OF DIFFERENT PROCESSING METHODS<br />
GRAZKA, E.; JASTRZEBSKA A.; GUT G.; UHRYNOWSKA-TYSZKIEWICZ I.; MAROWSKA J.; KAMINSKI A.<br />
National Centre for Tissue and Cell Banking, Warsaw, Poland.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Objectives:<br />
Electron beam (EB) irradiation of bone grafts is an alternative method of sterilization to gamma irradiation, with advantages<br />
including high dose rate and short exposure time on the expense of the approximately ten-times lower penetration into<br />
materials, requiring, in some cases, two-sided irradiation. Due to its characteristics, EB sterilization might result in low<br />
radiation damage of biological tissues, and, in consequence, low impairement of mechanical function. To adress this issue,<br />
we evaluated mechanical effects in human cortical bone grafts, induced by irradiation with two doses of EB irradiation (25<br />
and 35 kGy) in different processing conditions*.<br />
Material and methods:<br />
Left and right femoral shafts procured from six male cadaveric donors (mean age 51+/-3 yrs) were cleaned of soft tissues,<br />
transversely cut into slices of 10 mm height from which bone marrow was removed, and assigned to eight experimental<br />
groups of twelve specimens each, according to different processing methods (defatted or non-defatted), EB irradiation dose<br />
(25 or 35 kGy, Electron Beam Accelerator LAE-10, 10,2 MeV, Institute of Nuclear Chemistry and Technology, Warsaw), as<br />
well as temperature conditions of irradiation (ambient temperature or dry ice). Specimens from control groups (defatted or<br />
non-defatted) were not irradiated. Prior to mechanical testing for compression, cross-sectional area of each bone slice was<br />
measured using computer tomography (Toshiba Aquilion CT scan system TSX-101A). The compressive strength of bone<br />
rings was measured in the wet state with the speed of compression of 1mm/s, using Material Testing Machine Z250<br />
(Zwick/Roell, Germany), and mechanical properties calculated from the load-deformation curve.<br />
Results:<br />
We did not find any statistically significant differences in strength at fracture and Young’s modulus of EB-irradiated cortical<br />
bone rings as compared to control ones (non-irradiated), irrespective of EB dose applied (25 or 35 kGy), temperature of<br />
irradiation (ambient temperature or dry ice), as well as processing procedure (defatting or non-defatting) prior to irradiation.<br />
Conclusion:<br />
Electron beam sterilization of compact bone graft at the doses applied seems not to impair their mechanical competence<br />
under compression within elastic strain region, irrespective of irradiation temperature and processing method. * The study<br />
was partially supported by International programme „Safety and Optimisation of Radiation Sterilization in Tissue Banking:<br />
Studies on Functional Properties of Irradiated Tissue Grafts (CRP E3.10.06), IAEA Research Contract No.16114/R0.
ORAL - NOVEMBER 10TH - MUSCULOSKELETAL II<br />
ROOM 1 - 17.00-18-40<br />
ORAL<br />
PRESENTATIONS<br />
O-33<br />
BENEFITS OF USING A DECONTAMINATION METHOD IN BONE PROCESSING<br />
FARIÑAS, O. 1 , VILARRODONA, A. 1 , VITO, S. 1 , TABERA, J. 1 , HINOJOSA, M. A. 1 , SAVIO, A. 1 , SEGUR, J. M. 2 ,<br />
SUSO, S. 2 , TRÍAS, E. 1<br />
1 - Transplant Services Foundation - Hospital Clínic. Barcelona. Spain., 2 - Hospital Clínic. Barcelona. Spain.<br />
Abstract/Introduction:<br />
The main objective of a tissue establishment is to minimize the most disease transmission through allografts transplantation.<br />
This aim is achieved establishing a strict donor selection, a protocolized recovery system and a controlled tissue processing.<br />
In addition to bone processing into monitored cleanrooms areas, the use of a decontamination method on the grafts<br />
decreases significantly the risk of infectious disease transmission. It has been published that bacteries and fungi are mainly<br />
located inside the musculoskeletal tissue grafts. The function of some decontamination methods is to remove the blood, fat<br />
and bone marrow remains from inside the grafts affecting also the presence of microorganisms. Aim To evaluate the influence<br />
of using a decontamination method in musculoskeletal tissue processing from 2007 to 2010, comparing the contamination<br />
rates with the results obtained without using it from 2004 to 2006.<br />
Materials and Methods:<br />
This retrospective study evaluates the musculoskeletal tissue processings performed in our tissue establishment from 2004 to<br />
2010. Two different time periods have been differenciated: 2004 to 2006 when no decontamination method was used, and<br />
2007 to 2010 when we started using it. The analysis focused on the number of grafts obtained after processing, the positive<br />
cultures obtained, and the rate of grafts discarded because of microbiological contamination. The results obtained were<br />
compared between the two different periods with and without using decontamination method.<br />
Results:<br />
The number of musculoskeletal tissue donors have been increasing from last seven years (31 to 145 donors per year). In a<br />
parallel way the number of allografts obtained after processing have increased too (541 grafts in 2004, 4413 grafts in<br />
2009). During the period from 2004 to 2006 the musculoskeletal allografts were only subjected to a superficial clean<br />
process with sterile water. The rate of positive cultures after processing ranged 10.53-16.46%. The average rate of allografts<br />
discarded due to positive microbiological culture ranged 7.21-8.13%. In 2007 we started using a decontamination method<br />
that consisted in a mix of mechanical and chemical procedures to eliminate the remains of blood, fat and bone marrow from<br />
inside the grafts. The rate of positive cultures after processing ranged 0.45-3.16%. The average rate of allografts discarded<br />
due to positive microbiological culture ranged 0.23-1.58%.<br />
Conclusions:<br />
The use of a decontamination method in musculoskeletal tissue processing (independently from a terminal sterilization<br />
method) decreases the risk of infectious disease transmission. Other benefit obtained is to increase the availability of allograft<br />
to transplant due to its effect on the number of tissues discarded because of a positive microbiological culture.
O-34<br />
EVIDENCE BASED BONE AND TISSUE BANK PROCESSES STANDARDIZATION<br />
NAVAS, J.; MIETH K.; SOTO C.; GONZÁLEZ J. Fundación Cosme y Damián. Colombia.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
Standardization is essential to improve quality in every step of the bone and tissue chain of action, from donor selection to<br />
graft distribution. Objective Using the Fundacion Cosme y Damian bone bank experience we show a state of the art evidence<br />
based view of how, who, when and why should do a standardize process including the whole spectrum of activities of a<br />
bone and tissue bank. Material and methods There are 7 steps in any standardization process: To establish and entrance<br />
and an exit point, to build the process, define the desire outcomes, determine the key questions to obtain the outcomes,<br />
answer the questions based in current evidence, standardize and measure the results.<br />
Conclusions:<br />
Standardization improves quality in every level of the organization, it is a way of detection and correction of process errors<br />
and a form of allow to repeat the right steps in the same way every time. However, it is of paramount importance that<br />
before an activity is standardizing it must be validated. It would not make sense to standardize processes that are not<br />
legitimate.<br />
O-35<br />
COMPARISON OF FROZEN AND FREEZE-DRIED CANCELLOUS BONE GRAFT IN<br />
CAVITARY DEFECTS<br />
CAMACHO CARRASCO, P.; SEGUR VILALTA J.M.; GARCÍA OLTRA E.; GARCÍA ELVIRA R.; TORNER PIFARRÉ P.; FARIÑAS<br />
BARBERÁ Ó.; SUSO VERGARA S. Hospital Clínic of Barcelona. Spain.<br />
Abstract:<br />
Bone graft is the second most common transplanted tissue just behind blood. Autogenous grafts harvested from iliac crest<br />
remain the gold standard. However, their use is associated to rates of morbidity that range from 8.5 to 20%. Bone allografts<br />
are the most frequently chosen bone substitute and are mainly processed by freezing and freeze-drying. Few studies are<br />
reported in the literature that compared biological behaviour of frozen and freeze-dried bone allografts. We performed a<br />
histological and histomorphometric study to evaluate and compare the incorporation of cancellous grafts in cavitary defects.<br />
Forty-eight New Zealand rabbits weighing 3.5-4.5 kg were used in this experiment as it involved skeletal maturity. According<br />
to the method described by Katthagen in 1984, which is a modification of Maatz’s, a 6 mm defect was created in medial<br />
femoral condyle. This diameter restricts the spontaneous regenerative capacity as it represents more than a half of the<br />
condyle. Four treatment groups were established, control (without any implant), autologous, frozen allograft and freeze-dried<br />
allograft, and each of them was subdivided attending to the sacrifice time, at 4 or 12 weeks. After sacrifice bone specimens<br />
were processed for the study of non-decalcified bone. Sections were stained with von Kossa’s stain, which offered a great<br />
contrast that eased histomorphometric analysis. Histomorphometric measurements, such as implant surface, trabecular area,<br />
osteoid surface and osteoblast surface, were obtained directly from the digitalized microscopic images. Histomorphometric<br />
parameters derived from histomorphometric measurements and were specific trabecular bone surface (Sv), relative bone<br />
formation surface (Sf), osteoblastic-osteoid surface (OBOID) and mean osteoid seam thickness (MOST). These parameters<br />
informed about structure (Sv) and osteoblastic activity (Sf, OBOID, MOST). The histological analysis revealed minimum bone<br />
formation in periphery without spontaneous regeneration in the majority of the defect in control group. Autologous group
ORAL<br />
PRESENTATIONS<br />
showed bone trabeculae surrounded by abundant osteoid seams homogeneously distributed and osteoblasts. The frozen<br />
allograft group presented a more heterogenous distribution of bone trabeculae and a smaller quantity of osteoid and<br />
osteoblasts. The freeze-dried allograft group also exhibited the histological findings of frozen allograft group, but the most<br />
striking feature was the large number of osteoclasts. In global histomorphometric analysis related to treatment groups<br />
autologous graft showed best results, with statistically significant differences in OBOID. Overall frozen and freeze-dried<br />
allografts were comparable. One hand there was not statistically significant differences in Sf and MOST and on other hand<br />
there was found statistically significant differences in Sv and OBOID (greater values of Sv in freeze-dried group and of<br />
OBOID in frozen group).<br />
O-36<br />
VALIDITY OF AN AUTOMATIC MEASURE PROTOCOL IN DISTAL FEMUR FOR<br />
ALLOGRAFT SELECTION FROM A THREEDIMENSIONAL VIRTUAL BONE BANK<br />
SYSTEM<br />
APONTE-TINAO ALBERTO, L.; MILANO EDGARDO F.; FARFALLI LUIS G.; RITACCO EDUARDO L.; SCHWINT O.; SEILER<br />
C.; REYES M. Italian Hospital of Buenos Aires. Argentina.<br />
Abstract/Background:<br />
Ostearticular allograft is one of the possible treatments after wide surgical resections in large bone defects. Performing best<br />
allograft selection is of great relevance for optimal exploitation of the bone databank, good surgery outcome and patient's<br />
recovery. Current approaches are, however, very time consuming hindering these points in practice. We present a validation<br />
study of a software able to perform automatic bone measurements used to automatically assess the distal femur sizes across<br />
a databank, from six pre-defined anatomical landmarks.<br />
Methods:<br />
170 distal femur surfaces were reconstructed from CT data and measured manually using a measure protocol taking into<br />
account the transepicondyle distance (A), anterior-posterior distance in medial condyle (B) and lateral condyle (C). Intra-,<br />
and inter-observer studies were conducted and regarded as ground truth measurements. Manual and automatic measures<br />
were compared using, a statistic description (means, maximal and minimal differences), intraclass correlation coeficient.<br />
Results:<br />
A single operator was tested for intraobserver repeatability while using the above-mentioned A-B-C protocol twice on the<br />
bone surfaces, obtaining an intraclass correlation coefficient of 0.99 for all measures. Interobserver consistency of two<br />
separate observers was quantified as well for the same cohort, leading to an intraclass correlation coefficient of 0.99 for A<br />
measure, and of 0.98 for B and C measures. For the automatic measurements, the correlation coefficients between observer<br />
one and automatic method, were of 0.99 for A measure and 0.96 for B and C measures. The average time needed to<br />
perform the measurements was of 16 hrs for both manual measurements, and of 3 minutes for the automatic method.<br />
Conclusion:<br />
The proposed methodology is presented as a key element towards effective and fast allograft selection, advancing the state<br />
of the art over current time-consuming and labor-intensive solutions. The results demonstrate the high reliability and, most<br />
importantly, high repeatability of the proposed approach, and considerable speed-up on the planning.
O-37<br />
WHOLE ACETABULUM ALLOGRAFTS : A TWENTY YEARS EXPERIENCE<br />
RIBAS, M.; VILARRUBIAS J.; GINEBREDA I.; DE LA TORRE B.; BELLOTTI V.; DE MEO F.; CAVALIERE P.<br />
University Hospital Dexeus. Spain.<br />
Abstract/Introduction:<br />
Even today there is still no absolute consensus in the reconstruction of the severe columnar defects in hip revision. It is<br />
precisely in these cases where bone reconstruction provides by far lower survival, and therefore the trend is the early revision<br />
at the first sign of any eventual osteolysis induced loosening, that the patient may present. But historically this was not the<br />
case. During the seventies and eighties many cases have been associated to massive bone loss. In our case this condition<br />
led us in 1988 to use massive acetabular allografts that reproduced faithfully the bone stock and bony anatomy so far. This<br />
study presents a historical series that allows us to keep in mind how these allografts behave in the medium and long term.<br />
Material and methods:<br />
We present a series of 44 transplants acetabulum with a mean of 16.2 years (range 9-22). The mean patient age was 58.6<br />
years (range 19-83). According to the Classification of Gross 26 cases had type III acetabular defect, while 18 had type<br />
IV. The evaluation included Merle D'Aubigné score and radiological evaluation of the graft and the acetabular implant was<br />
performed according to radiological Engh criteria (JBJS, 1994).<br />
Results:<br />
The homogenization of the trabecular radiological pattern was observed in 42 of 44 cases (95.4%). There were 3 infections<br />
and 8 cases of aseptic loosening (18.1%), which were revised with only a new cup implantation. To date none of these eight<br />
cases have shown more signs of loosening. According to the Kaplan-Meier the overall survivorship rate ,- endpoint hip<br />
revision for any given reason -, was 76.4% at 15 years in the type III cases, but in cases of pelvic discontinuity (type IV),<br />
survival was significantly higher ( 85.7%, p = 0.018). There was a remarkable improvement in Merle d'Aubigné score up<br />
(preoperative 2.2 to 4.9 during follow-up, p = 0.021) and pain (2.5 preoperatively - 5.4 during follow-up, p = 0032).<br />
Conclusions:<br />
In spite of some articles published by different authors with high incidence of failures in the midterm with massive structural<br />
allografts in this series clearly it has been shown, that an original acetabular allograft can provide an acceptable result along<br />
the time and allows in the most severe cases restoration of acetabular bone stock, making possible that further reconstruction<br />
is possible in these patients.<br />
O-38<br />
ONLAY CORTICAL STRUT ALLOGRAFTING IN TOTAL HIP ARTHROPLASTY (THA)<br />
REVISION<br />
MEDRANO, C.; BORI G.; GALLART X.; SEGUR J. M.; FERNÁNDEZ-VALENCIA J.; RIBA J.; GARCÍA S.<br />
Hospital Clínic Barcelona. Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Objective:<br />
To report our experience in onlay cortical strut allografting in revision Total Hip Arthroplasty (THA). Material and methods:<br />
Retrospective, observational study on 13 patients treated with revision THA that were performed in our center and used a<br />
cortical strut allograft, from January 2001 to April 2009. For all the patients we determine before the surgery the following<br />
data: age, sex, comorbidities, age of the prosthesis, protein C reactive (PCR) and globular sedimentation velocity (VSG),
corporal mass index (IMC), bony scintygraphy with 99mTc-HMPAO leukocytes and TC hip scan in some cases. In all the cases<br />
samples from the periprosthesic material were taken for microbiological studies and polimorfonucleates count. We classify<br />
the type of femoral defects according to Paprosky, the type of allograft used and the prosthesis. We also evaluate<br />
complications and the functional evaluation by Merle d'Aubignè and Harris Hip Score.<br />
Results:<br />
14 cases were identified (13 patients: a case was bilateral); 12 cases were aseptic revisions and 2 cases were the second<br />
stage of a septic revision HA; 11 women and 2 men, the average age was 72.5 (rank 60-77) and the average prosthesis<br />
age was about 4 years (rank 2-11). The scintygraphy with 99mTc-HMPAO leukocytes was negative in the 12 aseptic cases<br />
and it was not performed in the 2 cases of the second stage of a septic procedure. The femoral defects were all type IIIA<br />
Paprosky. Two cases presented positive microbiological studies for ECN but the PMN count was not more than 5pmn/pc.<br />
In these 2 cases the antibiotic treatment was mantained 6 weeks postoperatively. The radiological results are: 9 cases with<br />
total integration of allograft, 4 cases of total resorption and 1 case of nonunion. No cases of implant failure was observed.<br />
The average follow-up has been of 31.57 months (rank 12-97). The functional evaluation Merle d'Aubigné score was about<br />
14, 53 (rank 11-17) and the Harris Hip Score was 66.21 (rank 43-91,80).<br />
Discussion:<br />
The use of onlay cortical strut allograft in revision hip surgery can be appropiate in those cases with radiological criteria of<br />
femoral defects and bone loss to prevent intraoperative periprosthesic fractures and to increase bone stock for successive<br />
surgeries if needed.<br />
O-39<br />
TREATMENT OF BENIGN BONE TUMORS WITH DEMINERALIZED BONE MATRIX<br />
AFTER CURETAGGE<br />
SOTO, C.; SOTO C.; NAVAS J.; MIETH K.; GONZÁLEZ J. Fundación Cosme y Damián. Colombia.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
Curettage has been the gold standard for treatment of benign bone tumors stages 2 and 2-3. It is important to fill the residual<br />
bone defect in order to recover the bone structure and mechanical properties. Therefore, a number of materials like<br />
autografts, demineralized bone matrix, and synthetic composites have been used to fill such defects.<br />
Objectives:<br />
To determine the feasibility and efectiveness of demineralized bone matrix (DMO) to fill bone defects after intralesional<br />
treatment of benign bone tumours.<br />
Methods:<br />
From December 2002 to March 2010, eighty nine (n=89) consecutive patients with benign bone tumours were treated with<br />
curettage and high speed burring. We use demineralized bone matrix to fill residual bone defects. The patients were followed<br />
every four months with simple X rays for an average of 12 months, time when radiographic signs of healing were evaluated.<br />
Results:<br />
The average of healing and allograft incorporation was 6.2 months. We report tumoral recurrence in four patients. No<br />
patient had a pathologic fracture during the early bone healing stage. We obtained 96% rate of success, measured as no<br />
tumor recurrence and restoration of the normal intramedular pattern using the Neer radiologic assessment tool.<br />
Conclusion:<br />
The use of demineralized bone matrix to fill residual bone defects is an effective osteogenic material in the intralesional<br />
treatment of bening bone tumors.
O-40<br />
AN OVERVIEW OF THE OPERATIONS, TRACEABILITY AND SAFETY ASPECTS OF<br />
HUMAN BONE ALLOGRAFTS AT THE CENTRE FOR TISSUE ENGINEERING – BONE<br />
BANK IN SOUTH AFRICA<br />
KARAKATSANIS, E. Centre for Tissue Engineering - Bone Bank (Tshwane University of Technology). South Africa.<br />
Abstract:<br />
Purpose of study The Bone Bank provides more than 18 000 allografts to the medical fraternity across South Africa annually.<br />
Therefore, the purpose of the study is to familiarise the end user of the various stringent critical quality control (QC) points<br />
that have been established in the processing of human bone allografts. The QC points are initiated prior to the procurement<br />
of tissue right through to the processing of the tissue.<br />
Description of methods:<br />
Musculoskeletal allografts provide the best solution to particular surgical procedures including revision surgery where an<br />
autograft is unavailable or as a supplement when an ample supply of autograft tissue is not available. Safety of tissue<br />
allografts remains priority, hence the thorough pre-donor selection criteria questionnaire and serology and microbiology (preprocurement,<br />
in-processing and post-sterilisation) testing and verification. Aseptic processing is a common method of<br />
allograft processing and aims to minimise contamination of the allograft tissue from the environment, processing personnel<br />
and equipment. Hence, the implementation of strict audited operating protocols and the maintenance of clean room facilities<br />
that minimise contamination. Traceability of tissue from procurement to the utilizable allograft is ensured and recorded at<br />
all critical control points of the process. Gamma irradiation (terminal sterilization) is utilised for the sterilisation of allografts.<br />
Summary of results:<br />
The quality and safety of our products is achieved through effective planning and monitoring which governs every step of<br />
all the required processes to minimise any possibility of product failure. Donor statistics, risk factors and trend analysis have<br />
been identified and understanding these results allows for more effective donor consent rates and the improvement of<br />
operational protocols.<br />
Conclusion:<br />
Implementing and maintaining ISO 9001 (Quality Management System) and ISO 13485 (Medical Device) allows us to<br />
measure the effectiveness and consequences of our work, which is continually monitored against defined process and quality<br />
objectives, which are seen as the foundation to high-quality process management.<br />
O-41<br />
TEST SYSTEM FOR OSTEOINDUCTIVE ACTIVITY OF MATERIALS CONTAINING<br />
RECOMBINANT MORPHOGENETIC PROTEIN RHBMP-2<br />
AKATOV V.; CHEKANOV A.; LEKISHVILI M.; FADEEVA I.; SKLYANCHUK E.; GURIEV V.; RYABOV, A.<br />
Institute of theoretical and experimental biophysics of RAS, Poushchico. Russian Federation.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Development of materials including recombinant morphogenetic proteins(BMPs) is one of the promising areas of<br />
traumatology, orthopedics and maxillofacial surgery. BMP proteins are key factors in the remodeling and regeneration of<br />
bone tissue. These proteins have a powerful osteoinductive effect and can stimulate new bone formation through
differentiation of mesenchymal cells to osteoblasts. This circumstance serves as the basis for application of recombinant<br />
human morphogenetic proteins (rhBMPs) in tissue-engineered materials to enhance their osteoinductive properties. Since<br />
2002, the material under the trade mark Infuse containing rhBMP-2 is used widely. Now the engineering of new materials<br />
including rhBMPs, in particulare rhBMP-2, is active developing field. To evaluate the osteoinductive activity of the materials,<br />
different test systems are applied including cell assays in vitro, ectopic implantation models, and models of orthotopic<br />
implantation in experimental animals. This report presents the results of a comparative evaluation of osteoinductive properties<br />
of the material produced using recombinant morphogenetic protein rhBMP-2 or without rhBMP-2 in the model of ectopic<br />
implantation in experimental animals. We used in our study a composite material including demineralized bone matrics,<br />
wich was filled with alginate gel containing rhВМР-2 or without rhВМР-2. Osteoinductive activity of materials was evaluated<br />
in a model of subcutaneous implantation in Wistar rats for a period of 1.5 months. Inclusion of rhBMP in material drastically<br />
increased the mineralization of the implants, stimulated the formation of a structured collagen. In particular, after the<br />
implantation, the content of mineralized calcium in fragments, which were previously treated with guanidine and then loaded<br />
with rhBMP-2 containing gel, was of 150 ± 30 mg / g of dry weight of the fragment, whereas in the same material, which<br />
were loaded without rhBMP-2, the calcium content was of 1 ± 1 mg / g of dry weight. Demineralized bone matrix without<br />
guanidine treatment (without removal of bone BMPs) and without the inclusion of rhBMP-2 was detected of 15 ± 5 mg / g<br />
of dry weight of explanted material. Histological analysis showed destruction of collagen and its replacement by disorganized<br />
collagen (scar) in explanted materials containing no rhBMP-2, whereas the formation of a structured collagen was detected<br />
in the implants containing rhBMP-2. The results obtained show the effectiveness of subcutaneous implantation model as test<br />
system of osteoinductive activity of materials containing rhBMP-2, and point to the prospects of the material tested for<br />
practical applications. This work was supported by the Ministry of Education and Science of Russian Federation, contracts<br />
№ 2.1.1/11708, № 02.740.11.0710 and № P609 and was made using devices of the Regional Center for Collective Use<br />
at the Pushchino Institute of Theoretical and Experimental Biophysics.<br />
O-42<br />
FREEZE-DRIED HUMAN SERUM ALBUMIN IMPROVES THE ADHERENCE AND<br />
PROLIFERATION OF MESENCHYMAL STEM CELLS ON MINERALIZED HUMAN BONE<br />
ALLOGRAFTS<br />
CSÖNGE , L.; SKALICZKY G.; KLARA T.; WESZL M.; SCHANDL K.; SZENDROI M.; LACZA Z.<br />
West Hungarian Regional Tissue Bank. Hungary.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Mineralized scaffolds are widely used as bone grafts with the assumption that bone marrow derived cells colonize and<br />
remodel them. This process is slow and often unreliable so we aimed to improve the biocompatibility of bone grafts by preseeding<br />
them with human mesenchymal stem cells from either bone marrow or dental pulp. Under standard cell culture<br />
conditions very low number of seeded cells remained on the surface of freeze-dried human or bovine bone graft or<br />
hydroxyapatite. Coating the scaffolds with fibronectin or collagenimproved seeding efficiency but the cells failed to grow<br />
on the surface until the 18th day. In contrast, human albumin was a very potent facilitator of both seeding and proliferation<br />
on allografts which was further improved by culturing in a rotating bioreactor. Electron microscopy revealed that cells do<br />
not form a monolayer but span the pores, emphasizing the importance of pore size and microstructure. Albumin coated bone<br />
chips were able to unite a rat femoral segmental defect, while uncoated ones did not. Micro-hardness measurements<br />
confirmed that albumin coating does not influence the physical characteristics of the scaffold, so it is possible to introduce<br />
albumin coating into the manufacturing process of lyophilized bone allografts.
ORAL - NOVEMBER 10TH - SKIN & AMNIOTIC MEMBRANE I<br />
ROOM 3 - 15.30-17.30<br />
O-43<br />
SKIN DECONTAMINATION AND IMPACT OF ANTIBIOTIC RESIDUES ON<br />
MICROBIOLOGICAL TESTS<br />
VASSANELLI, A.; BOSINELLI A.; RIZZI S.; LUCCHINI E.; GATTO C.; GIURGOLA L.; TOTHOVA J.<br />
Tissue Bank of Verona. Italy.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
The aim of the present study was to evaluate the performances of the BASE.128 medical in comparison with the antibiotic<br />
solution currently used by the Tissue bank of Verona and validate the time and temperature conditions of skin<br />
decontamination. Methods. Skin samples from five different donors were retrieved by the Skin Bank of Verona. Samples were<br />
divided in two halves of 100 cm2 each and processed in parallel using an antibiotic cocktail containing Penicillin,<br />
Streptomycin, Amphotericin B, Bactrim and Gentamicin prepared by the bank or BASE.128 containing Cefotaxime,<br />
Gentamicin, Vancomycin, Amphotericin B (AL.CHI.MI.A., Italy). Samples were decontaminated with the bank cocktail or<br />
BASE.128 at 22°C for 90 min. followed by three consecutive rinsing of 5 min. each with NaCl 0,9% or BASE (AL.CHI.MI.A.,<br />
Italy), respectively. All samples were cryopreserved in BASE containing 10% CRYO.ON (AL.CHI.MI.A., Italy) and stored at<br />
-80°C. Bacteriological tests were performed on donor skin swabs, tissue samples and transport and rinsing liquids using<br />
thioglycollate medium incubated at 37°C for 7 days. After thawing, the presence of antibiotic residues was evaluated on<br />
tissue homogenates by disk diffusion and sterility test performed according to the European Pharmacopeia after removing<br />
potential interfering antibiotics with a specific resin mixture. Contaminants were genetically identified by ribosomal RNA<br />
sequencing. Vitality assay with MTT colorimetric method was performed on each tissue before processing and 10 days after<br />
thawing.<br />
Results:<br />
No bacterial or fungal contamination was detected in the microbiological tests performed on donor skin swabs, tissue<br />
samples before and after processing, and transport and rinsing liquids. The sterility test performed on thawed tissues after<br />
removal of the antibiotic showed bacterial (Staphylococcus spp.) and fungal (Penicillinum spp., Candida spp.) contamination<br />
in all investigated tissues. The sterility test performed without removing the residual antibiotics showed the absence of<br />
contaminants, thus indicating a possible interference of residual antibiotics with bacterial growth during test. 6- and 2-fold<br />
greater S. Aureus and P. Aeruginosa inhibition zones were observed in the disk diffusion test of tissues treated with the<br />
bank cocktail as compared to tissues decontaminated with BASE.128. No inhibition of Candida Albicans was observed in<br />
tissues treated with both solutions, thus indicating the absence of antifungal residues. The vitality assay showed an average<br />
of 40% metabolically active cells in cryopreserved tissues decontaminated with both solutions as compared to fresh donor<br />
skin.<br />
Conclusions:<br />
The presence of antibiotic residues after tissue decontamination can interfere with the microorganism growth during sterility<br />
test and result in false negative. The amount of antibiotic residues can vary significantly with the antibiotic cocktail.<br />
Consequently, microbiological analysis shall only be performed after an accurate removal of the antibiotic residues. The time<br />
and temperature conditions, used for both decontamination solutions were not effective eliminating contaminants from donor<br />
skin. Further studies evaluating different decontamination time and temperature conditions are in progress.
O-44<br />
INACTIVATION OF BORRELIA BURGDORFERI BY GLYCEROL<br />
RICHTER , C.; OEI A.; DE WEVER B.; HOVIUS J. Euro Skin Bank. The Netherlands.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
Lyme disease is caused by the spirochete Borrelia Burgdorferi and is transmitted by infected ticks to humans. The first clinical<br />
sign of Lyme disease is an erythematous expanding cutaneous lesion, designated erythema migrans, caused by an<br />
inflammatory reaction directed against the spirochetes residing in the skin. A significant increase in the number of patients<br />
is observed the last decade. Therefore, a recent tick bite is a contra-indication for skin donation. It has been shown that<br />
several bacteria species can be inactivated after incubation in glycerol at high concentration (85%). Storage of donor skin<br />
in 85% glycerol is used by skin banks as a method to decontaminate donor skin. In this study, we investigated the effect of<br />
glycerol on the survival of the Borrelia Burgdorferi.<br />
Method:<br />
Full thickness sheets of skin were injected subcutaneously with different concentrations (101 till 105 as enumerated by darkfield<br />
microscopy and a Petroff-Hausser counting chamber) of viable Borrelia Burgdorferi strain B31 (50 µl per biopsy). The<br />
skin was cultured for 16h at 33°C in RPMI medium. Thereafter, part of the biopsies were directly put in an assay to detect<br />
viable Borrelia spirochetes (culture method using modified BSK medium), part were used for DNA isolation and detection<br />
by semi-quantitative PCR. The other biopsies were first treated with glycerol according to the normal procedure used in the<br />
skin bank for donor skin. Briefly, the skin is put first for 24h in 50% glycerol with pen/strep and then for 3 weeks in 85%<br />
glycerol. After the glycerol treatment again part of the biopsies were tested in the culture assay to detect viable spirochetes<br />
and part was used for DNA extraction and PCR. Biopsies of the skin without any treatment and skin injected with only<br />
medium served as controls. In addition, non-viable Borrelia (incubated at 56°C for 30 min) were injected.<br />
Results:<br />
The lowest concentration of injected spirochetes that could be detected using this human skin model was 100, both in the<br />
culture test and the semi-quantitative PCR. All biopsies injected with spirochetes were negative after the glycerol treatment<br />
in the culture assay, no viable bacteria could be detected. The PCR results showed the presence of Borrelia DNA in the<br />
biopsies were spirochetes were injected, also in the biopsies treated with glycerol. Positive PCR results were obtained also<br />
in the biopsies injected with the heat inactivated non viable Borrelia.<br />
Conclusion:<br />
And discussion In biopsies of skin that were injected with viable spirochetes and treated with glycerol 85%, no bacteria could<br />
be detected using the culture method. Borrelia DNA could be detected using the PCR. Most probably, the bacteria are non<br />
viable but the DNA is still intact. In earlier studies, at has been shown glycerol at high concentration results in cell death with<br />
intact morphology. In addition, the biopsies injected with heat inactivated spirocytes were also positive in the PCR. The<br />
results with the culture assay indicated glycerol 85% inactivates for at least 3 weeks inactivates the Borrelia spirochetes. Since<br />
the sensitivity of this model is 100 bacteria, further risk assessment must be used to decide if donor skin from a donor with<br />
a tick bite without an erythema migrans can be accepted after treatment with glycerol.
O-45<br />
ESTABLISHMENT OF A RINSING PROCEDURE TO ELIMINATE ANTIBIOTIC AND<br />
GLYCEROL RESIDUES FROM CRYOPRESERVED SKIN DECONTAMINATED WITH<br />
BASE.128<br />
PIANIGIANI, E.; GATTO C.; GIURGOLA L.; IERARDI F.; D'AMATO TOTHOVA J. Siena Skin Bank. Italy.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Skin decontamination and cryopreservation can result in the presence of antibiotic and glycerol residues that shall be<br />
removed before transplantation in order to ensure tissue safety. Purpose To establish a rinsing procedure, to be performed<br />
before skin transplantation, with the aim of removing at least 80% of the antibiotic and glycerol residues from the<br />
cryopreserved skin. Methods Human skin samples from five donors were processed by the Tissue Bank of Siena. For each<br />
donor, 100 cm 2 of skin were decontaminated and cryopreserved in a single processing phase either with solution prepared<br />
at the bank (15% glycerol, Penicillin, Streptomycin, Gentamicin sulphate, Amphotericin B, and DMEM) or with the medical<br />
device BASE.128 (AL.CHI.MI.A s.r.l.) containing Vancomycin, Gentamicin, Cefotaxim and Amphotericin B with the addition<br />
of 15% glycerol. After thawing, all tissues were divided in three segments for different rinsing treatments. In the first group,<br />
the skin samples were not rinsed. The second group was rinsed twice with BASE (AL.CHI.MI.A s.r.l.) for 10 min. and the third<br />
group was rinsed three times with BASE for 10 min. After rinsing, skin homogenates were prepared and assessed semiquantitatively<br />
for antibiotic residue content by agar diffusion test using agar plates seeded with C. Albicans (CA), P.<br />
Aeruginosa (PA), S. Aureus (SA). For determination of glycerol concentration in the skin samples, tissue homogenates were<br />
centrifuged and supernatant was derivatizated with periodate and acetylacetone solution before injection in U-HPLC system<br />
(Dionex) equipped with reversed phase C18 column. Results Agar diffusion test showed inhibition areas on SA, PA and CA<br />
seeded plates in all skin samples not undergoing rinsing, thus indicating the presence of different antibiotic residues. Inhibition<br />
zones were significantly higher in bank solution treated tissues not undergoing rinsing as compared to tissues treated with<br />
BASE.128. Two rinses of 10 min. each with BASE allowed to reduce the inhibition areas on SA, PA and CA seeded plates<br />
of 32 %, 48% and 100% in tissue treated with bank prepared solutions and of 65%, 96% and 100% in tissues treated with<br />
BASE.128, respectively. In addition, a third rinse of 10 min. in BASE allowed to further reduce the inhibition zone on SA<br />
seeded plates in BASE.128 treated tissues, thus achieving an 80% reduction of antibiotic residual concentration. Conversely,<br />
the third rinse did not affect the tissues treated with bank prepared solution, indicating possible binding of the antibiotic to<br />
the tissue. HPLC analysis of tissues homogenates showed similar glycerol content (98,3 mg/g of tissue, on average) in tissues<br />
not undergoing washing, both treated with bank prepared solution and BASE.128. Three rinses of 10 min. each in BASE<br />
allowed removing, on average, 89% of the glycerol from the tissue.<br />
Conclusions:<br />
The efficacy of elimination of antibiotic residues from cryopreserved skin depends on the type of antibiotics. Specific rinsing<br />
procedures should be established and validated for each solution used for tissue processing. The present study proposes a<br />
rinsing procedure, consisting of 3 rinses of 10 min. each with BASE, resulting in effective removal of all antibiotic and<br />
glycerol residues from cryopreserved skin after decontamination in BASE.128.
O-46<br />
THE INFLUENCE OF RADIATION STERILIZATION AND PRESERVATION ON CELL<br />
MORPHOLOGY OF HUMAN<br />
SUZINA, S. A. H 1 , KAMALIA, Z. 1 , YUSOF, N. 2 , ASNAH, H. 2<br />
1 - Tissue Bank, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia<br />
2 - Nuclear Malaysia Agency, Bangi, 43000 Kajang, Selangor, Malaysia. Indonesia.<br />
Abstract:<br />
The application of gamma doses lower than 25 kGy for terminal sterilization is a new approach in tissue banking for<br />
minimising the radiation effect. This is due to gamma radiation in combination with tissue preservation may have a significant<br />
effect on the morphology of human amniotic membrane (HAM) hence physiological condition. The aim of this study was to<br />
investigate the influence of different doses of gamma irradiation after different preservation techniques on cell morphology<br />
of HAM under Scanning Electron Microscope (SEM). HAM was processed and then preserved by either air drying (ADHAM)<br />
or submerged in glycerol (GPHAM). The sterilization of HAM was carried out using gamma irradiation from Cobalt-60<br />
source at 0 kGy, 15 kGy, 25 kGy and 35 kGy. In the control group (fresh HAM) no preservation and sterilisation were<br />
performed. The samples were fixed in 2.5% glutaraldehyde and the surface morphology was analysed under the SEM. The<br />
SEM examinations revealed some alteration in cell morphology of ADHAM and GPHAM as compared to fresh HAM. In<br />
addition, the cell morphology of ADHAM was more affected than GPHAM. The structure of HAM was not distinctively clear<br />
at low magnification (×250). However, at higher magnifications (more than ×1000) the intercellular channels and the cell<br />
surface covered with microvilli were clearly observed in both ADHAM and GPHAM. The cell structure was more preserved<br />
when stored in glycerol, the cells were beautifully arranged, homogonous, tended to round up at 25 kGy and started to have<br />
a gap between intercellular channels at 35 kGy. As for ADHAM, the cells seemed to form flat sheet of polygonal cells and<br />
the cytoplasmic strands were flattened and condensed at all of the sterilization doses. While in the fresh HAM the membrane<br />
was composed of polygonal cells forming mosaic pattern. At ×10000 magnifications, the SEM showed the microvilli covered<br />
the whole surface, causing intercellular channels in fresh HAM less clear. There were significant changes in the HAM surface<br />
after processing/preservation and sterilization. GPHAM was found to cause less morphological changes than ADHAM at<br />
doses lower than 25 kGy due to glycerol as a radioprotectant. The intercellular channels of ADHAM were not visible because<br />
the cells collapsed during air-drying and likely further damaged when irradiated. High dose of gamma irradiation at 35<br />
kGy led to marked changes in the cell morphology and the intercellular channels of HAM, fortunately the influences were<br />
less pronounced in GPHAM. In conclusion, this study revealed that the different preservation methods followed by sterilization<br />
by gamma irradiation caused changes in cell morphology of HAM. The findings recommended the use of glycerol<br />
preservation in combination with irradiation dose lower than 25 kGy for processing of HAM.<br />
O-47<br />
EFFECT OF INCUBATION TEMPERATURE ON VIABLE SKIN DECONTAMINATION<br />
RODRÍGUEZ, L.; TARRAGONA E.; GOMÁRIZ E.; ORTEGA I.; RODRÍGUEZ V.; GENÍS X.; NAVARRO A.<br />
Banc de Sang i Teixits. Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Background:<br />
Viable skin decontamination by grafts incubation in antibiotics is routinely performed in Tissue Establishments (TE). Major<br />
parameters defining effective decontamination protocols include, but not limited to; initial graft bioburden, type and<br />
concentration of antibiotic/antimicotic cocktails, incubation times and working temperature during decontamination process.
Purpose:<br />
The effect of antibiotic incubation temperature was evaluated for skin grafts arrived to our TE processing facility after<br />
procurement. Several grafts were incubated at 4ºC (n=36) or 37ºC (n=70) using the next decontamination cocktail<br />
composition (CPLVA): 240 µg/mL Cefoxitin; 123 µg/mL PolimixinB; 102 µg/mL Lincomycin; 50 µg/mL Vancomycin and 5<br />
µg/mL AmphotericinB. Final concentrations in culture medium M199 described.<br />
Materials and Methods:<br />
Biopsy (3-4 small pieces), antibiotic free transport and cryopreservation medium samples (5mL/each) were used to determine<br />
microbiological contamination status before and after decontamination. Tissues microbiological status acceptable for clinical<br />
transplantation, including those grafts positive for non pathogenic skin contamination, served as the evaluation criteria<br />
regarding the effect of temperature on decontamination.<br />
Results:<br />
Microbiological species isolated in skin grafts under study were mainly; Staphilococci sp including S.Aureus, Enterococci,<br />
E.Coli and Bacillus sp. Before decontamination samples taken resulted microbiologically out of specifications for 42% and<br />
54.3% of the grafts included in the 4ºC and 37ºC study groups respectively. In the 4ºC group, after decontamination, 61%<br />
and 44% of the evaluated grafts passed the clinical criteria for transplantation when biopsy or supernatant samples<br />
microbiological results were evaluated. Those proportions raised up to 94% and 74% for the above mentioned kind of<br />
samples when decontamination was performed at 37ºC. Incubation times ranged 19-23 hours and 11-24 hours in the 4ºC<br />
and 37ºC groups respectively.<br />
Conclusions:<br />
Incubation temperature is an important parameter for viable skin decontamination effectiveness. Rising temperature from<br />
4ºC to 37ºC during incubation resulted in a 30% increase on medical accepted for transplantation skin grafts for both biopsy<br />
and supernatant samples. Microbiology test results varied regarding the kind of analyzed sample (biopsy vs supernatant)<br />
in terms of microbial detection (sensitivity). As expected, medium samples showed a major proportion of positive results (up<br />
to 20%) when compared to biopsy samples, irrespective the incubation temperature during decontamination, suggesting<br />
major representation of those samples.<br />
O-48<br />
GAMMA RADIATION STERILIZED AMNIOS: RECENT CLINICAL APPLICATIONS IN<br />
MEXICO<br />
MARTÍNEZ-PARDO, M.; VALENCIA-FONSECA ELENA L.; LEÓN-TÉLLEZ Y.; VÁZQUEZ-MAYA L.<br />
Instituto Nacional de Investigaciones Nucleares. Mexico.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
The use of radiosterilized amnios in recent different clinical applications in Mexico is presented. The Banco de Tejidos<br />
Radioesterilizados del Instituto Nacional de Investigaciones Nucleares (Radiosterilized Tissue Bank at the National Institute<br />
of Nuclear Research BTR-ININ) have been collaborating with public health institutions, mainly with Ophthalmology Service,<br />
to treat affected patients with amnios esterilized with gamma radiation. The results have proven to be excellent as much for<br />
cosmetic purposes as for functional ones. Radiosterilized amnios do not provoke rejection, adverse reaction or transmit an<br />
infective disease and improve the quality of life of the patients.<br />
Introduction:<br />
In Mexico, the Banco de Tejidos Radioesterilizados del Instituto Nacional de Investigaciones Nucleares (Radiosterilized<br />
Tissue Bank at the National Institute of Nuclear Research BTR-ININ) was established in 1999, thanks to strong support of
ORAL<br />
PRESENTATIONS<br />
the International Atomic Energy Agency (IAEA). Since that time, the BTR, which is a nonprofit tissue bank, processes and<br />
sterilizes with 60Co gamma radiation tissues such as amnion and pig skin. These tissues have been used mainly as biological<br />
wound dressings in patients with first, second and third degree burns, ulcers, epidermolysis bullosa, bloody areas, and in<br />
wounds difficult to heal. The Secretaría de Salud (Mexican Ministry of Health) issued the sanitary license No. 1062000001<br />
to the BTR-ININ on July 7, 1999 [1]. The Quality Management System QMS of the BTR-ININ was certified by ISO 9001:2000<br />
on August 1, 2003 [2]. At present, the ISO 9001:2008 certification is kept. To offer more and diverse sterilized tissues to<br />
the medical community, the bank is now validating the process of human skin and musculoskeletal tissues, both from<br />
cadaveric donors. For that, the ININ has signed an agreement with the Instituto de Salud del Estado de México ISEM (Health<br />
Institute of Mexico State) since June, 2007, to obtain amnion, human skin and musculoskeletal tissues. Amnios Processing:<br />
Amnios from healthy mothers, with written consent and rigorous screening for HIV 1, 2, Hepatitis B and C, Syphilis and<br />
Chagas and clinical history, among others, were procured at an authorized ISEM hospital. The selected hospital is also<br />
responsible of preliminary cleaning of the tissue and temporary storage at refrigeration temperature, preserved in saline<br />
solution. Then the tissue and its documents were sent to the BTR-ININ. After first quality control inspection (QCI), the amnios<br />
are processed at the bank. The following activities are performed at the BTR in its facilities at the Nuclear Centre, according<br />
to its QMS: Tissue Reception, Checking of documentation, Washing, Drying, Cutting at desired size, Packaging (primary<br />
and secondary package) under laminar flow conditions, Labeling to identify the tissue bank, Microbiological control<br />
performed at the Departamento de Biología (Biology Dept.), to determine average initial bioburden, verification dose and<br />
sterilization dose. After second QCI, the processed tissues are sent to the Departamento del Irradiador Gamma (Gamma<br />
Irradiator Dept.). Sterilization is done at room temperature with cobalt-60 gamma radiation at a minimum dose of 25 kGy.<br />
Then the irradiated product is sent to the Departmento de Materiales Radiactivos (Radioactive Mtls. Dept.) for Final product<br />
sterility test. Again at BTR facilities, the Storage of irradiated tissues at room temperature, waiting for negative second<br />
serology results and Distribution of high quality tissues for clinical application are performed. Irradiated tissues are distributed<br />
mainly to public hospitals, after the corresponding quality control approval and the tissues are delivered with an instruction<br />
sheet and a follow-up form [1].<br />
Clinical applications:<br />
Radiosterilized amnios processed at the BTR have been used in Mexico to treat patients with burns, damaged ocular surface<br />
and dystrophic epidermolysis bullosa DEB (dominant form), among others[3]. Written and informed consent was obtained<br />
from all patients that were treated with the irradiated tissue. The Epidermolysis Bullosa is a rare hereditary and genetic<br />
illness due to a failure of connection between the epidermis and the dermis. The skin is very fragile and to blister easily. DEB<br />
is one of the major forms of epidermolysis bullosa. Mutations in the COL7A1 gene cause all three major forms of DEB. This<br />
gene provides instructions for making a protein that is used to assemble type VII collagen. Type VII collagen plays an<br />
important role in strengthening and stabilizing the skin. When type VII collagen is abnormal or missing, friction or other minor<br />
trauma can cause the two skin layers, epidermis and dermis, to separate. This separation leads to the formation of blisters,<br />
which can cause extensive scarring as they heal[4]. One-month old patient suffering Dystrophic Epidermolysis Bullosa (DEB)<br />
was treated with radiosterilized amnios in a private hospital at Pachuca city. At the patient´s birth, April 2010, the DEB<br />
affected his lower extremities, feet and thorax. One month later, the mayoritie of his injuries were in remission, except the<br />
feet. That is why on May 19, 2010, the amnios were placed in both to prevent anemia and to improve the lesions. The use<br />
of this irradiated amnios in Ophthalmology started in the country in 2005, either as a graft to replace the damaged ocular<br />
surface, or as a patch to prevent unwanted inflammatory reactions. Patients from the public Hospital General de México<br />
(HGM, Mexico City), suffering diverse pathologies such as keratoconjunctivitis, recurrent pterygium associated with<br />
symblepharon, corneal neurotrophic ulcers, chemical and thermal burns, and corneal thinnings, had been successfully<br />
treated with irradiated amnion. In the HGM, a clinical prospective study on lesions of the ocular surface of 17 eyes from 15<br />
patients, affected with the mentioned above pathologies, was successful in 88.2%[5]. In the Instituto Mexicano del Seguro<br />
Social in Metepec, Méx. (IMSS-UMAA 231), a controlled clinical randomized trial with 108 eyes from 100 patients, affected<br />
with primary nasal pterygium, was performed in 2009. The degree for the pterygium was classified in I for less than 1 mm,<br />
II for the range of 1 to 3 mm, and III for more than 3 mm. Fifty four eyes (Group A) were treated with radiosterilized amnion<br />
and intraoperative Mitomycin C to prevent recurrence after excision of the primary pterygium. Group B consisted of 54 eyes<br />
from 49 patients, was subjected to conjunctival autologous graft with intraoperative mitomycin C. Topical mitomycin C at<br />
0.05% concentration was used in this research [6]. In the IMSS-Centro Médico del Bajío, Unidad Médica de Alta<br />
Especialidad in León City, in the center of the country, a two year-old boy patient was addmitted for management of bilateral<br />
chemical burn with alkali on January 7, 2011. After initial exploration, the diagnosis was bilateral chemical burn grade II
ORAL<br />
PRESENTATIONS<br />
in right eye and grade III in left eye, according Hughes- Roper- Hall Clasiffication of chemical burns. Five days after treatment<br />
with antibiotics, analgesic, antiinflammatory and lubricant, pain and inflammation were getting better, and corneal<br />
epithelialization was started. Right eye with epithelial abrasion, 20% of total corneal suface (tcs), and a paracental leucoma.<br />
Left eye had epithelial abrasion, 60% of tcs, with epithelial blebs and stroma still hazy, affecting visual axis. Radiosterilized<br />
amnios were applied in both eyes covering cornea wound and fixed on clear cornea and conjunctiva, under general<br />
anesthesia and autolougus platelet rich plasma injected subconjunctival. Finally both eyes were covered with a therapeutic<br />
bandage contact lens. Results.-The results have proven to be excellent as much for cosmetic purposes as for functional ones.<br />
Gamma radiation sterilized amnion (A) represents an effective and secure treatment of the damaged ocular surface, with<br />
the advantage of total absence of bacteria, due to a SAL of 10-6. The tissue does not provoke rejection or transmit an<br />
infective disease from donor to recipient. Irradiated (A) does not require refrigeration, it can be stored at room temperature<br />
without any deterioration of its properties. The tissue has easy handling & low cost. The use of irradiated amnion in the<br />
paediatric patient affected with DEB gave him a better quality of life. Four months after (A) treatment, the appareance of<br />
his feet skin was dramaticalley improved. Unfortunately, this rare disease has a bad prognosis. The outlook depends on the<br />
severity of the illness. In the case of the paediatric patient affected with chemical burn in both eyes, four months lather, right<br />
eye cornea was completely clear. Left eye cornea only with a small and mild leucoma in the nasal edge of the pupil, which<br />
not affects visual axis. Due to the lack of corneal tissue suitable for transplantation (Tx), the irradiated (A) gives an extra time<br />
for those eyes waiting for cornea or before Tx, to improve the prognosis.<br />
Conclusions:<br />
Irradiated amnios can be satisfactory used in the treatment of the affections mentioned above. Without the treatment, the<br />
patients could have suffered a healing after-effect or loss of sight. The inflammation and pain were significantly reduced.<br />
Radiosterilized amnios do not provoke rejection, adverse reaction or transmit an infective disease and improve the quality<br />
of life of the patients.<br />
References<br />
1.- Martínez ME, Reyes ML, 2003, The tissue bank at the National Nuclear Research Institute in Mexico, Cell and Tissue<br />
Banking 4, p. 163-168.<br />
2.- Martínez-Pardo ME, Mariano-Magaña D, 2006, The tissue bank at the Instituto Nacional de Investigaciones Nucleares:<br />
ISO 9001:2000 certification of its quality management system, Cell and Tissue Banking 8, p. 221-231.<br />
3.- Martínez ME, Ley E, Reyes ML, Rodríguez P, Vázquez L, Salazar MA, 2007, Biological wound dressings sterilized with<br />
gamma radiation: Mexican clinical experience, Radiat. Phys. Chem. 76, p. 1771-1774.<br />
4.- http://www.<br />
5.- Vázquez L, Salazar MA, Martínez ME, 2009, Use of amniotic membrane radiosterilized with cobalt-60 for reconstruction<br />
of ocular surface, Revista Médica del Hospital General de México 72(1), p. 7-15.<br />
6.- León Y, Martínez ME, Vázquez L, 2009, Eficacia del trasplante de membrana amniótica radioesterilizada en cirugía de<br />
pterigión, Contacto Nuclear 55 p. 26-29.
O-49<br />
MICROBIOLOGICAL CONTROLS IN SKIN TISSUE BANK<br />
ORAL<br />
PRESENTATIONS<br />
PÉREZ, M. L. 1 , VILARRODONA, A 1 , SAVIO, A. 1 , OTERO, N. 1 , MARTÍNEZ-CONESA, E. 1 , NUÑEZ, V. 1 , AGUSTÍ, E. 1 ,<br />
TRÍAS, E. 1 .<br />
1 - Transplant Services Foundation-Hospital Clínic. Spain.<br />
Abstract/Introduction:<br />
Skin grafts from cadaveric donor have become one of the best options treating great burns, traumatisms, surgeries and diseases<br />
entailing the loss of cutaneous tissue. To ensure the viability of these grafts the tissue banks facilitate low temperature storage<br />
between procurement and delivery, this storage period providing the time necessary for microbiological assessment of the skin<br />
prior to grafting. In our tissue bank two basic preservation methods are carried out: glycerol preservation and cryopreservation.<br />
After processing the skin is maintained in quarantine, waiting for the blood culture results and the processing culture results.<br />
Looking for the highest quality of the grafts a new microbiological culture test was added to the protocol of skin procurement<br />
and processing. Thus, since middle 2009 a sample of skin is taken during retrieval for microbiological testing, as well as blood<br />
culture and culture during processing. Moreover in some cases we were able to collect a sample before the last preservation<br />
presentation, a post processing culture, in the case of glycelorized skin. Aim To evaluate the contamination rate of blood cultures<br />
and skin cultures from donors banked in the period between May 2009 and December 2010, assess the influence of any<br />
contamination in the viability of the tissues processed in the bank, and compare the numbers with the data obtained with the<br />
donors banked during 2009, with or without the extra microbiological information.<br />
Methodology:<br />
The study was conducted on 220 skin donors available to the skin bank from May 2009 to December 2010. Blood cultures<br />
were carried out to each donor, one (217), two (185) or three (68), as well as cultures of cutaneous tissue from each<br />
preserved fragment, both cryopreserved (426) and glycerolized (216). Moreover, a skin sample was taken during retrieval<br />
of all of these donors (220). Results The viability rate obtained with the 140 donors processed in 2009 was 85%, whereas<br />
the one obtained with the 220 donors with the extra information of the result culture of the skin taken during retrieval was<br />
almost the same, 84%. Thirty-five out of the total 220 donors were discarded as viable donors, 18 of them due to<br />
microbiological contamination (positive blood culture, skin culture or both of them). Focus on the results obtained with the<br />
new culture of the skin taken during retrieval, there are 19 donors whose cryopreserved skin was discarded but yet it was<br />
possible to distribute the glycerolized skin, due to the virucidal and batericidal potential of glycerol.<br />
Conclusions:<br />
The accomplishment of several cultures pre and post processing contributes to the safer skin grafts generation. Our<br />
decontamination method during processing has demonstrated to be highly effective. Both cryopreservation and<br />
glycerolization are suitable methods to assure the viability of the skin grafts distributed.
O-50<br />
AMNIOTIC MEMBRANE IN USE FOR VARIOUS PRECLINICAL AND CLINICAL<br />
INDICATIONS<br />
HENNERBICHLER, S.; PETERBAUER-SCHERB A.; PETTER-PUCHNER A.; KESTING M.; REDL H.; GABRIEL C.<br />
Red Cross Blood Transfusion Service of Upper Austria, Linz/ Austrian Cluster for Tissue Regeneration. Austria.<br />
Abstract:<br />
Preserved amniotic membrane is used in the field of ophthalmology and wound care due to its supporting properties. The<br />
Multi-Tissue-Bank of the Red Cross Blood Transfusion Service of Upper Austria supplies local and international hospitals with<br />
cryopreserved amniotic membranes for both indications. A less-known possible field of application could be hernia repair,<br />
as we could demonstrate in an experimental intraperitoneal onlay mesh technique in rats that cryopreserved, but still viable<br />
amniotic membrane as antiadhesive mesh coating reduces adhesions to the used mesh and suture materials. Furthermore,<br />
data of a clinical study applying amniotic membrane in oral surgery to cover split-skin donor sites will be presented. Within<br />
this study amniotic membrane was compared to conventionally used materials, particularly with regard to exudation, change<br />
of wound dressing, pain, pruritus, comfort and healing progress.<br />
ORAL - NOVEMBER 10TH - LEARNING FROM BIOVIGILANCE<br />
AUDITORI - 09.55-10.05<br />
O-51<br />
CATALONIAN SURVEILLANCE REGISTER<br />
BARRIO, R.; FÉLIX JESÚS M.; ARAN B.; DEULOFEU R. - OCATT. Barceona. Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Surveillance was inserted in the current Spanish legislation in 2006 by the RD 1301/2006, based on the European Law<br />
2004/23/CE. This law settles the requirements of quality and security in every tissue and cell procedure. Catalonian<br />
Surveillance Register was set up in June 2008. Before that, a manual containing all the reporting system and the different facts<br />
to notify, was published and distributed among all the professionals involved. In Catalonia, there are almost 230 authorized<br />
activities related to tissues and cells. There are nearly 150 teams authorized to graft different types of tissues and cells. Forty<br />
centers are authorized to obtain cord blood, 26 to retrieve different types of tissues and cells and 6 tissue banks. Two of them<br />
are multi-tissue and 4 are monographic (cord blood, hepatocytes, ocular and ovarian). Due to the large number of authorized<br />
centers, in 2010 more than 5800 tissue and cell implants were performed. Although the current law recognizes two different<br />
types of notifications, Serious Adverse Effects (SAEs) and Serious Adverse Reactions (SARs), we have differentiated three events.<br />
On one hand there are the Serious Adverse Effects (SAEs). Although the law does not differentiate, we have split SAEs into two<br />
sections. The first takes into account tissues grafted prior to detection, this is known as clear SAEs. The second section refers to<br />
detection before grafting. In this case we speak of incidents. On the other hand, there are the Serious Adverse Reactions (SARs).<br />
It was agreed to notify every fact detected at one stage of the process, but which had been overlooked at the previous one.<br />
From June 2008 till December 2010, the Catalonian Surveillance Register has received 63 notifications. 22 were clear Serious<br />
Adverse Effects, 22 incidents, 7 Serious Adverse Reactions and 2 medical alerts. 3 of the Serious Adverse Reactions were<br />
reported in living donors and the other 4 in recipients. All of them recovered without further complication.
O-52<br />
NATIONAL HISTOVIGILANCE EDUCATION SYSTEM IMPROVES PROFESSIONAL<br />
UNDERSTANDING OF SEVERE ADVERSE REACTIONS AND EVENTS<br />
CEBULC, G.; AVSEC D. Slovenija-transplant. Slovenia.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
To review the implementation of notification of severe adverse events and reactions (SARE) under Slovenian Act and Rules<br />
but Directives 2004/23/EC and 2006/86/EC as well. A national histovigilance education model is suggested. Background:<br />
After adoption of the Directives 2004/23/EC, 2006/17EC and 2006/86/EC, next step forward was to prepare national<br />
Rules for histovigilance (HV). First national report on histovigilance was done in 2009 but no reports from tissue<br />
establishments (TEs) were received to Competent Authority (CA) Slovenija Transplant (ST). In next year were collected 5<br />
cases only. Despite ST delivered EUSTITE vigilance and surveillance (V&S) tools to all accredited TEs within country. At the<br />
end of 2010 ST decided to organize professional meeting with well defined timetable for professionals from tissue and cells<br />
field. Second annual national meeting for histovigilance in May 2011 was performed.<br />
Objective:<br />
Despite Directives’ provisions related to HV were placed nothing happened in practice. As mentioned there were no expected<br />
reports of SARE in 2008. Following year some deficient reports were received by ST. EUSTITE V&S tools were translated<br />
and delivered by ST within all responsible persons of TEs in Slovenia. The need of education was huge and by decision of<br />
ST first national meeting for HV was performed in November 2010. It was measured that annual meeting including<br />
workshops and high skilled tutors are necessary.<br />
Methods:<br />
ST prepared one day education meeting model. Target groups were TEs responsible persons, transplant coordinators,<br />
clinicians and other professionals working in tissue and cells field. In Slovenia are registered 6 donor hospital and 16 TEs<br />
at the moment. Lectures included legislation V&S tools basis. Second part was consisted of interactive group workshop,<br />
whereas 5 to 7 participants from different background. Evaluation of the model was performed by questionnaire covered<br />
major aspects of the model including content relevance of lectures and workshop, presentation and sufficiency of lecturers’<br />
answers, organization and accessibility of the event as well. Presented were 55 participants in 2010 and 27 in 2011. Every<br />
participant had to fulfill test with 10 questions related to the presented topics. The average score of the test was 8,76 of 10<br />
in 2010 and 7,81 of 10 in 2011.<br />
Conclusions:<br />
National HV education system was established. International tutor from EUSTITE project was presented and guided practical<br />
part of the meeting. The huge necessity for HV in practice was confirmed by number of participants and feedback where<br />
the first meeting was scored with 4,28 of 5 (85,6%) comparing 4,06 of 5 (81,2%) in 2011. Additionally the average score<br />
of importance of HV education was 3,94 of 5 in 2010 comparing 3,76 of 5 in 2011. It was suggested one day professional<br />
meeting with HV workshop annually.
ORAL - NOVEMBER 10TH - GLIMPSE INTO THE FUTURE; ADVANCED<br />
THERAPIES - AUDITORI AND ROOM 3 - 14.10-15.30<br />
O-53<br />
MICROBIOLOGICAL CONTAMINATION MONITORING OF CLEANROOM FACILITIES<br />
IN A TISSUE AND CELL ESTABLISHMENT<br />
VILARRODONA, A.<br />
Transplant Services Foundation-Hospital Clínic. Barcelona, Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
Routine control of environmental microbiological contamination in cleanroom facilities is a critical part of tissue and cell<br />
establishments. Security of tissues and cells which are processed in cleanrooms may be compromised by the presence of<br />
microorganisms in the environment or surfaces of these facilities. Tissues and cells are processed in grade A area surrounded<br />
by grade B with adjacent grade C areas.<br />
Aim:<br />
The aim of this study is to evaluate the bioburden load in the controlled areas of our tissue and cell establishment (5<br />
cleanrooms, 1 corridor, 2 changing rooms and 1 material air-lock).<br />
Methods:<br />
According to the guidelines of Good Manufacturing Practices (GMP) our tissue and cell establishment has developed a<br />
monitoring program to control microbiological contamination of cleanroom areas. Between December 2009 and June 2011<br />
with a variable periodicity of 14 or 21 days, defined sampling points and airborne particles were collected using Tryptone<br />
Soy Agar (TSA) and Sabouraud Chloramphenicol (SC) contact and settle plates respectively. The plates were incubated at<br />
30-35ºC for bacteria and 20-25ºC for fungi during 72 h and subsequently 4 days at room temperature. Sampling was<br />
conducted in an at rest state (which is the condition where the facility is installed and operating complete with production<br />
equipment but with no operating personnel), the day before a radical sanitation and after the routine and daily cleaning<br />
procedures of the cleanrooms because it was considered the worst situation.<br />
Results:<br />
The obtained results are summarized in the following table (including working and non working areas): Negative results<br />
Positive results In of specs Out of specs Surface areas 2507 278 247 31 Air control 128 32 30 2 Total 2635 310 277 33<br />
From the total amount of microbiological cultures (2945 samples) the 10.53% resulted positives and the 10.65% from them<br />
were out of specifications. The most common microorganisms identified were Micrococcus sp (55.10%), Bacillus sp (28.23%)<br />
and Gram positive cocci (12.92%). The following table shows the distribution of the positive results on the different working<br />
areas: CR1 CR2 CR3 CR4 CR5 Air Class B B B B B In specs Air 8 5 7 6 4 Surface 5 5 1 12 3 Out specs Air 1 0 0 1 0 Surface<br />
0 1 0 0 5 None of the out of specifications results were obtained from working surfaces, and most of them correspond to<br />
door handles and floor surfaces. In reference to the accomplishment of GMP guidelines we observed a 99.88% of results<br />
within the specifications.<br />
Conclusions:<br />
In terms of traceability, it is important to highlight the bioburden knowledge to determine traceability from donor to final<br />
recipient considering the possibility of cleanroom facility influence. Despite the effectiveness of the decontamination process<br />
coming out from the corresponding validation study, the necessity of developing a microbial monitoring program and a<br />
periodical review of the validation study is concluded as highly advisable to guarantee the quality and safety of the tissues<br />
and cells with clinical purposes. Due to the major part of the microorganisms detected become from normal skin flora, the<br />
measures taken to achieved lower contamination rates must be focused on personnel hygienic practices.
O-54<br />
PRESENTATION OF THE PROJECT OF THE SUPPORT OFFICE TO THE CLINICAL<br />
RESEARCH IN ADVANCED THERAPIES<br />
ARAN, B.; FELIX M.; BARRIO R.; DEULOFEU R. Organització Catalana de Trasplantaments. Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Advanced Therapies (AT) involve cell therapy, gene therapy and tissue engineering. All of these therapies comprise<br />
regenerative medicine and try to restore or replace cells and tissues when cell function has been lost. AT are moving forward<br />
and lots of researchers are working to know how to improve the results. However, most of the treatments are still in an<br />
experimental phase. The AT are regulated guided by the Regulation 1394/2007. According to this regulation, cells and<br />
tissues used in AT must be considered medicinal products with all the requirements and conditions involved (GMP conditions,<br />
quality controls, etc). Cells and tissues are considered medicinal products if they have suffered a substantial manipulation<br />
before transplant, if the initial and final place in the human body is not the same or if the cell function changes. In Spain,<br />
the Medicament Spanish Agency (AEMPS) is in charge of approving the clinical trials with AT and also the industrial<br />
production of AT medicinal products. The requirements are the same in both conditions; although the situation is very<br />
different. Nevertheless, sometimes it is difficult to know if a therapy is an AT or a transplant, and the law is different in each<br />
case. Furthermore, researchers sometimes ignore technical and administrative prerequisites before a clinical trial is put in<br />
place. For these reasons, setting up of a Support Office to the Clinical Research in Advanced Therapies is proposed. It will<br />
be created by the Catalan Transplant Organization (OCATT) through an instruction from the Health Department and an<br />
agreement between the Catsalut-OCATT and the Center of Regenerative Medicine in Barcelona (CMRB). The office’s principal<br />
aims would be to create a registry including all the AT clinical trials taking place in Catalonia and to offer help and support<br />
to all researchers who need some advice before putting an AT clinical trial in place. The office will provide information<br />
about the requirements needed to perform the assay (available facilities, paperwork management, etc) proceeding as a link<br />
between researchers and the AEMPS. Other tasks of this office will be to approach basic and clinical research facilitating<br />
initiatives and creating relationships between both of them, as well as, to promote the diffusion of the results and to search<br />
calls and grants to fund the projects. An Advisory Commission will counsel and support the office by working groups. The<br />
Support Office to the Clinical Research in Advanced Therapies wishes to be a useful tool to help and support clinical<br />
researchers’ task. The office wants to establish relationships with them to know their needs and pushing up the research<br />
towards the clinic avoiding duplications.<br />
O-55<br />
BIOMECHANICAL PROPERTIES CAMPARATION BETWEEN FREEZE DRIED FLEXOR<br />
TENDON AND COMPOSITE OF FREEZE DRIED FLEXOR TENDON – BONE MARROW<br />
MESENCHYMAL STEM CELL IN RABBIT MODEL<br />
SUROTO, H.; FERDIANSYAH F.; RANTAM ABDUL F.; ROESHADI D. Biomaterial Center and Tissue Bank - Dr Soetomo<br />
General Hospital. Indonesia.<br />
Objective:<br />
To evaluate biomechanical properties of freeze dried flexor tendon and reconstitution of freeze dried flexor tendon seeded<br />
with bone marrow mesenchymal stem cell in rabbit model.<br />
Design:<br />
The post test only control group design of rabbit freeze dried flexor tendon and rabbit bone marrow mesenchymal stem cell.
Subject:<br />
We studied New Zealand White rabbit bone marrow mesenchymal stem cell cultured and expanded. 5 fresh and 5 freeze<br />
dried flexor tendon of NZW rabbit.<br />
Methods:<br />
Five flexor carpi ulnaris tendon was haversted from adult, male New Zealand White Rabbit. They were divided into three<br />
groups according to processing: group1,fresh flexor tendon as control group; group2, freeze dried flexor tendon; group3,<br />
composite freeze dried flexor tendon – bone marrow mesenchymal stem cell. All processing was performed at Biomaterial<br />
installation and tissue bank of dr Soetomo Hospital. Each specimen was tested by tension load using autograft shimatzu<br />
machine. The parameters of maximal load, tensile strength, tensile strain and modulus elasticity were obtained from the<br />
load-elongation curves and the stress-strain curves. Stastical analyses were performed using one-way analysis of variance<br />
(ANOVA) to compared maximal load, tensile strength and modulus elasticity. And to analyse tensile strain comparation we<br />
performed non parametric Kruskall-wallis test. Subsequent post-hoc comparisons were performed to detect significant<br />
differences (p
ORAL<br />
PRESENTATIONS<br />
Results:<br />
1. After the enzymatic isolation the cells started to grow within 24 hours and the confluent layer was reached within 7-14<br />
days.<br />
2. The cells from the explant culture started to grow after 2- 3 days and the monolayer was formed after 10- 14 days The<br />
sufficient number of the cells from both parallel cultivations was obtained within 10- 14 days without any passages of the cells.<br />
To obtain 2, 5 x 106 – 4x 106 cells using standard method take more than 28 day and higher number of passages is needed.<br />
This number of the cells is adequate for seeding on the amniotic membrane for transplantation to the patient.<br />
Conclusions:<br />
The utilization of the remnants of the biopsy allows reduce cultivation time to the half without passages and risk of changing<br />
of the limbal phenotype. Reduced cultivation time is more profitable for the patients.<br />
O-57<br />
TESTING ON CELL CULTURES OF BIOGENIC AND SYNTHETIC MATERIALS APPLIED IN<br />
DENTISTRY<br />
VOLOVA L.T., ROSSINSKAYA V.V., SHAROVATOVA A.U., MILYUKOVA M.N., KUPRYAHIN S.G. Russia, Samara State<br />
Medical University, Samara Tissue Bank. Russian Federation.<br />
Abstract/Introduction:<br />
Hydroxyapatite-containing materials are used in dental surgery and implantation. But their application in clinical practice<br />
doesn’t always lead to the required results. The Objective. To conduct the testing on cell cultures of such materials for<br />
cytotoxicity and biocompatibility. Materials and methods. The investigations were conducted on human dermal fibroblasts<br />
cultures. Russian synthetic osteoconductive preparatuions KollapAn-G, Kollapol, as well as allogenic hydroxyapatite (HAP)<br />
and spongiosa of “Lioplast®”series and samples of titanium rods with and without synthetic HAP spraying have been used<br />
as target test-objects. Investigations have been carried out using morphological, morphometrical, histochemical, biochemical<br />
methods of analysis as well as scanning electron (raster) microscopy.(SEM).Results. KollapAn-G and Kollapol preparations<br />
, containing xenogenic collagen and synthetic HAP, possess weak cytotoxicity (Kollapol cytotoxicity is more). Monomolecular<br />
layer cells proliferation processes inhibition, cells and nuclear form changes, cytoplasm vacuolization have been notes since<br />
the 3rd day of the experiment. The death of 67.3±4.2 % of monomalecular layer cells have been registered on the 7th day<br />
of Kollapol testing, the majority of the survived cells contain fat inclusions in their cytoplasm, which are normally absent in<br />
fibroblasts. Biochemical methods of cultural growth assessment confirm the damaging effect of these materials; by the 9th<br />
day of the experiment the number of living cells has reduced in 2.8 times in KollapAn-G testing and in 4.8 times in Kollapol<br />
testing. Living cells in these cases have been observed only up to the 7th day of experiment. Allogenic HAP and mineralized<br />
spongiosa of “Lioplast®” series testing did not reveal any cytotoxic effect and demonstrated their evident biological effect (<br />
cells proliferation stimulation in the culture). The presence of titanium sample with spraying results in adhesion decrease and<br />
the appearance of cells with vacuole dystrophy. Titanium samples without spraying are not toxic. Conclusion. Allogenic<br />
HAP and spongiosa of “Lioplat®” series as well as titanium sample without spraying are biologically compatible, not<br />
cytotoxic, stimulate the cells proliferation in the culture. KollapAn-G, Kollapol and titanium sample with synthetic HAP<br />
spraying are slightly cytotoxic: they cause vacuole and fatty fibroblasts dystrophy, reduce their proliferation activity and<br />
adhesion.
O-58<br />
BMP-2 AND BMP-7 CONTENT IN DBM PUTTY PRODUCED BY TISSUELAB IN A GMP<br />
PHARMACEUTICAL PLANT<br />
RAMELLI, M.; DELLA VALLE E.; MARINIELLO R. Tissuelab S.P.A. Italy.<br />
Abstract/Introduction:<br />
BMP-2 and BMP-7 are bone morphogenetic proteins, that have a putative role as osteogenic factors in vivo in the healing<br />
of bone defects . They have been implicated as a bone-stimulating agent for spinal fusion therapy and the treatment of nonunion<br />
fractures. This study investigates the capacity of processing method developed by Tissuelab to safeguard the presence<br />
of these proteins. Material: The cortical bone is obtained from 50 bone donations processed for Florence and Treviso Tissue<br />
Banks (Italy) and BISLIFE (The Netherlands). Quantikine immunoassay kits for BMP-2 and BMP-7 were purchased (R&D<br />
Systems) Methods: DBM putty is produced by mixing DBM powder with a carrier composed by PEG and glycerol. The paste<br />
is successively sterilised with gamma radiation at low temperatures (-80°C). On every batch Tissuelab carries out an ELISA<br />
assay to quantify the content of BMP-2 and 7. The BMPs were extracted from DBM powder with 4 M guanidine-HCl in 0,5<br />
M sodium acetate and then dialysed with an ipotonic solution. The samples were analyzed with the immunoassay kits and<br />
spectrophotometric detection (450 nm).<br />
Results:<br />
The ELISA analysis detected BMP-2 and BMP-7 in all 50 samples of DBM product. The average content of BMP-2 was 36,2<br />
ng/g of tissue with a coefficient of variation of 79,4%; the average concentration for BMP-7 was 99,6 ng/g of tissue with<br />
CV%= 56,4% Discussion and conclusion: The bone morphogenetic proteins (BMPs) make up a subgroup of the transforming<br />
growth factor β (TGF-β) superfamily. They were originally identified as regulators of cartilage and bone formation. They have<br />
also been implicated in embryogenesis and morphogenesis of various tissues and organs. According to literature there is<br />
an high variability related to characteristic of donors, such as gender, age and presence of metabolic bone disease. The<br />
obtained values are in compliance with the BMPs content calculated for similar products by Hyun W. Bae et al., (2006). The<br />
BMP values are measured by ELISA assay before and after Tissuelab processing and BMP content is preserved for about<br />
80%, both for BMP-2 and BMP-7 The Tissuelab process minimally affects BMP values in putty and their content is closely<br />
related to the donor characteristic. References: 1) Hyun W. Bae et al., Spine (2006); 12:1299-1306. 2) Blum N. et al.,<br />
Orthopedics 2004; 2: 161-5 3) Alanay A. et al., Spine J. 2008 Sep-Oct;8(5):789-95.<br />
O-59<br />
DIFFERENTIATION OF PORCINE DERMAL CELLS INSIDE AUTOLOGOUS FIBRIN<br />
SCAFFOLDS<br />
PUENTE, P.; LUDEÑA D.; LÓPEZ M.; RAMOS J.; ARANDA LUIS J.; VARELA G.; IGLESIAS J.<br />
Tissue Bank San Francisco Clinic Foundation. US.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
Mesenchymal stem cells (MSCs) are an ideal candidate cell type for tissue engineering and regenerative medicine. Our aim<br />
has been to evaluate multilineage potential of porcine dermal cells to differentiate inside autologous fibrin scaffolds. Fibrin<br />
scaffold provide a 3D structure for the cells to adhere, proliferate and differentiate. Porcine Dermal Cells have showed the<br />
ability of differentiation in vitro towards adipocytes, osteoblasts, and chondrocytes inside autologous fibrin scaffolds. Oil<br />
Red O staining confirmed the presence of intracellular red lipid droplets in adipogenic differentiated cells, Von-Kossa staining<br />
showed an evident mineralization of the matrix with calcium deposits in osteoblasts differentiated cells and osteogenic
ORAL<br />
PRESENTATIONS<br />
marker osteopontin was detected, and Alcian Blue staining revealed mucopolysaccharides synthesized and<br />
immunohistochemistry of collagen type II evidenced the production of collagen fibers by chondrogenic differentiated cells.<br />
In approximately a month we have been able to obtained differentiated cells in a 3D scaffold, which it is totally autologous,<br />
biocompatible and implantable. Autologous fibrin scaffolds represent a suitable structure for cell growth and tri-lineage<br />
mesenchymal differentiation, suggesting that fibrin scaffolds may be useful for tissue engineering.<br />
O-60<br />
COMPARISION OF DIFFERENT SEEDING STRATEGIES TO ENHANCE FIBROBLAST<br />
PENETRATION WITHIN A HUMAN ACELLULAR DERMIS FOR SOFT TISSUE<br />
AUGMENTATION<br />
VITACOLONNA , M.; SMITH M.; HOHENBERGER P.; ROESSNER E. Universitätsmedizin Mannheim ; Chirurgische Klinik.<br />
Germany.<br />
Abstract/Introduction:<br />
Effective cell seeding often determines the success of tissue-engineering products. To create a stable soft tissue replacement,<br />
it would be desirable to achieve a maximum seeding efficiency, but also a homogenous cell distribution throughout the<br />
ADM. Natural matrices such as acellular dermis have the disadvantage of low permeability, due to their dense network,<br />
compared to synthetic materials with larger pore size. The purpose of this investigation was to compare different cell seeding<br />
methods regarding their seeding efficiency, homogeneity, infiltration depth and proliferation within an acellular dermis.<br />
Methods The examined methods can be divided into static, dynamic seeding techniques and a combination of both optional<br />
with PDGF as mitogen. Static seeding techniques include surface seeding, pretreatment of ADM with collagenase, direct<br />
injection of cell suspension by a syringe, cutting the matrix to increase the surface and diffusion and application of lowpressure<br />
and ultrasonic bath to remove trapped air. Dynamic seeding methods include an orbital shaker and the use of<br />
centrifugal force with different rotational speed and duration. After seeding, ADMs were incubated for up to 12 days and<br />
analyzed at day 0, 4, 8 and 12. At each corresponding time point, seeded ADMs were fixed, embedded vertically in<br />
paraffin, histologically sectioned and stained with propidium- iodide to analyze the cell distribution and penetration depth.<br />
Furthermore, cell proliferation, seeding efficiency and survival was evaluated by a MTT assay.<br />
Results:<br />
When using static methods without low- pressure pretreatment, cells were deposited on the surface as a single layer and no<br />
penetration into the matrix could be detected. However, after degassing the matrix, we were able to detect a significant<br />
improvement in penetration and proliferation. Dynamic seeding using a centrifuge increases the initial number of entrapped<br />
cells into the ADM; nevertheless we could neither demonstrate a high proliferation nor find any cells in the central areas.<br />
Whereas centrifugal force combined with low- pressure forces significantly more cells inside the ADM and increases the cell<br />
mass and homogeneity within 12 days than compared to the other methods.<br />
Discussion:<br />
As we have shown, the air in the pores significantly impeded the proliferation and therefore the penetration. Thus, the use<br />
of a single conventional method results in relatively inefficient colonization results when trying to colonize a dense matrix.<br />
We could archive the highest seeding efficiency, homogeneity, infiltration depth and proliferation by using low- pressure and<br />
centrifugation at 300g for 3x 1Min in addition with PDGF. Thus, we conclude that this combination is the most effective to<br />
repopulate dense natural matrices for soft- tissue augmentation.
ORAL - NOVEMBER 10TH - CORNEA<br />
ROOM 1 - 15.15-16-55<br />
O-61<br />
CORNEAL ASSESSMENT USING LIGHT MICROSCOPY: THE DIFFERENCE BETWEEN<br />
HEALTHY AND PATHOLOGICAL TISSUE<br />
JIRSOVA, K. Ocular Tissue Bank and Laboratory of the Biology and Pathology of the Eye, Institute of Inherited Metabolic Disorders,<br />
General and Teaching Hospital and 1st Faculty of Medicine, Charles University in Prague, Czech Republic.<br />
Organ culture is the method of choice for the preservation of a human donor cornea for transplantation throughout Europe.<br />
This method is often closely connected with the assessment of the cornea using light microscopy.<br />
Photographs from light microscopy (both phase contrast and bright field) will be presented. The individual corneal layers<br />
(the corneal epithelium, the stroma and the endothelium) obtained from both normal and pathological corneas will be<br />
shown.<br />
The presentation will be focused particularly on qualitative and quantitative changes of the corneal endothelium, such as<br />
endothelial cell density, polymeghatism, pleomorphism, and other abnormal endothelial morphologies, including nuclear<br />
abnormalities. Epithelial and stromal opacities will be presented.<br />
Photographs taken before and after corneal storage in organ culture will be compared and discussed in relation to the<br />
changes in endothelial morphology and the swelling of the intercellular spaces. Bacterial and fungal contamination of the<br />
corneas will be shown.<br />
Among corneal pathologies, cornea guttata, corneal dystrophies (Fuchs endothelial corneal dystrophy, posterior<br />
polymorphous corneal dystrophy) and crystalline keratopathy will be shown using pathological explants. Moreover, the<br />
corneal pathologies will be correlated with histological findings. Finally, bacterial and fungal contamination of the cornea<br />
during organ culture will be shown.<br />
O-62<br />
CORNEAL PROCUREMENT, BANKING AND TRANSPLANTATION IN FRANCE<br />
MARTINEACHE, I. France.<br />
O-63<br />
PARAMETERS INFLUENCING MICROBIOLOGICAL SAFETY OF CORNEAS<br />
VAN GEYT, C. Belgium.<br />
ORAL<br />
PRESENTATIONS
O-64<br />
QUALITY MANAGEMENT WITHIN THE EYE BANK<br />
GAREISS-LOK, A. CEBT CEO, Hornhautbank Muenchen GmbH, Munich. Germany.<br />
Purpose:<br />
Establishment of a reproducible and traceable quality management system within the Eye<br />
Eye Bank to release corneal tissues for all purpose of surgery.<br />
Methods:<br />
Standardized evaluation of corneal tissues with slitlamp- and endothelium microscopy<br />
within a closed system.<br />
Corneal tissues are retrieved by enucleation of whole eye (preparation of corneal button is performed in eye bank under<br />
sterile conditions) or in-situ-excision of corneal button. Every corneal tissue is stored within corneal viewing chambers – no<br />
matter if organ culture or hypothermic storage is performed which allows every necessary examination within a ‘closed<br />
system’. All examinations are documented and filed within the database system of eye bank. Special programmed<br />
authorization and lodged quality criteria will avoid wrong usage of evaluated tissues.<br />
Results:<br />
The developed and established way of evaluation in combination with an exclusively programmed data base system reduced<br />
the failure rate of wrong documentation which may lead to wrong usage of corneal tissues tremendously (failure rate <<br />
1%).<br />
Conclusion:<br />
The establishment of a unique customized database system in combination with evaluation of corneal tissue leads to a<br />
reproducible and traceable documentation of the tissue evaluation within the eye bank.<br />
O-65<br />
PRE-CUTTING OF GRAFTS FOR POSTERIOR LAMELLAR KERATOPLASTY<br />
ORAL<br />
PRESENTATIONS<br />
JESPER HJORTDAL, MD, PhD, The Danish Eye Bank, Department of Opthalmology, Aarhus University Hospital, Nørrebrogade<br />
44, 8000 Aarhus C, Denmark.<br />
Today, the preferred treatment of primary and secondary corneal endothelial failure is posterior lamellar keratoplasty. In this<br />
technique, a human donor cornea is split in two parts. The inner part consisting of endothelial cells, Descemets membrane, and<br />
a thin part of stromal lamellae is inserted into the anterior chamber of the patient’s eye and pressed towards the inner side of the<br />
patient’s cornea by air.<br />
Preparation of the posterior lamellar graft is typically done in the operating theatre by the surgeon. This process is time-consuming<br />
and failure in the preparation may result in cancellation of surgery.<br />
Preparation of the posterior lamellar graft in the eye bank one or more days before surgery is possible. In the United States such<br />
pre-cutting of corneal donor tissue before cold storage in Optisol is very common. In Europe, organ culture storage is the most<br />
common storage method and there is very little published experience with pre-cutting of organ cultivated donor corneas.<br />
In the presentation, a summary of studies on the use of pre-cut corneal donor tissue will be given, along with the authors experience<br />
of using pre-cut organ cultivated tissue for posterior lamellar keratoplasty.
O-66<br />
SEROLOGY AND MICROBIOLOGICAL TESTING IN EYE BANKS:<br />
LEGAL ASPECTS IN EUROPE<br />
MAIER, P. University Eye Hospital Freiburg. Germany.<br />
ORAL<br />
PRESENTATIONS<br />
When the EU Directives 200423EC, 200617EC and 200686EC were transformed into the national legislation of the EU member<br />
states many procedures including the serological testing of the donor and the microbiological testing of the culture media were<br />
regulated in a new way.<br />
To minimize the risk of transmitting an infectious disease of the donor testing for HIV antibodies, hepatitis B antigen, hepatitis C<br />
antibodies, and Lues antibodies have already been standard procedures in most European eye banks. According to the EU<br />
directives tests for Hepatitis B core antibodies were added for donor screening and some countries even integrated NAT testing<br />
for HIV, HBC and HCV in their national test algorithm. However, it is not clear, whether these additional blood tests might help to<br />
improve patients’ safety as there are only two suspected transmissions of Hepatitis B by corneal grafts that happened when no<br />
routine testing was done at all. The major problem of the EU directives however was the decision of the European commission to<br />
limit the time frame for taking blood samples from deceased donors for serological and NAT testing to 24 hours whereas the cornea<br />
can be procured up to 48 or even 72 hours after death. This so called 24 hour rule lead to a significant reduction in corneal<br />
donation in many European countries although the reason for this limitation remains unclear as there is no scientific evidence that<br />
the 24 rule leads to increased safety of the tissue. Besides the problem of the 24 hour rule there are no validated serological and<br />
NAT tests for postmortal blood samples so that in some regions blood from deceased donors is not accepted at all. To overcome<br />
these problems a clinical trial has been performed that proved that serological and NAT test values regarding HIV, HBV and HCV<br />
remain stable for at least 48 hours after death of the donor and a validation for the necessary tests was also done. Therefore, we<br />
hope that it will be possible to change the European directive regarding the 24 hour rule in the future.<br />
Besides testing of the donor for infectious diseases the media of the tissue need to be tested microbiologically in order to detect<br />
contaminations of the tissue to avoid transmissions of bacterial infections by the graft. In this context many different testing methods<br />
exist ranging from simple agar plates to membrane filtration techniques. The European Directives do not give any detailed<br />
information on how the sterility tests should be performed so that each member state has to define its own regulations. In Germany<br />
for example there is no specific tissue law, but all tissues are included under the pharmaceutical law. This leads to various problems<br />
where one major issue is the sterile testing of the culture media. According the European Pharmacopoeia membrane filtration is<br />
the only suitable method for sterility testing of solutions like the organ culture media containing antibiotics and antimycotics.<br />
However, this method is very time and cost consuming compared to the currently most common method using blood culture bottles<br />
for sterility testing. Furthermore, as for serological tests there are no validated methods for sterility testing of culture media from<br />
corneal organ culture. Besides the cost factors one has to keep in mind that the cornea itself is never sterile, so if testing becomes<br />
too sensible many tests may give positive results leading to an even more reduced availability of donor tissue.
O-67<br />
IMPLEMENTATION OF EUROPEAN LEGISLATION IN EYE BANKING: THE CZECH<br />
EXPERIENCE<br />
KRABCOVA I., ZLACKA D., JIRSOVA K., Laboratory of the Biology and Pathology of the Eye, Institute of Inherited Metabolic Disorders<br />
and Ocular Tissue Bank, General Teaching Hospital and 1st Faculty of Medicine, Charles University in Prague, Czech<br />
Republic.<br />
Purpose:<br />
To draw attention to problems arising from the implementation of European legislation in the Czech Republic.<br />
ORAL<br />
PRESENTATIONS<br />
Introduction:<br />
Act No 296/2008 Coll. (Act on Human Tissues and Cells) and Decree No 422/2008 Coll. (concerning the detailed requirements<br />
for the safeguarding of the quality and safety of human tissues and cells) have been enacted in the Czech Republic implementing<br />
the European legislation: Commission Directives 2004/23/EC, 2006/17/EC and 2006/86/EC.<br />
Czech eye banks need accreditation from the State Institute for Drug Control (SIDC) as the competent authority. During inspections<br />
and evaluations with regards to legislation, these directives have started to be strictly enforced.<br />
We would like to draw attention to difficulties with the observance of the rules. Some of the critical material routinely used for tissue<br />
culturing, defined as In Vitro Diagnostic Medical devices (Directive 98/79/EC), is no longer allowed to be used and must be<br />
replaced by material based on Directive 93/42/EEC (Medical Devices). Thus, materials such as minimal essential medium,<br />
phosphate buffer and antibiotic solution (AppliChem, Germany or Gibco/Invitrogen, UK) have to be replaced by commercial<br />
solutions (Eurobio, France or Alchimia, Italy). This situation especially affects the storage of corneas in organ culture. No commercial<br />
solutions are offered for the storage of human amniotic membranes. Similar problems arise with the use of storage plastics.<br />
The resulting increase in the cost of the tissue is not matched by an increase in the reimbursement offered by insurance companies.<br />
The changes required by Czech legislation can not be introduced without appropriate clinical studies, and their impact on tissue<br />
quality and safety has yet to be assessed.
O-68<br />
PROSPECTIVE DEVELOPMENT OF EYE BANKING IN THE RUSSIAN FEDERATION<br />
BORZENOK, S.A., KOMAK. Russian Federation.<br />
O-69<br />
ORGANOTIPIC CULTURE PRESERVATION AS STRATEGY TO INCREASE THE NUMBER<br />
OF CORNEAS FOR TRANSPLANTS<br />
*OTERO, N. 1 , VILARRODONA, A. 1 , *MARTÍNEZ-CONESA, E. M. 1 , NÚÑEZ, V. 1 , PÉREZ, M. L. 1 , AGUSTÍ, E. 1 ,<br />
CASAROLI-MARANO, R. 2 , TRÍAS, E. 1<br />
Transplant Services Foundation-Hospital Clínic<br />
2.- Universidad de Barcelona<br />
*Contributed equally to the work. Spain.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract/Introduction:<br />
Our Tissue Bank is using simultaneously two methods for preserving corneas from retrieval to transplant: Preservation in<br />
Optisol® at 4-8ºC with a maximum expiration date of 8 days (fresh corneas) and organotipic culture preservation during<br />
14-28 days. Corneas in organotipic culture method from January 2010 till April 2011 were included in this study. In that<br />
period 33.2% of the retrieved corneas were organ-cultured. The final suitability for transplantation was 76% for fresh corneas<br />
and 62% for cultured ones.<br />
Purpose:<br />
To evaluate the quality and microbiological status of organ cultured corneal grafts from donors who have died mainly from<br />
septic multi-organ failure. Methods: 390 corneal grafts (205 donors) from septic and non-septic donors were stored in<br />
organ culture method at 31°C for 14-28 days. Evaluation of cultured corneas was carried out by specular microscope, slit<br />
lamp microscope and optic microscope examination. From each cornea donor one blood culture for aerobic and aerobic<br />
microorganisms was taken at the moment of clinical screening and a second blood culture during the tissue retrieval. Three<br />
microbiological tests have been performed at different preservation stages, the first microbiological sample was taken from<br />
the medium after 5 days of culture, the second one corresponded to the moment that tissue is transferred to the deturgescent<br />
medium and the final one 24 hours after. Corneal grafts with cell density values above 2000 cells/mm 2 were transplanted.<br />
Follow-up and traceability of transplanted corneas were analyzed taking into special consideration on satisfactory clinic<br />
evolution complications and microbiological controls.<br />
Results:<br />
In the period of study 390 corneas were included in the organotipic culture group in which 323 (82.8%) corneal grafts were<br />
finally put in organ-culture from 205 donors and 67 (15.9%) were discarded due to don’t achieve corneal quality criteria<br />
or donor history. The average of donor age was 67.8 years (± 13.9). From the group of cultured corneas 244 (75.5%) were<br />
suitable for transplantation and 79 (24.4%) were non-suitable from which 11(3.4%) were discarded due to contamination<br />
medium. In contrast, 158 corneal grafts (40.51%) had positive blood cultures. In the follow-up forms analyzed 97.8% had<br />
a satisfactory evolution versus 2.2% that had not a good evolution because of persistent epithelial defect with overinfection<br />
and corneal oedema during 2 months.
O-70<br />
MICROBIAL RISK ASSESSMENT IN EYE BANKING<br />
MAJORE, I.; SCHEIDER U.; WITTMERSHAUS I.; BÖRGEL M.; BLOMBERG L.; KNIPPER A.<br />
German Society of Tissue Transplantation. Germany.<br />
ORAL<br />
PRESENTATIONS<br />
Abstract:<br />
The manufacture of human cornea transplants is regulated by comprehensive rules to ensure that sound, high quality practices<br />
are followed to reduce the risk of tissue contamination and of communicable disease transmission to recipients. A central part<br />
of the manufacturing process in a GMP facility is a thorough risk assessment process. Even the healthy human eye as part of our<br />
body is steadily colonized by microorganisms. Therefore, the preparation of corneal tissues, even under rigorous conditions of<br />
good manufacturing practice (GMP), inevitably harbors a risk of pathogen transmission into the manufacturing plant. The main<br />
objectives of this study were (1) to determine the extent and specificity of the initial contamination of donated eyes upon arrival<br />
in the eye bank, (2) to examine the specificity and extent of pathogen transfer to the surrounding environment during corneoscleral<br />
disc preparation, (3) to develop a rational approach for microbial monitoring of corneoscleral disc processing. GMP-compliant<br />
manufacturing of cornea transplants was done in the grade A area (laminar air flow cabinet (LAF) of the clean room with the<br />
background environment of grade B. The work space of the LAF was divided in three zones - a tissue decontamination zone,<br />
an in-between zone and a zone for corneoscleral disc processing. Airborne microbial contaminations of these zones during the<br />
manufacturing process within the LAF were analyzed by use of settle and contact plates. The initial microbial flora of donated<br />
eyes was investigated through swab approaches. Subsequently, eyes were decontaminated in 5% povidone iodine solution for<br />
5 min. Prepared corneoscleral discs were stored in MEM supplemented with 2% FBS, antibiotics (penicillin, streptomycin) and<br />
antimycotics (amphotericin) at 37° C in a closed culture system without exposure to carbon dioxide. At day 4 to 5 the presence<br />
of contaminants in the organ culture medium was studied via BACTEC Plus blood culture systems. The results of this study show<br />
that (1) the PVP decontamination treatment during the eye procurement process, moistening the eye surface with antibiotics<br />
(gentamicin) and adherence to temperatures between 2 and 10 during transport to the eye bank result in a significant reduction<br />
in microbial contaminations; (2) the manufacturing process does not notably increase the risk of pathogen transmission; (3) the<br />
enrichment of organ culture medium with common antibiotics and antimycotics effectively eliminate most of the remaining<br />
contaminants. However, a strict separation of work space under the LAF during corneoscleral disc processing and a continuous<br />
microbial monitoring are strongly recommended for a safe manufacturing process.
<strong>POSTERS</strong>
INTERNATIONAL EXPERIENCE ON TISSUE BANKING<br />
P-01<br />
ACTIVITY REPORT OF THE MULTI-TISSUE BANK OF THE RED CROSS BLOOD<br />
TRANSFUSION SERVICE OF UPPER AUSTRIA<br />
HENNERBICHLER, S.; PETERBAUER-SCHERB, A.; PLÖDERL, K.; GABRIEL, C., Red Cross Blood Transfusion Service of Upper<br />
Austria, Linz/ Austrian Cluster for Tissue Regeneration. Austria.<br />
Abstract:<br />
In 2003 the Red Cross Blood Transfusion Service of Upper Austria started cord blood banking. It turned out as starting point<br />
for our multi-tissue bank. In 2007 all provisions were made for the first Austrian ISO 9001:2000- and GMP-certified multitissue<br />
bank situated at the new building of the Red Cross Blood Transfusion Service of Upper Austria. Since then our institution<br />
is specialized on several tissue banking products asides blood products. At this time we started to introduce heart valves and<br />
amniotic membrane as our first products besides cord blood. In 2008 the activity of the tissue bank was further expanded<br />
to the preparation of cornea transplants. Subsequently, in 2009 the tissue bank of the Red Cross Blood Transfusion Service<br />
was the first multi-tissue bank in Austria certified according to EC-Directives 2004/23/EC, 2006/17/EC, and 2006/86/EC<br />
and is now certified for procurement, testing, processing, storage and distribution of: * Cord blood * Heart valves and<br />
vessels * Amniotic membranes * Cornea * Ovarian tissue * Femoral heads, calvarium * Autologous platelet lysate * Serum<br />
eye drops * Semen and testes (in preparation).<br />
P-03<br />
ISO 9001:2008 AUDITS AND NONCONFORMITY REPORTING IN A TISSUE BANK<br />
KALAYDJIEVA, V.; POPOVA, P., Tissue Bank "Osteocentre Bulgaria". Sofia. Bulgaria.<br />
<strong>POSTERS</strong><br />
Abstract:<br />
The main ISO principles can be properly adopted in the quality management of a tissue bank. The improvement of the<br />
quality system can be maintained and measured by performance of internal audits following the ISO requirements.<br />
Objetive:<br />
The study describes the benefits of performing ISO-complied internal audits in our tissue bank and the maintenance of an<br />
adequate system for nonconformity reporting. Such approach is necessary to achieve the quality requirements and to review<br />
the effectiveness of the arrangements for quality assurance.<br />
Results:<br />
All organizational processes were identified and managed in compliance to ISO standards. Quality metrics were introduced.<br />
The internal audits conducted aimed at verifying the compliance of processes performed throughout the organization with<br />
respective ISO 9001:2008 standards. An internal quality auditing program was established after an appropriate training<br />
of employees. Numerous internal audits of all organizational units and processes throughout the year have been conducted.<br />
The internal audits were carried out on the basis of a prediscribed method in SOP. The methods used were as follows:<br />
interviews with employees; examination of records and documentation; complete trace of individual processes; checking of<br />
actual compliance with requirements; reporting nonconformities. Through internal audits we covered all ISO areas of<br />
requirements thus evaluating among all the state of the referring system, management of donor cases and monitoring the
<strong>POSTERS</strong><br />
reliability and accuracy of donor documentation. Focused audits have been conducted in parallel to monitor critical areas<br />
and when problems with quality have been identified. Improvements in management, training, donation and recovery<br />
processes supervisions, and statistical reporting became evident during internal audits. Although the results showed a lack<br />
of significant non-conformities, all detected deviations were reported according to the established nonconformity reporting<br />
system. A formal corrective action system has been developed and implemented. The periodical analysis of the type and<br />
severity of nonconformities was of great importance for continuous improvement of the quality system.<br />
Conclusions:<br />
Internal audit is a helpful tool for identification of problematic areas and opportunities for improvement in tissue banking.<br />
Positive effects on organizational quality culture has been evaluated during management review meetings. The conduction<br />
of internal audits according to ISO requirements is complementary to the audits performed to review the compliance with<br />
tissue banking standards. The implementation of a proper system for nonconformity reporting helped changing the concept<br />
of quality management from detection to prevention.<br />
P-04<br />
A 6 YEAR RETROSPECTIVE REVIEW OF THE KALEIDOSCOPE OF CHALLENGES<br />
ENCOUNTERED BY A SOUTH AFRICAN TISSUE BANK AND THEIR INFLUENCE ON<br />
DONOR STATISTICS AND ALLOGRAFT AVAILABILITY<br />
KARAKATSANIS, E., Centre for Tissue Engineering - Bone Bank (Tshwane University of Technology). South Africa.<br />
Abstract:<br />
Purpose of study The estimated South African population stands at approximately 50 million and the Centre for Tissue<br />
Engineering - Bone Bank provides more than 16 000 bone and soft tissue allografts to the medical fraternity across South<br />
Africa annually. As a developing third world country, the study highlights the various challenges (cultural diversities, language<br />
barriers, education, public awareness and the lack of standards), that influence the concept of donation, which in turn have<br />
an effect on the availability of safe transplantable tissue to the recipient. Description of methods Donor data and results were<br />
systematically accumulated over a period 6 years (2005 – 2010), from the three main divisions in the Bone Bank – namely,<br />
Procurement, Quality Control and Production. Summary of results The challenges include the lack of standards and<br />
regulations, outdated government policies/acts relating to tissue donation and transplantation, education, public awareness,<br />
cultural and language barriers have had a crippling and curtailed consequence in the availability of tissues in South Africa.<br />
Despite these overwhelming challenges, the Bone Bank has been able to maintain an adequate monthly retrieval rate which<br />
has translated to an increase of 41.27% transplantable tissue from 2005 to 2010. A high percentage of the donors were<br />
from the caucasian population group, while the lowest donor group was identified from the African population group.<br />
Within the 6 year period, 983 QC passed tissue donors, were processed into 96 325 safe quality allografts for the much<br />
needed South African public. Conclusion In spite of these challenges, the Bone Bank has maintained excellent working<br />
relationships with the Organ Donor Foundation (ODF), Eye Cornea Bank and numerous hospital groups, such as Netcare.<br />
The continuous in-house policy, procedural and process improvements and with the implementation of both ISO 9001 and<br />
ISO 13485, the Bone Bank has optimised its production processes and significantly improved allograft delivery to its South<br />
African recipients, despite the challenges. It is the responsibility of every health care professional and healthy individual in<br />
South Africa to become conscious for the need of solid organs, corneas, bone, heart valves and skin. Therefore, it needs to<br />
become a collective obligation in a two-fold manner a) to sensitise decision makers and government to this national challenge<br />
and b) encourage and inform patients and next-of-kin to be organ and tissue donors.
P-05<br />
COMMUNICATION & COSTUMER SERVICE IN A TISSUE BANK<br />
<strong>POSTERS</strong><br />
SOBREVILLA, E.; MERAYO, N.; GALLARDO, G.; REDONDO, M.; MIRANDA, I.; JARDÍ, F.; TRIAS, E., Transplant Services<br />
Foundation. Hospital Clínic of Barcelona. Spain.<br />
Abstract:<br />
Human tissues are a scarce commodity but in many cases essential for the healing of patients. Therefore, good coordination<br />
between the demand for human tissue and allograft availability is a key factor in the processes of tissue banks. Control all<br />
stages of the procedure from donation to distribution in the hospital, along with a thorough internal and external<br />
communication are the keys to success to improve the effectiveness and efficiency of the tissue bank. Fluid communication<br />
with the surgeons allows the tissue needs analysis to provide the most appropriate, depending on availability and timing<br />
needs. This is more relevant in cases where the demand is much higher than allograft availability as is the case in paediatric<br />
surgery Aim The goal is to analyze how a good and well organize communication & customer service can improve the<br />
effectiveness and efficiency of a tissue bank, managing something with limited availability as the human tissues are. This<br />
situation is even more important when we are speaking about paediatric tissues or paediatric necessities. Study Methods<br />
We have analyze all the procedures from tissue harvesting to allograft implantation, information generated by different<br />
steps and the communication necessary to provide the surgeon with the allograft more suitable for his surgical needs, and<br />
we have correlate it with the surgeon satisfaction on the Tissue bank services. To study an specific area, we have done<br />
comparative analyses between the demand of paediatric tissues and the distributed tissue, taking into account the waiting<br />
list. It has also been studied in which cases adult tissue can be adapted for paediatrics us and when the need of tissue<br />
transplantation is crucial for life saving. Results We studied all grafts provided since January 2010 and analyzed the<br />
adequacy of what served the needs of the clinical case by: - Surgeon's Feedback (complaints, thanks, communications) -<br />
Internal communication files - Currently, in many cases, we have to adapt adult tissue for use in paediatrics Conclusions A<br />
good internal& external communication are essential to optimize the results and offer the surgeon the best opportunity<br />
available to your patient. The recovery and distribution of adult and paediatric tissue is a need in growing demand. It is a<br />
must to organize the activities and design systems to increase the effectiveness in the tissue assign in order to optimize the<br />
tissues availability.<br />
P-06<br />
TISSUE PROCESSING RESULTS IN 2010 IN THE BANK TISSUE AREA OF BLOOD AND<br />
TISSUE BANK OF ARAGON<br />
MARTÍNEZ-LORENZO, M.; PIÑERO PIMPINELA, E.; IBÁÑEZ SOLLA, L.; VICENTE BORRUEL, O.; FERNÁNDEZ-PACHECO,<br />
J.; CALLÉN SEVILLA, L., Banco de Sangre y Tejidos de Aragón. Zaragoza. Spain.<br />
Abstract:<br />
The Blood and Tissue Bank of Aragon carries out activities relating to processing, preservation, storage, conservation and<br />
distribution of human tissues and blood components. The Area of Tissue Bank began operations in 2009. This area handles<br />
the processing, preservation, storage and distribution of tissues from living donors and cadaver donors that will be intended<br />
for clinical application in humans is regulated by the RD1301/2006. The aim of this study is to analyze the results for tissue<br />
donation, processing activity in the Tissue Bank Area and the implants performed in hospitals of the Autonomous Community<br />
of Aragon. To determine the suitability of tissue were used AEBT standards. All tissues met the requirements of the<br />
RD1301/2006. Tissue processing was carried out in Class B clean rooms and in class A laminar flow. To ensure the<br />
adequacy of tissue, we performed all the necessary microbiological controls (both the tissue processed as environmental
control and surface clean rooms used in processing). During 2010, the Tissue Bank Area processed 28 cadaveric donations.<br />
Of these, 28 donated eye tissue and 13 bone tissue. We obtained from bone tissue, cancellous bone, femoral condyle,<br />
proximal tibia, diaphysis and tendons (Achilles, patellar, tibial and semitendinous). In the case of eye tissue, maintenance<br />
of the corneas was performed at 4 º C or 31 º C, depending on the needs of the implanted equipment. The Tissue Bank also<br />
has pieces of amniotic membrane for ocular defects obtained from placenta. The number of implants in the community of<br />
Aragon, with tissue provided by our bank was 211 (74.8% bone tissue, 22.7% eye tissue and 4.2% amniotic membrane<br />
which led to 307 pieces of tissue. Only 2.6% of the implanted tissues were requested from other regional banks. The Tissue<br />
Bank area also performs serological and microbiological studies of implants. The Bank Tissue of Aragon has improved the<br />
accessibility of tissues to different hospital authorized to implant in the Community of Aragon. During 2010, we have<br />
supplied more than 97.4% of the tissue needs of the Community of Aragon. For these reasons, the control of the implants<br />
in the different hospitals (serology and microbiological serum bank of the implants) has also been greatly improved.<br />
SPECIFIC CHALLENGES OF DONATION; PROCESSING AND CLINICAL<br />
APPLICATION IN PAEDIATRICS<br />
P-07<br />
EFFECTS OF NEW EU DIRECTIVE ON POSTMORTEM DONATION ACTIVITY IN<br />
SWEDEN<br />
WALLBERG, L. 2 *, ODÖ, B. 2 , JOSEPHSSON, S. 2 , DAHLBERG, A. 2 , ALKASS, K. 2 , DRUID, H. 1<br />
1 - Karolinska Institutet, Stockholm 2 - Swedish National Board of Forensic Medicine. Sweden.<br />
<strong>POSTERS</strong><br />
Abstract:<br />
The EU directive 2004/23/EG implies that today, a blood sample from deceased donors must be collected within 24 hours<br />
after death for analysis of possible contagious agents in order to reduce the risk that serious diseases are transmitted to<br />
recipients during a transplantation procedure. This directive has become a Swedish law (2008:286), which require a more<br />
urgent handling of possible donors. Given the many obstacles during the identification of relatives and the procedures to<br />
obtain an informed consent in addition to logistic problems regarding the transport of a possible donor, it was expected that<br />
this directive would reduce the number of donation procedures. Hence, the Swedish National Tissue Project was started, and<br />
involved the Swedish National Board of Medicine, since this is a national agency running all forensic medicine departments<br />
in the country, at which most of the postmortem tissues are collected. To determine the impact of increased resources on the<br />
success rates, one department with a good track record of many donors, and two with a dormant activity were given support<br />
for extra personnel and compensation for on-call duty to collect blood samples within 24 h. A fourth forensic medicine<br />
department given no extra resources was included as a control site. During the project period, Sept 2009-Aug 2010, tissues<br />
from 94 donors were collected at the forensic medicine departments, including 40 donations of heart valves and 70 corneas,<br />
and in addition, from mostly the same subjects, tissues for research purposes could be obtained in 46 cases. The preceding<br />
12-month-period the corresponding figures were 23, 36 and 29, respectively. The increase was largest in Stockholm, where<br />
heart valve and cornea donors increased from 4 to 18 and from 13 to 45, respectively. At the department in Lund, not<br />
obtaining any extra resources, heart valve and cornea donors decreased slightly from 19 to 16 and from 18 to 16,<br />
respectively, partly as an effect of the EU directive´s requirement for blood sample collection within 24 hours. It is concluded<br />
that increased resources are necessary to compensate for the reduction of formally acceptable donors caused by the EU<br />
directive. Studies are running to examine the rationale for the 24 hour rule by comparing blood test results at different time<br />
points after death. Preliminary observations suggest that there are no significant differences in false negative results regarding<br />
HIV and HCV detections between samples collected before and after 24 hours.
P-08<br />
LEGAL MEDICINE VERSUS HEART VALVE DONATION<br />
BRAUN, C.; WITTMANN, G.; JECKLE, J.; SCHRAMM, W.; MATTHIAS, G., Institute for Legal Medicine Munich. Germany.<br />
Abstract:<br />
As human heart valves are in high demand for certain surgical indications and the use of hearts from non beating heart<br />
cadavers is possible, institutes of legal medicine present a interesting source of potential donors. In Germany, the Institute<br />
of Legal Medicine in Munich has a tradition of heart valve explantation since 1982. In 2009, the Bavarian Tissue bank was<br />
founded as a response to the demands of the German Tissue Law. Further conditions to be met are formulated by the<br />
controlling government agency, the Paul Ehrlich Institute. With higher requirements, the difficulties to integrate a program<br />
for heart valve donation in the special setting of an institute for legal medicine are rising as well. This concerns e.g.<br />
organisational matters, quality management and especially extensive disinfection/sterility measures during explantation to<br />
ensure recipient safety against the background of a high daily workload of postmortem examinations in the setting of a<br />
forensic autopsy room. We will present our procedure to balance the requirements of legal medicine on one hand against<br />
those of heart valve donation on the other, and discuss assets and drawbacks to this approach.<br />
P-09<br />
DONATION FOR RESEARCH USING A PROSPECTIVE STRATEGY<br />
ALKASS, K.,* BÄCKLIN, S, DRUID, H. KAROLINSKA Institutet, Stockholm, Sweden.<br />
<strong>POSTERS</strong><br />
Abstract:<br />
In 2005, a core facility for donation for research, KI Donation, was founded at Karolinska Institutet, Stockholm, and today<br />
tissues from 218 deceased donors have been collected and used for different research projects, resulting in important<br />
scientific contributions. The activities of KI Donatum are integrated with both the activities for donation for transplantation,<br />
and the routine forensic casework. Several donors have signed up both for donation for transplantation and for research<br />
(or the relatives have given consent for both purposes), and the collection procedures can often be performed in parallel.<br />
KI Donatum has a prospective strategy, implying that tissue regions of interest are carefully dissected out, and handled and<br />
processed according to protocols designed by the researcher responsible for each particular project. It also means that<br />
specific information, critical for the project, is gathered from different sources and carefully evaluated. Hence, a project<br />
regarding cardiomyocyte turnover requires detailed information about previous heart problems, and careful cardiac<br />
pathology examination whereas a project regarding neuropharmacological changes due to heroin abuse necessitates a<br />
thorough review of the drug use history, from relatives and medical charts, and comprehensive toxicological analyses of<br />
blood, urine and hair. The concept of KI Donatum also implies some ancillary services, e.g. certain tissue processing (from<br />
simple centrifugation to advanced procedures such as cell sorting), immunohistochemistry and specific biochemical analyses,<br />
in order to further characterize the individuals. Today research on postmortem tissues is typically performed on material<br />
available at tissue banks, but then the researchers have to accept that certain information important to the particular project<br />
may be missing or uncertain. Therefore a few tissue banks have started prospective sampling in parallel with their routine<br />
donation procedures. To improve the quality of research on postmortem tissues, we recommend tissue facilities to consider<br />
the strategy outlined above when procuring material for research purposes. Below are some selected publications based on<br />
the activities of KI Donatum. 1. Philipova et al. Differential forms of p53 in medulloblastoma tumor, cell lines and xenografts.<br />
Int J Oncol. 2011 Mar;38(3):843-9 2. Brennan et al. Antibody-based proteomics: fast-tracking molecular diagnostics in<br />
oncology. Nat Rev Cancer. 2010 Sep;10(9):605-17 3. Uhlen et al. Towards a knowledge-based Human Protein Atlas. Nat<br />
Biotechnol. 2010 Dec;28(12):1248-50 4. Bergmann et al. Evidence for cardiomyocyte renewal in humans. Science<br />
2009;324(5923):98-102 5. Bhardway et al. Neocortical neurogenesis in humans is restricted to development. PNAS<br />
2006;103:12564-8 6. Spalding et al. Retrospective birth dating of cells in humans. Cell. 2005;122:133-43.
P-10<br />
DESIGN OF AN INTEGRATED CAMPAIGN FOR DONATION AND PROCUREMENT OF<br />
SKIN AND TISSUES THROUGH SOCIAL NETWORKING AND MICROBLOGGING<br />
SERVICES: THE MEXICAN EXPERIENCE OF THE NATIONAL RESEARCH CENTER AND<br />
BURN CARE (CENIAQ)<br />
MARTÍNEZ-F, F.; SANDOVAL-Z, H., Skin and Tissue Bank of the National Institute of Rehabilitation. Ministry of Health,<br />
Mexico<br />
Abstract:<br />
Mexico and other emerging countries worry about a fragile culture for the donation of organs and tissues. Loopholes,<br />
idiosyncratic problems, inappropriate public policies, ethical dilemmas, private interests and insufficient campaigns in mass<br />
media, combine with public budget deficits of the health system. All this negative background represents a considerable<br />
shortfall in the procurement and subsequent transplantation of organs and tissues such as human skin, necessary for the care,<br />
treatment and rehabilitation after severe burns. In order to find alternatives, the Skin and Tissue Bank in CENIAQ has the<br />
task of designing a new campaign through social networking and microblogging services (Facebook, Twitter, Google+<br />
LinkedIn and others), with the aim of reaching young population groups with higher levels of education and income as<br />
potential future donors.The object is to develop open, public, horizontal, participatory and democratic networks, as opposed<br />
to limited success of institutional networks that operate in closed, vertical and hierarchical bureaucratic systems. The goals<br />
for the campaign are: 1) Take advantage of lower costs and mass diffusion associated with the dissemination of messages<br />
through social networking and microblogging services; 2) Develop integral ongoing campaigns to raise awareness of<br />
potential donors; 3) Use educational techniques through micro-stories and other massive intervention methodologies, and<br />
4) Create a database --with appropriate safeguards for privacy-- where all the members can sign up as potential donors<br />
of organs and tissues in the near future. The campaign, divided in four phases to complete in a maximum period of one<br />
year, includes the design and dissemination of intervention techniques and feedback from participants. We expect to get<br />
international financial resources and specialized advice from global NGOs and private enterprises like Google. Once<br />
implemented, the model will be available to any Latin American public health institution under the canons of a Creative<br />
Commons license. This strategy aims to replicate the methodologies and increasing the rate of altruistic donation of organs<br />
and tissues for therapeutic purposes in public health systems.<br />
P-11<br />
THE CHALLENGE TO COORDINATE DEATH IN THE NAME OF LIFE<br />
POPOVA, P.; KALAYDJIEVA, V., Tissue bank "Osteocenre Bulgaria". Sofia. Bulgaria.<br />
<strong>POSTERS</strong><br />
Abstract:<br />
The tissue donation starts from ”the end” of one’s life in order to donate new “beginning” of other life or to get better it’s<br />
quality. The successful donor coordination requires strict observance of the fundamental principle in the medical philosophy<br />
“Primum non nocere”. This principle faces each doctor to the challenge to remain faithful to the Hippocratic oath. The donor<br />
coordination is complicate and difficult process. Each hospital coordinator is obliged to be acquainted with and observe the<br />
following regulations and requests: 1. Strict observation of the donor medical suitability criteria. 2. Observation of the<br />
current legislation and the moral and ethical norms. 3. Compassion and psychological support to the donor’s family. 4.<br />
Confidentiality about the mystery of donation. 5. The donated tissue is priceless gift of life, granted only to the patients in<br />
need as a gift. 6. Equal standing for donation- irreversible right to everyone. The comprehensive approach of donor<br />
evaluation determines the best way for the donor medical assessment. The selection of the potential donors should follow
the maxim “maximal benefit and minimal risk” for the potential recipients, needed transplantation The coordination of death<br />
in the name of life requires: many knowledge, developed sense of partnership, strong believe in donor mission, determination<br />
to win in the fight for standing up for the right to live. The donor coordination requires one dedicated team of partners. The<br />
fundamental columns in each donor process are: hospital coordinator, clinical psychologist, hospital surgical recovery<br />
specialist and in house donor coordinator on call in the Tissue Bank. The basic stages in donor coordination are: management<br />
of the donor referral system, report for each potential donor case – discussion, timely contact with the relatives and inform<br />
about the opportunity of donation, serological screening, signing of a consent form, recovery procedure with observation<br />
of all medical criteria for septic and antiseptic, diligent reconstruction of the body, properly final completion of the documents,<br />
storage of the recovered tissue. The coordination of the combined organ and tissue donors is even greater challenge for the<br />
hospital coordinators. The simultaneously representation of the opportunity for organ and tissue donation is the most correct<br />
and gentle approach toward the relatives and is the key to success in such donor cases. The strict observation of the moral<br />
and ethical norms during the implementation of medical occupation ensures the positive relation of the patients, their relatives<br />
and the society as a whole to the Healthcare and donation. Donation gives recipients last chance for treatment as a present.<br />
For that reason it is very important to have “opened for donation” National Health policy. The donor coordination is a great<br />
challenge to remain a doctor, even after the death of his patient.<br />
P-12<br />
IMPACT ASSESSMENT OF SKIN DONATION PROGRAM IN COSTUMERS OF A<br />
EMERGENCY ROOM AND UNIT OF INTENSIVE CARE<br />
<strong>POSTERS</strong><br />
MARTÍNEZ-F, F.; GUZMAN-M, K.; LOMELI-R, C.; MADINAVEITIA-V, J.; ELIZONDO, B.; CARRILLO, E., Skin and Tissue<br />
Bank of CENIAQ. National Institute of Rehabilitation. Mexico.<br />
Abstract / Background:<br />
In a study by fattum BTL to 40 public transport users in Mexico City during 2010 to assess the knowledge of the donation<br />
of skin and its therapeutic use, important results were obtained indicating 100% of respondents agree with the donation of<br />
organs and tissues and therefore be altruistic donors; 32% (13/40) respondents are unaware the skin is an organ that can<br />
be donated, however, 100% are willing to be a donor of skin after death. Therefore we assume that according to data<br />
obtained in 2010 more than 50% of respondents would know that the skin is an organ and can be donated but the<br />
vulnerability of the user in a hospital that has an impact on the acceptance of the donation.<br />
Material and Methods:<br />
We conducted a survey of 30 users of the emergency department and intensive care unit at the National Institute of<br />
Neurology and Neurosurgery (INNN) prior to an induction course on skin donation, which included 11 questions that were<br />
designed for data collection about the knowledge of donation of organs and tissues besides the willingness to accept the<br />
donation of skin in case of loss of a relative.<br />
Results:<br />
87% (26/30) agree with the donation of organs and tissues, 57% (17/30) know that the skin is an organ and therefore may<br />
be donated, 70% (21/30) are unaware of the use of donated skin, 73% (22/30) don’t know the process of donating skin;<br />
instead 30 % (9 / 30) is willing to accept the donation of skin when they suffer the loss of their relative.<br />
Conclusions:<br />
The results obtained by fattum BTL show that 100% of the population is willing to be a donor, but it is important to note that<br />
their emotional state is not subject to a time of stress and fear. However the population studied in the INNN indicates that<br />
30% of respondents are willing to donate if they suffer loss of a relative, taking into account that these are in a situation where<br />
their relative's health is compromised. It’s important to design a massive campaign where information is more extensive, for
<strong>POSTERS</strong><br />
the results show that 70% do not know the process and the use of skin that is donated. Additionally the education to fomenting<br />
culture of donation has been traditionally focused to Organs making relevant the necessity of a massive campaign and<br />
restructured program for tissues.<br />
P-13<br />
REVIEW OF THE TISSUE DONATION PROCESS AT THE TATA MEMORIAL HOSPITAL<br />
SAMANT CHANDRASHEKHAR, U.; GAJIWALA LOBO, A.; LIMA, C.; MAYEKAR, V., Tata Memorial Hospital. India.<br />
Abstract / Introduction:<br />
The Tata Memorial Hospital (TMH) Tissue Bank provides frozen and freeze-dried, irradiated bone grafts, and freeze-dried,<br />
irradiated amnion dressings. These tissues are obtained mainly from living donors. This study is a review of the tissue<br />
donation process from 2006-2010.<br />
Material and Methods:<br />
During the study period there were 3927 bone donors and 6444 amniotic membrane donors. 1088 (27.70%) of the bone<br />
donors and 992 (15.39%) of the amniotic membrane donors were rejected. 3112 bones and 5452 amnion were processed<br />
generating 5929 bone grafts and 9518 amnion dressings. Of the total grafts generated, 0.82% of the bone grafts were<br />
rejected due to machine failure and 0.50% of the amnion dressings were rejected during processing. From 2009-2010 a<br />
number of measures were used to reduce tissue donor rejections. These included intensified efforts to motivate potential<br />
donors, continuous follow up with donating surgeons to ensure blood samples or blood reports, collection of tissues at<br />
regular intervals to avoid tissue deterioration, conducting programmes to educate staff on the importance of donor screening,<br />
labeling and storage of donated tissues.<br />
Results:<br />
Rejection of the bone and amniotic membrane donors was due to inadequate screening (14.14%) because of unavailability<br />
of blood samples, non compliance with screening criteria (4.57%) including a recent blood transfusion, and improper<br />
storage of the tissue (1.25%). The number of rejections of amniotic membrane donors fell by more than 50% in 2009 and<br />
2010. Similar results were obtained with bone donors.<br />
Conclusion:<br />
Frequent and consistent communication between the tissue bank and the donating agencies, along with awareness<br />
programmes to educate staff, results in greater co-operation and compliance with the Tissue Bank’s requirements, with a<br />
consequent reduction in the number of rejected bone and amniotic membrane donors.
P-14<br />
CT SCAN IMAGING AS A TOOL IN TISSUE DONOR SCREENING PROCESS<br />
HERSON ROMA, M.; MORGAN, N.; FITZSIMMONS, A.; PONIATOWSKI, S.; O'DONNELL, C.; WOODFORD, N.; CORD-<br />
NER, S., Donor Tissue Bank of Victoria / Victorian Institute of Forensic Medicine. Australia.<br />
Abstract:<br />
The Victorian Institute of Forensic Medicine is a statutory body established to provide the Victorian Coroner with further<br />
information regarding cause of death of individuals deceased within the state of Victoria. Along with the State Coroner’s<br />
Office, the Institute (incorporating the Donor Tissue Bank) is one of the occupants of the Coronial Services Centre, located in<br />
Southbank, Melbourne, Australia. In excess of 4000 cases are admitted to the VIFM Mortuary facility every year. As part of<br />
the admission protocol, bodies will be photographed and submitted to a full body CT-scan. The circumstances of death provided<br />
by Police, the findings of the physical external examination and the admission images, in particular CT scan, will be used by<br />
VIFM forensic pathologists to inform the Coroner who will direct the further steps of the investigation of the cause of death.<br />
The specific information provided by the CT scan imaging quite often will influence in the need or not to proceed with a full<br />
autopsy. The privileged access to human tissue for transplantation led the VIFM to establish the Donor Tissue Bank of Victoria<br />
(DTBV) in 1989. The DTBV rely its multi banking operation (skin, muscle skeletal and cardiac tissue) on an “all in house”<br />
capacity including a team of Tissue Donor Coordinators. The DTBV benefits from the unique relationship with the Coroner’s<br />
Court of Victoria which allows for the early access to all referred cases to the Victorian Coroner as much as to the information<br />
collected by VIFM. The DTBV Tissue Coordinators will evaluate all cases within hours of admission for their potential as to tissue<br />
donation. Access to the CT scanned images has provided an increased capacity for the Tissue Donor Coordinators to establish<br />
in potential donors: Extent of chest trauma and cardiac tissue compromise, in particular after direct trauma (MVA) and post<br />
cardiac resuscitation. Extent of and precise anatomy of clinically identified bone trauma. Correlation between identified<br />
scars on physical examination and underlying soft and hard tissue involvement. Measurement of tissues to match customized<br />
graft requests (e.g. meniscus and tendons). Measurement of cardiac valves. Access to such information has critically<br />
contributed to the outcomes of tissue donation coordination. Firstly, it has avoided approaching donor families where<br />
preliminary findings in the CT scan preclude donation. Secondly, being able to evaluate the status of soft tissues in closed trauma<br />
has provided increased opportunities to request tissues in cases overlooked otherwise. Finally, there is the possibility to crossmatch<br />
specific tissue requests and progress with donation request. Provided cases, and images, illustrate the contribution of<br />
CT scan imaging in decision making within diverse scenarios and the further potential in tissue donation.<br />
CARDIOVASCULAR<br />
<strong>POSTERS</strong><br />
P-15<br />
IN VITRO SUSCEPTIBILITY OF HIGH VIRULENCE MICROORGANISMS ISOLATED IN<br />
HEART VALVE BANKING<br />
VILLALBA, R.; SOLIS, F.; FORNÉS, G.; JIMÉNEZ, A.; EISMAN, M.; GONZÁLEZ, A.; GÓMEZ VILLAGRÁN, J., Centro Regional<br />
de Transfusión Sanguínea. Spain.<br />
Abstract:<br />
Heart valves are not retrieved under conditions where sterility can be absolutely guaranteed. In this sense, measures such<br />
as antibiotic incubation are normally taken to eliminate microbial contamination. The aim of this study has been an evaluation<br />
of in vitro susceptibility of high virulence microorganisms, isolated in our Heart Valve Banking on basis to monitor its
<strong>POSTERS</strong><br />
susceptibility pattern as well as a basis to improve further antibiotics mixtures. The study included hearts retrieved from<br />
human donors who had been received in the Tissue Establishment of Córdoba (Spain) since 1996 to 2009. A total of four<br />
samples of each valve were obtained for microbiological analysis Sample 1. After dissection and previously to antibiotic<br />
immersion, a small tissue fragment (100 – 200 mg) of the subvalvular myocardium was obtained for microbiological analysis.<br />
Sample 2. After antibiotic disinfection 30 ml sample of cocktail antibiotic was drawn with a syringe and distributed for<br />
analysis . Sample 3. Of each heart valve that was approved as suitable for further processing, another small tissue fragment<br />
(100 – 200 mg) was obtained after incubation period. Sample 4. After putting the graft and the cryoprotective medium in<br />
a freezing bag, a remaining volume of 20 ml cryoprotective medium was used for microbiologic examination.<br />
Microbiological test were carried out under aseptic conditions. Samples were incubated in fluid thioglycollate medium,<br />
trypcase soy broth and sabouraud dextrose chloramphenicol agar. Incubation was establish for not less 14 days. Cultures<br />
were observed several times during incubation period. In vitro susceptibility to antibiotics were performed using WIDER<br />
automatic system (Soria Melguizo, Spain) under susceptibility and resistance criteria recommended by the MENSURA<br />
Spanish group. Between 1996 and 2009 a total of 849 valves were processed in the Tissue Establishment of Cordoba<br />
(Spain). A total of 92 positive cultures were obtained after microbiological analysis, 56.2% due to presence of low virulence<br />
microorganisms and the rest of 43.8% due to the presence of high virulence microorganisms. Aminoglicosides, such as<br />
Amikacin, Gentamicin and Tobramycin, in addition to Quinolone (Ciprofloxacin) and Imipene were more efficient drugs<br />
against gram negatives isolates. On the other hand Vancomycin remains as a effective antibiotic against Gram positives.<br />
Amoxicillin / Clavulanic and Teicoplanin could be alternatives to be used. On basis to these data, high virulence<br />
microorganisms isolates in Heart Valve Banking shown a pattern of susceptibility similar to those shown in other clinical<br />
circumstances and the most commonly used antibiotics regimes are useful to date. One antibiotic mixture with an<br />
aminoglicoside in addition to ciprofloxacin and vancomycin is efficient to be used. Further studies will be necessary to<br />
monitoring patterns changes in vitro susceptibility of microbiological isolates in tissue banking.<br />
P-16<br />
SECURING DONOR SAFETY AND OPTIMIZING RECIPIENT OUTCOMES IN LIVING<br />
DONOR LIVER TRANSPLANTATION: SYSTEMATIC SELECTION OF GRAFT TYPES AND<br />
USE OF CRYO-PRESERVED VEIN GRAFT<br />
TAMURA, S.; MOTOMURA, N.; SAITO, A.; ONO, M.; SUGAWARA, Y.; HASEGAWA, K.; KOKUDO, N., Tokyo University.<br />
Japan.<br />
Abstract / Introduction:<br />
Donor safety is the most important factor in living donor organ transplantation. In living donor liver transplantation,<br />
preservation of functional hepatic mass in the donor is crucial for securing donor safety. Handling of the middle hepatic vein<br />
[MHV], a large outflow route for both the right and left liver, has been the key issue in donor hepatectomy. In case in which<br />
left liver graft may not provide sufficient graft volume, a right liver graft becomes an option. However, resection of the right<br />
liver with the MHV may result in significant congestion of the remnant left lobe in the donor. In our adult to adult living<br />
donor liver program, we have continued to apply systematic selection of graft types and use of cryopreserved vein graft for<br />
outflow reconstruction of the middle hepatic vein [MHV]. In the current study, we evaluate the efficiency of our approach.<br />
Method:<br />
Outcomes of one hundred consecutive living donor liver transplantation cases were studied. Donor complication was assessed<br />
with Clavien system.<br />
Results:<br />
Among the 100 donors, right liver graft was procured in 67 cases (67%), and left liver graft in 33 cases (33%). Left liver<br />
graft with the caudate lobe was selected in 24 cases (73%).In the right liver graft group, 9(14%) were with the MHV, and
<strong>POSTERS</strong><br />
54(86%) were without the MHV. No grade IV or mortality was observed. Overall, complications were observed in 89%, 52%,<br />
and 57% of the donors who were retrieved right liver graft with the MHV, right liver graft without the MHV and Left liver<br />
graft with the caudate lobe, respectively. The severity of complications were, Grade I-II;78%, 39%, 44%, Grade IIIa;11%,<br />
13%, 9%, and Grade IIIb;0%, 0%, 4%, respectively. No complications were reported in 11%, 48%, and 48% of each group.<br />
Five year survival of the recipients with right liver graft with the MHV, right liver graft without the MHV and Left liver graft<br />
with the caudate lobe were, 78%, 88%、and 92%, respectively.<br />
Conclusion:<br />
Use of cryopreserved vein graft for venous outflow reconstruction allows avoidance of right liver graft with the MHV under<br />
algorithm based systematic selection of graft types. The approach may be effective in securing donor safety and optimizing<br />
recipient outcomes in living donor liver transplantation.<br />
P-17<br />
THE EFFECT OF FREEZING RATES ON MOISTURE CONTENT AND SURFACE TEXTURE<br />
OF FREEZE-DRIED BOVINE PERICARDIUM<br />
RUSIDAH, M.; ADAM, Z.; CHEE WEI, T.; SUHAILI, Y.; SUZINA, S., Tissue Bank, School of Medical Sciences, Health Campus,<br />
Universiti Sains Malaysia.<br />
Abstract:<br />
Bovine pericardium has been used as tissue graft application especially for soft tissue repair. The continuance of the<br />
characteristic of this tissue graft is important for a good tissue repair performance. Freeze-drying is a process used to remove<br />
water and make the tissue product relatively inactive and able to be stored at room temperature. The freeze-drying<br />
formulation is very important in controlling moisture content on tissue graft which will affect the storage duration and tissue<br />
structure after sterilization process. Freeze-drying comprises of three stages: freezing, primary drying and secondary drying.<br />
The freezing phase is considered as the most critical step in the freeze-drying process. The aim of this study was to investigate<br />
the effect of freezing rates on moisture content and surface texture of freeze-dried bovine pericardium. The bovine<br />
pericardium was trimmed into sheets after the removal of adherent fat. The pericardium was cleaned, disinfected in 0.05%<br />
sodium hypochlorite and stretched on perspex frame. The samples were subjected to either slow freezing (0.8ªC/min) or<br />
fast freezing (15ªC/min) until -50ªC. Primary drying was conducted at -5ªC and 100 mTorr and for secondary drying the<br />
temperature was increased to 25ªC at the same pressure for both groups. The residual moisture of the freeze-dried samples<br />
(6 cm x 4 cm) were analyzed using moisture analyzer at a constant temperature of 100ªC. The surface texture was observed<br />
by optical 3D measurement device. The average moisture content of the slow freezing samples was 3.27% compared to fast<br />
freezing samples which was 10.45%. The lower moisture content was caused by easy sublimation of larger ice crystal<br />
formed during slow freezing. Optical 3D observation revealed higher surface texture roughness on the slow freezing sample<br />
compared to fast freezing sample. These finding may be due to the lower moisture content of the samples. In addition<br />
rougher surface textures induce cells attachment. In conclusion this study showed that slow freezing rate led to low moisture<br />
content hence optimizing the storage conditions of freeze-dried bovine pericardium and suitable for tissue scaffold.
P-18<br />
ANATOMICALFRECUENT AANOMALIES IN CARDIAC VALVES. 15 YEARS OF<br />
EXPERIENCE ALVAREZ MV, ALVAREZ PV, BERTOLOTTI AM, FAVALORO RR.<br />
CARDIOVASCULAR TISSUE BANK, FAVALORO FOUNDATION, BUENOS AIRES<br />
ARGENTINA<br />
ÁLVAREZ VIVIANA, M.; ÁLVAREZ VERONICA, P.; FAVALORO RENE, R.; BERTOLOTTI MARIO, A., Favaloro´s Foudation.<br />
Buenos Aires. Argentina.<br />
Abstract / Introduction:<br />
During the heart valve cryopreservation process there are anatomical findings that determine which harvested tissue will be<br />
finally discarded. International literature provides little data on the prevalence of these anomalies.<br />
Objective:<br />
To analyze the number of heart valves discarded due to anatomical anomalies (AA) during the dissection stage at the<br />
Cardiovascular Tissue Bank of the Favaloro Foundation.<br />
Material and Methods:<br />
A retrospective analysis was made of all the hearts processed from September 1996 to April 2011. A classification was made<br />
of the causes that led to discard the valves according to the anomalies found. The anatomical findings were compared<br />
according to the type of valve: aortic (AoV) or pulmonary (PV). The AA were: biscuspid valve leaflets (Bi), >20% calcification<br />
(Cf), intimal peel (IP), hemorrhage with fibrosis (H) and fenestrations (Fe) with >1 cm diameters. The chi square method was<br />
used to compare qualitative variables.<br />
Results:<br />
During the period established for the study, 950 hearts were received and processed. From 1620 potential valves, 324<br />
(20%) were discarded for different causes: 89 (27.6%) for AA and 235 (72.4%) for other causes (+ serology, + control<br />
culture, technical errors). Of the valves discarded for AA, 61 (68.6%) were AoV and 28 (31.4%) were PV. The AA according<br />
to AoV or PV were: Cf 22 (37%) vs 3 (12.5%) P=.06; Fe 10 (17%) vs 16 (56.3%) P=.007; H 3 (3%) vs 2 (6%) P=.5; IP 5<br />
(8.6%) vs 3 (12.5%) P=.5; Bi 21 (34.3%) vs 4 (12.5%) P=.09.<br />
Conclusions:<br />
An accurate and thorough examination of the above mentioned AA during dissection proves that there is a higher percentage<br />
of AoV discarded for Bi or Cf, while in PV there is a prevalence of Fe. The examinacion of these anomalies they generate<br />
a better quality of it implants.<br />
P-19<br />
MECHANICAL PROPERTIES OF MITRAL ALLOGRAFTS ARE NOT REASONABLY<br />
INFLUENCED BY CRYOPRESERVATION<br />
<strong>POSTERS</strong><br />
SPATENKA, J.J.; HLUBOCKY, J.J.; NOVACEK, V.; MOKRACEK, A.; KOCHOVA, P.; HABRMANOVA, A., University Hospital<br />
Motol, Prague. Czech Republic.<br />
Abstract / Introduction:<br />
During last half century the heart valve surgery has been developed – recently, heart valves are repaired, if possible, or<br />
replaced. Both lines - mechanical valvular substitutes as well as biological ones underwent huge technological development.
<strong>POSTERS</strong><br />
Mechanical prostheses remain popular for their durability, while the biological (xenograft tissue or allograft tissue) valves<br />
are preferred for low thrombogenicity. Although first experiments with mitral allograft (MA) have been reported even earlier<br />
than those with aortic allografts, MA was never widely used in clinical practice. The durability of MA was very disappointing<br />
even when operated by very experienced surgeons. Many disadvantages of MA in mitral position disappear when it is<br />
implanted into tricuspid position, e.g. to low pressure system. Patients with tricuspid valve bacterial endocarditis, in particular,<br />
can theoretically benefit from MA transplantation, and that awakened our interest. We decided to evaluate methods of<br />
tricuspid valve replacement by MA in a sheep model. As a basic step we decided to use the same protocol, which our tissue<br />
bank uses for human aortic and pulmonary allografts. At experimental settings the simple tearing test performed by the<br />
surgeon proved quite reliable for determining mechanical tissue properties. Short term as well as long-term sheep<br />
experimental results proved to be promising. In aortic allografts no detectable differences were found between the mechanical<br />
behavior of the cryopreserved allograft aortic leaflets and fresh tissue . There is not much data concerning mechanical<br />
properties of MA.<br />
Material & Methods:<br />
A control group of 39 fresh sheep MA and a test group of 13 cryopreserved sheep MA were studied. Sterile surgical<br />
exposure for the MA harvesting was achieved via a right anterolateral thoracotomy under general, intravenous anesthesia.<br />
Afterwards the animals were put to death by intravenous administration of Thiopental (10mg/kg) and potassium chloride<br />
(20ml/kg). Their hearts were explanted, mitral valves were harvested with rims of left atrial and left ventricular muscle and<br />
with the entire subvalvular apparatus, including both papillary muscles. After harvesting, 39 fresh MA were stored in saline,<br />
at +5 to +7 oC, and were mechanically tested within 24 hours. Another 13 MAVs were processed according to the standard<br />
protocol of the Tissue Bank. They were deposited directly into the cultivation medium E 199 with the antibiotic cocktail -<br />
cefuroxime 0.2 mg/ml, (Zinacef, GlaxoWellcome) + piperacillin 0.2 mg/ml (Pipril, Lederle) + amikacin 0.1 mg/ml (Amikin,<br />
Bristol-Myers Squibb) + fluconazol 0.1 (Diflucan, Pfizer). After 24 hours storage at the temperature of 37.0 oC the valves<br />
were kept at + 5 to + 7 oC over the period of 3-5 days. Then they were transferred under sterile conditions into the<br />
cryoprotective solution (E 199 with 10% dimethylsulfoxide) in a laminar flow box, and sealed into plastic bags (Gambro<br />
Hemofreeze/Haemo bags NPBI BV DF 1200, the Netherlands) using two-layer technique. Finally, MA were programmed<br />
cooled (between temperatures of + 10 to - 60 oC at the rate of - 1 oC/min.) and than stored in liquid phase of liquid<br />
nitrogen (- 196 oC) in a separate container. Average storage time in tissue bank was 5.3month (from 3.5 to 12 month). Thirty<br />
minutes before the operation MA were removed from the container and thawed in a standard way (15 minutes in room<br />
temperature followed by 15 minutes in 37 oC water bath). In the experimental laboratory we used a similar shape of sample<br />
in all cases so we were measuring the mechanical properties of all parts of mitral valve at one time. Samples always<br />
contained the mitral annulus, part of the anterior leaflet, corresponding chordae tendineae and the postero-medial papillary<br />
muscle. For mechanical testing, the MA tissue was fixed to the traction machine jaws with mitral annulus and papillary<br />
muscle. Stepwise stress-relaxation measurements were made on all tissue samples. Zwick Roell Z050 (Zwick GmbH & Co,<br />
Germany, http://www.zwick.de) traction machine equipped with pneumatic grips and 200 N loading cell was used for the<br />
MA tissue testing. The specimens were fixed between the grips of the apparatus with a free length corresponding to the<br />
specimens' dimensions. The samples were stretched in steps of 1 millimeter every 5 minutes and the loading protocol<br />
consisted in six loading cycles. The time elapsed from the beginning of the step was chosen to reach approximately the steady<br />
state. The force-elongation curves were recorded. A five element generalized Maxwell model was used for the description<br />
of the relaxation behavior of the tissue. A simple Maxwell body includes a viscous element, η, and an elastic element, E,<br />
connected in series. Generalized Maxwell models consist of some simple Maxwell bodies coupled in parallel. In addition,<br />
an elastic element, EP, may be connected in parallel to them. In stress relaxation experiments, the applied stretch e(t) = ê is<br />
instantaneous and constant throughout the loading cycle. Constitutive equation of the generalized Maxwell model consisting<br />
in n Maxwell bodies with parallel elastic element EP may be written as: F(t) = ê EP + n∑i=1 êEi exp(-t/ τi), where the time<br />
constant τi = ηi/Ei represents the corresponding relaxation time for one step of stretching and ηi and Ei denote the viscous<br />
and elastic modulus of i-th Maxwell body, respectively. For five element Maxwell model with n = 2, the above equation is<br />
the final force-time relation with five material parameters EP, E1, E2, η1, and η2 to be identified.<br />
Results:<br />
The above equations describe a single step of stretching at constant values of the parameters EP, E1, E2, η1, and η2. Their<br />
values are different for each relaxation step. To identify these parameters, a direct exponential fitting to the experimental<br />
data was performed using statistical software R. The fitting process was the Gauss-Newton algorithm based on the nonlinear
<strong>POSTERS</strong><br />
least-squares method. A set of material parameters was determined for each stretching step and for each specimen of the<br />
control group of fresh tissue and the test group of thawed cryopreserved MA, respectively. Coefficient of correlation R of<br />
the fitting procedure was better than 0.99 for all specimens and every loading cycle. From the total number of 52 heart<br />
valves, the control group of fresh tissue contained 39 specimens, while the test group of cryopreserved MA was represented<br />
by 13 specimens. The fitted values of material parameters were examined cycle by cycle. For each loading cycle, the outliers<br />
were carefully detected using standard techniques of exploratory data analysis and they were removed from the statistical<br />
files. For each viscoelastic parameter and each loading cycle, null hypotheses were formulated stating that the means and<br />
the variances of the two groups under consideration do not differ significantly. To test the hypotheses, t-test and F-test were<br />
carried out in R statistical package. The assumed normality of the populations was tested and confirmed before the execution<br />
of the above mentioned tests. Both statistical tests were carried out at 95% confidence level. The mean values of identified<br />
elastic moduli EP, E1, and E2 do not manifest any pronounced dominancy of one group of specimens over the other. Order<br />
of magnitude of all three elastic moduli ranges from 102 to 103 N/m. The identified values of the parallel elastic modulus<br />
EP range from 0.44 ± 0.24 kN/m in the first loading cycle to 4.65 ± 1.70 kN/m in the last loading cycle for the control<br />
group and from 0.58 ± 0.19 kN/m to 3.91 ± 1.79 kN/m for the test group. In the first loading cycle, the modulus of the<br />
test group is slightly higher than that of the control group while its standard deviation remains lower. Control group modulus<br />
is then slightly higher in the next loading cycles. Serial elastic modulus E1 is lower than parallel elastic modulus EP but of<br />
the same order. For the control group, the values range from 0.29 ± 0.13 kN/m to 1.40 ± 0.43 kN/m, and for the test group<br />
from 0.36 ± 0.12 kN/m to 1.30 ± 0.56 kN/m. The values of the test group are slightly higher than the control group values<br />
up to the fourth loading cycle, almost equal in the fifth loading cycle, and finally little bit lower in the last loading cycle. The<br />
identified values of serial elastic modulus E2 are very similar to those of serial elastic coefficient E1. The identified data<br />
encompass the interval from 0.25 ± 0.12 kN/m in the first loading cycle to 1.28 ± 0.48 kN/m in the last loading cycle in<br />
the case of the control group and from 0.23 ± 0.12 kN/m to 1.18 ± 0.64 kN/m in the case of the test group. The mean<br />
values of the test group are higher than means of the control group in the second and third loading cycle. This difference,<br />
however, is less than 0.05 kN/m up to the fourth loading cycle. No significant difference (p>0.05) was found for elastic<br />
parameters between the two groups of specimens, neither between the mean values, nor between the variances. Test group<br />
serial viscous modulus η1 has higher values than that of the control group. Order of magnitude of viscous modulus η1 lies<br />
between 104 and 105 Ns/m. It is the only viscoelastic parameter determined with such strongly pronounced dominance of<br />
one specimen group. The values range from 33.75 ± 13.36 kNs/m to 159.99 ± 48.48 kNs/m for the control group and<br />
from 44.08 ± 15.19 kNs/m to 163.30 ± 90.85 kNs/m for the test group. No significant difference (p>0.05) was found<br />
between the mean values of the two groups of specimens. Significant difference in variances was observed in the third, fifth,<br />
and sixth loading cycle with p equal to 0.027, 0.042, and 0.004, respectively. Test group standard deviation is more than<br />
1.65 higher than control group standard deviation in these three critical loading cycles. As the equality of variances is one<br />
of the assumptions of the Student’s t-test, the relevance of its results in the third, fifth, and sixth loading cycle may be<br />
compromised. Nevertheless, the difference of the mean values expressed as percentage of the higher mean gives 23%,<br />
25%, 27%, 14%, 19%, and 2%. It is obvious that the differences 27% in the third cycle and 19% in the fifth cycle fall into<br />
reasonable limits. Note only 2% difference in means in the sixth loading cycle. In this respect, the difference in means<br />
between the two groups remains negligible. Viscous modulus η2 of the test group has lower values than that of the control<br />
group and it has values of order of magnitude 103 Ns/m. Identified values of the viscous modulus η2 encompass the<br />
interval from 1.63 ± 0.79 kNs/m to 7.57 ± 2.49 kNs/m in the control group and from 1.74 ± 1.13 kNs/m to 5.84 ± 3.56<br />
kNs/m in the test group. As well as in the previous cases, no significant difference (p>0.05) was found between the mean<br />
values of the two groups of specimens. Significant difference (p=0.031) in variances was observed in the fourth loading cycle.<br />
However, the difference between mean values in this loading cycle is practically negligible.<br />
Discussion:<br />
The lifelong postoperative stress on a heart valves tissue prosthesis is known to be enormous. That is why the mechanical<br />
testing of the commercial heart valve prostheses of all types became routine during the last 50 years. Special simulators were<br />
developed for mechanical as well as for biological prostheses, and precise, scientific methods of tissue mechanics and tensile<br />
strength measurements were introduced. As our experimental MA transplants into the tricuspid position on the sheep model<br />
were intended as a first step of clinical project we decided to examine allograft tissue quality objectively. In other words we<br />
wanted to know if our aortic & pulmonary allograft processing and cryopreservation method was feasible for MA as well,<br />
or if we have to develop a special MA processing protocol. For aortic and pulmonary allografts some data on mechanical<br />
properties are available. The ultrastructure and mechanics of fresh, cryopreserved, and cellular extracted porcine aortic valve
<strong>POSTERS</strong><br />
leaflets were tested. Reduction in the fracture tension and increased tissue extensibility were observed. Cellular extraction<br />
preserves matrix structure and mechanics over the physiological loading range. On the contrary, the combination of<br />
extraction and fixation may lead, according to these authors, to early degenerative failure. When the mechanics of fresh,<br />
refrigerated, and frozen porcine arterial tissue was measured and statistically compared the effects and impact of common<br />
storage protocols on tissue mechanics were revealed. Subfailure stress, ultimate stress, and Young's modulus decreased<br />
significantly in refrigerated specimens while physiological, subfailure, and ultimate failure mechanics between fresh and<br />
frozen specimens were not significantly different (Stemper et al. 2007). It was reported that cryopreservation of decellularized<br />
arteries does not affect the structure and mechanical properties of the rabbit carotid artery (Fonck et al. 2008). Exhaustive<br />
overview of the biomechanics of heart valves and their function including solid and fluid mechanics and fluid-structure<br />
interaction studies have been published (Sacks et al. 2009). This paper shows that most of the research concerned with heart<br />
valve mechanics has been conducted on aortic valves. Indeed, only a few studies deal with mitral valves and they are usually<br />
limited to leaflets with no interest to related heart structures. Surface strains were determined for anterior mitral valve leaflet<br />
(Sacks et al. 2002). Biaxial mechanical testing of porcine mitral valve leaflets revealed significant difference in material<br />
properties between the anterior and posterior leaflets (May-Newman and Yin 1995). In order to quantify the influence of<br />
the cryogenic treatment on tissue mechanics, a testing protocol was defined and viscoelastic parameters were determined<br />
for a control group of fresh tissue specimen and a test group of cryogenically processed samples. A five-element Maxwell<br />
viscoelastic model was applied to characterize the tissue’s mechanical behavior. The tissue mechanics were studied in terms<br />
of forces and elongations rather than in terms of stresses and strains since the specimens' geometrical form was very<br />
complicated and irregular that would cause the difficulties in assessing of geometrical characteristics like cross-section area.<br />
The actual configuration of the experimental device did not allow us to immerse the specimen during the test into any liquid<br />
media, e.g. saline. Thus, the testing conditions are somewhat removed from the situation in the animal’s body. In order to<br />
approach real conditions and to prevent dehydration of the specimen, the specimens were moistened during the test, the<br />
total testing time was reduced to 30 minutes and there was no preconditioning phase in the testing protocol. Preconditioning<br />
is recommended for stabilizing the internal structure of the tissue that should decrease variability and lead to interpretable<br />
results (Fung 1993). Freezing is considered as one type of preconditioning since it also may change the mechanical<br />
properties of soft tissue (Gao et al. 2010). According to the authors, results from the thawed liver tissue were generally<br />
consistent, interpretable and no other preconditioning was applied. The preconditioned state of the porcine aortic valve<br />
material is a function of the deformation history that has occurred before the preconditioning cycles and preconditioning<br />
without an adequate rest period between tests increases predictive errors (Carew et al. 2000). However, may introduce a<br />
loss of stiffness (Liao et al. 2009) or plastic deformation of tissue specimen. Anomalous decrease of stiffness of skin and<br />
myocardium tissue was attributed to the small number of cyclic loads and it was concluded that this behavior is a true<br />
phenomenon unique to load controlled deformations that results from the interplay of nonlinear effects and creep behavior<br />
(Giles et al. 2007). Test specimens are usually preconditioned by applying a cyclic load to reduce the viscoelastic effects<br />
(Ghaemi et al. 2009). However, the viscoelastic effects are of the utmost interest in our study as they introduce time-dependent<br />
phenomena like creep and/or relaxation. The aim of this study was to evaluate the overall viscoelastic behavior and to<br />
compare the influence of a specific treatment on the viscoelasticity of the tissue that is represented by a set of viscoelastic<br />
coefficients. We decided to omit the preconditioning phase as this allowed us to examine the inherent material properties<br />
of the tissue under consideration that are not modified by previously introduced deformations and/or stresses. There are only<br />
a few studies focused on the influence of freezing on tissue mechanical behavior. The effect of freezing on the mechanical<br />
properties of spleen tissue has been reported to be negligible (Davies et al. 2000). No significant gross histological damage<br />
was observed in frozen liver tissue samples that were loaded within the elastic regime. Histological changes due to<br />
mechanical stresses were associated with permanent plastic deformations related to structural irregularities such as the blood<br />
vessels and bile ducts (Rabin et al. 1997). Similarly, the results of the present study do not reveal any significant difference<br />
in viscoelastic properties between fresh and frozen heart tissue. In the present study, no significant difference between the<br />
control and testing group was found in mean values of determined parameters suggesting that the tissue processing and<br />
cryopreservation do not alter the tissue’s structural components that play crucial role in its viscoelasticity. Tissue elasticity<br />
compensating mostly for the instantaneously applied load may be attributed to collagenous tissue, more precisely to the<br />
anterior leaflet and especially to the corresponding chordae tendineae. The viscous response corresponding to the relaxation<br />
of the specimen would be dominant mostly in muscular tissue. Significant differences observed in the variances of viscous<br />
parameters in certain loading cycles reflect a wider range of values of the test group compared to the control group. This<br />
difference cannot be attributed to the lack of a precondition phase since it would affect the control group as well. The<br />
difference may be due to variability in the geometry of the papillary muscle that contributes to the viscous response of the
<strong>POSTERS</strong><br />
specimen and, so, to variability in its cross-section bearing the mechanical load. A similar size and geometrical shape of<br />
papillary muscle of harvested MA samples cannot be guaranteed since it depends on many factors including surgeon’s<br />
judgment and experience. Moreover, attaching the muscle between the grips of the testing device may considerably change<br />
the free size of the muscle tissue that is not squeezed between the jaws and that participates on the viscous response to the<br />
applied load. Together with relatively low number of specimens in testing group and different number of specimens in each<br />
group, this factor may play a role in statistical analysis. Nonetheless, the results of the present study are very encouraging<br />
as they show that the tissue processing and cryopreservation do not alter significantly the overall viscoelastic behavior and<br />
mechanical performance of the tissue.<br />
Conclusions:<br />
Our study shows that current allograft heart valve tissue processing and cryopreservation protocol could be applied on MA<br />
tissue as well. The mechanical properties of cryopreserved MA tissue do not differ significantly from the quality of the native<br />
mitral valve tissue in sheep model. On the basis of this experimental mechanical testing the standard allograft heart valve<br />
bank protocol will be used even for MA processing for clinical purposes. Acknowledgments: We would like to thank the<br />
laboratories of the University of West Bohemia, PILSEN, Czech Republic, namely Department of Mechanics and New<br />
Technologies Research Centre for enormous help during our experiments.<br />
P-20<br />
FACTORS AFFECTING THE EFFECTIVENESS OF A CARDIOVASCULAR RECOVERY<br />
TEAM<br />
VITO, S.; OLIVA, R.; LUQUE, S.; HINOJOSA, M.; MIRANDA, M.; CALAMARDO, M.; PÉREZ, M., Transplant Services Foundation<br />
(TSF). Hospital Clínic. Barcelona. Spain.<br />
Abstract / Introduction:<br />
Advances in cardiac surgery, such as the use of cardiovascular homograft in paediatric surgery to repair congenital diseases,<br />
and the increase of life expectancy have caused an increase in tissue demand. At the same time, the decrease in the number<br />
of donors due to a lesser degree of traffic and industrial accidents, and lower hospital economical resources is forcing us<br />
to optimize the cardiovascular tissue recovery. Aim: The aim of this study is to assess the recovery team effectiveness<br />
analyzing tissue characteristics and its viability, and if there is any relation with medical history, cause of death and the age<br />
of the donor. Methods: The study has been carried out with 112 heart donors, recovered and processed during 2010. 73<br />
% of the above mentioned hearts were recovered by our own retrieval team (Team A), who has been specially trained in<br />
multi tissue recovery, and the 27% left have been retrieved by organ transplantation teams (Team B). The following factors<br />
have been analyzed; tissue viability, presence or absence of aortic arch and pulmonary bifurcation and its relation with the<br />
recovery team. It has been also studied (i) types of homografts obtained (ii) the total homografts retrieved per donor, (iii)<br />
and the total homografts obtained by the different recovery teams. Other factors considered were; age, medical history<br />
(cardiovascular and neurological diseases, obesity, diabetes, and surgical history), cause of death of the donor and type of<br />
donor (non heart beating donors, exitus, or brain death donors).<br />
Results:<br />
It has been found out that there are no differences between recovery teams in relation to the total amount of recovered<br />
tissue per donor (mean: 1.6 allograft per each recovered heart) However, there is a difference between recovery teams in<br />
relation to the type of recovered allograft; the total amount of aortic archs and pulmonary bifurcations retrieved by Team A<br />
is higher than the one recovered by Team B; by a 19.8% of aortic archs and by a 27.2% in. pulmonary bifurcations.
<strong>POSTERS</strong><br />
Conclusions:<br />
The study shows the importance of having a trained recovery team in multi tissue recovery. This fact highlights this is the way<br />
to get a better quality tissue that can be used for different clinical applications. A longer pulmonary artery that includes a<br />
pulmonary bifurcation, allows us during processing, to split it in a pulmonary valve and a pulmonary artery. Nowadays this<br />
is one of the best options to increase the number of tissues availability for paediatric congenital diseases surgery.<br />
P-21<br />
MINIMISING INFECTION & MAXIMISING POTENTIAL DURING CARDIECTOMY OF<br />
OXFORD DONORS<br />
DAVIES, J., AUCKBURALLY, F. CHARLESWORTH, K., THOMAS, Y., RATNTUNGA, C. Oxford Heart Valve Bank. UK<br />
Abstract:<br />
Oxford Heart Valve bank has routinely cultured the fluid surrounding the retrieved Donor Hearts and compared this to<br />
samples of untreated tissue segments since 2004. The heart samples are consistently
The Oxford incoming heart tissue and fluid studies have shown that most infections are derived from the bowel of the donor’s<br />
body and not external contaminants. Any movement would thus accelerate the proliferation of bacteria. The Heart Valve Bank<br />
team will always ensure that the movement of the body is minimised until after cardiectomy (e.g. heart valves will always<br />
be first tissue to be retrieved). Organ teams may have to move the body and the heart whilst dissecting and removing the<br />
other organs. No significant difference has yet been observed. Donor’s who have suffered a significant road traffic collision<br />
have, however, been shown to have a significant incoming positive results.<br />
The evidence regarding movement of a donor’s body increasing risk of infection would therefore not appear to support the<br />
movement of bodies to a central clean facility.<br />
The Heart Valve Bank team have made a record of the incoming requests and compared this to outgoing distributed tissue,<br />
the tissue in stock and the waiting list. This data analysis has been used to update cardiectomy practice. Oxford have thus<br />
recently decided to increase the minimum age limit for baby donors to reflect the current UK demand. There is a surplus of<br />
mid size aortic valves in supply and so Oxford is choosing not to keep all aortic valves if, at cardiectomy, they appear to<br />
be substandard and mid size. Instead of processing them in the tissue bank where they are unlikely to be used, the decision<br />
has been made to leave them in the donor’s body.<br />
Organ teams do not usually retrieve organs from baby donors less than two years of age. Oxford Heart Valve Bank however,<br />
is referred baby donors from various areas around the UK. Many babies are transferred prior to death to a hospice.<br />
Cardiectomy is usually performed in a cold room in the hospice. One of the main risks with baby donors is getting a good<br />
quality and sufficient blood sample for testing. A maternal sample has always to be taken and a repeat at one hundred and<br />
eighty days. As a result, these baby donor valves (which are very much in demand) are not discarded due to lack of a blood<br />
sample.<br />
In conclusion, there are many ways to try to minimise infection which have been investigated in Oxford, some of which are<br />
already having an effect. The Heart Valve Bank team and the Oxford Organ teams are working very closely together to set<br />
up a standard procedure for cardiectomy for UK Organ teams which will increase the number of bifurcated pulmonary valves<br />
in future.<br />
P-22<br />
RECIPIENT OUTCOME STUDIES IN OXFORD<br />
<strong>POSTERS</strong><br />
DAVIES, J.; CHARLESWORTH, K.; FOSTER, R.; THOMAS, Y.; AUCKBURALLY, F.; RATNTUNGA, C., Oxford Heart Valve<br />
Bank. UK.<br />
Abstract:<br />
Routine assessment of clinical efficacy by evaluation of surrogate markers is common but the risk is that the markers chosen<br />
may be misleading. Variations exist in Cardiovascular Tissue processing methods which may be due to designs to maximise<br />
viability of the tissue. Quality assessment however, is most pertinent if it evaluates a property of the tissue which is a proven<br />
key performance indicator. Viable cells may not be required for long term tissue function. Recipient outcome studies are direct<br />
evaluation methods and measure patient mortality or morbidity which may be more relevant markers. However, these studies<br />
are time consuming and costly. There are no Tissue Banking Association guidance for recipient follow up studies however<br />
many lessons have been learnt with respect to follow up studies of recipients of bioprosthetic & prosthetic valves. Currently<br />
in the UK, there is a clinical database – Central Cardiac Audit Database (CCAD) to which surgeons submit data following<br />
each operation. Reports are issued every two – three years and remain anonomysed. There is also an administrative dataset<br />
or coding system – Hospital Episode Statistics (HES). However, volunteered data sometimes over estimates survival by 20%<br />
thus it is has become important in UK to validate data by link to National statistics (e.g. to NHS number). There are lessons<br />
to be learnt from UK studies when analysing cardiovascular tissue data. To minimise the effect of varying case mix when<br />
comparing recipient outcomes in different centres, benchmark procedures should be used. One year survival is a better<br />
performance indicator than peri-operative mortality. Freedom from re-intervention is a less crude indicator than patient<br />
survival. This paper summarises different follow up studies performed in Oxford. 1704 individual tissues despatched by<br />
Oxford were followed up with written surgeon surveys. 1180 surveys were returned. Surgeons were verbally interviewed
about remaining tissues. 0% post operative infections were reported. A more detailed retrospective study of mortality and<br />
morbidity of 77 patients in one surgical centre have been followed up for 11 years. Oxford distributed 214 tissues in 2010.<br />
Detailed prospective outcome study forms have been sent out to each surgeon to complete relevant data such as NHYA<br />
grades and echocardiography results. Oxford has started a valve registry for comparison of all types of valves at yearly<br />
intervals and also a valve explant study both of which include cardiovascular tissue valves/patches. These follow up outcome<br />
studies may collectively be a more accurate marker of the Oxford tissue quality.<br />
REPRODUCTIVE<br />
P-23<br />
OVARIAN TISSUE BANKING - THE UPPER AUSTRIAN NETWORK FERTISAVE<br />
<strong>POSTERS</strong><br />
HENNERBICHLER, S., Red Cross Blood Transfusion Service of Upper Austria, Linz/Austrian Cluster for Tissue Regeneration.<br />
Austria<br />
Abstract:<br />
Kinderwunschklinik Wels, Austria (Loimer L., Swoboda M.) General Hospital Linz, Austria (Oppelt P., Krause S.)<br />
Krankenhaus Barmherzige Schwestern Linz, Austria (Stummvoll W., Costamoling W.) Klinikum Wels-Grieskirchen, Austria<br />
(Reisenberger K., Teiche P.) Red Cross Blood Transfusion Service of Upper Austria, Linz, Austria (Gabriel C., Peterbauer-<br />
Scherb A. , Hennerbichler S.) Radiation or chemotherapy of oncological diseases often causes a follicular decrease as<br />
adverse reaction, which may lead to infertility and premature menopause. Therefore cryopreservation of ovarian tissue<br />
would represent a possibility for fertility protection therapy. Fertisave Upper Austria (www.fertisave.at) is a medical network<br />
with an interdisciplinary team of several Upper Austrian institutions. The aim of this network is to maintain fertility in<br />
malignant diseases. It is certified and registered within superior networks (e.g. FertiProtect) and offers ovarian tissue banking<br />
as a possible fertility protection therapy in Upper Austria. In the future also cryopreservation of testicular tissue is intended.<br />
P-24<br />
FREEZING OF SEMEN PEARL SAMPLES AND SPERMATOZOIDS FROM TESTICLE<br />
BIOPSIES<br />
GARCÍA GARCÍA, M. 1 , RENDAL VÁZQUEZ, M.2, LÓPEZ PIÑÓN, M.1, CARBALLAL RODRÍGUEZ, M. 1 , RODRÍGUEZ<br />
CABARCOS, M. 2 , SOLA RODRÍGUEZ, A. 1 , ANDIÓN NÚÑEZ, C. 2 .<br />
1 - Unit of In Vitro Fertilization Complejo Hospitalario Universitario ACoruña, 2 - Unit Of Criobiology Complejo<br />
Hospitalario Universitario ACoruña. Spain.<br />
Abstract / Introduction:<br />
When the production of spermatozoids is limited or they can only be recovered from the testicles by biopsy, in these cases<br />
it is necessary to optimize the cryopreservation technique of the sample. The semen sample is frozen in small pearls to allow<br />
for several attempts of in vitro fertilization from one unique sample.
Objetive:<br />
Semen pearl samples are compared using the techniques for freezing of dry ice and liquid nitrogen.<br />
<strong>POSTERS</strong><br />
Material and Methods:<br />
Samples were used that were obtained from autologous semen donors (N=15) after informed consent. A recount and<br />
viability of the sperm is carried out using the Sperm Class Analyzer (SCA). After dilution of the sample with the cryoprotectant<br />
(Sperm-Cryo tm, Cryos) the sample was divided into two groups for study: Freezing was carried out on pearls directly in<br />
dry ice (group 1) or on pearls in liquid nitrogen (group 2). In the case of the pearls frozen in dry ice, small holes are<br />
hollowed out in the dry ice and by using a pipette a drop is allowed to fall from the semen suspension (15 microlites) on<br />
these same holes. After a few minutes two or three pearls are stored per cryotube. In the case of the pearls frozen in liquid<br />
nitrogen a drop is allowed to fall directly from the suspension into the liquid nitrogen. After a few minutes two or three<br />
pearls are stored per cryotube. After a month of storage in a nitrogen tank at a temperature below -150ºC, the samples were<br />
thawed by immersion of the same at 37ºC. After the elimination of cryoprotectant by dilution and centrifugation, count and<br />
viability of the sperm samples were carried out in SCA.<br />
Results:<br />
After thawing, a similar recount of spermatozoids is observed in both samples. In the samples frozen in pearls carried out<br />
with dry ice a greater number of moving spermatozoids are observed than in the pearl samples carried out using liquid<br />
nitrogen.<br />
Conclusions:<br />
The most effective protocol must be chosen for the cryopreservation of small semen samples that allow for the use of multiple<br />
cycles of intracytoplasmatic microinjection (ICSI).<br />
P-25<br />
OPTIMUM CRYOPRESERVATION OF SEMEN AND SPERMATOZOIDES FROM TESTICLE<br />
BIOPSIES<br />
GARCÍA GARCÍA, M.; RENDAL VÁZQUEZ, M.; CARBALLAL RODRÍGUEZ, M.; LÓPEZ PIÑÓN, M.; RODRÍGUEZ CABAR-<br />
COS, M.; SOLA RODRÍGUEZ, A., Complejo Hospitalario Universitario A Coruña. Spain.<br />
Abstract / Introduction:<br />
Sperm cryopreservation is an ancient method of preserving male fertility in a way that will maintain sperm viability for a<br />
long period of time. Sperm quality and quantity are the two main factors that can have an effect on the success of sperm<br />
cryopreservation.<br />
Objetive:<br />
Comparison between the classic technique of a slow-freezing method and preservation in liquid nitrogen vapour for the<br />
cryopreservation of normal sperm samples.<br />
Material and Methods:<br />
Samples from autologous donors of sperm (N=5) after informed consent were used. Count and viability of the sample were<br />
analyzed in the Sperm Class Analyzer. After dilution of the sample with the cryoprotectant (Sperm-Cryo tm, Cryos) the<br />
sample was divided into three groups for study. The samples were stored in cryotubes (samples of 0.25-050ml): Group 1:<br />
Programmed freezing was carried out using a freezing ramp (CM2000) with index of freezing of 3.1ºC/min until -34ºC<br />
and 15ºC/min until -120ºC or with index of freezing of 0.79ºC/min until -30ºC and 7.3ºC until -120ºC (group 2). Group
<strong>POSTERS</strong><br />
3: Freezing in vapour phase of liquid nitrogen placing the cryotubes directly on a floating platform in a polystyrene box<br />
with liquid nitrogen, avoiding the immersion. It is moved gently and after 30 minutes the cryotubes are submerged in liquid<br />
nitrogen. After a month of storage in a nitrogen tank at a temperature below -150ºC, the samples were thawed by immersion<br />
of the same at 37ºC. After the elimination of cryoprotectant by dilution and centrifugation, count and viability of the sperm<br />
samples were carried out in SCA.<br />
Results:<br />
After thawing, a similar recount of spermatozoids was observed in both samples although in the samples frozen in a<br />
controlled program a greater number of moving spermatozoids were observed than in samples frozen in vapor phase.<br />
Differences were observed as to mobility between the two curves used to carry out the programmed freezing. There being<br />
a greater mobility observed in the samples of programmed freezing of group 1.<br />
Conclusions:<br />
It is necessary to optimize the protocol of cryopreservation to diminish the damage caused to sperm on freezing.<br />
P-26<br />
SYSTEM FOR THE STORAGE AT ULTRALOW TEMPERATURES OF BIOLOGIC MATERIAL<br />
FOR IVF USE<br />
ANDION NÚÑEZ, C. 1 , RENDAL VÁZQUEZ, M. 1 , RODRÍGUEZ CABARCOS, M. 1 , FERNÁNDEZ LAGO, C. 2<br />
1 - Tissue Bank Complejo Hospitalario Universitario ACoruña, 2 - Hematology Service Complejo Hospitalario Universitario<br />
ACoruña. Spain.<br />
Abstract:<br />
The need to have at our disposal safe storage systems that allow for the optimization of space in mechanical freezers and<br />
liquid nitrogen tanks has made it necessary for the tissue bank of the CHUAC to develop its own classification and storage<br />
system which is characterized by its low economic cost, safety and versatility. The storage system is based on polycarbonate<br />
cassettes composed of cells which allow for the introduction of different sized containers used frequently in IVF, labs and tissue<br />
bank. Polycarbonate is a material which tolerates very well ultra-low temperatures and temperature changes and whose<br />
transparency also allows for a simple visualization of the content of each cell. At the beginning designed to be used with<br />
the usual straws containing embryos or sperm, which are normally used in IVF, they can also be used for other purposes<br />
where it might be necessary to store small samples of biological material. In the case of the straws the system can be easily<br />
adapted to different lengths of straw by simply displacing the base of the cell. The system allows for the perfect handling<br />
and identification of diverse materials which are susceptible of being classified by colors and the system is protected by<br />
copyright.
P-27<br />
CASE REPORT: THE USE OF PLATELET-RICH PLASMA IN ORTHOTOPIC<br />
CRYOPRESERVED OVARIAN TISSUE TRANSPLANTATION<br />
<strong>POSTERS</strong><br />
MURCIA, N.; TAUSTE, C.; RODRÍGUEZ, L.; GONZÁLEZ, S.; ALMEIDA, L.; SALVADOR, C.; CALLEJO, J., Hospital Sant<br />
Joan de Déu. Spain.<br />
Abstract / Introduction:<br />
Transplantation of viable cryopreserved ovarian tissue is a promising clinical option, mainly for young women, to restore<br />
fertility after gonadal dysfunction resulting from cancer therapy. We report here the case of a female patient with bilateral<br />
ooforectomy undergoing autotransplantation of viable cryopreserved ovarian tissue pre-treated with platelet-rich plasma<br />
(PRP) suspension to improve the ovarian function restoration by diminishing ischemic length period after implantation.<br />
Material and Methods:<br />
Female patient with previous unilateral ooforectomy underwent contralateral ooforectomy in 2001 due to dermoid cyst,<br />
when she was 20 year old. At the same surgical time, as iatrogenic menopause was going to be induced, little remanent<br />
ovarian tissue cryopreservation was performed in order to preserve her fertility. To cryopreserve the ovarian tissue, ovarian<br />
cortex was isolated from the medular and cut it up into small pieces of 2 mm thickness. Ovarian tissue was then transported<br />
in Flushing medium at 4ºC (specific medium for the conservation of oocytes that contains heparin, human albumin solution,<br />
recombinant human insulin and gentamicin sulphate) and cryopreserved with a controlled slow cooling method in the Tissues<br />
Bank. The woman has been treated with hormonal substitute treatment until the age of 30, when the patient undergoes<br />
laparoscopic orthotopic transplantation of the cryopreserved ovarian tissue into a bilateral peritoneal pocket in the pelvic<br />
peritoneum of the ovarian pit. Transplantation of ovarian grafts without vascular pedicle requires the establishment of a<br />
new blood supply that takes, at least, 5 days, and leads to a substantial loss of follicles. We use platelet-rich plasma (PRP)<br />
to improve the mechanisms leading to graft re-perfusion with the aim of reducing the avascular period. PRP is becoming a<br />
new application in tissue engineering and a developing area for clinicians and researchers because it is a natural source<br />
of growth factors, many of which can accelerate and promote angiogenesis.<br />
Results:<br />
For now this patient is being monitored with monthly estradiol and FSH determinations to evaluate how her ovarian function<br />
is being restored.<br />
Discussion:<br />
As a number of successful human pregnancies have been possible with viable cryopreserved ovarian tissue, the protocol<br />
described here gives a real hope to patients who need an urgent treatment when ovarian function failure occurs. Although<br />
heterothopic transplantation is less complicated to perform and allows an easier monitoring and access to the tissue when<br />
assisted reproduction techniques are underwent, orthotopic technique gives the possibility of spontaneous pregnancies and<br />
when it has been performed by experienced surgeons, intervention time is low and patient recovery is extremely satisfactory.<br />
The use of PRP is still uncertain but many good results have been obtained when used in other tissues.
MUSCULOESKELETAL<br />
<strong>POSTERS</strong><br />
P-28<br />
APOPTOSIS MEDIATED CELL LOSS AFTER HUMAN MENISCI CRYOPRESERVATION<br />
VILLALBA, R. PEÑA, J. NAVARRO P., LUQUE, E., JIMENA, I. ROMERO, A. GÓMEZ VILLAGRÁN, J. Centro Regional de<br />
Transfusión Sanguínea. Córdoba. Spain.<br />
Introduction:<br />
Removal of the meniscus leads to progressive degenerative arthritis of the knee on a long term basis, therefore meniscal<br />
allograft transplantation has been proposed as an alternative to menisectomy. Preservation methods are required to build<br />
up operational stocks and to provide living grafts of a practical size at the right time for patients.<br />
Methods for meniscus preservation have been published and relevant literature confirm that using standard cryopreservation,<br />
the chondrocyte survival in situ is inadequate and extremely variable and the cryoinjury mechanisms are not completely<br />
established. The aim of the present study is to further investigate possible cellular injury caused by cryopreservation by<br />
analyzing apoptosis and ultrastructural damage to menisci.<br />
Materials and Methods:<br />
Materials used were seven human menisci which were cryopreserved by standard method. All tissue samples were processed<br />
simultaneously for routine light microscopy, scanning and transmission electron microscopy as well apoptosis assessment by<br />
the use of ISOL method.<br />
Results:<br />
We observed significant differences (p
<strong>POSTERS</strong><br />
P-29<br />
DEVELOPMENT AND EVALUATION OF THE NEW METHOD OF GLUCOCORTICOID<br />
OSTEOPOROSIS TREATMENT<br />
VOLOVA L.T., SRADLER E.P., PISAREVA E.V., VLASOV M.U., NEFEDOVA I.F., Samara State Medical University, Samara<br />
Russia. Russian Federation.<br />
Abstract:<br />
The reduction of osseous tissue mineral density while administrating corticosteroids is a serious clinical problem requiring<br />
timely diagnostics and treatment. Severe secondary and, what is more serious, glucocorticoid osteoporosis requires<br />
optimization of treatment by correcting steroid therapy. The Experimental Medicine and Biology Institute of Samara State<br />
Medical University was the first in the world to develop and suggest the technology of obtaining “allogenic hydroxyapatite”<br />
as a waste-free, ecologically safe production of implants from a biological tissue. This material is related to bionanomaterials.<br />
60-70 % of its composition consists of the particles the size of which is less than 100 nm. It contains the whole complex of<br />
osseous tissue microelements and an organic component in addition to Ca and P. At the pre-clinical stage in vivo<br />
investigations have been carried out on white lab rats (1000 animals weighing 220 gm). Morphological, biochemical,<br />
chemical, and physical scientific methods including transmission and scanning electronic microscopy have been used to<br />
evaluate the material composition and osseous tissue metabolism. The testing of the preparation has been done for<br />
hyperglucocorticoid model. The animals were injected hydrocortizone in the dose of 40 mg/kg. In 28 days they developed<br />
the processes of osseous tissue resorption, reduction of collagen biosynthesis intensity, a marked level decrease of one of<br />
the osseous remodeling markers – protein-bound oxyproline, serum free oxyproline content increase. Histologically the<br />
preparations demonstrated resorption of Haversian (central) canals osseous cell walls, thinning and destroying of spongiosa<br />
osseous beams and the appearance of numerous osteoclasts in resorption gaps. Intramuscular injection of allogenic<br />
hydroxyapatite suspension in the dose of 40 mg/ml at the background of created osteoresorption resulted in the<br />
normalization of protein-bound and free oxyproline content. Spongy substance revealed osteoblasts, compact substance<br />
demonstrated prominent periosteum microcirculation, with well seen dilated plethoric vessels in it. We have noted osseous<br />
tissue structure restoration. Immunogenesis peripheric organ investigations did not reveal any immunogenic features in this<br />
preparation. The obtained data provide evidence of osseous tissue resorption process decrease as well as its remodeling.<br />
Such medical technology presents an effective method of secondary osteoporosis treatment.<br />
P-30<br />
DOES THE MENISCUS TRANSPLANT PREVENT OSTEOARTHRITIS? FUNCTIONAL<br />
OUTCOME AND RADIOGRAPHIC 5-YEAR FOLLOW-UP<br />
VALENCIA-GARCÍA, H.; FAHANDEZH-SADDI, H.; LÓPEZ-HUALDA, Á.; MARTÍNEZ-MARÍN, J.; TORREJÓN-DE LA CAL,<br />
M.; MARÍN-AGUADO, M.; VAQUERO-MATEO, J., Hospital Universitario Fundación Alcorcón. Spain.<br />
Abstract / Introduction:<br />
Partial meniscectomy is a very common procedure for treatment of meniscal injuries. The absence meniscal promoting the<br />
progression of chondral degeneration and as a result of osteoarthritis with joint space narrowing. Meniscal transplantation<br />
has been proposed for the symptomatic relief after meniscectomy, but has not yet been established whether slowing the<br />
onset of degenerative changes.
<strong>POSTERS</strong><br />
Material:<br />
We retrospectively reviewed meniscus transplants performed in our center. Inclusion criteria included at least 5 years followup<br />
and no other surgical maneuvers (on anterior cruciate ligament, osteotomy or chondral injuries). Finally we analyzed<br />
the results of 10 patients with mean age 34.5 years (21-45). The technique used is frozen at -80 º unirradiated meniscal<br />
allograft and transplantation without bone blocks.<br />
Results:<br />
6 were lateral and 4 medial meniscus. In all scales compared Lysholm, Tegner and VAS and joint space was measured on<br />
a preoperative radiograph charge and 5 years, and the radiological progression as Ahlbäck criteria. We obtained a<br />
satisfaction rate of 78% at 5 years, with an improvement in all scales (average preoperative Lysholm 55 to 85 to 5 years,<br />
Tegner from 3.5 to 6 and VAS from 6.8 to 2). We do not see a significant joint space narrowing (mean preoperative of 3.09<br />
mm and 3.01 mm at 5 years) with 2 cases of improvement of the same (cases 2 and 7).<br />
Discussion and Conclusions:<br />
The meniscus transplant is effective for pain control in symptomatic knee after meniscectomy, with a success rate of 60-88%.<br />
Also appears to decrease the short-term joint degeneration, but still do not know its importance in the long-term<br />
chondroprotective effect. The size and graft fixation are important prognostic factors transplant. The poor results are<br />
associated with graft irradiated limb malalignment or significant chondral degeneration. We did not find deterioration of<br />
affection compartment joint space at 5 years follow-up, but these results must be confirmed in longer term studies.<br />
P-31<br />
ALLOGRAFT SELECTION FOR TRANSEPIPHYSEAL TUMOR RESECTION AROUND THE<br />
KNEE USING THREE-DIMENSIONAL SURFACE REGISTRATION<br />
APONTE-TINAO ALBERTO, L.; SCHWINT, O.; RITACCO EDUARDO, L.; FARFALLI LUIS, G.; MILANO EDGARDO, F.; BOU-<br />
SLEIMAN, H.; REYES, M., Italian Hospital of Buenos Aires. Argentina.<br />
Abstract:<br />
Transepiphyseal tumor resection is a common surgical procedure in patients with malignant bone tumors. The aim of this<br />
study is to develop and validate a computerassisted method for selecting the most appropriate allograft from a cadaver bone<br />
bank. Fifty tibiae and femora were 3D reconstructed from computed tomography (CT) images. A transepiphyseal resection<br />
was applied to all of them in a virtual environment. A tool was developed and evaluated that compares each metaphyseal<br />
piece against all other bones in the data bank. This is done through a template matching process, where the template is<br />
extracted from the contralateral healthy bone of the same patient. The method was validated using surface distance metrics<br />
and statistical tests comparing it against manual methods. The developed algorithm was able to accurately detect the bone<br />
segment that best matches the patient’s anatomy. The automatic method showed improvement over the manual counterpart.<br />
The proposed method also substantially reduced computation time when compared to state-of-the-art methods as well as<br />
the manual selection. Our findings suggest that the accuracy, robustness, and speed of the developed method are suitable<br />
for clinical trials and that it can be readily applied for preoperative allograft selection.
P-32<br />
THREE-DIMENSIONAL MORPHOMETRIC ANALYSIS OF THE DISTAL FEMUR: A<br />
VALIDITY METHOD FOR ALLOGRAFT SELECTION USING A VIRTUAL BONE BANK<br />
APONTE-TINAO ALBERTO, L.; FARFALLI LUIS, G.; RITACCO EDUARDO, L.; MILANO EDGARDO, F.; SCHWINT, O.,<br />
Italian Hospital of Buenos Aires. Argentina<br />
Abstract:<br />
Tumor excision is the primary treatment of aggressive or recurrent benign bone tumors and malignant bone sarcomas. This<br />
requires a surgical resection with the potential for large residual osseous defects that could be reconstructed using fresh frozen<br />
allografts. Virtual bone banks enable the creation of databases allowing a 3D pre-surgery evaluation of such allografts,<br />
based on segmentation of DICOM-CT images. This study demonstrates the usefulness of patient specific 3D models for an<br />
accurate host–donor allograft match. We describe one way to select the best match according to size and shape. The results<br />
suggest that a robust and realiable technique has been established. Since it is difficult to plan an allograft on a distal femur<br />
deformed by the tumor, we propose to plan the surgery on the contralateral side. Our results support this limb symmetry<br />
hypothesis. The use of this measurement protocol enables accurate selection of allografts from a contralateral healthy femur<br />
3D CT model achieving the best match possible considering the geometry of available allograft candidate femur specimens.<br />
P-33<br />
DEEP-FROZEN FEMORAL HEADS: THE HIGHER THE WEIGHT, THE BETTER THE<br />
VIABILITY<br />
<strong>POSTERS</strong><br />
MIRABET, V.; SOLVES, P.; LARREA, L.; RÓDENAS, T.; PAMPLONA, T.; ALAVÉS, F.; ROIG, R., Banco de Células y Tejidos<br />
de la Comunidad Valenciana. Spain.<br />
Abstract:<br />
Cancellous bone is widely used in orthopedic surgery. So, femoral heads are stored in tissue banks as a source of this kind<br />
of bone. Deep freezing at -80ºC, without the use of cryoprotectant or controlled cooling rate, is a common method used for<br />
their storage. Thus, the osteoconductive and osteoinductive potential is maintained, which is important for their clinical<br />
efficiency. In addition, femoral heads are usually subject to long period of ischemia, without a nutrient solution, from surgical<br />
collection until processing. All of these conditions are widely accepted to have a significant detrimental effect on cell survival<br />
and, even more, for many authors, killing any cells that are present in the tissue. However, Heyligers and Klein-Nulend, in<br />
2005, and Simpson et al, in 2007, described the presence of osteocyte-like phenotype cells with a low proliferative capacity<br />
in culture, in deep-frozen femoral heads. In order to explain the reason for the presence of viable cells in this frozen bone,<br />
we have used an “in vitro” culture method to assess the differentiation potential of hematopoietic precursor cells from femoral<br />
head bone marrow. Once collected, the femoral head was kept in a kapton-teflon bag. No transport solution was used during<br />
ischemia (at 4ºC, maximum 12h). After 1h incubation in a disinfectant solution (vancomycin, tobramycin, co-trimoxazole<br />
and amphotericin B, all 50µg/ml in Hanks balanced salt solution) the tissue was frozen by placing it in a freezer at -80ºC,<br />
and stored at this temperature. We have processed 27 femoral heads from living donors undergoing hip surgery: gender,<br />
37% female and 63% male; pathology, 67% coxarthrosis and 33% other; mean age 67.5 years (range 51-87); mean<br />
storage 133 days (range 20-442); tissue mean weight 77.4g (range 28-125). Thawing was performed by immersion in a<br />
water bath at 37ºC. Afterwards, the tissue was extracted from the bag and the bone marrow was aspirated with a syringe<br />
using heparinized (50 IU/ml) medium M199. The suspension was filtered with a 100µm pore size and centrifuged (900g<br />
10min). The pellet was resuspended in Iscove’s medium, seeded in Methocult and incubated for 14 days at 37ºC, 5%CO2
in a humidified atmosphere, to assess the growth of hematopoietic colony forming units. Colonies, defined as aggregates<br />
of more than 40 cells, were counted under an inverted microscope. A thermocouple was inserted inside the femoral head,<br />
monitoring the cooling rate during freezing. Four ramps could be distinguish, which could be appropriate for cell survival<br />
in the case of heavier tissues (1.5ºC/min during the transition phase from liquid to solid). Results showed CFU-GM, CFU-<br />
GEMM and/or BFU-E growth in 63% of cases. A weight of the femoral head heavier than 79g was significantly associated<br />
to the presence of viable cells.<br />
P-34<br />
ALLOGENIC CARTILAGE APPLICATION IN RHINO- AND GENIOPLASTY<br />
<strong>POSTERS</strong><br />
PAVLUK-PAVLUCHENKO, L.; LEKISHVILI, M.; DUBININ, S.; RYABOV, A., Peoples’ Friendship University of Russian. Russian<br />
Federation.<br />
Abstract:<br />
Allogenic chondral implants application in surgery has more than centenary history. It’s positive properties proved by priority<br />
researches of domestic scientists in various areas of plastic surgery. Last 5 years (2006-2011) there were 125 patients under<br />
our observation with rhinoplasty operations (112 patients) and a chin surgery (17 patients), where have been applied implants<br />
from allogenic costal cartilage. allogenic implants were used at nose defects like a saddle deformations of posttraumatic and<br />
postoperative etiology, at defects of a backrest, tip part of a nose, a nasal septum. Hypognathia was the indication allogenic<br />
chondral implants application too. This method consisted in a restore of volumes and increasing of a nose and chin support.<br />
Implants getting from the preparations of 5-7 costal received from corpses on special technology were used. (The manufacturer<br />
- laboratory of CITO Tissue bank). Short technique of application: the cartilage was taken from sterile package and 30 minutes<br />
was exhibited at the sterile capacity filled of 500,0 ml of a normal saline solution. Then on a flat plastic board from rectilinear<br />
or curvilinear sites of a cartilage (under indications) by means of scalpels N.º 10, N.º 11, N.º 15 and rasps it was made<br />
implant of the settlement sizes and the form which after antiseptic processing, was entered into nose area (under periosteum,<br />
on a surface of cartilages or in soft tissues) or a chin (under periosteum). For preventive maintenance of secondary deformations<br />
chose a costal place, reinforced a cartilage threads, or PDS plates. Implants were fixed in a place in the various ways. The<br />
remote results are studied in terms from 1 till 4th years. Positive takes at a rhinoplasty are received in 96 % of observations.<br />
Secondary deformations are noticed in 3 %, a pyesis in 1 % of observations that has demanded repeated operations.<br />
Genioplasty has led to positive takes in all observations, except one, when the patient has demanded excision of implant on<br />
subjective motivation. Allogenic chondral implants application in plastic surgery of a nose and a chin – a reliable and effective<br />
way to restore of volume and increase tissue support, allowing to receive high esthetic results.<br />
P-35<br />
INFLUENCE OF BISPHOSPHONATES AS A PART OF A BIOCOMPOSITE MATERIAL<br />
ON AN OSTEOGENESIS<br />
LEKISHVILI, M., RODIONOVA, S. YUROSVA, Y., TORGASHIN, A., RYABOV, A., Priorov Central Institute of Traumatology and<br />
Orthopaedics (CITO) Tissue bank, Moscow. Russian Federation.<br />
Abstract:<br />
According to some information, application bisphosphonates, simultaneously with oppression of resorbtion reduces intensity<br />
of an osteal tissue formation. The research purpose: to estimate influence of Ibandronic acid "Bonviva" as a part of a
iocomposite material on osteogenesis process. � Materials and methods: experiment is spent on 20 females of white nonlinear<br />
rats who have been parted on 2 groups. In skilled group defect of a tibial bone was filled with non demineralized lyophilized<br />
bone graft, bridged to a material containing Ibandronic acid "Bonviva" 1 mg/ml, the biocomposite material didn't contain in<br />
control group bisphosphonate. Animals deduced from experiment for 90 days. The estimation of results was spent<br />
morphologically (light microscope Zeiss Axioskop 40). Preparations painted: a hematoxylin and eosine. � Intensity of an<br />
osteogenesis and character of changes in area of bone graft estimated in points: 1 point – a weak osteogenesis (the area of<br />
osteal defect is filled by a quaggy fibrous tissue and fragments of implant, the presented osteal beams without osteocytes), 2<br />
points – a moderate osteogenesis (in a defect projection there are centers of a neogenic mature osteal tissue round bone graft<br />
or a regional osteogenesis on the basis of a cartilaginous tissue with the rests of implant), 3 points – the expressed osteogenesis<br />
(the defect area is filled by a neogenic mature osteal tissue without the rests of implant). Statistical calculations carried out<br />
under program SPSS, with a significance value р
<strong>POSTERS</strong><br />
basic tissues of the recipient that promotes not only to prevention of development postoperative complications, but also<br />
restoration of physiological functions of a nose�<br />
P-37<br />
APPLICATION OF DEMINERALIZED XENOGENIC IMPLANTS IN ORAL SURGERY<br />
LEKISHVILI, M., YURASOVA, Y., REZNIKOVA, N. ,, RYABOV, A.<br />
N.N. Priorov Central Institute of Traumatology and Orthopaedics (CITO) Tissue bank, Moscow. Russian Federation.<br />
Abstract:<br />
Considering a difficult situation of morally-ethical character with application of allogenic materials in out-patient practice<br />
of oral surgeons, the increasing value is got by biomaterials made of a bone of animals. Using of grinded demineralized<br />
xenogenic implants (DXI), made by original licenced technology from a cortical layer of long tubular bones of a bull became<br />
one of variants of the decision of this problem. The purpose - the analysis of work with DXI during preparation and at<br />
carrying out of dental implants surgery, and also an estimation of the remote results of using DXI at patients with bone<br />
pathology in oral cavity. The technology of their manufacturing includes some stages. The fence and a choice of a donor<br />
material is carried out in full conformity with legislation demands. Cleared of soft tissues and blood elements osteal fragments<br />
freeze and then crush in the original equipment. A following stage is degreasing of an osteal crumb by an admixture of<br />
Chloroformium and ethyl alcohol. The subsequent demineralization of a crumb is spent by weak solutions of a hydrochloric<br />
acid. Demineralized material subject to additional processing for neutralization of the rests of acid with the subsequent fast<br />
cooling to -35C. Material preservation carry out by a lyophilization. As a rule, after a lyophilization a dry material by<br />
means of a set sieves part on fraction on the size of particles, pack up and pack into double plastic packages. The technology<br />
final stage is sterilization which spend in the radiative way, influencing a stream of fast electrons a dose of absorption within<br />
20-25 kGr. The size of the grinded chips ready to clinical use varies from 0.1 to 5 mm. In our observations the materials<br />
which size has made 0.5-2 mm have been used. At an experimental investigation phase high efficiency of the given material,<br />
including in comparison with the most widespread analogs in Russia that has made necessary its introduction in clinical<br />
practice has been confirmed. At a preclinical stage the number of patients which for surgical treatment used DXI in the form<br />
of chips included 30 patients with the blasted teeth which are subject to excision, defects of alveolar ridges (including after<br />
dental implants excision), cysts, chronic generalized periodontitis. A material admixed with a blood clot of the patient and<br />
placed in bone defect separately or round inserted earlier dental implant. The age of patients from 21 till 65 years. Inspection<br />
of patients was spent with use clinical, laboratory, radiological (including a computer tomography) methods. Postoperative<br />
terms of observation have made from 6 till 12 months. At a part of patients at surgical treatment used lyophilized collagenic<br />
membranes. Using of DXI in the form of chips with the size of particles from 0.5 to 2 mm has shown their high clinical<br />
efficiency at carrying out of surgery on preparation of alveolar ridges to dental implants insertion, and also at its immediate<br />
carrying out (including it is single-step with an tooth extraction). High degree osteoinductive and osteoconductive material<br />
potention, fast terms of formation of an osteal tissue in an implantation zone (3 to 4 monthes), absence of inflammatory<br />
reaction, and also fastness formed bone regenarate to resobtion after dental implants loading becomes perceptible.<br />
Application of the xenogenic parentage implants, made on the patented technology (“Newbone”) dilates possibilities of<br />
surgeons at treatment of a various pathology of an bone tissue in an oral cavity.
P-38<br />
OSTEOGENIC EFFECT OF TETRACYCLIN IN COMPOUND XENOGENEIC BONE<br />
IMPLANTS<br />
ZUNINO, J. H. 1 , FILOMENO, A.2, CIGLIUTTI, G. 2 , SEMIGLIA, G. 2<br />
1 - INDT, Tissue Bank, Hospital de Clínicas, 2 - Cátedra de Pequeños Animales, Orientación Quirúrgica, Facultad de Veterinaria.<br />
Uruguay.<br />
Abstract:<br />
Since about 7 years ago, we have been using -both experimentally and clinically- an antigen-free, freeze-dried bone<br />
implant from bovine source that retains the osteoinductive properties of the bone matrix.<br />
The current knowledge about the non-antibiotic properties of low-dose topic Tetracyclines , inhibiting bone resorption<br />
(inhibition of bone matrix metalloproteinases and osteoclastic activity as well) and stimulating osteogenesis (increase of<br />
Type I collagen expression and increase of the number and activity of osteoblasts), drove us to design a compound implant<br />
expressing both the osteoinductive properties of our antigen-free bovine bone implant and the osteogenic properties of<br />
Tetracyclines.<br />
We therein designed a double-blind experiment in rabbits, comparing the magnitude of the osteogeneic response achieved<br />
by two different orthotopic bone implants placed in critical defects at the skull: bovine bone alone and Tetraclyn-embedded<br />
bovine bone.<br />
Results demonstrated a larger, conspicuous and systematic osteogeneic response within the defects filled with Tetracyclinembedded<br />
bovine bone. Microscopically, those defects showed a significant reduction of the mononuclear infiltrate,<br />
osteoclast number and an increased rehabitation of osteoplasts and osteoid and woven bone formation.<br />
Keywords:<br />
Tetracyclines, xenoimplants, osteogenesis, bone banking.<br />
P-39<br />
COMPARATIVE STUDIES ON BIOMECHANICAL PROPERTIES OF IRRADIATED<br />
PORCINE TENDON<br />
SPINOSA, M. 1 , SANTORO, N. C. 1 , BLANGINO, A. E. 2 , KAIRIYAMA, E. 1<br />
1 - National Commission of Atomic Energy, 2 - Faculty of Engineering, University of Buenos Aires. Argentina.<br />
<strong>POSTERS</strong><br />
Abstract:<br />
Collagenous tissues are employed in many reconstructive procedures. Tendon is one of them and the mechanical contribution<br />
of tendons is determined by their mechanical properties. For reconstruction following injury, these data are necessary for<br />
selecting the appropriate graft material.<br />
Radiation sterilization after processing and packaging has been used to ensure sterility for a variety of allografts.<br />
In this study, forty two porcine’s tendons were used and the irradiation was carried out with a 60Co source. The<br />
biomechanical properties of the tendon were measured with two universal testing machines by uniaxial loading of<br />
longitudinal fibers of collagen.<br />
The aim of this work was to evaluate mechanical properties of irradiated tendon in an animal model.<br />
From fresh porcine feet were obtained the tendons. The animals were male pigs of 6 month, weighted around 150 kg. The<br />
tendons were harvested and dissected, placed in polyethylene bags of 100 µm thick and immediately preserved at -80 ºC<br />
(gradual freezing). The length of each tendon was 15 cm.<br />
The irradiation was carried out at -80 ºC. Fourteen samples were irradiated at 15 kGy, fifteen samples were irradiated at<br />
30 kGy and thirteen samples were preserved for control.
<strong>POSTERS</strong><br />
The biomechanical properties of the tendon were measured with two universal testing machines with 1000 N load. In order<br />
to do these tests, the specimen must be preconditioned by repeated cycling and must respond to the similarity law.<br />
Frozen tendons were thawed in saline solution (0,9 %) during 15 minutes at room temperature. The tensile speed in all tests<br />
was 0,25 mm/sec. Each specimen was subjected to an initial preload of 25 N, then three cycles at the same speed and finally<br />
the test itself.<br />
Statistical assessment was performed using an analysis of variance (ANOVA) to compare the three groups.<br />
The calibrated areas or cross section areas were measure from each tendon and were used to calculate the stress, strain<br />
and the elastic modulus of Young of the material. This cross section area was around 25 mm2. The Young´s Modulus<br />
calculated was around 91 ± 21 Mpa.<br />
Biomechanical testing on animal model of irradiated fresh frozen tendon through the Young's modulus of elasticity, do not<br />
shown significant differences between irradiated and non-irradiated specimens. According to these results the gamma<br />
irradiation do not produced mechanical changes in porcine tendon irradiated up to 30 kGy in frozen condition.<br />
P-40<br />
OSTEOINDUCTIVE AND OSTEOGENIC PROPERTIES OF TWO XENOGENEIC BONE<br />
MATRICES<br />
ZUNINO HAMLET, J.; SEMIGLIA, G.; DE PRO, C.; CARZOGLIO, J.; FILOMENO, A., INDT, Tissue Bank, Hospital de Clínicas.<br />
Uruguay.<br />
Abstract:<br />
Bone implants for biologic reconstruction of the skeletal system must ideally be osteoinductive, osteoconductive and antigenfree.<br />
Different Tissue Banks produce a wide variety of different bone-derived matrices, with inherent different biologic<br />
properties. Some decades ago, xenogeneic bone was used in several human trials, with controversial results. The processing<br />
methods therein used for rendering porcine and bovine bone antigen-free, destructed the matrix inductive proteins (BMPs).<br />
Since 2003, we process bovine bone preserving natural BMPs within the implants. Lacking prionic diseases (cattle and<br />
humans), the National Tissue Bank in our Country can safely produce and distribute processed bovine bone for biologic<br />
skeletal reconstruction purposes. In addition, bovine bone does not transmit human infectious diseases (HIV, Hepatitis C, etc.),<br />
is a low-cost material and has demonstrated a satisfactory biomechanic performance. Conversely, processing method for<br />
human bone at our Tissue Bank does not seem to preserve BMPs in the implants. To compare the osteoinductive potential of<br />
bovine and human processed bone, we designed a double-blind study in mice, using two processed, xenogeneic bone<br />
matrices (human and bovine), heterotopically implanted (quadriceps muscle). Results clearly demonstrate the benefits of<br />
processed bovine bone related to its osteoinductive and osteogeneic properties. Keywords: Osteoinduction, Osteogenesis,<br />
Xenogeneic bone, Tissue Banking.
P-41<br />
COMPUTER ASSISTED NAVIGATION FOR TUMOR RESECTION AND ALLOGRAFT<br />
TRANSPLANTATION<br />
APONTE-TINAO ALBERTO, L.; FARFALLI LUIS, G.; RITACCO EDUARDO, L.; MILANO, F.; AYERZA ANGEL, M.; MUSCOLO<br />
LUIS, D., Italian Hospital of Buenos Aires. Argentina.<br />
Abstract / Introduction:<br />
Wide-local resection with an adequate margin is a crucial step in the management of patients with a musculoskeletal<br />
malignancy. Early results in the use of image fusion for computer-assisted bone tumor surgery seem to have accurate results<br />
in bone tumor resections. The aim of this study was report our experience in preoperative planning with image fusion, tumor<br />
resection and allograft transplantation according to the desired plane using intraoperative navigation assistance.<br />
Methods:<br />
Thirty patients were 3D reconstructed in a virtual platform and planned determining the osteotomy position according to<br />
oncology margins in a CT-MRI image fusion. Tumor resections and allograft transplantation were performed using a computer<br />
navigation system according to previously planed. We analyzed the technical problems (crashes), time for navigation<br />
procedure during surgery, accuracy of the registration technique and surgical margins.<br />
Results:<br />
In four patients (13%) the navigation was not carried out due to technical problems. In two cases the crash was secondary<br />
to software problems, and in the remaining two cases the crash was secondary to hardware problems. Of the 26 cases where<br />
the navigation was performed, the mean registration error was 0.63 mm (range 0.3-1.1). The mean time for navigation<br />
procedures during surgery was 22 minutes (range 11-37). Histological examinations of all specimens showed a clear tumor<br />
margin in all patients.<br />
Discussion and conclusion:<br />
Our findings suggest that preoperative planning, tumor resection and allograft transplantation guided by navigation is<br />
accurate and useful method for bone tumor surgery. In our study, the navigation could not be performed in 13% of series.<br />
P-42<br />
ACHILLES ALLOGRAFT AS A NEW EXTENSOR SYSTEM OF THE KNEE<br />
<strong>POSTERS</strong><br />
MÉNDEZ GIL, A.; ARES RODRÍGUEZ, O.; LOZANO LIZARRAGA, L.; SEGUR VILALTA, J.M.; MARTÍNEZ PASTOR, J.; PO-<br />
PESCU, D.; GARCÍA OLTRA, E., Hospital Clínic Barcelona. Spain<br />
Abstract:<br />
AIM Acute or chronic injuries of the patellar tendon are a rare complication in knee arthroplasty. Prevalence between 0.17<br />
to 2.5% is reported in the literature. These require surgical repair to ensure proper functionality of the arthroplasty. There<br />
are several treatment options. We present our technique with Achilles allograft for the reconstruction of the extensor<br />
mechanism of the knee and the clinical results of five cases.
<strong>POSTERS</strong><br />
Material and Methods:<br />
All cases had a deficit in active extension with passive full extension. It is strictly necessary that the patient has full passive<br />
ROM. We performed an anterior approach with eversion of the patella and a trench at the tibial tuberosity was made in<br />
order to fix the calcaneus part of the allograft in the trench with two cannulated screws. The Achilles tendon was divided<br />
into 2 bundles. It is important to avoid the patella alta in order to keep the patella in a correct position. The lateral bundle<br />
had a transtendon quadriceps position whereas the medial bundle had a medial position. Both bundles were sutured with<br />
Ethibond 5. Knees were immobilized in extension for two months. After this period gradual rehabilitation started getting a<br />
right balance, functionality and ability to walk in all cases.<br />
Results:<br />
With this new technique we avoid the problems with the patella resurfacing made before or the use of patello femoral<br />
arthroplasties. Our patients have a better ROM (near full extension) than preoperative situation after one year FU. The<br />
fixation with two screws help us with the arthroplasty has a tibial stem. The deep frozen Achilles tendon is helpful for this<br />
technique.<br />
Conclusions:<br />
This surgical technique offers a solution to a very complex problem of great concern. Our cases and the reviewed studies<br />
provide an overview of the surgical technique and short-term follow-up. Despite the promising initial results, a long-term<br />
follow-up should be made in order to reach a conclusion.<br />
P-43<br />
IMPROVEMENT PLAN FOR USE OF GRAFT FEMORAL HEAD OF LIVING DONOR<br />
VALENCIA-GARCÍA, H.; MARÍN-AGUADO M.; TORREJÓN-DE LA CAL M.; LÓPEZ-HUALDA A.; MARTÍNEZ-MARTÍN J.;<br />
VAQUERO-MATEO J.; FAHANDEZH-SADDI H., Hospital Universitario Fundación Alcorcón. Spain.<br />
Abstract / Introduction:<br />
The use of bone graft in orthopedic surgery has increased in recent years throughout the world, especially in revision surgery<br />
and prosthetic spine surgery that requires contributions of cancellous bone. That has become the bone in the second most<br />
transplanted human tissue after blood and requires health authorities and surgeons to reduce to a minimum the risk of<br />
transmitting disease. Obtaining allogeneic bone can be a living donor or cadaveric multiorgan donations. In both cases it<br />
requires informed consent from the donor, the validation of the clinical history (presence of cancer, autoimmune diseases,<br />
blood transfusions in the past 6 months, treatment with steroids or growth hormone, radiation exposure or travel to Britain<br />
1980 to 1996, to give an example), the negativity in serological test set by regulatory agencies and the absence of pollutants.<br />
We present a review of procurement and feasibility of femoral heads obtained from living donor in the HUFA in the last 13<br />
years. In this study we propose a possible improvement plan for the use of femoral head obtained from living donors.<br />
Material:<br />
We use data for our work in retrospect of the past 13 years in the HUFA on femoral heads obtained from living donor<br />
surgeries implant primary hip arthroplasty. Were analyzed that were implanted, those that were discarded and the reason<br />
thereof and which still remain in the bone bank waiting to be used.<br />
Results:<br />
Of the 210 femoral heads obtained in the extraction, 42% was used (90 heads without complications during follow-up), 44%<br />
were discarded and 13% in reserve is suitable for distribution . Of the Heads discarded (94), one third of them (34 pieces,<br />
16% of total extracted) was due to contamination, 40% (38 parts, 18% of total extracted) with systemic disease (history of<br />
cancer, serology positive) or local (3 osteonecrosis) and 23% (22 parts, 10% of total extracted) fault logistics process (absence
<strong>POSTERS</strong><br />
of donor consent, absence of any serological or microbiological tests, absence of pathology). While the number of<br />
contamination is similar to the range of 5-20% in most series, we consider that 55% of parts not usable for other reasons<br />
(60 pieces, 28% of total extracted) are an important workload and cost should try to improve.<br />
Discussion and Conclusion:<br />
The ideal graft should be osteoinduction, osteoconductive, osteogenic and mechanical properties if the situation demands.<br />
Faced with increasing demand, tissue banks must seek new ways to obtain, but always ensuring a secure fabric.<br />
Complications of allogeneic transplantation are non-binding, bacterial or viral disease transmission and the responsible bank<br />
should try to avoid those that depend on the selection process, storage or distribution. Sometimes the removed parts do not<br />
meet all administrative requirements, serological or microbiological and should be discarded. Our improvement plan includes<br />
a demanding patient selection, surgical technique and strict aseptic packaging to ensure safe graft. The involvement of all<br />
staff involved in the process helps reduce the "failures of logistics" and therefore the costs incurred and make reasonable<br />
economic cost of the procedure.<br />
P-44<br />
AUDIT OF THE FIRST 13 YEARS OF EXISTENCE OF BONE AND TISSUE BANK OF<br />
HOSPITAL UNIVERSITARIO FUNDACIÓN ALCORCÓN<br />
VALENCIA-GARCÍA, H.; TORREJÓN-DE LA CAL, M.; MARÍN-AGUADO, M.; FAHANDEZH-SADDI, H.; MARTÍNEZ-MAR-<br />
TÍN, J.; VAQUERO-MATEO, J.; LÓPEZ-HUALDA, A., Hospital Universitario Fundación Alcorcón. Spain.<br />
Abstract / Introduction:<br />
The demand for bone graft has increased especially in prosthetic revision surgery, reconstructive, rachis, oncology and<br />
sports. That has become the bone in the 2 nd most transplanted human tissue after blood and requires health authorities<br />
and surgeons to reduce to a minimum the risk of transmitting diseases. Bone banks are nonprofit organizations that<br />
coordinate the selection, collection, storage and distribution of bone and osteoarticular tissue donated for transplantation<br />
human and its role is essential to provide the tissue with the maximum guarantees of quality and sterility. Periodic audits allow<br />
to optimize and improve all procedures performed in the process of tissue donation / transplantation of tissue. With this work<br />
we try to evaluate the results obtained in the first 13 years of the Bone and Osteoarticular Tissue Bank in the University<br />
Hospital Foundation Alcorcón.<br />
Material:<br />
Retrospective review of 13 years in the HUFA in obtaining bone allograft (living donor or cadaver), under the<br />
recommendations of the Council of Europe, the American Association of Tissue Banks and the European Association of<br />
Musculoskeletal Transplant. In all cases informed consent was sought from the donor, the validation of the clinical history<br />
(presence of cancer, autoimmune diseases, blood transfusions in the past 6 months, treatment with steroids or growth<br />
hormone, or radiation exposure trip to Britain 1980 to 1996, to give an example), the negativity in serological test set by<br />
regulatory agencies (syphilis, HBV, HCV, HIV 1 and 2, HTLV I and II and Cytomegalovirus) and the absence of contaminants<br />
in aerobic and anaerobic cultures samples obtained during the extraction.<br />
Results:<br />
In the first 13 years of the Bank has proceeded to obtain tissue from 140 deceased donors and 210 living donors (undergoing<br />
hip replacement). Thus shows the evolution of donors per year. It also analyzes the distribution of the number of pieces<br />
obtained (minimum 18, maximum 34, average 22), the distribution of frozen tissue implanted (total 1310 pieces), the<br />
distribution of implanted freeze-dried tissue (out of 337) and as the rate of contamination, and tissue tracking provided. We<br />
analyze the profile and indications of implant recipients through the diagnosis and cause of death of deceased donors to<br />
study their profile over the years.
Discussion and Conclusion:<br />
The ideal graft should be osteoinductive, osteoconductive, osteogenic and mechanical properties if the situation required.<br />
Faced with increasing demand, tissue banks must seek new ways to obtain, but always ensuring a secure fabric.<br />
Complications of allogeneic transplantation are non-binding, bacterial or viral disease transmission and the responsible bank<br />
should try to avoid those that depend on the selection process, storage or distribution. Any Bank should to audit bone activity<br />
to optimize their work while guaranteeing a safe bone.<br />
P-45<br />
THE INFLUENCE OF SINTERING TEMPERATURE ON COMPOSITION AND SURFACE<br />
MORPHOLOGY OF BOVINE BONE<br />
FIRDAUS, A.; ADAM, Z.; HAWANI, A.; CHEE WEI, T.; HIDAYU, C.; SUZINA, S., Tissue Bank, School of Medical Sciences<br />
Health Campus Universiti Sains Malaysia.<br />
Abstract:<br />
The use of hydroxyapatite as bone replacement has been widely discussed. The good biocompatibility, bioactivity, high<br />
osteoinductivity and osteroconductivity, noninflamitory behaviour and nonimmunogenicity properties of hydroxyapatite make<br />
it a good choice for hard tissue repair. There are various method used to synthesize these bioceramic such as ultrasonic<br />
irradiation, radio frequency (RF), thermal plasma and micromolecules. All of these processes are complicated and biologically<br />
unsafe. Recently hydroxyapatite has been extracted by normal calcinations of some biowaste such as bovine bones, pig<br />
bones and fish bones. Therefore, the aim of this study was to analyze the effect of sintering temperature on the composition<br />
and surface morphology of bovine bone. The cancellous part from the bovine femoral head were cleaned and cut into 10<br />
mm x 10 mm x 10 mm. The bones were sintered at 200C, 400C, 600C, 800C and 1000C using sintering furnace. The nonsintered<br />
bones were used as control group. The changes of the composition and surface morphology were analyzed at every<br />
temperature stage using Field Emission Scanning Electron Microscope (FESEM). FESEM showed non-sintered bones were<br />
covered by organic substance possibly protein and collagen. However the organic materials were completely removed at 600C<br />
making the surface appeared smooth and the pore structure remained intact. Typical EDX spectrum showed the decreased in<br />
the carbon element with the increment of the temperature. At 800C calcium and phosphate were markedly higher compared<br />
to the samples sintered at 600C. At this temperature there is a possibility of crystalline hydroxyapatite formation. In addition<br />
there was evidence of ruptured areas of the bones sintered at 800C. At this high temperature it is known that damage of pore<br />
structure may occur. The result of this study showed that the suitable sintering temperature of 800C was able to remove<br />
organic compound, maintain porous structure and form hydroxyapatite of cancellous bovine bone.<br />
P-46<br />
SEARCH FOR METHODS TO INCREASE THE NUMBER OF PROCUREMENTS IN A<br />
TEMPORAL BONE BANK<br />
CAREMANS, J.; MUYLLE, L.; HAMANS, E.; VAN DE HEYNING, P., University Hospital Antwerp. Belgium.<br />
<strong>POSTERS</strong><br />
Abstract / Introduction:<br />
Tympano-ossicular allografts (TOA) provide unique reconstructive capabilities, allowing reconstruction after radical removal<br />
of cholesteatoma and other middle ear pathology. To provide TOA, the University of Antwerp Temporal Bone Bank (UATB)<br />
was established in 1988. As the number of procurements has drastically decreased over the past years following the
implementation of the EU directives (e.g. exclusion of potential donors with history of malignant disease), we wanted to search<br />
methods to increase the number of procurements.<br />
Materials and Methods:<br />
Data of donors and potential donors reported have been captured in an Access database. Data were entered in the database<br />
on a daily basis right after procurement or after finding a contra-indication for a potential donor. The distribution of donors<br />
per age group and per sex and the distribution of contra-indications in 2009 and 2010 were analyzed.<br />
Results:<br />
More than half of the procured donors are over 75 years old (72.2% in 2009 and 51.9% in 2010). This correlates to the<br />
bigger death rate in this age category. Also, there is a big difference in distribution between male and female donors. In<br />
2009, 70.9% of all donors were female, 64.8% in 2010. This is explained by males often being rejected because of not<br />
having enough hair to allow a perfect reconstruction of the incision being made during the procurement procedure. The<br />
percentage of reported deaths that lead to a procurement is very low (6.3% in 2009 and 4.3% in 2010) and is concordant<br />
with the drop between 2006 and 2009 as shown by Van Rompaey et al (Cell and Tissue 2011). The 3 most important<br />
contra-indications in 2009 and 2010 were logistical reasons (26.6% in 2009, 43.7% in 2010), malignancies (27.2% and<br />
23.0% respectively) and risk of imperfect reconstruction (14.4% and 14.3% respectively). The main logistical reasons are:<br />
potential donors reported too late, no one available to perform the procurement, no access to pre-mortem blood samples<br />
in some hospitals and impossibility to collect a blood sample within 24 hours post-mortem, limited time slot to perform<br />
procurement because of visiting hours for family of donor.<br />
Conclusions:<br />
Despite the positive impact of the omission of the upper age limit of 75 years by the Superior Health Council in August 2008,<br />
the number of procurements has drastically decreased over the past years because of new regulations (exclusion of<br />
malignancies and uncontrolled systemic infections since June 2007).The search for solutions to increase the number of<br />
procurements should in the first place be focused on a maximum reduction of logistical problems and secondly in getting<br />
more reports of potential donors. Therefore ways of reporting potential donors are being evaluated and optimized where<br />
possible and new hospitals are being contacted to get approval for on-site procurements.<br />
P-47<br />
THE USE OF BONE ALLOGRAFT IN REVISION TOTAL KNEE ARTHROPLASTY<br />
<strong>POSTERS</strong><br />
LÓPEZ-HUALDA, A.; VALENCIA-GARCÍA, H.; MARTÍNEZ-MARTÍN, J.; FAHANDEZH-SADDI, H.; MARÍN-AGUADO, M.;<br />
TORREJÓN-DE LA CAL, M.; VAQUERO-MATEO, J., Hospital Universitario Fundación Alcorcón. Spain.<br />
Abstract / Introduction:<br />
Management of bone loss is a challenging problem in revision total knee arthroplasty. Options for reconstruction include:<br />
metal augments, autografts, allografts, tumor prostheses and porous metal. The purpose of the present study was to analyze<br />
the complications after revision total knee arthroplasty with bone allograft and the integration of the allograft.<br />
Methods:<br />
We reported a retrospective study which included revision of total knee arthroplasty with allograft, treated in our institution<br />
between 1999 and 2010. Thirteen knees were followed by clinical evaluation and periodic radiographs at a mean of 65.23<br />
months (range 12-125 months).<br />
Results:<br />
Three primaries total knee arthroplasties failed because of infection and ten had an aseptic loosening. After replacement<br />
using allograft, two cases had an aseptic loosening, and required a second replacement. No case had a septic loosening.
Bone allograft was used in all cases. In small defects we used morselized allograft (73%), and massive defects (type II-II of<br />
AORI classification) were solved with structural allograft. The allograft was preserved frozen in 86% and lyophilized in<br />
14%. Allograft incorporation was successful in 92 % of the cases, according to the knee society criterion. There was no<br />
fracture, collapse or total resorption of the allograft.<br />
Conclusions:<br />
Bone loss is commonly encountered at the time of revision total knee arthroplasty. Bone allograft is a safety option, with a<br />
low rate of complications and successful results. We believe that tissue bank provides a good solution for patients requiring<br />
revision total knee arthroplasty in the setting of massive bone loss.<br />
P-48<br />
INFLUENCE OF DONOR TYPE IN BONE AND TISSUE GRAFTS OUTCOMES IN THE<br />
QUARANTINE PERIOD<br />
MIETH, K.; GONZÁLEZ, J.; NAVAS, J.; SOTO, C., Fundación Cosme y Damián. Colombia.<br />
Abstract:<br />
Introduction In Colombia, there are two main ways of bone and tissue donation. We have voluntary donors from the national<br />
organ and tissue transplantation net, and donors from the national forensic institute. Grafts from the later are obtain thanks<br />
to a legal figure known as donation presumption. Donor selection criteria, grafts extraction protocols and quarantine<br />
standards are the same for both groups. We would like to determine if grafts outcomes in the quarantine period are similar.<br />
Objective:<br />
To determine weather or not there are differences in outcomes in the quarantine period of the grafts and donors of two<br />
different types of donation at the Fundacion Cosme y Damian bone and tissue bank in Bogota, Colombia.<br />
Material and Methods:<br />
We did a full review of our records in the last 5 years, from 2006 to 2010. In this retrospective cohort we divided our<br />
donors in the two above mention groups and determine in each one the following variables: Donor rejection, percentage<br />
of graft losses and loss causes (i.e.: positive cultures or positive serologic tests) For continue variables we use means and<br />
standard deviations, and for qualitative variables, proportions. We present the associations between the type of donation<br />
and the outcomes in the quarantine period after a multivariate analysis.<br />
P-49<br />
FUNDACIÓN COSME Y DAMIÁN: A MODEL OF A PRIVATE, NON FOR PROFIT<br />
SINGLE TISSUE BANK<br />
SOTO, C.; NAVAS, J.; MIETH, K.; GONZÁLEZ, J., Fundación Cosme y Damián. Colombia.<br />
<strong>POSTERS</strong><br />
Abstract:<br />
Our institution was open in 1990 being the first in Colombia and neighbor countries to procure bone and tendons from<br />
cadaveric donors and processing them for supplying the national requirements in Orthopedic Surgery. By that time there<br />
were not public projects for tissue banking. Our structure is based in academics and research since we are orthopedic
surgeons practicing in a private medical school facility. Our institution started the medical training in allograft´s reconstructive<br />
surgery programs and participated actively in the cration of legal regulations of tissue banking in our country, following the<br />
AATB and EATB standards. Therefore our institution is entirely dedicated to musculoskeletal allografts. From 1990 to date<br />
we have procured 2700 donors, have processed 120.000 allografts and distribute 90.000 for implantation.<br />
P-50<br />
OSSICULAR PROSTHESES FROM BANKED BONE VIA NUMERICALLY CONTROLLED<br />
MICROMILLING<br />
RAMELLI, M.; LISCIO, M.; CARRIAGGIO, F.; BERRETTINI, S.; DANTI, S.; MANCINI, I., Tissuelab S.p.a. Italy.<br />
<strong>POSTERS</strong><br />
Abstract / Intro:<br />
In otitis media surgery the auditory ossicles often have to be replaced. The reconstruction of the ossicular chain has been<br />
performed either by autograft or homograft ossicles or by synthetic prostheses. Starting from the late 1980s, homografts and<br />
also xenografts fell progressively out of use because of the fear of transmittable infectious disease following<br />
allotransplantation. For this reasons synthetic materials were introduced. Unfortunately the extrusion, although reduced, still<br />
represents an important clinical phenomenon, which may occur in up to 20% or more of cases in long term follow-up. The<br />
current achievements in industrial processing now potentially allow the banked bone prostheses to be produced, similar in<br />
shape and weight to hydroxyapatite prostheses, thus combining the most favourable aspects of both synthetic (reproducibility,<br />
convenience and biosafety) and biological replacement (total biocompatibility).<br />
Materials and Methods:<br />
Cortical bone from donations was supplied by Bone Tissue Bank of Tuscany Region (Florence Italy). The bone was processed<br />
by Tissuelab to ensure complete decellularisation and biosafety according to GMP pharmaceutical standard. Cortical blocks<br />
were preliminarly measured with digital callipers and then tightened in the lock-plate of the CNC ultraprecision micromilling<br />
machine (MDX40 – Roland) to be modelled as ossicular replacement prostheses (ORPs) through a subtractive rapid<br />
prototyping method. Technical drawings and milling parameters were uploaded by means of software (Rhinoceros 4.0 and<br />
Modela player 4, respectively). The ORPs were sterilised/virus inactivated with a 50 KGy dose of gamma rays. Samples<br />
were fixed, dehydrated with alcohols, embedded and sectioned with an ultratome and stained for hystological analysis.<br />
Results:<br />
Banked cortical bone PORPs and TORPs could be produced via CNC micromilling with high dimensional accuracy. The<br />
PORPs weights averaged 31,2 ±0,6 mg, and the TORPs averaged 69,3 ±0,7 mg. From a histological point of view, the ORP<br />
cortical bone was void of cells, and its extracellular matrix compactness even appeared superior to that of the auditory<br />
ossicles.<br />
Discussion and Conclusion:<br />
The availability of ORPs based on homologous compact bone may represents the optimal solution to prevent extrusion in<br />
ossiculoplasty. The whole production process was shown to be potentially performed in accordance with the GMP guidelines.<br />
The PORP and TORP weights were comparable to those of the auditory ossicles. GMP processing of homologous bone<br />
includes controlled industrial treatment and advanced sterilization procedures that render such a biological risk almost null.<br />
Homologous bone ORPs may finally reduce to zero the extrusion-related complications that persist in ossiculoplasty. At<br />
moment 14 of the preformed bone ossicular prostheses have been implanted, within the framework of a clinical study<br />
approved by the ethics committee, with positive clinical results. References: 1) Berrettini, MD et al.; Ann. Otol., Rhinol. &<br />
Laryngol. 2011, 120 (1): 9-16 2) Alanay A. et al., Spine J. 2008 Sep-Oct;8(5):789-95.
<strong>POSTERS</strong><br />
P-51<br />
NEW METHODS FOR CARTILAGE GRAFT EVALUATION: OPTICAL COHERENCE<br />
TOMOGRAPHY, POLARIZATION SENSITIVE OPTICAL COHERENCE TOMOGRAPHY<br />
AND THERMOGRAVIMETRIC ANALYSIS<br />
MARTINHO JUNIOR, C., A.; FREITAS ZANARDI, A.; BROCARDO DIVA MACHADO, L.; SANTIN PLUMERI, S.; SOARES<br />
AUGUSTO NEVES, F.; MATHOR BEATRIZ, M., Nuclear and Energy Research Institute. Brazil.<br />
Abstract:<br />
Since some tissue banks have adopted ionizing radiation as a secure method to sterilize allografts, the basic question about<br />
the real effects of radiosterilization on tissue structure remains in mind of medical groups. For cartilage allografts, until now,<br />
the evaluation methods include mechanical and microscopic tests that are destructive methods. In this work we evaluate<br />
cartilage allografts modification induced by radiosterilization by three new methods. For the first time we have able to<br />
qualify and quantify cartilage collagen network without any previous preparation as occurs with microscopy, using a relative<br />
new technology of Optical Coherence Tomography (OCT) and Polarization Sensitive Optical Coherence Tomography (PS-<br />
OCT), which avoid unnecessary loss of allograft. Moreover, we have applied Thermogravimetric Analysis (TGA) to detect<br />
the direct effects on water flow inside cartilage before and after radiosterilization, providing new data to better understand<br />
the effects of ionizing radiation on collagen network of allografts. Human costal cartilages were obtained from 15 cadaveric<br />
donors aged 18 to 45 years old. Right costal cartilage was preserved in high concentration of glycerol and the left costal<br />
cartilage was deep-frozen at -70 °C. Each sample was divided in four fragments. One of them was kept as control and the<br />
other three were irradiated by a Co-60 source with doses of 15, 25 and 50 kGy. Before the tests were carried out,<br />
glycerolized samples were rehydrated in sterile saline solution and deep-frozen samples were thawed at room temperature.<br />
OCT images were obtained from OCP930SR (Thorlabs, USA) and a homemade software was used to determine the total<br />
optical attenuation coefficient. PS-OCT images were obtained from OCS 1300 device (Thorlabs, USA) with a PSOCT 1300<br />
device coupled. Thermogravimetric curves were obtained from TGA-50 device (Shimadzu, Japan) set to warm the samples<br />
with a rate of 10 °C/min until 105 °C and the dehydration rate was calculated in the linear region of the curve. According<br />
our results, dose of 15 kGy promote crosslinking in collagen structure that cause an increase in total optical attenuation<br />
coefficient and a decrease in the dehydration rate of the samples for both glycerolized and deep-frozen cartilages. For 25<br />
and 50 kGy, for glycerolized samples no significant differences to control group for total optical attenuation coefficient and<br />
dehydration rate were found. However, deep-frozen cartilages irradiated with 50 kGy showed significant differences to<br />
control group for both total optical attenuation coefficient and dehydration rate. PS-OCT images had demonstrated that<br />
birefringence of collagen network is kept in all samples after radiosterilization. Thus, the dose of ionizing radiation for<br />
cartilage sterilization depends of preservation method, once glycerolized cartilages can receive doses of up to 50 kGy,<br />
which is not recommended for deep-frozen cartilages. Acknowledgements: IAEA, FAPESP and CNEN.
P-52<br />
THE USE OF THE OF SPONGY BONE IMPLANT IN THE CORRECTION OF THE<br />
IDIOPATIC SCOLIOSIS<br />
JACAS TORNE F, M.; GARCÍA MESA, N.; ÁLVAREZ CAMBRA, R., Complejo Científico Ortopédico Internacional "Frank<br />
Pais". Cuba.<br />
Abstract:<br />
In the last three years (April 2009-March 2011) forty patients were operated on of Idiopathic Scoliosis using the technique<br />
protocol in our Service of Column Surgery: Intervention after the 30 grades of deformity, artrodesis with Freeze-Dried bone<br />
spongy sterilized by means of Cobalt 60 Irradiation and Luke-Harrington Instrumentation. The bone autographs employment<br />
doesn't justify the risk-benefit mobility and discomfort for the patients. In our series the clinical results, mechanics and<br />
biological they were good, without bigger complications. The radiological signs of consolidation evident starting from the<br />
12 weeks with good correction of deformity, The results showed good correction of the curves, ranging 60% of dorsal curves<br />
and 52% of lumbar curves.<br />
P-53<br />
BONE ALLOGRAFT USE FOR SALVAGE OF FAILED TREATMENT OF<br />
INTERTROCHANTERIC HIP FRACTURES<br />
<strong>POSTERS</strong><br />
GARCÍA OLTRA, E.; ZUMBADO DÍJERES ALONSO, J.; CAMACHO CARRASCO, P.; MÉNDEZ GIL, A.; LÓPEZ ZABALA,<br />
I.; TORNERO DACASA, E.; SEGUR VILALTA, J.M. Hospital Clínic Barcelona. Spain<br />
Abstract / Introduction:<br />
Hip fractures in the elderly are frequent. Intertrochanteric hip fractures account for approximately half of all hip fractures in<br />
the elderly; of these, from 50% to 60% are classified as unstable. Unstable fracture patterns occur more commonly with<br />
increased age and low bone mineral density and they had been associated with comminution of posteromedial buttress which<br />
exceeds a simple lesser trochanteric fragment or subtrochanteric fracture lines. The most frequent mechanical complication<br />
in patients with unstable femoral fractures is migration of lag screw or blade through the femoral head that occurs more often<br />
in unstable fracture patterns, poor bone quality and suboptimal position of internal devices. The purpose of the current study<br />
was to evaluate the results of revision internal fixation and bone grafting for salvage of failed internal fixation of<br />
intertrochanteric hip fractures.<br />
Patients and Methods:<br />
Between may 2007 and november 2010 nine patients with intertrochanteric fractures that failed initial internal fixation<br />
were treated with revision internal fixation and bone grafting. There were five women and four men with an average age<br />
of 83.11 years (range, 76-95 years). All patients had acute failure fixation without extensive acetabular involvement and<br />
three of them associated with infection. Three patients had cut-in of the lag screw, five cut-out and one a cephalic screw<br />
disassembly. All patients were treated with implant removal, debridement, antibiotic prophylaxis, new intramedullary nail<br />
and bone allograft. In six cases a cylinder of structural frozen corticocancellous bone allograft was inserted to improve<br />
bone density and to reinforce the femoral head defect followed by morselized cancellous chips with an impactor and<br />
fluoroscopic control. In the other three cases freeze-dried morselized cancellous chips. Clinical and radiographic results<br />
were reviewed retrospectively.
<strong>POSTERS</strong><br />
Results:<br />
Using this rescue technique after six months of follow-up seven patients had radiological consolidation of the fractures<br />
allowing them to perform activities of daily living. One patient had nonunion and the internal fixation was removed and the<br />
other one died because of massive pulmonary thromboembolism. At follow up five patients had no pain and three mild pain.<br />
Discussion:<br />
The treatment of failed internal fixation of intertrochanteric hip fractures is challenging because of fracture pattern, bone loss,<br />
bone quality and possibility of associated infection. Treatment options include prosthetic replacement and revision internal<br />
fixation. This retrospective study showed a high rate of union and functional improvement with revision internal fixation<br />
and bone allograft with few complications.<br />
P-54<br />
BIOMECHANICAL CHARACTERIZATION OF A PROCESSED, ANTIGEN-FREE BONE<br />
XENOIMPLANT FOR SKELETAL RECONSTRUCTION<br />
ZUNINO HAMLET, J.; LANTERO CARLOS, J.; AGUIAR, S.; DERI, E., INDT, Tissue Bank, Hospital de Clínicas. Uruguay.<br />
Abstract:<br />
INDT, Tissue Bank, Hospital de Clínicas, Montevideo, Uruguay. Processed, antigen-free, freeze-dryed, irradiated bone<br />
implants from bovine source have been used in our Country since 2003, for skeletal reconstruction purposes; both<br />
experimentally and clinically, as well. Results from their osteoconductive and osteoinductive properties have proven<br />
successfull, both in animals and humans, and already published elsewhere. Bovine-derived processed bone is a safe and<br />
useful surgical tool for biologic skeletal reconstruction in our Country, where cattle and humans are free of prion diseases<br />
(Kuru, Jacob-Creutzfeldt, Fatal Familiar Insomnia, Gertsmann- Straussler-Scheiner Disease). Nevertheless, bone processing<br />
methods for implant purposes could modify its biomechanical properties. Thus, we design an experiment to compare basic<br />
biomechanical properties (distraction and bending) of fresh bovine bone compared to processed, freeze-dryed, irradiated<br />
bone for implants. According to previous similar biomechanical tests performed on human banked bone, we established<br />
maximum forces to be applied to bovine bone on an hydraulic press (Instron machine) for four-point flexion (bending) and<br />
for longitudinal traction. Density of bone probes was assessed for fresh and processed bone, before the essay. Results<br />
showed that fresh bone could bear double the four-point bending applied force compared to processed, freeze-dryed, nonhydrated,<br />
irradiated bone. Besides, both fresh and processed bovine bone performed similar to traction forces, without<br />
failure, up to 250 kg. Biomechanic characteristcs of processed bovine bone herein exposed -besides its osteoinductive and<br />
osteoconductive properties-, renders this material an interesting alternative to allogeneic and even autogeneic bone grafts<br />
for biologic skeletal reconstruction. Keywords: Biomechanics, Xenoimplants, Bone Banking.
<strong>POSTERS</strong><br />
P-55<br />
PROCESSING METHODS VALIDATION FOR PATELLAR TENDON ALLOGRAFT USING<br />
X-RAY DIFFRACTION<br />
PÉREZ CAMPOS, H.; WODOWOZ, O.; SALDIAS, M.; FACCIO, R.; MOMBRU, A.; ÁLVAREZ, I., Instituto Nacional de donación<br />
y trasplante de células, tejidos y órganos (INDT) - Laboratorio de Cristalografía Facultad de Química Udelar. Uruguay<br />
Abstract:<br />
AIM To validate, by X ray diffraction structural analysis, the most suitable processing method for patellar tendon allograft<br />
preservation, sterilized by gamma radiation -for clinical application. This research is in part being supported by IAEA<br />
Research Contract No: 15546/R0.<br />
Materials and Methods:<br />
Six patellar tendon allografts were harvested from six cadaveric donors, under informed consent. Three tendons were<br />
packed in a triple vacuum sealed sterile bag and storaged in a -80ºC freezer. Three tendons were previously glycerolated<br />
and storaged at the same temperature. All of them were irradiated at 17 kGy dose, and preserved for 30 days until their<br />
defrosting. Previously defrosted and deglycerolized, segments of 1cm2 surface and 2 mm thick, were obtained transverse<br />
to the longitudinal axis from each cathegory tendon: Frozen-Irradiated (FI), and Glycerolized- Frozen -Irradiated (GFI). X<br />
Ray Diffraction was performed (Diffraction System Device: Rigaku Ultima IV) on the samples and average diffractometric<br />
profiles were obtained from each cathegory. The planimetric surface under each obtained curve was calculated as indicator<br />
of the collagenic stroma molecular ordering. From the algebraic sum of both planimetric surface were obtained differencial<br />
positive and negative values, to define the Differential Planimetric Surface (DPS). Ordering Planimetric Coefficient (OPC) was<br />
obtained from the cocient between Σ DPS positive values and Σ DPS negative values. OPC is the indicator of the relative<br />
molecular ordering between both cathegories: FI and GFI. The comparative statistical analysis was performed by "t" test.<br />
Results:<br />
The avarage measures for the positive planimetric algebraic sum were 8803,00 (relative values), and for the negative sum<br />
8310,33 (relative values). The OPC resulting from the comparison between both tendon cathegories, FI and GFI, was 1,059 (n/s).<br />
Comments:<br />
The results obtained from the two different processing methods analysed, do not show significative differences related to<br />
molecular ordering modifications of the FI and GFI patellar tendon collagenic structures.<br />
P-56<br />
EVOLUTION OF THE ACL ALLOGRAFT IN THE ACL RECONSTRUCTION SURGERY.<br />
A 15 YEAR FOLLOW UP<br />
LÓPEZ ZABALA, I.; CLARET GARCÍA, G.; PEREIRA, E.; GRANADOS, N.; GARCÍA OLTRA, E.; SASTRE SOLSONA, S., Hospital<br />
Clínic. Barcelona. Spain.<br />
Abstract / Goal:<br />
Retrospective evaluation of ACL reconstruction with allograft throughout a 15 years follow up. Materials: 120 patients<br />
underwent surgery for ACL reconstruction using freshly frozen allograft from 1991 up to 1999. 19 patients with a mean<br />
age of 39 years were evaluated and underwent physical examination and Radiographies (Kellgren). Tegner, Lysholm and<br />
IKDC indexes were used in the evaluation. The follow up mean was 15 years.
<strong>POSTERS</strong><br />
Results:<br />
Out of 19 patients, all of them were restored back to their previous physical activities. Lysholm (78) and Tegner (5’2) tests<br />
overviewed satisfactory results. Kellgren mean punctuation of the injured knee was 2’5, while the healthy knee punctuated<br />
a 1’1 mean.<br />
Discussion:<br />
Frozen allograft has been widely used in the past. With the use of freshly frozen allograft in our series, stability of the injured<br />
knee was achieved and maintained.<br />
Conclusion:<br />
ACL reconstruction with allograft provides long term fulfilling results, which turns this procedure into an ensuring option to<br />
achieve ACL reconstruction.<br />
P-57<br />
TRANEXAMIC ACID AS A SUBSTITUTE OF APROTININ FOR CHONDROCYTE GRAFT<br />
PREPARATION<br />
BURSIG, H.; SITEK, P.; WYSOCKA, A.; KURZAK, A.; KRÓL, D.; DYLAG, S.; NOZYNSKI, J., Tissue Bank. Poland.<br />
Abstract:<br />
Three dimensional scaffolds are becoming increasingly in treatment of cartilage defect. The fibrin graft is stabilized by<br />
antifibrynolitic agent - aprotinin. Since 2007 aprotinin was withdrew from clinical use because of a few serious adverse<br />
reactions observed. The aim of study was to prepare grafts based on fibrinogen and tranexamic acid as an antifibrynolitic<br />
agent and confirm that TA is a good substitute for aprotinin.<br />
Materials and Methods:<br />
Human chondrocytes were expanded in monolayer culture for 3 weeks. 450 mL blood was collected to produce the<br />
fibrinogen from Regional Blood Center voluntary donor. To prepare gel-like grafts, chondrocytes were mixed with fibrinogen<br />
and then with thrombin in CaCl2. The dose of TA was 10mg/mL and the aprotinin was 5000KIU. There was used different<br />
concentration of fibrinogen to find which dose, together with TA, is sufficient to prepare stable graft. The quantity and quality<br />
methods were used to compare cells viability and ECM production. Grafts were cultivated for 4 weeks in vitro to evaluate<br />
and compare their disintegration.<br />
Results:<br />
The grafts were stable for the time of observation. No shrinkage of the grafts was observed. The examination showed that<br />
TA has no influence on cells viability and ECM production and that migration from the graft to the culture plate is observed<br />
Conclusions:<br />
Tranexamic acid can be used, instead of aprotinin, as a safe antifibrynolitic agent for chondrocyte graft preparation.
P-58<br />
AUTOLOGOUS CHONDROCYTE CULTURE – 5 YEARS EXPERIENCE<br />
BURSIG, H.; WYSOCKA, A.; SITEK, S.; KURZAK, A.; KEPSKI, F.; DYLAG, S.; GAZDZIK, T., Tissue Bank. Poland.<br />
Abstract:<br />
Tissue engineering has been shown to be a promising method for the treatment of articular cartilage defects. ACI is presently<br />
successfully adopted all over the world, and clinical and histopathological findings confirm the effectiveness of the method.<br />
Our Tissue Bank prepare autologous chondrocyte for orthopaedic clinics according the GMP principles, EU directives, and<br />
regulations applicable to Tissue Banks. Aim of study We performed chondrocyte culture for 120 patients for nine different<br />
hospitals.<br />
Material and Methods:<br />
Cartilage was taken from the patient's femoral condyle, as well as blood, for the purpose of preparing autologous serum.<br />
Chondrocyte were cultured about 3 weeks. 450 ml patient own blood was collected prior to transplantation to produce<br />
autologous fibrinogen, alternatively the allogenic fibrinogen was prepared from Regional Blood Center voluntary donors.<br />
Before surgery the chondrocyte suspension or gel-like fibrograft were prepared.<br />
Results:<br />
Within 5 years we prepared 120 autologous chondrocyte grafts for patients with cartilage defect: 42 in suspension, 32 in<br />
autologous fibrinogen, 14 in allogenic fibrinogen, 32 in commercial fibrin glue. The mean age of the patients was 31 years.<br />
The average defect size was 5,4 cm2 . The mean weight of harvest cartilage was 193 mg and the mean number of implanted<br />
cells were 2,8mln per cm2 of cartilage defect. Sterility control cultured cells were performed at different time points.<br />
Histological evaluation of biopsy reveal round-shape chondrocytes and proteoglycan presence. Gene expression profile<br />
confirmed chondrogenic phenotype of implanted cells.<br />
Conclusions:<br />
Chondrocyte (fibro)graft preparation in our Tissue Bank is a promising method for treatment of cartilage defect. A novelty<br />
in our work is the application of autologous fibrinogen in combination with autologous chondrocytes.<br />
P-59<br />
DEVELOPMENT OF AN EFFICIENT METHOD TO DEMINERALISE BONE<br />
<strong>POSTERS</strong><br />
CALAMARDO, M. A. 1 , VITO, S. 1 , FARIÑAS, O. 1 , LUQUE, S. 1 , VILARRODONA, A. 1 , TABERA, J. 1 , SEGUR, J. M. 2 , TRÍAS, E. 1<br />
1 Transplant Services Foundation - Hospital Clínic. Barcelona. Spain., 2 - Hospital Clínic. Barcelona. Spain.<br />
Abstract / Introduction:<br />
The objective of this study was to develop an easily reproducible method to obtain demineralised osteoinductive bone,<br />
including cancellous chips and cortical powder, obtained from cadaveric donors. A method of monitoring the<br />
demineralisation process was also established in order to obtain bone with a specific desired residual calcium level with the<br />
maximum osteoinductive activity (0.99). At the essays performed by pulse cycles (with the same conditions of temperature,<br />
stirring and volume of hydrochloric acid) we found that at 2% of residual calcium, the pH of the solution was near 0.9 and<br />
the time of demineralization was near 4 minutes. It has been corroborate the importance of the particles sizes in the<br />
demineralisation process, because our cancellous chips samples contained a wide range of particles sizes and some of our<br />
results do not correlate with the expected outcomes.
Conclusions:<br />
We observe that monitoring the demineralisation process by the eluent pH is better than doing that with the time of<br />
demineralisation as it also depends on the initial quantity of bone material and on the initial % of calcium. In addition, we<br />
also conclude that it would be necessary to homogenize bone particles prior to demineralisation. Finally, if both<br />
demineralising methods are compared, we can determine that the best process to demineralise bone is the method of<br />
pulse cycles of acid because it provides a slower rate of demineralisation that allows us to control more efficiently the<br />
process.<br />
P-60<br />
DISTAL TIBIA FAILURE IS THE MOST COMMON COMPLICATION IN MASSIVE<br />
INTERCALARY MASSIVE BONE. ALLOGRAFT RECONSTRUCTION IN PEDIATRIC<br />
TUMOR<br />
MARTANTO, T. W., EDWARD, M., Dept. of Orthopaedic Dr. Soetomo General Hospital/Airlangga University. Indonesia.<br />
Abstract:<br />
A Case Series Tri Wahyu Martanto 1, Mouli Edward 2, Ferdiansyah3,4, Sjahjenny Mustokoweni5, Paulus Rahardjo6 Sri<br />
Andreani7 Rosy Setyawati8 Abstract<br />
Background:<br />
Pediatric Tumor had special characteristic in management. Ability of regeneration, thickness of periosteun were benefit for<br />
treatment in pediatric pathology. Allograft widely used for reconstruction of extremity cause of tumor , trauma and other bone<br />
defect. The Expensive of Expandable mega prosthesis and refusal of patients for amputation makes us use massive allograft<br />
as a substitute for bone tumors, but we had problem with distal part failure in tibia allograft reconstruction<br />
Objective:<br />
To evaluate of the result of this procedure since 2005 until 2010.<br />
<strong>POSTERS</strong><br />
Subject and Method:<br />
10 subject after resection of bone tumor has been done and substitute with massive allograft. The result was evaluate<br />
clinically, radiological, and Function base to the ISOLS criteria.<br />
Result:<br />
10 cases,they all are intercalary. Intercalary allograft evaluated according radiological ISOLS criteria, 6 patients were<br />
excellent, 1 patients was fracture and healed by external support, 1 patients was implant loosening and he was done by<br />
minor change of the implant, 2 Patient failure in corporation in distal part of tibia allograft reconstruction. Function post<br />
reconstruction, 6 cases were excellent, and good in 1 cases, and 2 cases fair with external suport.<br />
Conclusion:<br />
Massive allograft reconstruction can be considered as the method to prevent amputation and maintain the limb after bone<br />
resection of bone tumor, with excellent result, with carefully selective case and carefully preservative the graft. Keyword :<br />
massive allograft, Pediatric, Intercalary, resection bone tumor 1. Consultant in Pediatric Orthopaedic, Dept Of. Orthopaedic<br />
& Traumatology, Dr. Soetomo General Hospital/ School of Medicine, Airlangga University Surabaya 2. Consultant in Tumor<br />
Musculoskeletal, Dept Of. Orthopaedic & Traumatology, Dr. Soetomo General Hospital/ School of Medicine, Airlangga<br />
University, Surabaya 3. Head Section of Tumor Musculoskeletal, Dept Of. Orthopaedic & Traumatology, Dr. Soetomo General<br />
Hospital/ School of Medicine, Airlangga University, Surabaya 4. Director of Dr. Soetomo Tissue Bank & Biomaterial, Dr.<br />
Soetomo General Hospital, Surabaya 5. Consultant in Musculoskeletal Pathology, Dept Of. Pathology, Dr. Soetomo General<br />
Hospital/ School of Medicine, Airlangga University, Surabaya 6,7,8 Consultant in Musculoskeletal Radiology, Dept Of.<br />
Radiology, Dr. Soetomo General Hospital/ School of Medicine, Airlangga University.
P-61<br />
INFECTION AND LOOSENING PROBLEM IN BONE ALLOGRAFT APPLICATIONS IN<br />
TUMOR CASES<br />
MARTANTO, T.; EDWARD, M., Dept. of Orthopaedic Dr. Soetomo General Hospital/Airlangga University. Indonesia.<br />
Abstract:<br />
A Case Series Mouli Edward 1 ,Tri Wahyu Martanto 2 ,Ferdiansyah 3,4 , Sjahjenny Mustokoweni 5 , Paulus Rahardjo 6 Sri Andreani 7<br />
Rosy Setyawati 8<br />
Abstract Background:<br />
Massive allograft reconstruction have been done in Surabaya in limb salving procedure to prevent amputation of the<br />
extremity in patient with bone defect post resection in bone tumor and to maintain the complete limb. As an un living bone<br />
allograft had risk for infection and losening.<br />
Objective:<br />
To evaluate of the result of this procedure since 2007 until2010.<br />
<strong>POSTERS</strong><br />
Subject and Method:<br />
12 subject after resection of bone tumor has been done and substitute with massive allograft. The result was evaluate<br />
clinically, radiological, and Function base to the ISOLS criteria.<br />
Result:<br />
12 cases, there are intercalary allograft (8 cases) and osteochondral allograft (4 cases). Intercalary allograft evaluated<br />
according radiological ISOLS criteria, 5 patients were excellent, 1 patients was infected, 2 cases fail in healed. Function post<br />
reconstruction, 5 cases were excellent, good in 1 cases, 2 fair in external support. Osteocondral allograft evaluated<br />
according radiological ISOLS criteria 4 patients were excellent (100%).Function post reconstruction, 4 cases were excellent<br />
for 2 years, and started loosening and broken in third year.<br />
Conclusion.<br />
Massive allograft reconstruction can be considered as the method to prevent amputation and maintain the limb after bone<br />
resection of bone tumor, with excellent result, with carefully selective case and carefully preservative the graft. Keyword :<br />
massive allograft, Intercalary, Osteochondral, resection , bone tumor 1. Consultant in tumor musculoskeletal, Dept Of.<br />
Orthopaedic & Traumatology, Dr. Soetomo General Hospital/ School of Medicine, Airlangga University 2. Consultant in<br />
Pediatric Orthopaedic, Dept Of. Orthopaedic & Traumatology, Dr. Soetomo General Hospital/ School of Medicine,<br />
Airlangga University 3. Head Section of tumor musculoskeletal, Dept Of. Orthopaedic & Traumatology, Dr. Soetomo General<br />
Hospital/ School of Medicine, Airlangga University 4. Director of Dr. Soetomo Tissue Bank & Biomaterial, Dr. Soetomo<br />
General Hospital, Surabaya 5. Consultant in Musculoskeletal Pathology, Dept Of. Pathology, Dr. Soetomo General Hospital/<br />
School of Medicine, Airlangga University 6,7,8Consultant in Musculoskeletal Radiology, Dept Of. Radiology, Dr. Soetomo<br />
General Hospital/ School of Medicine, Airlangga University.
P-62<br />
TENDON ALLOGRAFT FOR MULTILIGAMENT KNEE INSTABILITY<br />
MORALES, J., MORALES, J. D., POPESCU, D., SEGUR, J. M., Hospital Clínic. Barcelona. Spain.<br />
<strong>POSTERS</strong><br />
Abstract / Introduction:<br />
High energy trauma, like knee dislocation cause varus or valgus forced moments on the knee and produce multiligament<br />
disruption (anterior cruciate, posterior cruciate, lateral and medial sided capsuloligamentous). In multiligament injuries of<br />
the knee, several techniques for anatomic reconstruction have been used. The optimum management of these injuries remains<br />
controversial including timing, technique (repair versus reconstruction) and rehabilitation protocol. Variables to consider for<br />
repairing are: Anatomical individual variations Number of damaged structures Number of previous surgeries Image exams<br />
findings Allografts are a suitable resource for multiple ligaments reconstruction. We present 3 cases using multiple allografts<br />
for these injuries. Case 1 36 years old woman.<br />
Accident:<br />
Knee dislocation after doing sports with multiple ligament disruption. Picture 1 Surgical procedure: Picture 2,3 Table 1<br />
Surgery Procedure Allograft type First PCL* + PLC* reconstruction. Laprade technique Achilles tendon + Anterior tibialis<br />
tendon Second (6 months later) ACL reconstruction Anterior tibialis tendon<br />
Results:<br />
Balance 0-120. No knee fails or instability complaining. Case 2 27 years old man Problem: Chronic ACL + PLC disruption.<br />
Picture 4 Surgical Procedure: Picture 5,6 Table 2 Surgery Procedure Allograft type First time ACL reconstruction Anterior<br />
tibialis tendon Second time PLC recontruction Fanellis technique Anterior tibialis tendon Results: No knee instability complains.<br />
Patient keep on rehabilitation for muscle strengthening Case 3 42 years old man Problem: Knee dislocation after car accident.<br />
Multiple ligament disruption. Picture 6 Surgical Procedure: Picture 8,9 Table 3 Surgery Procedure Allograft type First PCL*<br />
+ PLC* reconstruction. Larson technique Achilles tendon + Anterior tibialis tendon Second (4 months later) ACL reconstruction<br />
Achilles tendon Results: No knee instability complains. Returned to daily activities. Not sportive activities.<br />
Discussion:<br />
Allograft has proved to be a good replacement for anatomic reconstruction of damaged ligaments in several studies 1,2 Ligament<br />
reconstruction results in short and long term studies using either allografts or autografts are similar. 3,4,5 Postoperative care is<br />
mainly determined by the type of reconstruction, but essentially allograft and autografts are treated using the same protocol. 6,7<br />
The main goal of the surgical technique is recreation of the central knee axis through anterior cruciate ligament and posterior<br />
cruciate ligament allograft reconstruction. This involves complete release and excision of scar tissue, use of the most reliable<br />
surgical technique and maintenance of postoperatively stability and functional motion with a functional brace.<br />
Conclusion:<br />
Allograft tendons provide a valuable alternative to host tissue in cases of multiple ligament disruption. Its low immunogenicity,<br />
low donor site morbidity, availability and biological safety make them reliable for surgery. Bibliography 1 Anatomic posterior<br />
cruciate ligament reconstruction with allograft. Stannard JP J. Knee Surg 2010 jun 23(2):81-7 2 Surgical Technique: Medial<br />
Collateral Ligament Reconstruction Using Achilles Allograft for Combined Knee Ligament Injury. Marx RG. Clin Orthop<br />
Relat Res 2011 jun10 3 Anterior cruciate ligament reconstruction: a multicenter prospective cohort study evaluating 3<br />
different grafts using same bone drilling method. Leal Blanquet J. Alentorn Geli E, Tuneu J. Clin J Sports Med 2011<br />
jul:21(4):294-300 4 Posterior cruciate ligament reconstruction using single-bundle patella tendon graft with tibial inlay<br />
fixation: 2- to 10-year follow-up Cooper DE, Stewart D. Am J Sports Med. 2004 Mar;32(2):346-60. 5 Comparison between<br />
hamstring autograft and free tendon Achilles allograft: minimum 2-year follow-up after anterior cruciate ligament<br />
reconstruction using EndoButton and Intrafix Noh JH, Yi SR, Song SJ, Kim SW, Kim W. Knee Surg Sports Traumatol Arthrosc.<br />
2011 May;19(5):816-22. Epub 2011 Feb 3. 6 Rehabilitation following knee dislocation with lateral side injury:<br />
implementation of the knee symmetry model. Kinzer A, Jenkins W, Urch SE, Shelbourne KD. N Am J Sports Phys Ther. 2010<br />
Sep;5(3):155-65. 7 Rehabilitation after multiple-ligament reconstruction of the knee. Edson CJ, Fanelli GC, Beck JD. Sports<br />
Med Arthrosc. 2011 Jun;19(2):162-6.
<strong>POSTERS</strong><br />
P-63<br />
EFFECTS OF CHONDROPROTECTIVE AGENTS: GLUCOSAMINE, CHONDROITIN<br />
SULFATE, AND HYALURONIC ACID ON CHONDROCYTES CULTURED FOR<br />
TRANSPLANTATION<br />
OLENDER, E.; UHRYNOWSKA-TYSZKIEWICZ, I.; KAMINSKI, A., Medical University of Warsaw; National Centre for Tissue<br />
and Cell Banking. Poland.<br />
Abstract / Background:<br />
In the conservative treatment of articular cartilage defects chondroprotective agents (also natural components of extracellular<br />
matrix) are administered. Autologous chondrocytes transplantation (ACI) is a modern method of surgical treatment of isolated<br />
cartilage defects. Chondrocytes are retrieved from a cartilage biopsy and propagated in culture. The success of the treatment<br />
depends on the number of chondrocytes transplanted and their ability to synthesize collagen type II, which is characteristic<br />
for healthy articular cartilage. Chondrocytes propagate poorly in culture and dedifferentiate during the culture which<br />
manifests itself in cessation of the synthesis of collagen type II and switching to the synthesis of collagen type I typical for<br />
scar tissue. In the experiment culture medium was supplemented with chondroprotective agents applied in the conservative<br />
treatment. The aim of the experiment was assessment of the effects of such supplementation on the propagation and collagen<br />
gene expression, and in practice, developing of new more effective culture media for chondrocytes cultured for ACI purposes.<br />
Methods:<br />
Cartilage fragments were digested enzymatically in order to isolate chondrocytes. Chondrocyte from each donor were<br />
divided into several groups and next cultured in a medium supplemented differently. Supplements used were: glucosamine,<br />
chondroitin sulfate, hyaluronic acid. Osmosity of supplemented media was assessed. Positive control was culture in medium<br />
supplemented with insulin growth factor-1 (stimulating chondrocytes to synthesize collagen type II). The results were compared<br />
with a culture in standard medium. After each passage (3) the number of chondrocytes was assessed as well as the<br />
expression of genes for collagen I and II (real time PCR).<br />
Results:<br />
In the experiment positive effects of the supplementation of culture media with chondroprotective agents on the proliferation<br />
and gene expression were observed.<br />
Conclusions:<br />
Chondroprotective agents applied in conservative treatment can be applied with positive effects as components of media<br />
for culturing of chondrocytes for transplantation.
P-64<br />
STRUCTURAL BONE ALLOGRAFT FOR RECONSTRUCTION OF THE ACETABULAR<br />
COMPONENT IN SEVERE HIP ARTHROPLASTY LOOSENING<br />
FERNÁNDEZ-VALENCIA, J.; MEDRANO, C.; TORNERO, E.; TOMÁS, X.; BORI, G.; SEGUR, J.M.; GALLART, X.<br />
Hospital Clínic. Barcelona. Spain.<br />
Abstract:<br />
Acetabular revision of total hip replacement is challenging when bone stock is severely deficient. In these cases, bulk structural<br />
allografts can be indicated. Although it is a well-known technical option, there are few series evaluating the results in the<br />
medium and long term. The present series describes the results of this technique for a 10 years period in our institution, with<br />
a follow-up of 4.2 years (range 2-11). 13 hips in 12 patients were identified. Two patients died at 2 years-follow-up due<br />
to causes not related with the surgery. The indication was aseptic loosening in 9 cases and the second stage treatment of a<br />
septic loosening in 4 cases. The arthroplasty mean duration was 10.8 years (range 1-30). Acetabular deficiency was defined<br />
as type IIIA in 9 cases, IIIB in 2 cases and IIC in 3 cases, according to Paprosky´s Classification. Radiographic analysis<br />
included a detailed study of implant migration, screw breakage and radiolucencies; 2 cases were considered as possibly<br />
loose according to the criteria by Gill et al. modified by Van der Linde. A minor resorbtion of the graft was observed in 4<br />
cases. Graft integration was evaluated by CT-Scan: a complete or more than 50% of integration of the graft was observed<br />
only in 4 cases; 4 cases were integrated less than 50% and in 3 cases no integration was present. No patients required<br />
revision of the allograft for problems related to the acetabular reconstruction. The functional result at the final follow-up visit<br />
was 14.63 (range 10-18) according to Merle d’Aubigné, and 65.19 (range 41-94.9) according to Harris Hip Score. The<br />
results showed in the present series, outline structural bone allograft as a satisfactory option to treat sever acetabular<br />
deficiencies. However, an incomplete integration was the most common finding in the CT-Scan. We consider that this<br />
technique should evolve in order to increase the integration rate, to guaranty a high survival rate of the reconstruction.<br />
P-65<br />
HOW OPTICAL COHERENCE TOMOGRAPHY CAN BE USED TO EVALUATE<br />
QUANTITATIVELY GRAFTS BEFORE ITS USE<br />
<strong>POSTERS</strong><br />
MATHOR BEATRIZ, M.; MARTINHO JUNIOR, C. A.; SANTIN PLUMERI, S.; SOARES AUGUSTO NEVES, F.; FREITAS ZA-<br />
NARDI, A., Nuclear and Energy Research Institute. São Paulo. Brazil.<br />
Abstract:<br />
For a long time, in order to evaluate graft safety before its use, tissue banks have been used destructive methods, as<br />
mechanical tests and microscopy, reducing the amount of tissue available for transplantation. Optical Coherence Tomography<br />
(OCT) is a non-destructive technique that provide high resolution images in real time. Because it is a contactless technique,<br />
it is possible evaluate grafts after radiosterilization process, once the light penetrate trough plastic bags. Analogous to<br />
ultrasound, OCT measure the backscattering intensity of infrared laser rather than sound. Once backscattering intensity<br />
cannot be measured directly due to the high speed of the light, the OCT use a technique known as low coherence<br />
interferometry. Basically, in an OCT system the broadband light is coupled into a fiber-optic Michelson interferometer and<br />
is divided in two light beams by a beam splitter. One beam is directed to a reference mirror that is precisely controlled by<br />
a computer and another light beam is directed to the sample. Light backscattered by the sample and the light reflected by<br />
the mirror are recombined at the beam splitter generating an interference pattern which is collected by the detector. OCT<br />
does not only generate images and the data contained in each pixel of the digital image may be used to evaluate
quantitatively the tissue. Indeed, our group have been used the “total optical attenuation coefficient” (TOAC) to determine<br />
mathematically the modifications induced by ionizing radiation in tissues as cartilage and bones after radiosterilization.<br />
TOAC is calculated according with exponential decay of the light when it penetrates in tissue. With a homemade software<br />
we can calculate the average value of pixel for each column and a final average for all columns of the interest region<br />
selected by user. The software also perform an exponential fit using the function f(x)= a exp(-b x) + c, where “a” is the<br />
amplitude of backscattering signal in the surface, “b” is the total optical attenuation coefficient inside the sample and “c” is<br />
a background level due to signal noise. Our first results shown that TOAC can be used to stipulate the mechanical tension<br />
property of cartilages, suggesting that TOAC is an indirect measure of the real state of internal structures in the tissue. Thus,<br />
OCT associated with TOAC can be very useful for tissue banks to determine the graft safety before its use in surgeries.<br />
Acknowledgements: International Atomic Energy Agency (IAEA - project RC16119), Foundation for Research Support of<br />
the State of São Paulo (FAPESP – project 2008/10437-9) and National Nuclear Energy Commission (CNEN).<br />
P-66<br />
ENDOGENOUS AND EXOGENOUS RESISTANCE FACTORS OF CONNECTIVE TISSUE<br />
TRANSPLANTS TO THE EXPOSURE OF THE RADIATIVE EMISSION<br />
SHANGINA, O., Russian Eye and Plastic Surgery Center. Russian Federation.<br />
<strong>POSTERS</strong><br />
Abstract:<br />
The scientific development of the radiation sterilization technologies which on the one hand would guarantee full sterility of<br />
donor tissues and on the other hand the preservation of their biological properties is one of the most topical problems of<br />
modern transplantology and restorative surgery. Allografts made of tendons, derma, fibrous capsule of the kidney,<br />
subcutaneous fat, hyaline cartilage have different degree of resistance to the radiative effect. Changes in the structure of the<br />
allograft fibrous framework following the radiation sterilization depend first of all upon the donor tissue fibroarchitectonics.<br />
The least resistant to the radiation effect are tendon allografts, the fibroarchitectonics of which are determined by a dense<br />
pouch of collagen fibre (CF) bundles oriented strictly in one direction. High radiation resistance of derma allograft is<br />
determined by two endogenous factors, the first being a complicated spatial fibroarchitectonis of the fibrous framework in<br />
which CF bundles have a spiral tract passing from one layer into another, one and the second factor being the presence of<br />
the hyaluronic acid. The inhomogeneity of layers are characteristic for the allograft of the kidney fibrous capsule. The<br />
external dense layer is of the maximum radiation resistance since it is presented by complicatedly twisted CF whereas the<br />
internal layer fibroarchitectonics has a maximum radiation resistance since it is presented by thin isolated collagen fibre<br />
bundles. High degree of the hyaline cartilage allograft radiation resistance is determined by the heterogeneity of its structure;<br />
i.e. the prevalence of II type collagen and high concentration of sulfated glycosaminoglycanes which are thought to be<br />
natural radiation protectors. The allograft structure of subcutaneous fat is connective tissue bands consisting of packed<br />
unidirectional collagen fibres which, in the isolated form, present a sufficiently radioresistant structure and fat cells containing<br />
neutral fats (triglycerides) which play a role of radioprotectors and increase radioresistance of the given allografts. Thus the<br />
allograft resistance to the radiation effect depends upon the peculiarities of their fibroarchitectonics and composition of<br />
noncollagenous components defining by us as endogenous factors of the allograft radiation resistance. There is still a number<br />
of exogenous factors that have an influence upon the allograft radiation resistance, i.e. their chemical and physical treatment.<br />
The radiation resistance of allografts, which underwent the lyophilization process, for example, substantially decreases<br />
compared with the allografts preserved in the liquid media. It is necessary to assume that the destruction process in<br />
lyophilized allografts takes place as a result of the ionizing radiation direct effect whereas the allografts preserved in liquid<br />
media are subjected to the indirect radiation effect. To exogenous radiation factors of allografts it is necessary to refer types<br />
(gamma radiation or high-speed electrons) and doses of the radiation effect. Thus allograft radioresistance is determined<br />
by a set of endogenous and exogenous factors. Endogenous factors are the so-called constant reflecting the initial structure<br />
of the donor tissue whereas exogenous factors may be modified depending upon the aims of the allograft practical use.
P-66 Bis<br />
LABRAL TRANSPLANTATION IN HIP INSTABILITY<br />
<strong>POSTERS</strong><br />
RIBAS, M.; TEY, M.; BELLOTTI, V.; ERQUICIA, J., ICATME – Institut Català de Traumatologìa i Medicina de l’Esport University<br />
Hospital Dexeus – USP, Barcelona. Spain<br />
Introduction:<br />
Acetabular labrum is important to maintain hip sealing and shock absorption. There is an increasing interest in hip-joint<br />
related procedures to preserve the labrum. Labral debridement can lead to increased joint degeneration when compared<br />
to reattachment.We present a 2 year follow up case report of a sportsman with hip pain and instability secondary to subtotal<br />
labral debridement due to a traumatic rupture. It was treated by allogenic labral transplantation.<br />
Case report:<br />
A 27 years old amateur basket ball player was operated in 2005 due to hip pain related to sport trauma. A hip arthroscopy<br />
was carried on, an anterior extensive labrum lesion was identified and regularized. Five months later, new hip arthroscopy<br />
was carried out for partial labrectomy.<br />
The patient could not return to sports due to mechanical symptoms. In 2006 a CAM lesion was identified and another hip<br />
arthroscopy was undertaken for femoral osteoplasty and subtotal labrectomy through indirect hip access technique. The<br />
patient was first visited at our clinic because of onsetting hip instability and pain. He was able to deal with daily live activities,<br />
but he couldn’t perform heavy works or even sports activities. X Ray showed still CAM lesion with grade 0 jointspace<br />
according to Tönnis hip degenerative classification. MR showed absence of anterior acetabular labrum from 10 to 1 o`clock.<br />
In 2007, after obtaining patient consent, allogenic labral transplantation and femoral re-osteoplasty was performed. A 50<br />
mms long labrum allograft was procured from a living donor 6 weeks before this procedure and preserved at -80º C. EATB<br />
protocol was used to manage all allografting process (standards in musculoskeletal tissue banking, EATB).<br />
A mini open anterior approach was used. Degenerative labrum around the defect was eliminated with a 35 mm final defect<br />
in the anterolateral area. Acetabular host bed at the rim was refreshed with a round burr. Labral defect was measured and<br />
transplant was fixed with 5 high fix bone anchors. Weight bearing was protected with crutches during 3 weeks. Rotations<br />
and flexion over 80º were avoided for 6 weeks. At 3rd month flat running was allowed and high demand sports was<br />
restricted for 8 months. At 18th month follow up the patient was pain free (WOMAC score 90,6% and NAHS 95% without<br />
any hip instability. The patient was able to return to sports at an inferior competitive league (UCLA score level 10).<br />
Discussion:<br />
Labral debridement has been in the past the gold standard technique for labrum pathology for many years. It was assumed,<br />
that labral resection did not significantly increase pressure on acetabular surface. Published results of labral debridement<br />
demonstrated initial pain relief. However pain control and physiological labral function are compromised in the midterm.<br />
New strategies have been developed in the recent years to treat labral lesions, like sutures with bone anchors to reconstruct<br />
and preserve labral function.<br />
In case of non reparable labral tears and subsequent labrectomy, hip instability has been reported. Such a condition has<br />
lead to look for another surgical alternatives like reconstruction with autologous Fascia Lata grafts.<br />
Labral allogenic transplantation can be a good option to treat defects or non reparable labrum tears. It can be managed<br />
with similar routine as for meniscal transplantation in a musculoskeletal tissue bank.
P-66 Bis 2<br />
INVETERATE SERIOUS DEFECTS OF ACHILLES TENDON.<br />
RECONSTRUCTION PROCEDURES<br />
M. NÚÑEZ-SAMPER. Clínica Virgen del Mar. Madrid. Spain<br />
<strong>POSTERS</strong><br />
Introduction and Objectives:<br />
We see with low frequency, patients with severe lost of substance in the Achilles tendon that they can not be repaired with<br />
termino-terminal suture or basic procedures.<br />
The origins of these lesions are repeated failures surgical treatment, misdiagnosis, or degenerative cystic tendinosis treated<br />
with multiple steroids infiltrations.<br />
This situation produce progressive elongation of the gap and dysfunction of own Achilles tendon with affection of achilleous<br />
calcaneous plantar system<br />
Over therapist view, we present our experience in these cases by Christensen surgical technique for defects less than 8 cms.<br />
And the cryopreserved massive tendon bone transplant from donor for cases with lost of substance with more than<br />
8 cms.<br />
Material and methods:<br />
Because there are very small serious diseases, we present a number of cases operated on by us in that we have used the<br />
techniques described in the previous section.<br />
Results:<br />
All patients have at least a following of one year. The clinical and functional results have been optimal, valued in individual<br />
way.<br />
Comments and Conclusions:<br />
We think that these techniques solve the problems posed by these serious injuries. Massive Transplantation Tendon, consider<br />
it an excellent solution for very specific cases, being the world bibliography very scarce for the latter procedure.
SKIN & AMNIOTIC MEMBRANE<br />
<strong>POSTERS</strong><br />
P-67<br />
IN VITRO RECONSTRUCTION OF AN ARTIFICIAL BILAMINAR MULTICELLULAR SKIN<br />
SÁNCHEZ-MUÑOZ, I.; GRANADOS, R.; SÁNCHEZ-TORIBIO, M.; CASARES, M., Hospital Universitario de Getafe. Spain.<br />
Abstract / Background:<br />
In recent years, the reconstruction of human skin by tissue engineering represents a clinical challenge and has offered a<br />
therapeutic alternative. Avascular engineered skin equivalents have been available for several years and used to treat<br />
wounds due to burns, non-healing ulcers and surgical excisions. These artificial skin equivalents are constituted by many<br />
types of cells and a three-dimensional structure that permits cultured cells to proliferate and to create three-dimensional<br />
tissues or tissue substitutes. The problem is that this artificial skin is the lack of blood supply, since the survival and proliferation<br />
of cells depends on an adequate supply of oxygen and nutrients and the removal of toxins. These functions are served by<br />
the vascular system, whenever is not present, the graft is rejected. We have produced a new model of endothelialized<br />
reconstructed dermis that promotes the formation of capillary-like structures. We have seeded human umbilical vein<br />
endothelial cells (HUVECs) with dermal fibroblasts and adipose derived mesenchymal stem cells (ADMSC) in a fibrin matrix.<br />
Dermal fibroblasts and ADMSC have been able to produce extracellular matrix and permit cells to proliferate and grow.<br />
ADMSC secrete significant quantities of angiogenic and antiapoptotic factors (VEGF, HGF). They are able to differentiate<br />
into endothelial cells in vitro and to maintain the optimal microenvironment to promote angiogenesis and participate in<br />
tissue repair and skin regeneration processes.<br />
Objectives:<br />
To describe a novel strategy to obtain vascularized skin substitutes in vitro. Methods. Vascularized artificial skin substitutes<br />
were prepared by seeding 4,5x105-1x106 dermal fibroblasts and ADMSC, and 2-3x106 HUVECs in a fibrin matrix along<br />
with other extracellular components secreted by the ADMSC. 1x106 keratinocytes were seeded on the dermis to make a<br />
complete skin. Artificial skin was maintained in culture for up to 15 days. The culture medium was changed three times a<br />
week. Afterwards, the artificial skin was collected for histological and immunohistochemical analysis.<br />
Results:<br />
We obtained an artificial skin with similar histological structure to normal skin, including dermis and epidermis. By<br />
immunohistochemistry, we demonstrated that endothelial cells (CD31 positive cells) grew, proliferated and organized<br />
themselves into capillary-like structures in the fibrin matrix. The epidermis showed a complete epithelization (cytokeratin<br />
positive cells) with keratohyalin granules, hyperkeratosis and parakeratosis.<br />
Conclusions:<br />
In this study, we have established a novel model of artificial complete skin based on the culture of keratinocytes, HUVECs,<br />
ADMSC and dermal fibroblasts. We have observed that this model has the capability of promoting a spontaneous formation<br />
of a capillary-like network. Our artificial skin model would provide a novel therapeutic approach to different pathologies<br />
and could be a useful tool for regenerative medicine.
P-68<br />
HUMAN AMNIOTIC MEMBRANE AS SCAFFOLD FOR KERATINOCYTES AND<br />
FIBROBLAST CULTURE<br />
SAN LUIS VERDES, A.; RENDAL VÁZQUEZ, M.; DOMENECH GARCÍA, N.; YEBRA-PIMENTEL VILAR, M.; GARCÍA BA-<br />
RREIRO, J.; ANDIÓN NÚÑEZ, C., Unit of Criobiology Complejo Hospitalario Universitario A Coruña. Spain.<br />
Abstract / Objective:<br />
To determine that the amniotic membrane can be used as support for the culture of fibroblasts and keratinocytes and that<br />
this will allow for the development of a model of artificial total skin for use in patients.<br />
Material and Methods:<br />
The placentas are obtained from cesarean sections. The amnio was separated from the chorion and was incubated in 199<br />
medium with antibiotic solution during 24h at 4ºC and after this was cryopreserved. From 5 biopsies of skin of patients the<br />
fibroblasts and the keratinocytes were obtained after a double digestion with trypsin and collagenase, respectively. The<br />
amnio was placed with the chorionic side or with the epithelial side upwards in a Cell Crown insert so that it was immobilized.<br />
The following models were performed: chorionic side without treatment, epithelial side without treatment, and epithelial<br />
side with treatment to eliminate the epithelium. For the elimination of the epithelium, the membrane was treated with the<br />
following treatments: trypsin 1%, 0.25% (10 or 30 minutes) and EDTA 0.1% during 30 min at 37ºC. The same number of<br />
fibroblasts and keratinocytes obtained for each biopsy were sowed in each model for its study. Histologic studies<br />
(hematoxylin-eosin) and immunohistochemic studies (AE1/AE3) were carried out.<br />
Results:<br />
A great number of fibroblasts and keratinocytes were obtained after double digestion. The fibroblasts adhere to and<br />
proliferate better on the chorionic side without treatment of the amnio and the keratinocytes on the basal side without<br />
epithelium. To remove the epithelium from the amnio the best treatment was with trypsin at 1% for 30 min. The amnio with<br />
keratinocytes and fibroblasts, showed a similar structure histologically to that of normal skin after seven days of culture.<br />
Conclusion:<br />
The amniotic membrane is a good support for the adhesion and expansion of fibroblasts and keratinocytes.<br />
P-69<br />
PRODUCTION OF RADIOSTERILIZED PIG SKIN AS BIOLOGICAL DRESSING<br />
REYES FRÍAS, M.; REBOYO BARRIOS, D., Instituto Nacional de Investigaciones Nucleares. Mexico.<br />
<strong>POSTERS</strong><br />
Abstract:<br />
Production of pig skin as a biological dressing in the Radiosterilized Tissue Bank at the National Institute of Nuclear Research<br />
of Mexico began in 2001. Pigs considered for skin retrieval are selected from two slaughterhouses. All pig specimens<br />
undergo veterinarian control prior to procurement, which is carried out at the slaughterhouse. In each harvest, skin from 2<br />
pigs is collected. In the Tissue Bank, porcine xenografts are processed under controlled conditions: the subcutaneous fat is<br />
removed, the skin is shaved, washed, disinfected and obtained using an electric dermatome. The grafts are double packed,<br />
labeled and frozen by placing into a freezer or they are freeze dried during 24 hours and then double packed and labeled.<br />
Both, frozen and freeze dried xenografts are sent to the Irradiator Department of the National Institute of Nuclear Research,<br />
where the grafts are sterilized with gamma radiation. Sterilized pig skin grafts are preserved at -80 °C or under ambient
conditions. Quality control is performed in all the steps of pig skin dressings production. From 2001 to 2010, the production<br />
of pig skin dressings reached a total of 301446 cm2. Pig skin xenografts have been provided by the Radiosterilized Tissue<br />
Bank to several hospitals located in different places of Mexico. The main user is the Burned Children Unit of the “Dr. Nicolás<br />
San Juan” Hospital. Processing of human skin in the Radiosterilized Tissue Bank began in 2007. Due to the number of<br />
donors is limited, the use of xenografts is an excellent alternative for many patients.<br />
P-70<br />
MORPHOMETRICS ANALYSIS OF IRRADIATED SKIN FROM TRANSMISSION<br />
ELECTRON MICROSCOPE IMAGES<br />
<strong>POSTERS</strong><br />
SPINOSA, M.; PACHADO ALBERTO, J.; HORAK INÉS, C.; KAIRIYAMA, E., National Commission of Atomic Energy.<br />
Buenos Aires. Argentina.<br />
Abstract:<br />
In the treatment of burns or accidental loss of skin, cadaveric skin allograft provides an alternative to cover temporarily the<br />
wounded area. The skin is a highly contaminated tissue, so is necessary to reduce it to safe level in order to avoid infection.<br />
Radiation is an excellent alternative and Skin Banks of Argentina are using this method of sterilization for around two<br />
decades and frozen irradiated skin allografts are successfully being applied in repair surgery. The aim of this work was to<br />
study, from images of transmission electron microscope, the effect of gamma rays on the ultrastructure of frozen and<br />
glycerolized human skin by measuring the thickness of the collagen fibers. Cadaveric human skin was processed for<br />
preservation by two methods. One of them, treated with glycerol 10 % was frozen at – 70 °C, and the other was glycerolized<br />
to 85 % and stored, at 4 °C. The processed skin allografts were treated with a 60Co source, at 25,0 kGy and 50,0 kGy;<br />
non irradiated samples were kept as control. The dose of 25,0 kGy is habitually used in tissue radiation sterilization and<br />
50 kGy was chosen in order to evaluate a maximum tolerated dose. Histological evaluation was carried out by observing<br />
the collagen structure under a transmission electron microscope, Zeiss EM 109T. The thickness of collagen fibers was<br />
analyzed by a morphometric technique. The software used for measuring the thickness was Image J 1.43u. Statistical<br />
assessment was performed using a factorial analysis of variance (Factorial ANOVA) and Tukey-Kramer Multiple- Comparison<br />
Test. Glycerolized (85%) skin samples showed no significant differences (p=0,05) in the thickness of collagen fibers between<br />
samples unirradiated and irradiated with 25 kGy, perhaps due to the protector effect of glycerol, a well-known scavenger<br />
of reactive species produced during the irradiation process. But differences were found between unirradiated and 50 kGy<br />
as well as between 25 and 50 kGy. The stored frozen skin samples showed differences between unirradiated and irradiated<br />
with 25 and 50 kGy, but no significant differences were found between irradiated samples with 25 kGy and with 50 kGy.<br />
In spite of morphometric differences observed between controls and irradiated frozen samples, evidences of excellent results<br />
in clinical application of irradiated frozen skin allografts have been obtained in Argentina. In the usual sterilization dose of<br />
25 kGy no significant differences in the thickness of collagen fibers were found between both conditions, glycerolized and<br />
frozen, thus these preservation methods would be suitable.
P-71<br />
DONOR PARAMETERS AFFECTING SKIN TISSUE RECOVERY<br />
SAVÍO, A. M., PÉREZ, M. L., FARIÑAS, O., ALEMANY, X., VITO, S., OLIVA, R., LUQUE, S., ZARDOYA M., AGUSTÍ E.,<br />
VILARRODONA A., AND TRÍAS E., Transplant Services Foundation-Hospital Clínic. Barcelona. Spain.<br />
Introduction:<br />
Nowadays advances in critical care and burn resuscitation have led to the improvement in survival with large complex<br />
burns. The increase in the population of survivors among these patients has forced to widen the sources to obtain products<br />
able of covering the body surface area burned. Lack of autologous split-thickness skin graft requires the use of temporary<br />
skin substitutes, being the skin from human deceased donors a good choice.<br />
Aim:<br />
To establish relation between the amount of square cm of suitable recovered skin from each donor and several factors related<br />
to skin recovery process, in order to assure the quantity and quality of the skin allograft available in our tissue bank.<br />
Methods:<br />
This prospective and retrospective study was conducted on 344 skin donors available in Transplant Services Foundation tissue<br />
bank during the period of January 2009-June 2011.<br />
All statistical analyses were performed using the Statistical Package for Social Sciences (SPSS) version 18.0 for Windows<br />
software package (SPSS Inc., Chicago, Illinois, USA).<br />
The statistical analysis involved the Spearman’s correlation coefficient for continuous variables (significance at the 0.05<br />
level) and for categorical ones the chi-squared test with significance at p
<strong>POSTERS</strong><br />
heart-beating (HB) donors, which was retrieved by the Skin bank of Siena. Microorganisms resistance to the antibiotics<br />
was determined by Kirby-Bauer method. Microorganisms were cryopreserved in growth broth with addition of 15%<br />
glycerol. For time-kill studies, microorganisms were cultivated under optimal growth conditions (Thioglycollate broth) at<br />
37°C overnight and inoculum concentration was determined by Mc Farland method. BASE.128 RED and BASE (growth<br />
control) were inoculated with 10(4) CFU/ml of the selected microorganisms, each assessed in triplicate, and then<br />
incubated at +4°C for 72h and +37°C for 24h. The number of living microorganisms was determined by dilution and<br />
spread plate technique at 0h, 3h, 6h, 9h, 12h (24h, 48h, 72h only for incubation at +4°C) spreading 1 ml of three<br />
successive dilutions on a solid surface (Fastidious agar and TSA). Each sample was assessed in triplicate, and colonies<br />
were counted after 24h.<br />
Results:<br />
All isolated strains were resistant to one or more antibiotics, mainly to Penicillin, Gentamicin, Cefoxitin, and Oxacillin. Timekill<br />
curves and growth curves of the three bacterial strains exhibited significantly different kinetics depending on the<br />
incubation temperature. All microorganisms showed a lag phase of approximately 6 h at +4°C, followed by a killing phase<br />
at 24h of 1.93, 1.23 and 3 log for S. Epidermidis, S. Capitis and P. Acnes, respectively. The killing curve of P. Acnes was<br />
linear and reached complete elimination (4 log killing) at 48h. The curves of the other two strains showed a plateau after<br />
24h. The growth curves at 4°C of all strains showed an initial lag phase of 6h, followed by 0,4 - 1 log growth. A plateau<br />
was reached at 24h for S. Epidermidis, S. Capitis and at 48h for P. Acnes, indicating overlapping between growth and killing<br />
phases. All microorganisms showed an exponential growth at +37°C, which reached approximately 4-6 Log at 12h. After<br />
a 3h lag phase for S. Epidermidis and S. Capitis and a 6h lag phase for P.Acnes, killing of 2.7, 3 and 4 log was observed<br />
at 12h for S. Epidermidis, S. Capitis and P. Acnes, respectively. When incubated at +37°C, complete elimination of all three<br />
strains (4 log killing) was observed within 24 hours.<br />
Conclusions<br />
Our results showed that BASE.128 eliminated effectively all investigated microorganisms and that the decontamination<br />
efficacy varied as a function of the incubation temperature. All investigated microorganisms were completely eliminated<br />
within 24 hours, when incubated at +37°C. Only P. Acnes was completely eliminated after incubation at +4°C. On the<br />
contrary, complete elimination of S. Epidermidis II and S. Capitis was not obtained even after 72h of incubation. Our results<br />
suggest that an exponential growth phase is required for efficient killing with BASE.128. Incubation at 37° might be<br />
considered as a part of the decontamination procedure in order to increase the skin decontamination effect.<br />
P-73<br />
UV LIGHT STIMULATES CELL PROLIFERATION BY EGR-1 ACTIVATION IN HUMAN<br />
PRIMARY DERMAL FIBROBLASTS OBTAINED FROM MULTIORGAN DONORS<br />
MARTÍNEZ-F, F.; BARRERA-L, A.; LOMELI-R, C.; MADINAVEITIA-V, J.; SANDOVAL-Z, H.; MACHUCA-R, C.; VILLEGAS-<br />
C, H., Skin and Tissue Bank of CENIAQ, National Institute of Rehabilitation. Mexico<br />
Abstract / Background:<br />
The EGR-1 protein is a finger zinc transcriptional factor with transcriptional activity on collagen type I and TGF-B gene in<br />
skin. This transcriptional activity could be is induced by UV, serum, hormones and drugs trough serum response elements<br />
(SRE), and CAMP response-like elements (CRE). Hereby we demonstrate conserved transcriptional regulation of Egr-1 in<br />
human dermal fibroblasts obtained from skinfoil allografts. Human cell culture: Human Primary Fibroblasts (HPF) were<br />
obtained from skingrafts procured on multiorgan donors. Samples of skin foil (0.5 cm2) were enzimatically digested isolated<br />
and cultured in standard conditions. Viral transduction efficiency was determinate on HPF at 25, 50, 75, 100, 125 MOI´s<br />
with an adenovector control (Ad-GFP), and using Laser Scanning Confocal Microscopy at 488 nm. Cells were starved by<br />
serum for 24 h and exposed to UV light at 254nm. Induction assay was performed in 1)FBS(+); 2)FBS(-); 3)FBS(-<br />
)/betamethasone(+). All conditions were grouped in A) UV(-) and; B). UV(+). The reporter activation was measured by<br />
luciferase activity assay at 3 and 6 hours and quantified by luminometry in a DTX50 Multilector.
<strong>POSTERS</strong><br />
Results:<br />
Optimal MOI of Ad-Egr-1-Luc7 in HFP was 50. The optimal dose of exposition to UV for induction of promotes transcriptional<br />
activity of AdEgr-1-Luc without cell death was 120”. In presence of UV light Ad-Egr-1-Luc7 (4.1X106LC), Luciferase<br />
maximum activity was observed at 6 hours (10 fold times more than Ad-CMV-Luc (4.5 X 105LC). Interestingly, the CMV<br />
activity is 2 fold times more over than Egr-1 in presence of SFB. For induction assays in the presence of betamethasone,<br />
luciferase activity shows a lag time to 3 hours.<br />
Conclusion:<br />
Hereby we demonstrate the functional activation of egr-1 promoter and correlates with cell proliferation, presumably induced<br />
by UV light in human primary fibroblast cultured from skin allografts. Additionally, UV light promotes cell proliferation at<br />
lower dose (15 seconds of exposure). Possible applications could be considered for preconditioning of tissues before<br />
preservation or therapeutically implantation on burned patients. Research granted by the Ministry of Health of Mexico and<br />
de ICYT-DF (PIFUT-08169).<br />
P-74<br />
MICROBIOLOGICAL ANALYSIS OF SKINGRAFTS PROCESSED BY THE SKIN & TISSUE<br />
BANK OF THE INR IN MÉXICO<br />
MARTÍNEZ-F, F.; LOMELI-R, C.; BARRERA-L, A.; VIZCAINO-D, A.; GUZMAN-M, K.; SANDOVAL-Z, H.; MADINAVEITIA-<br />
V, J., Skin and Tissue Bank of CENIAQ. National Institute of Rehabilitation. Mexico.<br />
Abstract / Background:<br />
The viable skingrafts is considered the best biological coverage in the treatment of severe burns. Unfortunately, common<br />
sterilization methods are not applicable for skin without compromise physiological, biological and structural properties.<br />
Previous reports about microbiological analysis have be shown contamination overmore of 50% of skin banked samples.<br />
Contaminant microorganism includes: E. coli, K. pneumoniae, S. aeurus, C. albicans and also the presence of microorganisms<br />
of slow growth. Hereby we present the analysis of microbiological screening of human skin foils allografts obtained and<br />
process according to the implemented process in the skin banking of the INR in México. Patients, tissues and methods. We<br />
analyzed 14 batches of skin foils procured from multiorganic donors under aseptic conditions into the surgical room. All<br />
procured tissues (0.5mm thick) were ablationated from the back body regions (Shoulder, gluteus and legs). Sequential and<br />
timed pre-surgical washing was used for decontamination process, based on clorhexidine, iodopovidone and isopropanol.<br />
Screening for microbiological control was performed for: 1) transport media, 2) washing solutions and 3) tissue samples for<br />
the detection of bacteria and fungi based on 14 days of culture in rich media for growth of aerobes, anaerobes and fungi<br />
(Thioglycollate broth, Sabouraud and blood agar). Visual test was used to report bacterial growth 1, 7 and 14 days.<br />
Results:<br />
1 of 14 batches (7.14 %) was positive for bacterial on blood agar plates after 24h of incubation. The resting 13 batches<br />
(92.86%) were negative for bacterial growth pattern (turbidity) at 7 and 14 days. However, the retrospective analysis to<br />
determine the possible infective focus, shows evidence of human field of a packaging process. A sample of a positive<br />
bacterial culture reports E coli as contaminant agent only in the washing solution. Positive batches was subjected in a second<br />
new microbiological assay with negative results.<br />
Conclusion:<br />
E. coli growth was due to contamination in the process of manipulation of the striatum of the plate and all tests were<br />
performed in triplicate and only one plaque was presented that colonial development, the sample was subjected to a second<br />
analysis without positive results. 99.7% of sample analyzed presented negative microbiological growth at 14 days. This<br />
preliminary analysis, shows a very low rate of contamination, and is possible due to: a) a carefully selection process for<br />
donors; 2) ablation of skingrafts like a surgical process in a surgical facilities and aseptic technique; 3) Effective<br />
decontamination process based on antibiotics and washing process may help to eradicate prolific bacterial load. Finally,<br />
the possible contamination in the handling of tissue during the process of cryopreservation is part of the learning curve<br />
process. Project granted by Ministry of Health of Mexico and the ICYTDF-PIFUT08-169.
<strong>POSTERS</strong><br />
P-75<br />
THE EXPERIENCE OF THE OPERATIVE MODEL THE SKIN AND TISSUE BANK OF INR<br />
IN MÉXICO<br />
MARTÍNEZ-F, F.; VIZCAÍNO-D, A.; LOMELI-R, C.; BARRERA-L, A.; ESTRADA-V, E.; VILLEGAS-C, H.; MADINAVEITIA-V, J.,<br />
Skin and Tissue Bank of CENIAQ. National Institute of Rehabilitation. Mexico.<br />
Abstract / Background:<br />
The tissue banking is a medical practice developed with scientific bases since 1960. However the international standards<br />
of quality for the operation on skin banks have been serious modified in the last decade due the technologic and scientific<br />
advance in cell biology, cryopreservation, molecular biology and biotechnology. In México the skin and tissue bank of<br />
National Institute of Rehabilitation (INR) founded in 2009 is the only one tissue bank that has the corresponding health<br />
license and the authorization for dispose organs, tissues and cells with therapeutic purpose in the extraction, preservation<br />
and release mode. Also being the only one in Latin America to implement a process focused to preserve cell viability of skin<br />
foil allograft from multiorganic donators, (less than 6 hours of death) using washing solutions, transport and preservation<br />
medium formulated by the own skin and tissue bank core facilities.<br />
Methods:<br />
The skin ablation process, like an others tissues is developed under aseptic conditions in a surgery room. The ablation zone<br />
is meticulously clean and desinfected before release the ablation. Them the obtained tissues are rinse in washing solution<br />
for 25 minutes (a set of buffers, antibiotics and antimicotics) and set in transport medium (high glucose iso-osmolar media<br />
and PH buffer agents) at 4ºC until translate it to the skin and tissue bank of the INR. Also blood samples are taken to be<br />
analyzed in the quality control process. In the bank, the tissues are processed and subjected to quality control by different<br />
testings that includes: The 14 days microbiological culture for tissue and transport medium to determinate presence of<br />
bacterial load and fungis. Biological safety test includes: a.) Hemoculture from the donator for aerobic and anaerobic<br />
microorganism, b) molecular probes based on Real Time-PCR ( to discard the presence of HIV, HBV, HCV, Citomegalovirus<br />
and Treponema pallidum) and c). Histopathological analysis. Once the tests are all negative and the biological security is<br />
guaranteed, the tissues are process and cryopreserved (Medium with glucose, osmolytes, buffers and antibiotics) at -80°C<br />
until be required. If not, the infected tissues are discarded. The cell viability assay is performed by the DCF method and<br />
analyzed by Laser Scanning Confocal Microscopy.<br />
Results:<br />
Viability assays recently performed to skin samples cryopreserved on 2009 shows more than 80% of alive tissue (n= 12).<br />
By other way there is not microbiological growth in the tissues as well as in the washing solutions.<br />
Conclusions:<br />
The correct selection of the donator and the ablation of the tissue within 6 hours after death guarantee high percentage of<br />
tissue´s viability in cryopreservation for a longer time. Low microbiological load in the tissues, has been reported with this<br />
operative model. Project granted by Ministry of Health of Mexico and the ICYTDF-PIFUT08-169.
P-76<br />
ACTIVITY IN THE RECENTLY ESTABLISH SKIN BANK OF TSF<br />
<strong>POSTERS</strong><br />
SAVÍO, A.; PÉREZ, M.; FARIÑAS, O.; HINOJOSA, M.; MIRANDA, M.; MANOTAS, C.; ZARDOYA, M., Tissue Bank, Transplant<br />
Services Foundation (TSF) - Hospital Clínic. Barcelona. Spain.<br />
Abstract / Introduction:<br />
Donor skin transplant is a valid therapy for skin loss due to extensive severe burns, trauma, surgery and several diseases<br />
such as bullous epidermolisis, diabetic and pressure ulcers. Deceased donor allograft skin provides temporary cover of the<br />
burn wound when autograft is not available. Lack of autologous split-thickness skin graft requires the use of temporary skin<br />
substitutes, being the human skin from deceased donors a good choice. Two preservation methods, glycerol and<br />
cryopreservation, are commonly used by tissue bankers for long-term storage, showing both to be effective. To evaluate our<br />
results this study has been carried out with the data available so far, composed by the returned follow ups forms of the skin<br />
distributed directly by our bank and part of the data collected from the intermediary banks. Aim: On one hand to know the<br />
general features of the skin donors, on the other hand to determinate if we are able to have available skin in stock to provide<br />
hospitals. For the last, we have considered the difference between the amount of recovered skin square centimeters (cm2)<br />
with the amount of the distributed one during 2009-1010. The last aim is to report the results obtained after transplantation<br />
of the skin delivered in the period 2009–2010.<br />
Methods:<br />
This retrospective study evaluates the amount of skin obtained and shipped to several hospitals in Spain and overseas. The<br />
general features of the shipped donors such as age, sex, type of donor, and mean body cooling time before retrieval have<br />
been reported. The clinical results from the recipients after transplantation were reviewed. The analysis was conducted in<br />
our tissue bank during the period of January 2009 -December 2010.<br />
Results:<br />
During the study period we have dispatched 590570 cm2 of skin from 229 donors, of which, 255700 were cryopreserved<br />
and 334870 were glycerol-preserved. At the time we have stored others 877666.40 (cm2) of suitable skin from 279 donors,<br />
of which, 363637.00 were cryopreserved and 514029.40 were glycerol-preserved. The viability rate obtained with these<br />
279 donors was 84%. Nowadays we have in stock more than 100000 cm2 of viable skin graft from deceased donor. Until<br />
now, we have never received any report of complications that could be related to the skin after transplantation.<br />
Conclusions:<br />
Skin recovered from human deceased donors, both, cryopreserved and glycerol-preserved, is a safe and effective choice<br />
to skin loss due to extensive severe burns and other diseases. Our institution is a high-volume skin graft provider and it is<br />
able to have in stock suitable skin graft to respond to the routine demand.
CORD BLOOD<br />
P-77<br />
USE OF AMNION IN THIRD DEGREE BURNS AND NON-HEALING WOUNDS<br />
LOBO GAJIWALA, A.; GAJIWALA, K., Tata Memorial Hospital. India.<br />
Abstract / Introduction:<br />
The use of freeze-dried, irradiated amnion in the management of second degree burns is well established. The present study<br />
assesses its possible role as a non-surgical option for the treatment of third degree burns and non-healing infected wounds.<br />
Materials and Method:<br />
Between June 2008 – 2011, 15 patients with third degree burns or infected, non healing wounds of various sizes were<br />
treated with freeze-dried, irradiated amnion obtained from the Tata Memorial Hospital Tissue Bank.<br />
Results:<br />
12 of the 15 patients managed with amnion dressings had complete wound healing. In the remaining 3 the wound size<br />
reduced. The dressing was painless, therefore avoided anesthesia. Reduced pain post dressing preempted the use of pain<br />
killers and allowed the patient to perform daily activities comfortably. One patient was hospitalized during the early treatment<br />
period. All the others were treated on an outpatient basis. Good healing was achieved within 28 to 60 days depending on<br />
the depth and extent of the wound. Hypertrophic scars were observed in 5 patients. Standard care of these patients would<br />
have involved surgical intervention entailing skin grafting with associated risks and costs. This was avoided.<br />
Conclusion:<br />
Freeze-dried, irradiated amnion dressing may provide an effective option to surgical intervention in wound healing,<br />
especially in patients who are either reluctant to or not fit for surgery.<br />
P-78<br />
EVOLUTION FROM 2007-2010 IN MALAGA CORD BLOOD BANK<br />
<strong>POSTERS</strong><br />
RIZO ALFARO, P.; PONCE VERDUGO, L.; GÓMEZ MALDONADO, P.; GALEOTE PADILLA, A.; HERNÁNDEZ LAMAS, M.;<br />
PRAT ARROJO, I., Málaga Cord Blood Bank. Spain.<br />
Background and objective:<br />
Over the years, in relation to the existence of Cord Blood Banks (CBB), quality parameters that measure the work system<br />
has been established, and they improve the own banks.<br />
The aim of this work is to define these parameters and apply them to CBB of Malaga to assess their current situation and<br />
optimize the available units for transplant.<br />
Material and methods:<br />
We have analyzed a retrospective study about the total sent units to Malaga Cord Blood Bank from 3 autonomous<br />
communities in Spain.<br />
In this study, we have analyzed several clinical parameters:<br />
– Received units.<br />
– Average of initial volume.
– Total Nucleated Cells (TNC).<br />
– Percentage recovery of TNC.<br />
– Shipped units for transplantation and their TNC.<br />
They settled on an annual average during 2007 to 2010.<br />
Results:<br />
Our results showed that the total received units were 27.822 and processed 13.286.<br />
The average of initial volume was 94.71 ml in 2007, 93.89 ml in 2008, 11.79 ml in 2009 and finally, 13.30 ml in 2010.<br />
The percentage recovery of TNC was 88.13% in 2007, 86.76% in 2008, 87.59% in 2009 and 87.79 in 2010.<br />
The shipped units for transplantation and their TNC were 18 units (2007), 23 units (2008), 34 units (2009) and 52 (2010)<br />
and average cellular were 13.3 x 108 (2007), 15.7 x 108 (2008), 17.7 x 108 (2009) 18.7 x 108 (2010).<br />
Conclusion:<br />
Malaga Cord Blood Bank has evolved positively.<br />
The transplant unit demand is keeping with more content units of TNC.<br />
It is necessary to continue optimizing the umbilical cord blood for transplantation.<br />
P-79<br />
VIABILITY AND FORMATION OF ERYTHROID, GRANULOCYTE/MACROPHAGE AND<br />
MEGAKARYOCYTIC COLONIES IN BLOOD HEMATOPOIETIC PROGENITORS AFTER<br />
20 YEARS STORED IN LÍQUID NITROGEN<br />
ROMERO VEGA, E.; MECA ROVAYO, M.; RIVERO ARIAS, G.; SALAT MARTÍ, A., Banco Sectorial de Tejidos de Cádiz.<br />
Spain.<br />
Introduction:<br />
Blood hematopoietic progenitor cells destined for allogenic transplantation, often are stored in liquid nitrogen (LN) for long<br />
periods of time.<br />
It is considered that in LN (-196ºC), any activity that would induce the degradation of these cells is paralyzed, remaining<br />
therefore intact their properties.<br />
At the same time, there has been demonstrated degradation of certain proteins at very low temperatures.<br />
Objectives:<br />
Study the viability and the capacity of proliferation of blood hematopoietic progenitors (BHP) stored for 20 years in LN.<br />
Materials and Method:<br />
We have tested ten samples of BHP criopreserved apheresis with DMSO to 10 % in the year 1990. In all samples were studied<br />
the viability (v) for two different methods, tripan blue and orange acridine; also erythoid (CFU-E), granulocyte/macrophage<br />
(CFU-GM) and megakaryocytic (CFU-MK) colonies that were obtained cultivating for duplicate 3x104 nucleate cells in<br />
MethoCultl ® and MegaCulttm-C ® mediums.<br />
In 2011, after more than 20 years stored, we have repeated the same studies, using the same methods.<br />
Results: Tables 1, 2 and 3<br />
<strong>POSTERS</strong>
Table 1. Results year 1990<br />
Sample V CFU-E CFU-GM CFU-MK<br />
1 78 36 25 14<br />
2 87 26 18 18<br />
3 79 25 17 22<br />
4 85 20 24 15<br />
5 74 42 23 16<br />
6 78 35 18 18<br />
7 82 37 18 14<br />
8 68 35 23 21<br />
9 76 38 21 17<br />
10 86 26 23 18<br />
9.3 32 21 17.3<br />
Table 2. Results year 2011<br />
Sample V CFU-E CFU-GM CFU-MK<br />
1 67 12 16 8<br />
2 82 18 13 13<br />
3 76 21 16 12<br />
4 86 22 18 5<br />
5 70 26 15 14<br />
6 79 25 13 16<br />
7 79 24 15 12<br />
8 67 22 3 16<br />
9 72 23 19 11<br />
10 76 21 10 9<br />
75.7 21.5 13.8 11.6<br />
<strong>POSTERS</strong><br />
Results table 3<br />
Average value 1990 Average value 2011 Difference p IC<br />
V 79.3 75.7 -3.6
P-80<br />
NEW GMP METHOD FOR CORD BLOOD CRYOPROCESSING WITH CLOSED TUBING<br />
SYSTEM AND PRE-COOLED GEL SLEEVES<br />
KUBESOVA, B.; KARKOSKOVA, L.; MATEJKOVA, E., Tissue Bank University Hospital Brno. Czech Republic.<br />
Abstract / Introduction:<br />
Precise proceeding of cryoconservation is fundamental for obtaining maximal numbers of thawed cells and it is very<br />
important especially in hematopoietic stem cell (HSC) transplantations. According to the actual European legislative new<br />
method of HSC processing is needed. We developed close system with pre-cooled gel sleeves for HSC manufacturing which<br />
is regardful of cells and in compliance with GMP standards. No clean rooms are required then.<br />
Material and Methods:<br />
For the process validation we used umbilical cord blood from eight healthy donors after the sign of informed consent. During<br />
the process we used only our designed system of bags and tubes, sterile tubing welder and gel sleeves for cooling the<br />
cryopreservative solution during the mixing with cell suspension.<br />
Results:<br />
We compared two methods of cord blood processing before freezing. In the first case, we used system of bags and tubes<br />
joined with three-way cocks. For cooling the cell suspension before adding the cryoprotectant (DMSO) we used standard<br />
water bath; the temperature inside the bag was 3.6-3.8°C, median 3.7°C). Via the temperature probe inside the bag, we<br />
noted significant increase in temperature during the addition of DMSO (12.4 – 12.7 °C, median 12.5°C). In the second case<br />
we used close system of bags and tubes joined with sterile welds. For cooling the cord blood (starting temperature: 3.5-3.8°C,<br />
median 3.6°C), we used pre-cooled gel sleeves. At the time of DMSO adding we logged out only slight temperature variation<br />
(4.2 – 4.8°C, median 4.7°C).<br />
Conclusions:<br />
In Department of Hematopoietic Stem Cells in Tissue Bank of UH Brno, new method of HSC proceeding before<br />
cryopreservation was developed. In this technique, specially designed gel sleeves for bags were used. Previous cooling of<br />
these cases minimizes the risk of warmth damage of the cells during the process of their mixing with cryoprotectant containing<br />
(DMSO). This method was approved with Czech national authority (SUKL).<br />
P-81<br />
INVENTORIES: MALAGA CORD BLOOD BANKING<br />
<strong>POSTERS</strong><br />
RIZO ALFARO, P.; PONCE VERDUGO, L.; GÓMEZ MALDONADO, P.; FALCÓN MORALES, F.; MAYORGA PASTRANA,<br />
C.; HERNÁNDEZ LAMAS, C. M.; PRAT ARROJO, I., Málaga Cord Blood Bank. Spain.<br />
Background and objective:<br />
Umbilical cord blood (UCB) is a source of hematopoietic precursors for transplantation. The creation of UCB banks (UCBB)<br />
in 1992 led to the storage of units for unrelated transplant. The distribution of the historical inventories is not homogenous<br />
concerning the cell content of the units and many units are not therefore suitable for adults. The aim of this study was to<br />
analyze the UCBB inventory and calculate the economic impact of the current process per unit of UCB stored, in order to<br />
determine future expectations.
Material and Methods:<br />
Three study periods were defined: the first, from 23 January 1996 to 9 January 2006 with TNC (Total Nuclear Cell)<br />
acceptance levels for processing of 4-6 x 10e8 and a manual processing system; the second, from 1 October 2006 to 30<br />
July 2010, with automated processing and variation in the number of TNC from 8-10 x 10e8; and the third, from 1 January<br />
2009 to 30 June 2010, with an automated Sepax-BioArchive procedure and starting TNC >10 x 10e8. Within each period<br />
various ranges of cryopreserved TNC units were established: A, >16.2 x 10e8; B1, from 12.5-16.1 x 10e8; B2, from 5,2-<br />
12.4 x 10e8; and C,
P-83<br />
INFLUENCE ON TIME TO CRYOPRESERVATION<br />
PONCE VERDUGO, L.; GÓMEZ MALDONADO, P.; RIZO ALFARO, P.; GALEOTE PADILLA, A.; MARTÍN BEJAR, T. M.;<br />
HERNÁNDEZ LAMAS, C. M.; PRAT ARROJO, I., Málaga Cord Blood Bank. Spain.<br />
Abstract / Background and objective:<br />
It is accepted that the Umbilical Cord Blood (UCB) units can be cryopreserved to 48 h after their harvest and the delay in<br />
this procedure influences the final quality. The aim of this study was to determine the influence on time to cryopreservation<br />
in Umbilical Cord Blood Units.<br />
Material and Methods:<br />
A selection of the 2134 samples arrived to the UCB Bank from 36 Andalusian linked hospitals was analyzed for a period<br />
of six months. The date and hour of collection, hospital of origin, time in the hospital, time of transport, time of arrival to the<br />
bank to cryopreservation, viability and CD34+ cells were determined.<br />
Results:<br />
Our results showed that the median time from harvest to cryopreservation was 27.9 h. However, a great variability was<br />
found (range: 5.3 to 47.6 h), which was dependent on transport variables: time from collection to leave the hospital<br />
(median 12.6 h, range: 0.1-28.8), time of transport from hospital to UCB Bank (median 1.8 h, range: 0.1-20.6) and time<br />
kept in the bank before freezing. Viability of cells were reduced directly proportional in samples that took longer to be<br />
cryopreserved (93.9 vs 96.6%, p=0.001), by contrast, CD34+ count did not differ significant at shorter or longer times of<br />
cryopreservation.<br />
Conclusion:<br />
The delay in transport time and in processing reduce the umbilical cord units quality, consequently, the time of transport and<br />
storage previous to cryopreservation must be reduced less than 36 h.<br />
P-84<br />
UMBILICAL CORD BLOOD:ECONOMIC STUDY<br />
<strong>POSTERS</strong><br />
PONCE VERDUGO, L.; HERNÁNDEZ LAMAS, C. M.; RIZO ALFARO, P.; PÉREZ PARRADO, S.; ESPAÑA RAMOS, T.; ARE-<br />
NAS AGUILAR, P.; PRAT ARROJO, I., Málaga Cord Blood Bank. Spain.<br />
Abstract / Background and objective:<br />
Malaga Cord Blood Bank started its activity in 1996, and this bank now receives UCB from 61 maternity units. The aim of<br />
this study was to analyze the cord blood inventory, calculate the economic impact of the current process per unit of stored<br />
UCB in order to establish future expectations.<br />
Material and Methods:<br />
A total of 20,762 units processed between 1996 and 2010 were studied. In 2006 a volume reduction system was initiated<br />
with HES, automated separation (Sepax®), programmed freezing and automated storage (BioArchive®).The cost of the UCB<br />
units processed by Sepax® was 720.41 euros per unit.
<strong>POSTERS</strong><br />
Results:<br />
The cost of the UCB units processed was 720.41 euros per unit, the collection cost was 17.1 euros, which included a<br />
Macopharma® collection bag, tubes, containers, forms and various collection material. The processing cost was 310.15<br />
euros, which included validation, separation by Sepax Biosafe®, cryopreservation and storage using BioArchive®, as well<br />
as administrative expenses. This heading also included sample storage (fetal red blood cells, fetal and maternal plasma bank,<br />
and tissue fragment). The quality control included HLA typing, flow cytometry (CD34+), viability, cell count blood cultures,<br />
ABO and Rh grouping, and testing for transmissible diseases (HCV, HBV, Lues, HIV, Chagas, Malaria). The data for the<br />
personnel were studied for each person involved throughout the process: midwife, laboratory technician, administrative<br />
staff, professional staff. The analysis did not include units that were not processed.<br />
Conclusion:<br />
The implantation of quality systems, economic impact and their legislation have all contributed to the continual improvement<br />
in the quality of stored UCB units. Umbilical cord blood is scarce and expensive, and the protocols for obtaining it should<br />
be optimized and involve careful selection of donors in order to reduce the number of units discarded.<br />
P-85<br />
MALAGA CORD BLOOD BANK: EVALUATION THE UNITS SENT FOR TRANSPLANT<br />
PONCE VERDUGO, L.; HERNÁNDEZ LAMAS, C. M.; RODRÍGUEZ RUIZ, Y.; NIETO CHUPS, V.; ROMAN MORILLAS, C.;<br />
TORNAY GÓMEZ, A. A.; PRAT ARROJO, I., Málaga Cord Blood Bank. Spain.<br />
Abstract / Background and objective:<br />
Umbilical cord blood (UCB) is a source for the transplant of hematopoietic precursors. The creation of UCB banks (UCBB)<br />
in 1992 led to the storage of units for unrelated transplant. Malaga Cord Blood Bank started its activity in 1966 and this<br />
bank now receives UCB from 61 maternity units. The aim of this study was to analyze the cord blood inventory, evaluate<br />
the units sent for transplant to establish future expectations.<br />
Material and Methods:<br />
A total of 20,762 units processed between 1996 and 2010 were studied and a total of 172 umbilical cord blood units were<br />
sent to transplant and distributed for all over the world.<br />
Results:<br />
The number of units sent for transplant has increased in proportion to the number of units stored and the increase in the<br />
number of TNC. The units were transplanted in Spain (23.7%), the rest of Europe (42.7%), USA (25.4%) and elsewhere<br />
(8.2%). The mean TNC of the units sent for transplant was 14 x 108 (4.6 x 10e8 to 36.5 x 10e8).<br />
Conclusion:<br />
The implantation of quality systems, their legislation and the use of practices based on scientific evidence have all contributed<br />
to the continual improvement in the quality of stored UCB units to transplant.
P-86<br />
CONTINUING EDUCATION MALAGA CORD BLOOD BANK<br />
<strong>POSTERS</strong><br />
GÓMEZ MALDONADO, P.; PONCE VERDUGO, L.; HERNÁNDEZ LAMAS, C. M.; RIZO ALFARO, P.; PRAT ARROJO, I.,<br />
Málaga Cord Blood Bank. Spain.<br />
Abstract / Background and objective:<br />
Malaga Cord Blood Bank received 9684 UCB (umbilical cord blood) units in 2010 and were accepted 40%. They were<br />
obtained in 59 linked maternities of three Spanish geographical areas (0-600 km). It is the responsibility of the Bank, within<br />
a system of quality management, initial and continuing training of the collection facilities depending on it. The training<br />
program began in 2006 and is developed by the responsible external quality. Since that time the program has continued<br />
to grow in scope and currently performs in 59 maternities. It consists of classroom training, developed in maternity and aimed<br />
at all health staff involved in donations of UCB. The main aim of this study was to analyze the period 2009-2010 and<br />
compared with the previous year. In 2009-2010 more than 600 people attended these courses.<br />
Material and Methods:<br />
The aim of this program is to prepare and recycle healthcare professionals involved in donations and have an update list<br />
of people trained according to standards of quality NETCORD and Joint Committee on Acreditation. The specific objectives<br />
are: ● To provide participants with the knowledge and skills necessary to obtain UCB. ● To promote and encourage<br />
donations. ● Get UCB under the procedures of Cord Blood Bank. ● Improve the quality of the units received. ● Implement<br />
a methodology of work under a quality system. - Audiovisual material: slides. - Support for material donation. Results: There<br />
was a tendency to increase in 11.3 ml volume of the units in 2010, increase the number of samples received (in 2007: 4922<br />
units, in 2008: 5068 units, 2009: 8148) and decreases the percentage of UCB discarded for reasons related to the hospital<br />
of origin. Conclusion: ● Improving the quality of the units received. ● Consistency of documentation. ● Awareness of<br />
teamwork. ● Promotion of communication links: -Audiovisual material: slides. -Support material donation.<br />
P-87<br />
THE CORD BLOOD BANK OF THE RED CROSS BLOOD TRANSFUSION SERVICE OF<br />
UPPER AUSTRIA<br />
HENNERBICHLER, S.; PETERBAUER-SCHERB, A.; PLÖDERL, K.; HAVLICEK-PÖTSCHER, M.; SÜSSNER, S.; GABRIEL, C., Red<br />
Cross Blood Transfusion Service of Upper Austria, Linz/ Austrian Cluster for Tissue Regeneration. Austria.<br />
Abstract:<br />
The Cord Blood Bank of the Red Cross Blood Transfusion Service of Upper Austria is the first public cord blood bank in<br />
Austria. It was founded in 2002 as part of the blood bank. The Red Cross Blood Transfusion Service of Upper Austria is<br />
certified according to the EC Directives 2004/23/EC, 2006/17/EC, and 2006/86/EC for cord blood and several tissues,<br />
as well as GMP- and ISO9001:2008- approved. At the beginning cord blood units were obtained from a single hospital<br />
next to the cord blood bank. In the meantime donations are already sent to the cord blood bank from eight hospitals located<br />
in Upper Austria. Initially, processing of the donations was done manually according to the method by Rubinstein et al.<br />
(1998) but since 2004 cord blood units are processed automatically on the Sepax System (Biosafe, Switzerland). Freezing<br />
and storage is performed with the BioArchive System (Thermogenesis, USA). In 2010 a total of 911 donations arrived at<br />
the cord blood bank. From these obtained donations 211 (23.16%) fulfilled the quality criteria for processing which resulted<br />
in 183 cord blood units (20,09%) that were finally stored in the BioArchive. Since 2002 about 1,400 units were stored in<br />
our cord blood bank. Based on our participation in the Austrian Stem Cell Register “Geben für Leben”<br />
(www.stammzellspende.at) since 2007, three products were already released and shipped for transplantation. Furthermore<br />
we are currently completing our participation in Cytolon (www.cytolon.de).
P-88<br />
SEMI-AUTOMATED HIGH-THROUGHPUT NEXT GENERATION SEQUENCING BASED<br />
HLA-TYPING FOR CORD BLOOD SAMPLES<br />
HENNERBICHLER, S.; STABENTHEINER, S.; NIKLAS, N.; HOFER, K.; PRÖLL, J.; DANZER, M.; GABRIEL, C., Red Cross<br />
Blood Transfusion Service of Upper Austria, Linz/ Austrian Cluster for Tissue Regeneration. Austria.<br />
Abstract:<br />
To overcome some of the intrinsic HLA-typing problems, we established a new, sequence-based method using Roche / 454<br />
Life Science’s sequencing technology. Applying this high-throughput technology offers the possibility for ambiguity-free<br />
typing and high level automation. The use of the clonal pyrosequencing approach with approximately 450 base pairs long<br />
sequence reads covering the whole exons made a straightforward amplicon strategy feasible. For exon 1-7, where<br />
appropriate, we developed HLA-A, -B, -C, -DP, -DQ and -DR locus specific primers. For amplicon generation target specific<br />
primers and GS-FLX specific adaptors were elongated with sample-identifying barcode sequences to trace each of the 40<br />
samples which represent the current maximum loading capacity of one single GS-FLX sequencing run. Sequencing protocols<br />
were performed according to supplier’s instructions without modifications. But, we set up protocols for Hamilton robotics<br />
based amplicon production and post emulsion-PCR bead preparation and present correspondent HLA-typing results.<br />
Although the great number of samples was identified and typed correctly we currently work to establish improved protocols<br />
for problematic allele combinations and automated HLA allele assignment with less manual editing needed and to deepen<br />
our data pool consequently. The presented system can be used for low or high allele resolution in dependence of the used<br />
amplicons for high-throughput typing of cord blood samples.<br />
GLIMPSE INTO THE FUTURE; ADVANCES THERAPIES<br />
P-89<br />
HEPATIC CELLULAR THERAPY UNIT, IIS-HOSPITAL LA FE: ESTABLISHMENT OF<br />
HEPATIC CELL BANK FOR CELLULAR TRANSPLANTATION<br />
<strong>POSTERS</strong><br />
BONORA-CENTELLES, A.; PAREJA, E.; CORTÉS, M.; MIRABET, V.; MIR, J.; CASTELL, J.; GÓMEZ-LECHÓN, M., Instituto<br />
de Investigación Sanitaria - Fundación para la Investigación Hospital La Fe. Spain.<br />
Abstract:<br />
The creation of a regulatory-compliant cell bank is an essential element in the production of uniform biological products.<br />
Cell banking systems assure that a uniform population of cells is preserved, that their integrity is maintained and that a<br />
sufficient supply of material is readily available for the life of the product. Of equal importance is the maintenance of these<br />
banks in a secure, controlled and monitored storage environment. Hepatocyte transplantation is an alternative therapy to<br />
orthotopic liver transplantation for the treatment of liver diseases. The application of hepatocytes for transplantation purposes<br />
demands high-quality cell preparations, which depend to a large extent on the nature of the tissue used for cell harvesting.<br />
However, isolation, cryopreservation, and thawing processes can seriously impair hepatocyte viability and functionality.<br />
The Hepatic Cellular Therapy Unit-IIS La Fe (UTCH) began its activity in 2007, and since then it has processed more than<br />
forty livers under ISO standards. The main aim of this unit is to offer excellent batches of cells that allow restoring hepatic<br />
function loss or maintaining hepatic function while the patient is on the waiting list. In the present work, we summarize the<br />
recent effort of the UTCH at our hospital for the purpose of establishing a cell bank of cryopreserved human hepatocytes<br />
for clinical application. To achieve this goal we focused on: a) expanding the liver donor pool by exploring the suitability
of livers from non-heart-beating donors (NHBDs) and from neonates as a potential source of hepatocytes for transplantation;<br />
b) analyzing the influence of the length of the cold ischemia time on the outcome of isolated hepatocytes; c) improving the<br />
cryopreservation protocol; and finally, d) assessing the functionality of hepatocyte preparations with a view to promoting<br />
customized cell preparation for each receptor. For this purpose, viability, attachment efficiency and metabolic functionality<br />
(ureogenic capability, cytochrome P450 and phase II activities) were assayed prior to clinical cell infusion to determine the<br />
quality of hepatocytes. Moreover, the evaluation of urea synthesis from ammonia and UDP-glucuronosyltransferase 1A1<br />
activity, a newly developed assay using beta-estradiol as substrate, allows the possibility of customizing cell preparation for<br />
receptors with urea cycle disorders or Crigler-Najjar Syndrome type I. Sources of human liver and factors derived from the<br />
procurement of the liver sample (warm and cold ischemia) have also been investigated. The results show that grafts with a<br />
cold ischemia time exceeding 15 h and steatosis should not be accepted for hepatocyte transplantation. Finally, livers from<br />
NHBDs and neonates are a potential suitable source of hepatocytes which could enlarge the liver donor pool.<br />
P-90<br />
CLINICAL TRIAL COMPARING PLATELET-RICH PLASMA TO BETAMETASONA AND<br />
BUPIVACAIN SUBACROMIAL INJECTION IN ROTATOR CUFF TENDYNOSIS.<br />
PRELIMINARY RESULTS<br />
PACHA, D.; PACHA VICENTE, D.; LLUSÁ, M.; NARDI, J.; NAVARRO, A.; RODRÍGUEZ, L.; GENIS, X.,<br />
Vall d'Hebron Hospital. Spain.<br />
Abstract:<br />
The platelet-rich plasma (PRP) is an ultraconcentrate of peripheral blood containing growth factors that, as some studies<br />
demonstrate, favor the healing process of soft tissues. The PRP has been used to treat tendinitis in some areas of the human<br />
body, epicondylitis, patelar tendinitis, aquilean tendinitis … The chronic tendinitis of the rotador cuff, pathology very<br />
prevalent among the active population, also could to be treated with PRP. The present prospective, randomized and double<br />
blind clinical trial, tries to compare the therapeutic effect of two alternative treatments in a group of patients with subacromial<br />
syndrome due to chronic tendinitis without rotator cuff tear. The new treatment will consist on subacromial injection of<br />
autologous PRP and the other one will consist on subacromial injection of betametasona acetate and bupivacaina, as it is<br />
used to do in the clinical practice while its reported little efect. PRP is prepared following the Anitua method. The injection<br />
is guided with ultrasounds. We show the preliminary results which support PRP usage with the first 30 patients.<br />
P-91<br />
ESTABLISHING THE SNBTS ISLET ISOLATION LABORATORY<br />
<strong>POSTERS</strong><br />
MITCHELL LEIGH, D.; MCQUILLAN, T.; DRAIN, J.; CASEY, J.; GALEA, G.; MCGOWAN, N., SNBTS Tissues & Cells Directorate.<br />
Edinburg, UK.<br />
Abstract:<br />
SNBTS Tissues and Cells in conjunction with NHS Lothian recently received funding from the Scottish Government to establish<br />
a pancreatic islet cell transplantation service for Type I diabetic patients presenting with poor glycaemic control and/or<br />
awareness, often associated with numerous hypoglycaemic events. Over the last 18 months SNBTS Tissues and Cells have<br />
installed and commissioned a new Grade B/C processing facility and established the islet isolation service (based on the<br />
Edmonton Protocol), to the appropriate GMP and legislative standards, creating in excess of 150 controlled documents and
completing over 20 individual validations. The process of islet cell isolation (as described in the 'Edmonton' protocol, Shapiro<br />
et al. NEJM. 2000) is laborious and requires several members of staff working concurrently. Once the organ is received it<br />
is decontaminated before being cannulated and perfused with collagenase enzyme until visibly distended. The organ is<br />
then cut into pieces and placed within a chamber in the presence of warm (active) collagenase, which is recirculated through<br />
the chamber and associated circuit. The combination of mechanical and enzymatic digestion results in a crude islet<br />
preparation that can then be purified on the basis of density using a COBE 2991 cell processor. Purified islets are quantified<br />
before being placed in culture for a period of 24-48 hours, enabling additional tests/checks to be completed (endotoxin,<br />
viability), prior to clinical/QA release. Islets are then transplanted into the portal vein of the patients' liver under local<br />
anaesthetic. The clinical service was established in December 2010 and the first transplant took place in February 2011,<br />
the first of its kind in Scotland and one of just over 20 in the UK. Early indications suggest the graft was extremely successful<br />
as evidenced by a reduction in insulin requirements, reduced hypoglycaemic events, restoration of glycaemic awareness and<br />
production of c-peptide, a marker of endogenous insulin production. A second transplant in the same patient 6 weeks later<br />
has led to insulin independence.<br />
P-92<br />
HUMAN AMNIOTIC MEMBRANE AS A SOURCE OF PROGENITOR CELLS FOR<br />
HUMAN ARTICULAR CARTILAGE REPAIR<br />
<strong>POSTERS</strong><br />
RENDAL VÁZQUEZ, ME. 1 , MUIÑOS LÓPEZ, E. 2 , DÍAZ PRADO, S. 2,3 , HÉRMIDA GÓMEZ, T. 2 , SANLUIS VERDES, A. 1 ,<br />
FUENTES BOQUETE, I.2,3 , DE TORO, FJ 2,3 , ANDIÓN NÚÑEZ, C. 1 , BLANCO GARCÍA, FJ. 2 , 1 - Cryobiology Unit Complejo<br />
Hospitalario Universitario ACoruña, 2 - INIBIC Complejo Hospitalario Universitario ACoruña, 3 - INIBIC Complejo<br />
Hospitalario Universitario ACoruña INIBIC University of ACoruña. Spain.<br />
Abstract / Purpose:<br />
There is an increasing interest about human amniotic membrane (HAM) for its use in the field of regenerative medicine. Two<br />
kinds of progenitor cells can be removed from the amniotic membrane, the amniotic mesenchymal stromal cells (hAMSC)<br />
and amniotic epithelial cells (hAEC). Both cell types are capable of differentiating towards three germinal cell lines. It has<br />
large advantages over the other sources of progenitor cells. The aim of this study is to determinate the usefulness of the<br />
hAMSC and the hAEC for regenerating human joint cartilage in an in vitro model.<br />
Methods:<br />
HAM was used as support for the culture of hAMSCs or hAECs. Focal injuries in human joint cartilage biopsies were done.<br />
Later, a pellet of cells (hAMSCs or hAECs, depending on the repair model developed) was implanted into the focal defects<br />
of cartilage. HAM, with the cells grown on it, was placed in direct contact with the cartilage surface to be repaired. These<br />
implants were cultured in DMEM+10% FBS for 8 weeks. The repair tissues were analyzed by histological and histochemistry<br />
analysis considering the ICRS macroscopic evaluation of cartilage repair.<br />
Results:<br />
hAMSCs and hAECs cultured on HAM and transplanted onto focal injuries of cartilage penetrated into the nearby surface<br />
of the chondral defect. The Hematoxylin and Eosin staining showed that the hAMSC or hAECs pellet filled the chondral<br />
defect. There was a good integration between the repair tissue and native cartilage. Type II collagen and aggrecan stainings<br />
of repair tissue were slightly positive on the extracellular matrix, and positive inside the cytoplasm of the cells. The safranin<br />
O staining expressed the presence of proteoglycans. Finally, type I collagen stainings were weak or totally negative. We<br />
realized an ICRS macroscopic evaluation of cartilage repair to compare both kinds of progenitor cells in the in vitro model.<br />
The hAMSCs displayed better degree of defect repair, greatest integration to border zone and, in general, higher repair<br />
assessment.
<strong>POSTERS</strong><br />
Conclusion:<br />
We get a reduction of the area of the defect with quality integration. The morphology of the repair tissue showed a<br />
fibrocartilaginous appearance and a high cellularity. The hAMSCs showed better results considering the ICRS macroscopic<br />
evaluation of cartilage repair.<br />
P-93<br />
COMPARISON OF DIFFERENT METHODS OF GENOMIC DNA EXTRACTION AND<br />
PURIFICATION FROM TISSUE<br />
ISIDRO MARRÓN, P.; ASTUDILLO GONZÁLEZ, A.; MARTÍNEZ CAMBLOR, P.; VALLINA ÁLVAREZ, A.; FERNÁNDEZ JUÁ-<br />
REZ, L., BioBanco Hospital Universitario Central de Asturias-Oficina de Investigación Biosanitaria del Principado de Asturias<br />
(OIB). Spain.<br />
Abstract / Introduction:<br />
In a Tissue Biobank, genomic DNA (gDNA) extraction and purification from tissue are indispensable. A method’s<br />
optimization for this extraction and purification to find a reproducible procedure are necessary to obtain high performance<br />
and elevated concentration and hereby improve the samples quality. OBJECTE Evaluate the amount and quality of gDNA<br />
obtained by different extraction and purification methods from frozen lung tissue.<br />
Material and Methods:<br />
In this study, five gDNA extraction and purification methods were compared: three methods based on silica membrane<br />
columns: gDNA from Tissue by MACHEREY-NAGEL (method 1), ReliaPrepTM gDNA Miniprep System by PROMEGA<br />
(method 2), QIAAMP® DNA Mini and Blood Mini Kit by QIAGEN (method 3); and two methods based on alcohol<br />
precipitation: Gentra® Pueregene® by QIAGEN (method 4) and ArchivePure DNA Purification by 5 PRIME (method 5). We<br />
tested 10 samples for method and each sample was adjusted to 30 mg wet weight before storage at -80°C. All samples<br />
were extracted from an only donor’s surgery. Chemical (Proteinase K digestion) and mechanical (shaking at 56°C for 24<br />
hours) pre-treatment are used to facilitate an efficient homogenizing and lysis. gDNA obtained are eluted or dissolved in<br />
100 µL of Buffer Solution and shaking for 2 days at room temperature. To evaluate the amount and quality we use<br />
spectrophotometer Nanodrop® by ThermoFisher. Express gDNA concentration on ng/µL. The absorbance ratios at 260 and<br />
280nm (A260/280) and at 260 and 230nm (A260/230) are used to assess the purity of nucleic acids. For pure gDNA,<br />
A260/280 and A260/230 are larger than 1.8. Continuous variables are described by medians and non-parametric tests<br />
are used in order to compare them.<br />
Results:<br />
Concentration: methods 1 and 2 are homogeneous (P-value 0.123 and medians 160.60 and 199.27 ng/µL); methods 3,<br />
4 y 5 are homogeneous too (P-value 0.285 and medians 48.12, 48.02 and 54.80 ng/µL). Medians of methods 1 and 2<br />
are clearly larger than medians of methods 3, 4 and 5. A260/230 ratio: similar than Concentration; methods 1 and 2<br />
homogeneous (P-value 0.165 and medians 2.17 and 2.11) and methods 3, 4 and 5 homogeneous too (P-value 0.295 and<br />
medians 0.85, 0.78 and 0.85). A280/230 ratio: for this criterion, methods 1 and 3 are homogeneous (P-value 0.739 and<br />
medians 1.86 and 1.84) and methods 4 and 5 too (P-value 0.909 and medians 1.99 and 1.98). The method 2 obtained<br />
the higher median (2.06).<br />
Conclusion:<br />
Due to the observed medians of the ratio A260/280 were bigger than 1.8 (usual cut-off value), all considered methods yield<br />
High Quality gDNA. However, methods 1 and 2 obtained concentrations and A260/230 -ratios larger than the ones<br />
derived from the methods 3, 4 and 5. Therefore, it can be concluded that the best method among the five studied ones was<br />
the 2. The results get for this method are clearly the best ones for all considered criteria.
<strong>POSTERS</strong><br />
P-94<br />
HUMAN MESENCHYMAL CELLS OF ADIPOSE TISSUE INHIBITS THE PRODUCTION OF<br />
PGE2 IN HUMAN MONOCYTES<br />
PLATAS, J.; MIRABET, V.; VILLAMÓN, E.; OLIVAR, T.; ALCARAZ, J. M.; GUILLÉN, I. M., Dpt. Pharmacology, University of<br />
Valencia and IDM. Spain.<br />
Objective:<br />
t is known that mesenchymal stem cells have immunomodulatory capabilities. However there is very little documentation about<br />
what mechanisms may be involved. Prostaglandin E2 (PGE2) is a lipid mediator that acts by enhancing the inflammatory<br />
response. Numerous cell types produce PGE2 in response to other inflammatory mediators. In this work we studied the<br />
effect of mesenchymal stem cells from human adipose tissue (CMTAh) on PGE2 production in human monocytes from<br />
peripheral blood.<br />
Methods:<br />
CMTAh were obtained from 3 healthy individuals lipectomy by digestion with collagenase I. The resulting cell suspension<br />
was incubated until the confluence with growth medium (GM: DMEM/F12 containing 15% human serum and antibiotics).<br />
Phenotypic characterization of the cell population was performed by flow cytometry using the following markers: CD45,<br />
CD34, CD90, CD105 and CD13. A different time, the culture medium (MA) was collected, centrifuged and frozen at - 80<br />
°C. Human monocytes were isolated from 5 buffy-coats by Ficoll density gradient. The suspension of nucleated cells obtained<br />
was seeded at a density of 106 cells / ml with medium DMEM/F12 containing 15% human serum and antibiotics. Monocytes<br />
were selected for their ability to adhere to culture support. Phenotypic characterization was performed using markers CD11c<br />
and CD14. After 12 h of incubation, monocytes were stimulated with LPS (1ng/ml) with GM or MA for 24 hours. PGE2<br />
production was measured by radioimmunoassay.<br />
Results:<br />
Phenotypic characterization showed that CMTAh are CD13, CD90 and CD 105 positive cells, being variable in the case of<br />
CD34 (which decreases with time in culture), and lack CD45. Monocytes were characterized as cells positive for CD45,<br />
CD11c and CD14. In monocytes incubated with DMEM/F12 and treated with LPS there was a significant increase in PGE2<br />
production compared to the basal cells (23 ± 2 ng/ml vs. 1.3 ± 0.7 ng/ml, p
P-95<br />
HEPATOCYTE ISOLATION FROM LIVERS NOT SUITABLE FOR WHOLE ORGAN<br />
TRANSPLANTION. LIVER CELL THERAPY<br />
<strong>POSTERS</strong><br />
M. MANYALICH 1-2 ; J. MEYBURG 3 , WOLFGANG RUEDINGER 4 ; MP GÓMEZ 5 ; 1. Research Biomedical Foundation.<br />
Barcelona Clínic Hospital. Barcelona, Spain. 2 Transplant Coordinations Service. Barcelona Clínic Hospital.<br />
Barcelona Spain. 3 Department of General Pediatrics, University Children’s Hospital, Heidelberg. Germany.<br />
4 Cytonet GmbH&Co KG, Weinheim, Germany. 5 Donation & Transplantation Institute DTI. Barcelona Spain<br />
Abstract / Introduction:<br />
The development and use of new clinical cell therapies from llivers not suitable for whole organ transplantation could improve<br />
the problem of scarcity of donor livers. Urea Cycle Disorders (UCD) in children is a rare group of inherited metabolic<br />
diseases that have a poor prognosis despite optimal conservative treatment. Early liver transplantation before the onset of<br />
neurological damage may cure the disease. However, the obtainment of livers compatible for this kind of recipient is difficult<br />
due to the small size of the recipients. Liver cell transplantation (LCT) may be a good alternative to early liver transplantation,<br />
as it is less invasive and can be performed during the first weeks of life.<br />
Material and Methods:<br />
A liver network was created with defined rules and criteria for organ selection. Collaborative centers were coordinated and<br />
logistic and communication processes were established. Two steps were defined to link hospitals to the network. LCT was<br />
performed on four patients with severe UCD with neonatal onset and poor prognosis.<br />
Results:<br />
The network started in 2003 and linked hospitals in Spain, Germany, Italy and Portugal. So far, 99 livers have been obtained<br />
and a total of 250,3 billon cells with an average cell viability of 77% have been isolated. The Cryopreserved human liver<br />
cells were transplanted in the four children through the portal vein after thawing. Metabolic stabilisation was shown in all<br />
patients. However, one patient died after four months from a fatal decompensation triggered by infection and poor<br />
compliance regarding immunosuppression. In the remaining three children, metabolic crises vanished after LCT, and<br />
ammonia levels remained in the normal range. One girl is doing well on conservative therapy 28 months after LCT. In the<br />
two other boys, subsequent liver transplantation was performed 10 and 15 months after LCT. Both host organs were<br />
investigated after retrieval. Whereas the activities of the affected enzymes were around 0% prior to LCT, total enzyme<br />
activities of 4.5% and 15.6% of healthy controls were found in the explanted livers. These enzyme activities are in the range<br />
of mildly affected heterozygotes.<br />
Discussion:<br />
The promising results of the first clinical series of liver cell transplantation show that this innovative method may be a suitable<br />
option for the treatment of various hepatic-based diseases. It is possible to obtain viable cells from livers rejected for<br />
transplantation. Because of the implication involved in using organs for research or other uses other than transplant, it is<br />
necessary to clearly identify and define the steps involved in organ procurement and distribution.
P-96<br />
DIFFERENCES IN SURFACE MARKERS EXPRESSION AND CHONDROGENIC<br />
POTENTIAL AMONG DIFFERENT TISSUE-DERIVED MESENCHYMAL CELLS FROM<br />
ELDERLY PATIENTS WITH OSTEOARTHRITIS<br />
SANZ MORENO, J.; DESPORTES BIELSA, P.; ALEGRE AGUARÓN, E.; GARCÍA ÁLVAREZ, F.; MARTÍNEZ LORENZO, M.,<br />
Servicio de Inmunología del Hospital Clínico Universitario Lozano Blesa. Spain.<br />
Abstract:<br />
Mesenchymal stem cells (MSCs) are self-renewing cells with multipotential capacity that could be used to repair or regenerate<br />
cartilage, which is damaged by some diseases such as osteoarthritis (OA). These cells are present in some adult and fetal<br />
tissues, including bone marrow [Prockop, 1997], adipose tissue [Zuk et al., 2002], synovium [De Bari et al., 2001],<br />
peripheral blood [Zvaifler et al., 2000], umbilical cord blood [Erices et al., 2000; Mareschi et al., 2001] and term placenta<br />
[Fukuchi et al., 2004]. In this study, we have used bone marrow, adipose tissue (from articular and subcutaneous locations)<br />
and synovial fluid samples from 18 patients with knee OA in order to find an alternative source of bone marrow for the<br />
isolation of MSCs with a high chondrogenic potential. MSCs from all the tissues showed a fibroblastic morphology, but their<br />
rate of proliferation was different: while subcutaneous fat-derived MSCs proliferated faster than bone marrow and Hoffa´s<br />
fat pad-derived MSCs, the growth rate of synovial fluid-derived MSCs was slower. The phenotype of the MSCs from the four<br />
different analyzed tissues was similar, with some isolated differences such as CD36 and CD54 expression. The highest<br />
expression for these surface markers was in subcutaneous fat-derived MSCs, which differentiated poorly towards hyaline<br />
cartilage. Synovial fluid-derived MSCs presented only a medium chondrogenic differentiation capacity, while Hoffa´s fat padderived<br />
MSCs showed a great chondrogenic potential. In conclusion, MSCs from elderly patients with osteoarthritis still<br />
showed a suitable chondrogenic potential, but it depended strongly on their tissue of origin. In conclusion, from all our<br />
results, bone marrow-derived MSCs showed a proliferation pattern and chondrogenic potential similar to that of Hoffa´s fat<br />
pad-derived MSCs in elderly patients with osteoarthritis, while subcutaneous fat-derived MSCs differentiated poorly toward<br />
hyaline cartilage and synovial fluid-derived MSCs proliferated slower.<br />
P-97<br />
ADIPOSE TISSUE BANKING FUTURE PROSPECTS: FIRST STEP, SET UP OF<br />
CRYOPRESERVATION METHODOLOGY<br />
<strong>POSTERS</strong><br />
AGUSTI, E.; BRUGADA, P.; VILARRODONA, A.; MARTÍNEZ, L.; PÉREZ, L. M.; MARTÍNEZ-CONESA, E.; TRÍAS, E.,<br />
Transplant Services Foundation-Hospital Clínic. Barcelona. Spain.<br />
Abstract / Background:<br />
Nowadays, adipose aspirates are widely used in plastic surgery for conducting reconstructive interventions. Fat transfer is<br />
defined as harvesting fat from parts of the body where it accumulates for transferring into other body locations in the same<br />
patient. Unfortunately, inflammation and congestion of the treated area often do not allow using all lipo-aspirated tissue. In<br />
addition, some part of the transferred material is reabsorbed and more interventions are needed for achieving the expected<br />
results. The leftover fat has to be thrown away if no preservative method is used. There are several published studies of<br />
adipose tissue potential applications like the obtainment and culture of adipose-derived stem cells (ADSC) and the isolation<br />
of extra cellular matrix (ECM) for tissue engineering.
Objectives:<br />
The aim of this study was to set up an optimal, safe, reliable and cost effective lipoplasty-derived adipose aspirates<br />
cryoprotective methodology that ensures the viability of the tissue after cryopreservation.<br />
Methods<br />
The study was mainly conducted in female patients undergoing reconstructive surgery after suffering breast cancer. Raw<br />
adipose material obtained from conventional lipoplasty was transported to the laboratories at room temperature. When it<br />
arrived, a microbiological control was done. The intermediate phase of adipose aspirates was collected after centrifugation<br />
and each specimen was randomized into 4 groups: control group (fresh adipose aspirates without preservation); simple<br />
cryopreservation group (adipose aspirates preserved with our established cooling and thawing protocol); complex<br />
preservation group 1 (adipose aspirates preserved with our protocol using 0,35M Trehalose as cryoprotective agent (CPA));<br />
complex preservation group 2 (adipose aspirates preserved with our protocol using 0,25M Trehalose + 0,5M DMSO as CPA<br />
solution). Before next step, another microbiological control was done to asses the processing methodology. Cryopreservation<br />
of adipose aspirates was conducted with controlled slow cooling and fast rewarming rates. Fresh or cryopreserved adipose<br />
aspirates in each group were evaluated by viable adipocyte counts, glycerol-3-phosphate dehydrogenase (G3PDH) assay,<br />
and routine histology.<br />
Results:<br />
At this moment the project are in an embryonic stage. Since we have started, we have been setting up the optimal controlled<br />
slow cooling program for two kinds of cryopreserving containers (cryovials and cryobags), counting viable adipocytes from<br />
fresh samples and making microbiological controls for the assessment of the arriving samples.<br />
Conclusion:<br />
More samples are needed to achieve statistically reliable results. However, our first experiences are encouraging. In terms<br />
of cellular viability, results show that room temperature is the best condition to transport samples; also we can say that our<br />
established cooling and thawing protocol seems to treat with care adipose tissue.<br />
P-98<br />
INTRA-ARTICULAR KNEE INJECTION OF PLATELETS RICH PLASMA TO TREAT<br />
OSTEOARTHRITIS<br />
<strong>POSTERS</strong><br />
RODRÍGUEZ, L.; GENIS, X.; TARRAGONA, E.; ORTEGA, I.; GOMARIZ, E.; NAVARRO, A.; JOSHI, N., Banc de Sang i Teixits.<br />
Spain.<br />
Abstract / Introduction:<br />
Platelets rich plasma (PRP) is autologous plasma containing thrombocytes in a concentration above normal blood counts.<br />
Due to its high platelets concentration and therefore of growth factors, cytokines and plasmatic bioactive proteins, PRP is<br />
considered a promising and useful biological product in wound healing and tissue regeneration. In orthopedic surgery, PRP<br />
has been largely used for tendon or ligament injury treatments as well as to medicate lesions of muscular tissue. Nevertheless,<br />
the benefit of PRP in cartilage injuries, specifically those with a degenerative origin, has not been clearly determined.<br />
Hypothesis PRP intra-articular injections in osteoarthritic knees reduce pain and improve knee function.<br />
Objective:<br />
To determine the clinical usefulness of PRP in the knee osteoarthritis treatment regarding pain, joint function, and quality of<br />
life improvement. Methodology Thirty patients with painful knee osteoarthritis were treated with three intra-articular PRP<br />
injections. Quality life score (SF36), knee function score (KOOS) and visual analogue pain score (VAS) were used for clinical<br />
evaluation. Complications, adverse events and patient satisfaction were also recorded.
Results:<br />
We evaluated 30 patients, 10 men and 20 women, with mean age of 62,63 years. The mean BMI (body mass index) was<br />
29, most of the patients presented degenerative chondral lesion Kellgren 3. All of the evaluated parameters improved at 1<br />
and 3 months follow-up visits. Quantitatively, VAS improved from 7,13 points pre-treatment to 4 and 3,7 at 1 and 3 months<br />
post-treatment respectively. SF36 improved from 55 points to 60 and 59 for the same time points of study. Additionally all<br />
the 5 items evaluated in KOOS also improved.<br />
Conclusions:<br />
Our results indicate that administration of PRP injections can reduce pain and improve knee function and quality of life at<br />
short-term. Further studies are needed to find out which platelet concentration and which frequency of infiltrations provides<br />
better and more durable results.<br />
P-99<br />
COLD STORAGE OF PRIMARY HEPATOCYTES IN TIPROTEC VARIANTS<br />
<strong>POSTERS</strong><br />
PLESS-PETIG, G. 1 , SAUER, I. M. 2 , RAUEN, U. 1<br />
1 - Institut für Physiologische Chemie, Universitätsklinikum Essen, 2 - Klinik für Allgemein-, Viszeral- und Transplantationschirurgie,<br />
Charité, Berlin. Germany<br />
Abstract:<br />
For the use of primary hepatocytes in liver cell transplantation or extracorporeal liver support, short term storage options<br />
are required. However, diverse cells are very vulnerable towards cold-induced cell injury. Previous research in adherent rat<br />
hepatocytes and other cell types showed that cold-induced injury is iron dependent and that rat hepatocytes in addition suffer<br />
a chloride-dependent injury. Based on studies in adherent cells and isolated vessels we recently developed the tissue<br />
preservation solution TiProtec®, which is chloride-rich, contains several amino acids and two iron chelators. Here, we tested<br />
whether this solution or variants thereof can be used for the cold storage of adherent human hepatocytes and whether it might<br />
also be useful for the cold storage of hepatocyte suspensions. Adherent rat and human hepatocyte cultures and rat hepatocyte<br />
suspensions were stored in diverse variants of TiProtec®, cell culture medium or organ preservation solutions (UW, HTK) at<br />
4° C, rewarmed/seeded in cell culture medium, and cell injury, cell attachment and metabolic function (resazurin conversion,<br />
gluconeogenesis) were assessed. Survival after cold storage and rewarming was significantly improved in adherent rat and<br />
human hepatocytes in variants of the solution TiProtec®. However, while rat hepatocytes survived best in a chloride-poor<br />
variant of TiProtec®, human hepatocytes survived best in the chloride-rich TiProtec®, both with an increased deferoxamine<br />
concentration compared to TiProtec®. In this solution, human adherent hepatocytes maintained > 60 % of their viability (LDH<br />
retention) after 3 weeks of storage, largely exceeding results in UW solution or cell culture medium (< 10 % viability after<br />
2 weeks of storage). After 1 week of storage in TiProtec with the higher deferoxamine concentration, metabolic activities were<br />
similar to unstored control cells, after 2 weeks of storage 60-80% of control cells. Rat hepatocytes were then used to test the<br />
potential of these solutions for the storage of cell suspensions. The chloride-poor variant also proved best in suspensions of<br />
these cells. After one week of cold storage in this solution cell attachment rate was 76 ± 24% of control cells, far better than<br />
after storage in cell culture medium (1 ± 2%), UW solution (4 ± 2%) or HTK solution (2 ± 6%). The main protective effects<br />
could be allotted to glycine/alanine which inhibited injury during cold storage, and iron chelators and absence of chloride<br />
which improved cell attachment. Attached cells displayed normal morphology and metabolic function was similar to unstored<br />
adherent control cells. In summary, it is possible to significantly improve viability of adherent human hepatocytes and<br />
attachment ability and post-attachment function in rat hepatocyte suspensions using variants of TiProtec®. Current studies<br />
assess the use of TiProtec® or variants for the storage of human hepatocyte suspensions.
<strong>POSTERS</strong><br />
P-100<br />
BIOLOGICAL PROPERTIES OF POROUS BIOMATERIALS OF THE CONNECTIVE TISSUE<br />
ORIGIN AND POTENTIALS OF THEIR USE IN SURGERY<br />
MUSLIMOV, S.; KHASANOV, R.; SHANGINA, O.; MUSINA, L.; KORNILAEVA, G.; KHASANOVA, Y., Russian Eye and<br />
Plastic Surgery Center. Russian Federation.<br />
Abstract:<br />
One of the possible ways to expand the allogeneic biomaterial spectrum for restorative surgery is a structural modification<br />
of donor tissues. We have developed a technology of the physico-chemical treatment which allows to obtain porous<br />
biomaterials as well as to increase the period of their biodegradation following the implantation thanks to the structure<br />
modification of the initial tissues. Aim of the investigation. To study the biological properties of allogeneic porous biomaterials<br />
and outline the prospects for their use in restorative surgery.<br />
Materials and Methods:<br />
Two series of the experiments were carried out to achieve this goal. In the first series two types of the biomaterials varying<br />
in resistance to collagenolytic enzymes were subcutaneously implanted to 48 Vistar rates. In the second series the<br />
reconstruction of the eye anterior chamber angle drainage zone was performed with the aid of the porous biomaterial on<br />
36 grey rats with modeled glaucoma. To study the dynamics of the morphological changes in the operated on zone, the rats<br />
were being sacrificed after 14, 30, 60, 90, 180 and 360 days following the surgery. The histological sections were<br />
hematoxilin-eosin stained as per Mallory and Van Giezon. There were also used transmission and scanning electron<br />
microscopy methods. In the second series tonographic methods of the investigation were as well used.<br />
Results:<br />
In case of the subcutaneous implantation during the early period after the surgery, the biomaterials were infiltrated my<br />
macrophages and in the following the collagenous fibres were subjected to a gradual enzymatic lysis and synchronically<br />
were substituted by the newly-formed ones. There is established a biomaterial degradation period dependence of the forming<br />
regenerate structure. In the first group of the animals a full substitution of the biomaterial was observed after 90 days<br />
following the implantation which led to the formation of the regenerate with a relatively loose collagen fibrous framework.<br />
In the second group the regenerate was being formed in later periods (120 days) and had a dense fibroarchitectonics. In<br />
the second series of the experiments the porous biomaterial in the reconstructed drainage zone of the eye anterior chamber<br />
angle was being impregnated in the early period by the aqueous humour which led to the normalization of the intraocular<br />
pressure. Thereafter the collagen fibres of that part of the biomaterial which faced the eye anterior chamber were not<br />
subjected to biodegradation. The walls of the biomaterial cells were being gradually covered by the regenerating endothelium<br />
and thus a drainage zone of full value was being formed. At the back part of the biomaterial there were taking place a<br />
degradation of collagen fibres and the substitution by the newly-formed tissue with a well-developed venous channel. The<br />
afore-said morphological changes led to the ophthalmotonus normalization and restoration of the optic nerve structure.<br />
Conclusion:<br />
The use of porous biomaterials exhibits promise for the development of new high effective restorative surgeries in different<br />
fields of surgery and ophthalmology.
P-101<br />
A DECELLULARIZATION PROCESS IN TISSUE BANKING<br />
RAMOS CARRO, J.; IGLESIAS MUÑOZ, F.; LÓPEZ LAGUNA, M.; DE LA PUENTE GARCÍA, P. M.; VÁZQUEZ REYERO, J.<br />
J.; FERNÁNDEZ GONZÁLEZ, A., Tissue Establishment San Francisco Clinic Foundation. Spain.<br />
Abstract:<br />
Decellularization is an important tool when we consider the tissue engineering area but it can also plays a significant role<br />
to improve the service offered in a tissue bank. This process is based in the complete elimination of the cellular component<br />
although maintaining the extracellular matrix as intact as possible. Most of the decellularization processes are linked to the<br />
use of a detergent, as a chemical method to ensure cell death, and they are followed by a washing step to remove the<br />
cellular material. It has been described that putative remains of the detergent in the decelllularized scaffold could have an<br />
adverse effect in cellular recolonization. We have optimized a decellularization protocol replacing the use of a detergent<br />
component by a combination of hypotonic and hypertonic solutions. Histological techniques and viability assays have shown<br />
that it could be possible to get a complete decellularization avoiding the use of detergents. This modified protocol has been<br />
applied in several tissues such as cornea, muscle, heart valve, tendon, pericardium, bone, sclera… According to the sample<br />
size, the time in the decellularization solutions has varied between short times for smaller and longer times for bigger ones.<br />
This protocol has kept the same decellularization results on the different tissue sources. In the next step, it’s needed to evaluate<br />
the recellularization potencial of these acellular scaffold obtained. The main goal is that the implanted decellularized tissue<br />
has biological activity and mechanical integrity to support the cell migration from the surrounding tissues. An additional<br />
option it’s to maintain the acellular scaffold “in vitro” with a cell culture obtained from the own tissue receptor. A tissue bank<br />
has to adequate the new science tools to improve his service in order to ensure the quality of his products. We have found<br />
that this decellularization process may possibly be the key to the most immediate future.<br />
CORNEA<br />
P-102<br />
ASSESSMENT OF SUBJECTIVE PARAMETERS OF PATIENTS WITH AUTOLOGOUS VS<br />
HETEROLOGOUS EYE DROPS TREATMENT IN OCULAR PATHOLOGY<br />
GUIJARRO, L.; GUIJARRO-HERNÁNDEZ, L.; BENÍTEZ DEL CASTILLO, J.; PONCE-VERDUGO, L.; GÓMEZ-MALDONADO,<br />
P.; HERNÁNDEZ-LAMAS, M.; PRAT-ARROJO, I., Hospital de Jerez de la Frontera. Spain.<br />
Introduction:<br />
Autologous serum is an effective method to stimulate viability of corneal-conjunctival epithelial cells,due to its content in<br />
growth factors, which are shortfall in dry eye syndrome associated with ocular surface diseases (OSD). It’s considered the<br />
last therapeutic stage after numerous pharmacological options. Because of the impossibility of extraction in some patients,<br />
it might be done with donor’s heterologous serum.<br />
Objective:<br />
Evaluation of the effectiveness of the autologous Vs heterologous eye drops in patients suffering OSD.<br />
<strong>POSTERS</strong><br />
Material and Method:<br />
Retrospective observational study using a validated OSD status self-assessment questionnaire, completed for the each patient<br />
and comparing before and after the treatment score, of 18 differents items (red eyes, lid inflammation, flakes in lashes, sticky
<strong>POSTERS</strong><br />
eyes in the morning, discharge, dry eye, sandy and foreign body sensations, burning, itching, discomfort, pricking, tearing,<br />
teary eyes, photophobia, transitory blurring vision, lid reddening and heavy sensation).<br />
Results:<br />
Study sample has two groups: heterologous serum eye drops (28 patients: Sjögren syndrome 16 and neurotrophic<br />
Keratopathy 12) and autologous serum eye drops (33 patients: Sjögren syndrome 23 and neurotrophic Keratopathy 10)<br />
Mean age is 68,36 (SD= 14,66) years in heterologous group and 61,67 (SD= 15,94) in autologous group. Proportion<br />
according to sex is 6/28 men and 22/28 women in heterologous group and 5/33 men and 28/33 women in autologous<br />
group. Mean questionnaire scoring before the treatment for each group is 35,18 (SD= 11,33) in heterologous group and<br />
41,24 (SD= 16,95) in autologous group. There are not statistycal significant differences between groups for these variables.<br />
Mean administrated vials is 21,86 (SD= 22,35) in heterologous group and 55,68 (SD= 58,79) in autologous group, (p=<br />
0.0058).<br />
Exclusion causes for autologous serum eyes drops elaboration are: 15 cases of impossibility for extraction (53,57 %); 7 HCV+<br />
(25 %); 3 HBV+ (10,71%), 1 HIV+ (3,57%) and 2 cases of hematologic pathology (7,14%)<br />
Subjective improvement defined as the difference between the mean punctuation before and after the treatment results<br />
statistically significant with p
<strong>POSTERS</strong><br />
normative legal base of cadaver tissue donor regulations and cell technologies, an ophthalmic care in the field of surgical<br />
treatment of corneal pathology and anterior segment of the eye. The Section is organized with the purpose to realize basic<br />
trends of development for the Russian ophthalmic and transplantation science and practice in the field of tissue donor<br />
selection and transplantation, cell technologies and surgical treatment of corneal pathology and anterior segment of the eye.<br />
P-104<br />
MARKERS TO IDENTIFY LIMBAL STEM CELLS ON HUMAN AMNIOTIC MEMBRANE<br />
RENDAL VÁZQUEZ, M.; SANLUIS VERDES, A.; YEBRA-PIMENTEL VILAR, M.; LÓPEZ RODRÍGUEZ, I.; DOMENECH GAR-<br />
CÍA, N.; ANDIÓN NÚÑEZ, C.; BLANCO GARCÍA, F., Unit of Criobiology Complejo Hospitalario Universitario A Coruña.<br />
Spain.<br />
Abstract / Objectives:<br />
To evaluate the culture of a limbal biopsy on human amniotic membrane (HAM): directly on the chorionic side and on intact<br />
epithelium, and the expression of the stem cell associated markers: ABCG2, p63.<br />
Material and Methods:<br />
Eyes are collected within 24 hours post-mortem from human cadavers, with permission. The limbal epithelial fraction is<br />
scraped of and collected and sent to the unit of cryobiology in antibiotic solution (DMEM, 1% Penicillin and Streptomycin).<br />
Human placentas are obtained from Caesarean Section. The amnio is separated from the chorion of the placenta and then<br />
incubated in 199 medium with antibiotic solution for 24h at 4ºC and cryopreserved. Two biopsies are carried out directly<br />
in fresh after the extraction of the limbal biopsy. In 13 biopsies, the biopsy of the limbus is placed in a culture dish and<br />
exposed for 5-10 minutes to Dispase (1.2U/ml) at 37ºC. The amnio is placed on a Cell Crown insert that permits<br />
immobilization of the sample. Then the biopsy is placed in the middle of the amnio directly on the epithelial side (9 biopsies)<br />
or on the chorionic side (4 biopsies). Histological and Immunohistochemical studies are performed to evaluate certain<br />
markers: p63 and ABCG2, directly in fresh limbal biopsies and in limbal biopsies after expansion on HAM.<br />
Results:<br />
Fresh limbal samples show that the limbus had a multilayered epithelium with the basal layer arranged in a palisade pattern<br />
and that the palisades of Vogt in the superior limbus are structured with papilla-like columns. Histological studies show that<br />
both the chorionic and epithelial sides of the HAM showed a characteristic pattern with 1-2 layers of cell growth. However,<br />
on the chorionic side the monolayer of cells is sometimes not attached directly to the membrane causing the release of limbal<br />
epithelial cells cultured upon it. In our study, in fresh and after 3-4 weeks, the p63 protein was immunodetected in the nuclei<br />
of the limbal epithelial basal layer, but not in most limbal suprabasal layers and the ABCG2 transporter protein is primarily<br />
immunodetected in the cell membrane and cytoplasm of certain limbal basal epithelial cells, but not in most limbal suprabasal<br />
cells.<br />
Discussion:<br />
Our analysis confirmed that a normal limbal epithelium is formed on the HAM. This epithelium is 1-2 cell layers thick. The<br />
basal layer of cells shows high expression of the putative limbal stem cells markers p63, ABCG2 and intact HAM is a good<br />
substrate to culture limbal stem cells but on the epithelial side preferentially and not on the chorionic side.
P-105<br />
CIRCUIT FOR IMPLEMENTING A DONATION OF EYE TISSUE<br />
CABREJAS, A., CANO, M. P., Hospital de Bellvitge. Spain.<br />
<strong>POSTERS</strong><br />
Abstract / Introduction:<br />
To avoid possible loss of donor corneas in our hospital developed a system for detecting all exitus. This defined a channel<br />
circuit to the warnings of the patients who died in asystole and the family does not know the donation.<br />
Objectives:<br />
Increase cornea donation in a tertiary hospital. - Identify potential loss of organ donors METHODOLOGY We designed a<br />
circuit to give notice to the coordination of transplant nurses, in charge of the program on a schedule (Monday to Friday<br />
from 8h to 15h) and providing the number of seeks. Also designed a brochure that was included in the folder of the patient<br />
for more knowledge. The circuit was circulated at the meeting to supervisory and emergency room attendants who provide<br />
death certificates in our hospital. The supervisors explained the circuit to staff their units both physicians and nurses. The<br />
nurse transplant coordination to notice, assess the case and if there is no contraindication to the interview should be familiar<br />
to the application of the will of corneal donation. If the family agrees the documentation is completed, blood samples are<br />
obtained and alerts the ophthalmologist for removal.<br />
Results:<br />
In one year (June 2010 - June 2011) of operation of the circuit is detected 40 warnings, 37.5% accepted the donation and<br />
62.5% did not become a donor for several reasons: a refusal rate of 35% and 27.5% according to our protocol of<br />
contraindications for donor selection of fabrics: 2 older than 80 years and multimorbidity, 6 hematology diseases, 1<br />
pseudomonas colonized by HIV and MDR-1. Notice I made the flight attendants 77%, 10% nurses, medical staff 7.5% and<br />
5% nursing supervision. For slots, from 9am to 11am, 57%, from 11h to 13h 12% 13h to 15h 30% of the ads. Were obtained<br />
from 15 donors and 30 corneas taken real.<br />
Conclusion:<br />
Activation of a multidisciplinary program detection alerts the patient died in-hospital avoids the loss of potential cases and<br />
increase the donor pool because many families unaware of the possibility of donation by various criteria such as age,<br />
underlying disease or other.<br />
P-106<br />
EVOLUTION OF OCULAR TISSUE COLLECTION IN AN ACUTE CARE HOSPITAL FROM<br />
1995 TO PRESENT DAY<br />
MARIN FERNANDEZ, I.; VALLEJO OLEA, P.; TOBARUELA RASCÓN, P.; RUSIÑOL BOIXADERA, J.; AZAGRA LEDESMA,<br />
M.; QUINTANA RIERA, S., Hospital Universitari Mútua Terrassa, Social Work. Spain.<br />
Introduction:<br />
Social workers meet with the families after a death that has occurred at our centre. This means that other of their<br />
responsibilities is to ask about eye tissue donation when there are not medical contradictions.<br />
There have been changes in the criteria for acceptability of eyes tissue causing more rejections of the potential donors.
Objective:<br />
To analyse differences between 4 periods based on changes in the acceptance criteria of eye tissue and socioeconomic and<br />
cultural changes that may affect the donation, with the hypothesis that despite using more restrictive criteria have achieved<br />
more donations.<br />
Material and Method:<br />
nine months of 1995, 1999, from April 2005 to March 2006, and the first half of 2011 (daytime only) make up the four<br />
periods mentioned. Age and sex from the prospective donor, effective donation, and type of ceremony (burial vs. cremation)<br />
were collected. We used descriptive statistics, Chi2 and ANOVA.<br />
Results:<br />
There have been collected data from 1277 requests for corneas of 1946 deaths, which were obtained in 4 periods totalling<br />
16 years. The mean age of potential donors was 73 years (SD 15), with differences between the fourth quarter with the<br />
previous periods (74 vs. 71, p = 0.001). Tendency to increase the number of women 40% in the first period to 48% in the<br />
last (p = ns). By changes in selection criteria, they asked 100% in the first period, 78% in the second, and 33% and 42% in<br />
the third and fourth respectively (p
variable frame analysis method. The final ECD was the mean of all ECDs. We made a retrospective analysis of keratoplasty<br />
procedures performed in one hospital by three surgeons from the years 2008 to 2010. Statistical evaluation of baseline ECD<br />
among donor age groups was performed using Pearson correlation test.<br />
Results:<br />
Between January 2005 and December 2010, 840 corneas were received and evaluated in our eye bank. Eligible corneas<br />
were from donors 2 to 80 years old with a measured endothelial cell density from 1235 to 4505 cells/mm 2 . Corneas from<br />
2- to 60-years old donors had a median baseline cell density (2731 cells/mm 2 ) higher than that of corneas from donors 66<br />
to 80 (2404 cells/mm 2 ). Median ECD was 2516 cells/mm 2 for the graft failure cases and 2641 cells/mm 2 for the nonfailure<br />
cases (p=0.17). Although donor age correlated significantly with endothelial cell density (r=- 0.56), this association appears<br />
to have no effect statistically significant on the transplant outcome.<br />
Conclusions:<br />
The ECD declines with age in the normal cornea, however cornea ECD and donor age are unrelated to graft failure. Our<br />
results indicate that older donor corneas could be considered suitable for transplant.<br />
P-108<br />
ANALYSIS OF FACTORS AFFECTING THE VIABILITY OF CONEAL TISSUE<br />
<strong>POSTERS</strong><br />
GENÍS, X., RODRÍGUEZ, L., TARRAGONA, E., ORTEGA, I., GOMARIZ, E., NAVARRO, A. Banc de Sang i Teixits. Spain.<br />
Abstract / Introduction:<br />
Allogenic corneal tissue transplantation is an effective therapy for restoring vision in patients affected for inherited or acquired<br />
cornea diseases. Medical, technical and scientific activity of tissue banks can be considered, after donation, the main actor<br />
to provide corneas for this medical treatment. Major concern in tissues bank is to make better use of donated tissues<br />
minimizing the proportion of grafts rejected for transplantation. Banc Teixits is the division responsible for the procurement,<br />
processing and distribution of tissues of the Banc de Sang i Teixits (BST). Purpose Examine exclusion causes which determined<br />
corneal graft rejection for transplantation in the BST. Identify critical points to increase the number of viable tissues for clinical<br />
use.<br />
Material and Methods:<br />
We performed a retrospective review of the corneas received in BST from January 2009 until June 2011. We have evaluated<br />
the following parameters: age, ischemia time, whole-globe enucleating versus cornea retrieval and causes of rejection. The<br />
causes of rejection were divided into: medical history, serologies, microbiology, technical incidence and tissue quality control.<br />
Results.<br />
The number of corneal tissue donors were: 202 in 2009 (394 total corneas obtained with a 26.9% rejection), 245 in 2010<br />
(472 corneas with a 30.7%) and 134 donors during the period January-June 2011 (266 corneas with a 25.1% of rejection).<br />
The median age of cornea donors was 67 years (2-81) and total ischemia time was 6 hours (1-8) in average. Whole-globe<br />
enucleating versus cornea has varied from 49 % in 2009, 62% in 2010 to 76% in 2011. Regarding the causes of rejection,<br />
which has suffered major variation is tissue quality control (QC), representing the 48 % of rejections in 2009 (51 corneas),<br />
36% on 2010 (53 corneas) and 33% in 2011 (22 corneas). In the period studied during the 2011, QC rejection causes were:<br />
40% stromal edema, 26% endothelial damage, 22% epithelium defects, 7% inadequate free diameter and 5% injury during<br />
retrieval.<br />
Conclusion:<br />
During the period 2009-2011 our organization has been able to increase the number of transplanted corneas. Analyzing<br />
this improvement we identified the decrease of rejection by QC as a major cause although not having a significant higher
number of cornea donors. Among all the variables studied; age and ischemia time variables were constant parameters.<br />
Increased whole globe enucleation in front of cornea button excision seemed to directly influence the enhancement of viable<br />
corneas registered. Finally, protocolized cornea culture, optimized cornea allocation avoiding caducity as well as the<br />
advantage offered by new surgical techniques has permitted the transplant of a higher number of corneas. We believe that<br />
improvements at any step of the process including screening of the donor, retrieval, processing and client services can<br />
reduce the number of corneas rejected and thus increase the number of transplants.<br />
P-109<br />
COMMUNICATION BETWEEN TISSUE BANK AND IMPLANTING SURGEON:<br />
SHORT-TERM FOLLOW-UP OF TISSUES FOR OCULAR SURGERIES<br />
<strong>POSTERS</strong><br />
NÚÑEZ, V., AGUSTÍ, E., MARTÍNEZ, E., OTERO, N., PÉREZ, M., CASAROLI-MARANO, R., VILARRODONA, A., TRÍAS, E.<br />
Transplant Services Foundation. Hospital Clínic de Barcelona. Universidad de Barcelona. Spain.<br />
Abstract / Introduction:<br />
Traceability of tissue grafts in recipients is a crucial tool to asses the quality of distributed tissues by tissue banks. Follow up<br />
forms result extremely useful to know the evolution of patients and if possible incidents such as adverse effects or reactions<br />
occur in a short term basis. Through follow up forms the tissue bank is also informed about other parameters such as clinical<br />
diagnosis, characteristics of the recipients and microbiological issues which are also of a great importance for both<br />
traceability and for the tissue bank.<br />
Purpose:<br />
This study is focused on a short term monitoring of ocular tissue transplanted. To conduct this study, specific follow up forms<br />
have been sent together with the distributed graft. Follow up forms have been reported back by transplant centers to the tissue<br />
bank and data has been collected for analysis between April 2010 and April 2011.<br />
Results:<br />
During this period, 46 scleral grafts, 312 amniotic membrane grafts and 644 sclero-corneal buttons have been distributed.<br />
Response through follow up forms was 21.7% for scleral tissue, 15.7 % for amniotic membrane and 40.4 % for corneal<br />
buttons. All scleral grafts forms reported refer to patients with glaucoma to which grafts have been used to covering drainage<br />
tube surgery devices. Regarding amniotic membrane grafts, main indications were ulcer, chemical burns and corneal<br />
perforation. In corneal buttons, Penetrating Keratoplasty was the main use reported. However, during 2011, Anterior<br />
Lamellar Keratoplasty has significantly increased. Transplant indications were mainly corneal edema, different type of<br />
corneal dystrophies, corneal opacities and keratoconus. Microbiological control was varied depending on the reported<br />
tissue: no evidence of microbiological tests regarding scleral grafts, in amniotic membrane tissue 32 % of the centers reported<br />
negative microbiological tests while 52 % did not conduct any test or did not answer. For corneal buttons, 67 % reported<br />
negative results, 5.7 % were positive and 27.4 % did not conduct any test or did not answer. Postsurgical information about<br />
the evolution of the patients was very similar for the three studied tissues resulting in 98.46 % of reported satisfying evolution.<br />
Conclusions:<br />
The analysis of follow-up forms is very helpful to improve traceability of transplanted grafts. Unfortunately, it has been<br />
observed that only 31.5% of the follow-up forms have been reported. This fact indicates that communication between<br />
transplant centers and tissue banks need to improve in order to achieve a complete feedback of transplanted tissues and,<br />
therefore, permit tissue banks to offer adequate, safer and high quality tissues.
<strong>POSTERS</strong><br />
P-110<br />
A COMPARISON OF DIFFERENT CULTURE MEDIA FOR THE STORAGE OF HUMAN<br />
DONOR CORNEAS FOR GRAFTING<br />
ZLACKA D.; KRABCOVA, I.; KORTUSOVA, J. AND JIRSOVA, K., Laboratory of the Biology and Pathology of the Eye, Institute<br />
of Inherited Metabolic Disorders and Ocular Tissue Bank, General Teaching Hospital and 1st Faculty of Medicine,<br />
Charles University in Prague, Czech Republic.<br />
Abstract / Purpose:<br />
To compare the influence of different culture media on the qualitative and quantitative endothelial parameters of human donor<br />
corneas stored in organ culture for grafting.<br />
Methods:<br />
Thirty-one paired human corneas obtained from the Ocular Tissue Bank Prague were stored in organ culture for 28 days<br />
at 31°C in the following media: Minimal Essential Medium with Earle's salts and 2% Foetal Bovine Serum (EMEM)/EMEM<br />
with 5% dextran prepared by the bank staff, commercial media adhering to Directive 93/42/EEC: a) TissueC/CarryC<br />
(Alchimia, Italy) and CorneaMax/CorneaJet (Eurobio, France). Comparisons of EMEM vs. Alchimia (G1) and EMEM vs.<br />
Eurobio (G2) were performed on paired corneas. Corneal transparency, endothelial cell density (ECD), live endothelial cell<br />
density (LECD), the percentage of dead endothelial cells (% DC), their hexagonality (6A) and coefficient of variation (CV)<br />
were assessed. The assessment was performed before storage (1st assessment), at day 28, i.e. before transfer to deswelling<br />
medium (2nd assessment), and after 24 hours in deswelling medium (3rd assessment). The evaluations were performed<br />
from phase contrast and bright field photographs using a semiautomatic computer analysis system. A two-tailed Student's<br />
t-test was used for the statistical analysis. P values
1ª Jornada Iberoamericana<br />
de Bancos de Tejidos y Células<br />
Barcelona, 8 de noviembre 2011<br />
Asociación Española<br />
de Bancos de Tejidos<br />
Asociación Lationoamericana<br />
de Banco de Tejidos
1ª Jornada Iberoamericana de Bancos de Tejidos y Células<br />
Auditorio Centre Salut Mental. Sant Boi de Llobregat. Barcelona<br />
COMITÉS<br />
Comité organizador<br />
Elba Agustí<br />
Oscar Fariñas<br />
Josep M. Segur<br />
Esteve Trias<br />
Anna Vilarrodona<br />
María Zardoya<br />
8 de noviembre 2011<br />
Asociacó n Española<br />
de Bancos de Tejidos<br />
Comité científico<br />
PRESIDENTE<br />
Rafael Matesanz<br />
Margarida Amil<br />
Elba Agustí<br />
Miguel Casares<br />
Rafael Matesanz<br />
José Navas<br />
Rosana Reis Nothen<br />
Oscar Schwint<br />
Josep M. Segur<br />
Carlos Soratti<br />
Helder Trindade<br />
Asociación Lationoamericana<br />
de Banco de Tejidos
1ª Jornada Iberoamericana de Bancos de Tejidos y Células<br />
Auditorio Centre Salut Mental. Sant Boi de Llobregat. Barcelona<br />
8 de noviembre 2011<br />
Asociación Españ ola<br />
de Bancos de Tejidos<br />
Asociación Lationoamericana<br />
de Banco de Tejidos<br />
PROGRAMA CIENTÍFICO - 8 de noviembre de 2011<br />
09:00 h Presentación de la Jornada<br />
Josep M Segur (Presidente AEBT) - Oscar Schwint (Presidente ALABAT)<br />
Moderadores: Rafael Villalba (Espa a) / Helder Trindade (Portugal)<br />
09:15-10:00 h Situación actual de los Bancos de Tejidos y Céulas<br />
Oscar Schwint (Argentina) Estado actual y futuro de los Bancos de Tejidos en Améica Latina<br />
Margarida Amil (Portugal) Situación actual de los Bancos de Tejidos en Portugal<br />
Gregorio Garrido (España) Evolución de los Bancos de Tejidos en España<br />
10:00-10:30 h Inauguración<br />
Rafael Matesanz (Director ONT) Cooperación española-latinoamericana en el campo de los tejidos<br />
10:30-11:00 h Coffee-Break<br />
Moderadores: Antoni Gaya / Liliana Bisignano<br />
11:00- 11:45 h Enfermedades emergentes por reas geogr ficas en un entorno global. Experiencia a compartir.<br />
Asunció Moreno (España)<br />
Tom s Pumarola (España)<br />
Chagas<br />
HTLV<br />
Moderadores: Gregorio Garrido / Jose Navas<br />
11:45-12: 45 h Diferentes modelos de gestión y organización<br />
- Desarrollo de los bancos del sistema osteoarticular en la Argentina<br />
Horacio Bazán (Argentina)<br />
- Bancos públicos /Bancos privados<br />
Jose Navas (Colombia)<br />
- Bancos de Tejidos y Terapias Celulares<br />
Jacinto Sánchez (España)<br />
12:45-13:30 h Herramientas de optimización en los Bancos de Tejidos y Células<br />
- Sistemas informáticos como herramienta imprescindible en el trabajo diario de los Bancos de Tejidos<br />
Liliana Bisignano (Argentina)<br />
- Plataforma de apoyo a las Terapias Avanzadas en Catalunya<br />
Begoña Aran (España)<br />
- Aplicación de Lean Healthcare a los Bancos de Tejidos<br />
Xavier Serigó (España)<br />
13:30-15:00 h Comida<br />
Visita instalaciones TSF (2 grupos x 15 minutos)<br />
Moderadores: Miguel Casares / Josep M. Segur<br />
15:00-16:00 h Estrategias para mejorar la seguridad de los injertos durante el procesamiento<br />
- Irradiación<br />
Eulogia Kairiyama (Argentina)<br />
- Controles microbiológicos de los tejidos: Cómo, cuándo, donde?<br />
Vicente Mirabet (España)<br />
- Procesamiento en un entorno controlado: Salas Blancas<br />
Anna Vilarrodona / Manuel Fernández (España)<br />
16:00-17:00 h Coffee-Break<br />
Visita instalaciones TSF (2 grupos x 15 minutos)<br />
17:00 h Autocar para No Miembros AEBT<br />
17:00 h Asamblea AEBT (Sant Boi)<br />
18:00 h Autocar miembros AEBT
SITUACIÓN ACTUAL DE LOS BANCOS DE TEJIDOS EN PORTUGAL<br />
PONENCIAS<br />
M. Amil, R. Piteira, Autoridade para os Serviços de Sangue e da Transplantação, Ministério da Saúde, Portugal.<br />
En 2009 la Ley n.º12/2009 ha transpuesto la Directivas Europeas relativas a los requisitos legales en materia de calidad<br />
y seguranza de tejidos y células de origen humana y regulan la actividad de las unidades de extracción, banco de células<br />
y tejidos y de las diferentes unidades de aplicación.<br />
La Ley también establece un período de 12 meses para la adaptación de los Bancos de Tejidos, a los nuevos requisitos<br />
impuesto, y presentación del pedido de autorización a ASST.<br />
Durante el periodo de adaptación, fueran realizados 2 workshops, dirigidos a formación de los profesionales que participan<br />
en las actividades de los bancos de tejidos sobre los nuevos requisitos técnicos y legales.<br />
Después de la fase de adaptación, se ha empezado a llevar a cabo inspecciones para la concesión de la autorización.<br />
Destáquese que los servicios, cuyas actividades empezaran antes de la publicación de la legislación nacional, y que hayan<br />
presentado el pedido de autorización a ASST en el período legal, continuaron su actividad hasta la inspección.<br />
Las inspecciones a las unidades de la extracción, los bancos de tejidos y células, y las unidades de aplicación normalmente<br />
se llevan a cabo en colaboración con expertos invitados de otros Estados Miembros, en particular, Afssaps, Transplant<br />
Service Foundation o la Fundación Carreras, con probada experiencia en el área de tejidos y células.<br />
Desde la publicación de la Ley n º 12/2009, fueran presentados 137 pedidos de autorización, de 94 instituciones, para<br />
las actividades de las unidades de extracción (67), bancos de tejidos (51), y unidades de aplicación (79). Actualmente, se<br />
concedió un total de 33 autorizaciones, basados en las inspecciones y análisis de los pedidos evaluados por los expertos.<br />
La evaluación de los pedidos, precede a la inspección, sin embargo, muy a menudo la información sobre los documentos<br />
presentados no coinciden con las observaciones hechas. Esto crea obstáculos y dudas a la concesión de la autorización de<br />
basarse únicamente en el análisis documental, por lo que este tipo de autorización está disponible sólo para las unidades<br />
de extracción y aplicación, y está sujeto a la inspección.<br />
En el curso de la inspección había problemas comunes a muchos servicios, incluyendo:<br />
– Desconocimiento de la necesidad de instalaciones clasificadas (por ley), o la ignorancia de las necesidades particulares<br />
de estas zonas;<br />
– Falta de entrenamiento de los responsables (que determinan las necesidades del servicio) y de los que trabajan en áreas<br />
clasificadas;<br />
– Falta de validación y cualificación de instalaciones y equipos;<br />
– Los circuitos no son adecuados para las actividades definidas;<br />
– Falta de procedimientos de limpieza para áreas específicas de riesgo en contacto directo con los productos;<br />
– Contratos con terceros, incluyendo responsables de la limpieza y desinfección, transporte o extracción, están ausentes o<br />
muy incompletos, dejando por definir los procedimientos y responsabilidades de ambas partes;<br />
– Registros inapropiados, como falta de documento relativos a los productos almacenados;<br />
– Inapropiada evaluación de la calidad de los tejidos y selección de donantes;<br />
– Falta de definición de los criterios de aceptación.<br />
Frecuentemente la gravedad del incumplimiento ha obligado a la suspensión de las actividades, y a la posterior reinspección<br />
y presentación a ASST de la documentación de las acciones correctivas tomadas. A pesar de la gravedad de<br />
los problemas detectados, y si son comunes en muchas instituciones inspeccionadas, de manera general los servicios fueron<br />
capaces de adaptarse y responder a las observaciones formuladas. Por lo tanto, se considera que las inspecciones<br />
permitirán a los servicios evolucir para satisfacer los requisitos legales y las buenas prácticas establecidas.
PONENCIAS<br />
Con relación a la circulación y importación de tejidos, Portugal difiere un poco de la mayoría de los demás Estados<br />
miembros, desde que ha definido en su legislación nacional requisitos de calidad más estrictos, en particular, la realización<br />
de ensayos de ácidos nucleicos para las enfermedades transmisibles a todos los donantes de tejidos y células. Por lo tanto,<br />
ya se trate de bancos Europeos o procedentes de terceros países, deben demostrar el cumplimiento de los requisitos de la<br />
legislación nacional con el fin de obtener la autorización de la ASST para la distribución de tejidos en el país.<br />
Dada la facilidad de interacción con algunas autoridades competentes de otros Estados miembros, en relación con el control<br />
de la calidad y seguridad de los tejidos para Portugal, la estrategia fue la circulación/importación de los tejidos por un<br />
banco de tejidos único autorizado para esta actividad. En la actualidad se recibe en Portugal con total seguridad y calidad<br />
tejidos de Francia, Italia y España.<br />
También con respecto a la exportación de tejidos nacionales la ley establece que: "se permite sólo cuando haya la suficiente<br />
disponibilidad de tejidos y células en los bancos nacionales o por razones de compatibilidad justificado", por lo que los<br />
bancos de tejidos sólo pueden exportar / enviar a los Estados miembros tejidos para los que Portugal es autosuficiente.<br />
En el caso particular de los bancos de tejidos y células privados, que sólo desean almacenar estos productos para uso<br />
autologo, la exportación / movimiento de los tejidos extraídos en Portugal sólo se autoriza cuando:<br />
– El banco de tejidos está autorizado por autoridad competente homologa;<br />
– La Autoridad homologa confirma que los productos recibidos de otros países están sujetos a los mismos estándares de<br />
calidad y seguridad de otros tejidos extraídos en el banco del país de origen;<br />
– Son establecidos protocolos con las unidades de extracción autorizadas en Portugal;<br />
– Se verifica el cumplimiento de las buenas prácticas en materia de transporte (máximo, control y monitorización de la<br />
temperatura, indicación de "sin irradiar") y selección de los donantes.
DESARROLLO DE BANCOS DEL SISTEMA OSTEOARTICULAR EN LA ARGENTINA<br />
Dr. Horacio Bazán, ECODAIC.<br />
Hay algunas consideraciones que se tiene que tener en cuenta para entender el desarrollo de los Bancos del Sistema<br />
Osteoarticular en Argentina.<br />
El país tiene 3 millones de Km cuadrados de extensión con una población de 41millones de habitantes, de los cuales el<br />
47 % se encuentran concentrados en el 10% del territorio y el 70% en el 30 % de la superficie.<br />
La financiación del sistema de Salud básicamente se encuentra dividida en tres:<br />
1) Medicina Prepaga con aportes voluntarios<br />
2) Obras Sociales aportes obligatorios del trabajador y el empleador<br />
3) Cobertura estatal quienes no tienen ninguna cobertura<br />
Esta situación determinó que surjan dos tipos de Instituciones Públicas y Privadas.<br />
De la misma manera los Banco de Tejido Osteoarticular se desarrollaron principalmente en las instituciones privadas y en<br />
menor medida en las públicas.<br />
Esto generó que algunos de los bancos privados funcionaran en forma cerrada con poca liberación de los tejidos y<br />
quedando el banco público como proveedor de una demanda que lo superaba ampliamente.<br />
Las provincias generadoras de donantes del tejido óseo coincidían con las que poseían banco, y si había un donante en<br />
otra provincia por la distancia y los costos del traslado no se podía ablacionar.<br />
Analizando esta situación se modifica las Resolución sobre los Bancos de Tejidos en la cuál establece criterios de calidad<br />
elevados similares en la resolución europea , tanto en las condiciones edilicias acomo recursos humanos, y el área de<br />
procesamiento solo puede ser utilizada por los bancos de tejidos y no para otra actividad de esta manera todas las<br />
Instituciones privadas que utilizan el quirófano como área de procesamiento deberán construir un área exclusiva para los<br />
tejidos o bien dejar de funcionar como bancos.<br />
Acompañando a esta resolución se modifica la asignación de los donantes de tejidos osteoarticular. Si el donante provenían<br />
de un Hospital Público se lo asigna a un banco público provincial, si no tiene o no está operativo al público regional y sino<br />
a un privado provincial y luego regional.<br />
El 80% de los donantes provienen de Hospitales Públicos.<br />
PONENCIAS<br />
Conjuntamente se pusieron a disposición los fondos necesarios para el desarrollo de los bancos públicos con un criterio<br />
regional o provincial de acuerdo a la actividad de procuración y la densidad demográfica.<br />
Como consecuencias de estas acciones es que se produjo un incremento significativo del 20 % entre el 2006 y 2010 en la<br />
cantidad de donantes del sistema osteoarticular principalmente a través de donantes a corazón batientes y en menor<br />
medida de donantes a corazón parado y este año la curva continuará en ascenso.<br />
Actualmente los bancos desarrollan:<br />
– huesos con fines estructural (fresco congelado), tendón hueso, menisco.<br />
– huesos liofilizado e irradiado con fin de relleno de utilización en forma masiva por parte de los odontólosgos, tablas, chips,<br />
granulado, polvo.
BANCOS PÚBLICOS / BANCOS PRIVADOS<br />
José Navas, M.D.<br />
Partiendo de la premisa que la Donación es un acto altruista que busca el beneficio de un receptor se derivan 3 principios<br />
fundamentales: el primero que la donación la hace el donante solo al receptor, segundo que por ello solo estos dos sujetos<br />
son los actores principales y tercero que los demás actores son secundarios y deben favorecer el acto altruista.<br />
Dado que al momento de la donación el donante desconoce el receptor, podría decirse que el acto de donación se hace<br />
en forma genérica y a priori. Por el contrario, la recepción del tejido (alotrasplante) se hace en forma específica y a<br />
posteriori. En otras palabras, la óptica del donante busca el beneficio social a través de una decisión altruista y por<br />
motivación propia, mientras que la óptica de receptor busca un beneficio personal con visión individual y es reactiva a una<br />
circunstancia particular.<br />
De aquí puede derivarse que los tejidos donados podrían considerarse un bien público porque se originan en un acto de<br />
solidaridad social y deben estar disponibles para quien los requiera.<br />
Parecería entonces que existe un conflicto entre dos visiones potencialmente opuestas: beneficio social y beneficio individual,<br />
pero que obligatoriamente deben armonizarse.<br />
Se han mencionado los actores principales; donante y receptor, pero existen otros actores claves en el proceso quienes son<br />
los responsables de conseguir esta armonización de intereses y que normalmente incluyen al Estado, los bancos, los<br />
hospitales y los médicos. Estos, por supuesto, son actores secundarios que tienen la responsabilidad de hacer efectiva esta<br />
donación, de tal forma que las dos visiones (donante-receptor) no solamente no sean antagónicas sino que se complementen<br />
para cumplir los objetivos de cada una.<br />
En este proceso de armonización los actores secundarios tienen además responsabilidades particulares.<br />
PONENCIAS<br />
Las funciones del Estado están alineadas primordialmente con la visión altruista y de beneficio social del donante, como<br />
sería:<br />
1. El fomento de la donación para suplir las necesidades de injertos de la sociedad, además porque, es el único que tiene<br />
los elementos necesarios para lograrlo.<br />
2. Reglamentar, controlar y vigilar la actividad de donación y trasplante de tal forma que cumpla con los todos los requisitos<br />
sociales de calidad, equidad y eficiencia.<br />
Por el contrario, la función principal de los hospitales y de los médicos está mas cercana a la visión personal del receptor<br />
al exigir la disponibilidad del tejido necesario para cada paciente individual que lo requiera , previa definición de<br />
pertinencia, sin que puedan eludir la responsabilidad de facilitar toda la actividad de donación y trasplante.<br />
Por último, la función primordial de los bancos debe estar centrada en instrumentar la convergencia de las dos visiones de<br />
tal forma que se cumplan los objetivos sociales del donante (Estado) y los personales del receptor (Paciente).<br />
Para lograrlo, los bancos, públicos o privados, debe tener claridad de que su función es adecuar un bien de beneficio común<br />
(Tejido) para que sea utilizable por quien lo requiera. De aquí surgen dos consideraciones: qué condiciones tiene el manejo<br />
de un bien de beneficio común y qué requisitos tiene los bancos para hacer aplicables estos tejidos.<br />
Por bien público se entiende, en filosofía, aquello que es compartido por y de beneficio para todos los miembros de una<br />
comunidad y existen dos principios que lo hacen bien común:
PONENCIAS<br />
Principio de No Rivalidad. La no rivalidad en el consumo de un producto implica que el consumo por parte de un individuo<br />
no impide el uso por otros y que en el caso de los bancos hace referencia a disponibilidad.<br />
Principio de No Exclusión. La no exclusión tiene que ver con la imposibilidad de excluir del consumo de un producto a<br />
determinadas personas y que en nuestro caso estaría representado por la equidad en el acceso.<br />
Para lograr que los pacientes individuales obtengan los beneficios de los tejidos donados, los bancos están obligados a<br />
garantizar:<br />
1. Bioseguridad de los injertos.<br />
2. Equidad en la distribución.<br />
3. Eficiencia tanto en términos de oportunidad como costo.<br />
Los términos particulares de estas obligaciones normalmente deben ser establecidos por el Estado y siempre acorde con el<br />
desarrollo del conocimiento.<br />
Basados en todos los conceptos anteriores, parecería indiferente que la naturaleza de los Bancos de Tejido fuera privada<br />
o pública, ya que todas los condicionamientos señalados serían de forzoso cumplimiento para ambos y el Estado estaría<br />
obligado a exigirlos por igual.<br />
Sin embargo, con frecuencia se confunde el significado de “Bien Público” entendido como de “beneficio para todos” con<br />
el de “propiedad de todos” y equivocadamente se piensa que por el hecho de ser “Privado” no puede ser de beneficio<br />
público. En Colombia los Bancos de Tejido, privados o públicos, están legalmente obligados a ser de beneficio para todas.<br />
De igual manera, con frecuencia se considera que todo lo privado se hace por lucro (repartición de excedentes financieros)<br />
y consecuentemente alguien distinto al receptor se beneficia de la donación. Esto tampoco es cierto en Colombia en donde<br />
la normatividad expresamente lo prohíbe y controla. Sin embargo, para garantizar la supervivencia de los Bancos y la<br />
disponibilidad duradera de este recurso terapéutico que permite mejorar la cantidad y/o calidad de vida de los enfermos,<br />
los Bancos, privados o públicos, deben ser autosuficientes y por supuesto disponer de excedentes que permitan mejorar el<br />
cumplimiento de su objetivo social. Paradójicamente, mientras la normatividad usualmente obliga a los bancos privados a<br />
reinvertir los excedentes exclusivamente en su objetivos misionales (sin ánimo de lucro) para los bancos públicos no es<br />
obligatorio. Adicionalmente y aunque no pueda existir una diferencia misional entre ambos, si existe una diferencia<br />
motivacional: los bancos privados existen por convicción mientras los públicos lo deben hacer por obligación.<br />
Como ejemplo de la aplicación de esta conceptualidad, se presenta la experiencia de un banco privado de tejido<br />
osteomuscular que en Colombia ha suplido las necesidades de tejido durante los últimos 25 años.
BANCO DE TEJIDOS Y TERAPIA CELULAR<br />
Jacinto Sánchez Ibáñez. Director de la Oficina de Coordinación de Trasplantes de Galicia.<br />
PONENCIAS<br />
España fue uno de los primeros países en disponer de una legislación específica sobre tejidos humanos, vio la luz en 1996,<br />
el ya famoso Real decreto 411/1996. La distinción entre órganos tejidos y células estaba bien definida, se procesaban los<br />
llamados tejidos clásicos, responsabilidad de los llamados Bancos de tejidos. El apoyo y el impulso que en aquel momento<br />
se le dio por parte de la red de coordinadores de trasplantes fue clave en la creación de dicho marco legislativo.<br />
A nivel europeo aunque si que existía una actividad mas o menos desarrollada en determinados países, la tónica general<br />
era una ausencia de marco legal en la gran mayoría de los países. Varias iniciativas ponían de manifiesto la necesidad de<br />
disponer en Europa de un marco legal común. Es por ello que bajo presidencia española en el año 2000 se acuerda<br />
promover la creación de una directiva europea sobre células y tejidos, que finalmente vio la luz en el 2004.<br />
Durante su redacción y elaboración solo algunos Bancos de Tejidos realizaban cultivos celulares. En el año 2003 sale a la<br />
luz una Directiva Europea sobre medicamentos de uso humano y en el anexo 1 se refería a los medicamentos de terapia<br />
avanzada que incluía a la terapia génica, terapia celular somática tanto humanas como xenogénicas. De la noche a la<br />
mañana mientras que se discutía la futura Directiva de Células y Tejidos , los cultivos celulares que se venían realizando en<br />
los Bancos de tejidos pasaron a ser considerados medicamentos.<br />
Ese año el Observatorio Europeo de Ciencia y tecnología de la Comisión Europea publicó un informe sobre los productos<br />
de ingeniería tisular, su mercado actual y sus perspectivas de futuro. Era el informe de la gallina de los huevos de oro. Se<br />
hablaba de un mercado potencial de varios cientos de millones de euros para cada tipo de terapia celular.<br />
Se puso de manifiesto que era imposible que los Bancos de tejidos en aquella época ni los establecimientos de tejidos<br />
podían de la noche a la mañana adecuarse a unos procedimientos que estaban lejos de los medicamentos clásicos y que<br />
al menos en España, los centros que venían realizando alguna actividad tendrían que cesarla, el costo para la sanidad<br />
pública española si quisiera disponer de ellos sería elevadísima y en ocasiones imposibles de obtener.<br />
El punto clave lo determinaba lo que precisamente definía la terapia celular somática, células alteradas substancialmente<br />
como resultado de su manipulación y en las que se incluía la expansión o activación de poblaciones celulares autólogas<br />
ex vivo, la utilización de células alogénicas y xenogénicas asociadas con productos sanitarios empleados ex vivo o in vivo.<br />
Se tenían que producir bajo condiciones GMP y se tenía que registrar el producto en la Agencia Española del Medicamento<br />
En el 2007 ante las numerosas dudas y preguntas que se estaban planteando ve la luz un nuevo reglamento del Parlamento<br />
Europeo y el Consejo que en España entro en vigor en Diciembre del 2008. Lo importante de este Reglamento 1394/2007<br />
es que “se excluyen los medicamentos de terapia avanzadas preparados ocasionalmente, de acuerdo con normas de<br />
calidad especificas y empleados en un mismo Estado Miembro, en un Hospital y bajo la responsabilidad profesional<br />
exclusiva de un medico colegiado, con el fin de cumplir una prescripción facultativa individual de un producto hecho a<br />
medida destinado a un solo paciente, asegurando al mismo tiempo, que no se menoscaban las normas comunitarias<br />
relativas a la calidad y la seguridad”. Y otro punto clave es que al menos define que no es una manipulación sustancial:<br />
Corte, Trituración, Moldeo, Centrifugación, Inhibición en disolución antibiótica, Esterilización, Irradiación, Filtrado,<br />
Liofilización, Congelación, Criopreservación, Vitrificación, Concentración o purificación celular y Separación.<br />
En nuestro país se introdujo el concepto de terapia celular consolidada, aquella que la bilbiografia ha demostrado un<br />
beneficio clínico demostrado y se acordó que eran el cultivo de condrocitos, de queratinocitos y las células del limbo<br />
corneal. El resto de terapias celulares tendrían que plantearse como un ensayo clínico.<br />
En el año 2000 se comenzó a trabajar en el Complejo Hospitalario Universitario de A Coruña para trasladar desde el<br />
campo de la investigación un programa de aplicación clínica de cultivo de condrocitos. Se trabajó en disponer de un
PONENCIAS<br />
manual de procedimientos, instaurar un programa de calidad, disponer de hojas de trabajo, validar procedimientos etc.<br />
No hay que olvidar que en ese momento había que literalmente diseñar absolutamente todo y desde la investigación llegar<br />
a la aplicación clínica. El Hospital participó activamente en el diseño y seguimiento del protocolo de uso tutelado de la<br />
aplicación de condrocitos y era uno de los pocos centros donde se desarrollaba dicha actividad.<br />
Desde el marco legal del 2007 el cultivo de condrocitos para uso clínico ha disminuido su actividad de manera drástica<br />
en parte porque el interés que despertó como una terapia que solucionaba muchos problemas ha decaído a casi cero y<br />
por otra parte cierta ambigüedad en cuanto a los requerimientos que se solicitan y que autoridad competente regula este<br />
tipo de cultivo.<br />
Todo el esfuerzo que han llevado a cabo ciertos establecimientos de tejidos en relación a la terapia celular, gran reducción<br />
de costos frente a empresas farmacéuticas, la inversión de I+D+I que permitió poner a punto las diversas técnicas junto con<br />
la asimilación de estos productos a productos farmacéuticos ha llevado a que prácticamente, salvo en un caso, las terapias<br />
celulares hayan tenido que salir de los establecimientos de tejidos clásicos y que el futuro de estas terapias sea incierto.
PLATAFORMA DE APOYO A LAS TERAPIAS AVANZADAS EN CATALUNYA<br />
PONENCIAS<br />
Begoña Aran, Organització Catalana de Trasplantaments. (OCATT), Centre de Medicina Regenerativa de Barcelona<br />
(CMRB).<br />
Situación de las Terapias Avanzadas (TA) en Catalunya<br />
La posibilidad de reparar o sustituir tejidos y órganos dañados por enfermedades o lesiones ha generado que muchos<br />
grupos de investigación básica y clínica se planteen la utilización de células como una posible opción para el tratamiento<br />
de enfermedades producidas por la pérdida de la función celular.<br />
Las TA, Terapia Celular, Terapia Génica e Ingeniería de Tejidos, elaboradas con tejidos i/o células de origen humano, son<br />
consideradas medicamentos y están reguladas por el Reglamento CE nº1394/2007. Este reglamento deriva de una serie<br />
de directivas comunitarias transpuestas en nuestro país (1-6) a diferentes niveles y en función de la fase del proceso de<br />
trasformación de los tejidos y células destinados a elaborar estos medicamentos de TA.<br />
Actualmente la mayoría de los proyectos con TA están en fase de investigación y en menor número en fase de ensayo<br />
clínico. Para realizar un ensayo clínico, cada proyecto debe ser aprobado por la Agencia Española del Medicamento y<br />
Productos Sanitarios (AEMPS), y las CCAA no tienen competencias en su aprobación, aunque si las tienen (y es obligatorio)<br />
en la autorización de los centros para la obtención/extracción y evaluación de tejidos y células utilizados en cada proyecto<br />
(RD 1301/2006). Dicha legislación establece los mismos requisitos y normas tanto para los ensayos clínicos de TA<br />
realizados en los hospitales como para la producción industrial de los productos resultantes, a pesar de que la situación<br />
en cuanto a la finalidad del proyecto, tamaño de muestras, medios económicos, etc., puede ser muy diferente.<br />
Aunque la aprobación de los ensayos clínicos es competencia de la AEMPS, estos se realizan en los hospitales públicos y<br />
privados de Catalunya, pero dada la dinámica establecida, las autoridades sanitarias de la CA desconocen el número y<br />
las características, así como la finalidad y los resultados de los mismos.<br />
Por otro lado, dada la complejidad del marco legislativo, existe cierto desconcierto entre los investigadores básicos y<br />
clínicos en cuanto a los requisitos administrativos necesarios para llevar a cabo estos ensayos e incluso, en algunos casos,<br />
en la interpretación de si una terapia debe considerarse TA o trasplante de tejido o células, y por tanto, en que marco<br />
legislativo debe considerarse.<br />
Propuesta de ordenación del ámbito de las TA<br />
La tradición de la Organización Catalana de Trasplantes (OCATT) en el ámbito de la donación y el trasplante, así como su<br />
metodología transparente, favorecen que los profesionales depositen su confianza en la Organización y pidan consejo al<br />
poner en marcha su proyecto. A pesar de ello, la acción de la OCATT es únicamente facilitadora y canalizadora, ya que<br />
no tiene competencias en este ámbito. Sería muy útil para el Departament de Salut conocer los ensayos clínicos de TA que<br />
se están realizando en Catalunya, por lo que se propone crear una Plataforma de Apoyo a las TA que permita:<br />
1. Conocer el contenido del ensayo clínico previsto previa presentación a la AEMPS, con el fin de evitar errores iniciales y<br />
facilitar su aceptación.<br />
2. Disponer de toda la información, actualizada y real, sobre los ensayos aprobados en Catalunya o en vías de aprobación,<br />
realizando un seguimiento periódico de los mismos.<br />
3. Canalizar iniciativas facilitando el paso de la investigación básica al ensayo clínico.<br />
4. Ofrecer el apoyo del Departament de Salut, a través de la OCATT, a los profesionales de la investigación clínica,<br />
facilitando información y gestión de trámites.<br />
5. Potenciar las iniciativas de mayor impacto dentro del ámbito de la salud y coordinarlas.<br />
6. Propiciar e impulsar sinergias en el ámbito de los ensayos clínicos canalizándolos de una manera ordenada y<br />
multidisciplinaria que permita la optimización de recursos.<br />
7. Buscar convocatorias y subvenciones de programas de ayuda internacional para la financiación de proyectos en<br />
Catalunya.
8. Buscar financiación, pública y privada, para los proyectos que el Departament de Salut considere prioritarios para el<br />
tratamiento de pacientes.<br />
9. Facilitar la obtención de patentes y la colaboración entre clínicos y empresas biotecnológicas.<br />
La Plataforma de Apoyo a las TA constará de:<br />
a) Un Registro de Ensayos Clínicos de TA en Catalunya: este registro debería incluir no sólo los ensayos clínicos aprobados,<br />
si no también los pendientes de aprobación, así como los presentados y rechazados y las causas. Sería conveniente<br />
también recoger un resumen de los resultados de los ensayos finalizados.<br />
b) Una Comisión Asesora de TA, formada por profesionales expertos en el ámbito de la investigación biomédica que pueda<br />
formar grupos de trabajo específicos en función de las materias a tratar. Las funciones de esta Comisión serían las<br />
siguientes:<br />
b.1) Analizar, evaluar, hacer el seguimiento y conocer los resultados de los ensayos clínicos presentados.<br />
b.2) Coordinar y promover la coordinación entre equipos.<br />
b.3) Proponer mejoras en la gestión y desarrollo de los proyectos.<br />
b.4) Orientar posibles sinergias entre investigadores clínicos y centros.<br />
b.5) Asesorar al Departament de Salut en la materia.<br />
b.6) Coordinación interdepartamental<br />
b.7) Promover la difusión y la formación de profesionales de los diferentes niveles que intervienen en los ensayos clínicos<br />
en TA.<br />
b.8) Elaboración de informes científicos y técnicos y una memoria anual.<br />
c) Un portal WEB que recoja todas las actividades de la Plataforma.<br />
PONENCIAS<br />
Hasta el momento se han registrado 26 ensayos clínicos; 3 con terapia génica y 23 con terapia celular, 10 de los cuales<br />
se realizan con células de la médula ósea, 4 con células mesenquimales de tejido adiposo, 6 con distintos tipos celulares<br />
de sangre periférica, 1 con progenitores de sangre de cordón umbilical, 1 con queratinocitos y 1 con células hepáticas.<br />
Es necesario que el Departament de Salut pueda conocer y centralizar la actividad en TA que se está haciendo en Catalunya.<br />
La Plataforma de Apoyo a las TA facilitaría una análisis cuantitativo y cualitativo de la misma, evitando duplicaciones y<br />
creando sinergias de colaboración entre los diferentes colectivos implicados (grupos de investigación, universidades,<br />
centros hospitalarios, etc.), facilitando gestiones a los profesionales y favoreciendo un buen posicionamiento de Catalunya<br />
en esta campo a nivel internacional. De esta manera también se podría favorecer aquellas terapias con mayores<br />
posibilidades de éxito en la práctica clínica.<br />
Referencias<br />
(1) R. D 223/2004 por el que se regulan los ensayos clínicos con medicamentos.<br />
(2) RD 2183/2004 por el que se desarrolla y regula el régimen de autorización de los laboratorios farmacéuticos……<br />
(3) Ley 29/2006 del garantías y uso racional de medicamentos y productos sanitarios.<br />
(4) RD 1301/2006 por el que se establecen las normas de calidad y seguridad … de células y Tejidos para su uso en<br />
humanos.<br />
(5) Ley 14/2007 de investigación Biomédica.<br />
(6) Reglamento CE nº 1394/2007 del parlamento Europeo y del Consejo sobre medicamentos de terapia avanzada.
ESTRATEGIAS PARA MEJORAR LA SEGURIDAD DE LOS INJERTOS DURANTE<br />
EL PROCESAMIENTO<br />
Eulogia Kairiyama, Irradiación<br />
PONENCIAS<br />
Resumen:<br />
Las radiaciones ionizantes se utilizan para tratar una gran diversidad de productos con distintos objetivos. Entre las<br />
principales aplicaciones se puede mencionar la esterilización de productos de uso médico, productos farmacéuticos y<br />
envases, entre otros. La esterilización por irradiación de tejidos humanos para injerto se puede considerar como una<br />
consecuencia de la tecnología de esterilización por irradiación de productos médicos establecida hace más de 50 años.<br />
Es un proceso confiable y seguro. No deja residuos. Los tejidos procesados pueden ser esterilizados en su envase final, y<br />
ser utilizados en forma inmediata. Asimismo, durante el proceso no hay aumento significativo de la temperatura, por lo<br />
que pueden tratarse productos sensibles al calor y en diferentes condiciones físicas, tales como congelados, deshidratados,<br />
en diferentes medios, así como productos de tamaños y diseños muy variados.<br />
El proceso de esterilización de tejidos para injerto es una etapa de su procesamiento, pudiéndose aplicar la radiación luego<br />
de procurado el tejido previo al ingreso a la sala de procesamiento aséptico, o como esterilización final del tejido producido<br />
en su envase final previo al almacenamiento o distribución.<br />
Las radiaciones que se utilizan en estos procesos son ya sea electromagnéticas provenientes de fuentes industriales de los<br />
radionucleidos cobalto-60 o cesio-137, y también rayos X (energía < a 5 MeV); como asimismo radiaciones particuladas<br />
producidas por máquinas aceleradoras de electrones (< a 10 MeV). La energía de las radiaciones es absorbida por el<br />
material expuesto produciendo ionización y excitación de átomos y moléculas, como resultado del efecto directo de las<br />
radiaciones al interactuar con los electrones orbitales de los átomos. Las reacciones secundarias se producen entre las<br />
especies altamente reactivas producidas por la radiación con otros compuestos del medio; esto se conoce también como<br />
efecto indirecto de la radiación.<br />
Estos efectos producen cambios químicos de los componentes de los organismos vivos y del medio en que se encuentran,<br />
conllevando a una serie de eventos que conducen a cambios biológicos.<br />
El efecto de la radiación sobre las células es atribuible a la deposición de la energía en estructuras críticas o blancos, que<br />
causan desorden metabólico que producirán su muerte o sobrevida. El blanco principal para la inactivación de los<br />
microorganismos por radiación es la molécula de ácido desoxirribonucleico (ADN), debido al rol que juega en la división<br />
celular. Asimismo se producen cambios en la membrana, ribosomas y otros elementos de la célula. Los efectos de las<br />
radiaciones dependerán principalmente de las condiciones en que se encuentran los microorganismos en el tejido<br />
(congelado, liofilizado, glicerolado u otro estado), dosis absorbida, su estado fisiológico, su capacidad intrínseca de<br />
reparación de la molécula de ADN, entre otros.<br />
El objetivo de la esterilización es la inactivación de los microorganismos contaminantes del producto. La cinética de<br />
inactivación de un cultivo puro de microorganismos por agentes físicos y/o químicos puede ser descripta por una relación<br />
exponencial entre el número de microorganismos y la intensidad del tratamiento con el agente esterilizante. Existe una<br />
probabilidad finita de que un microorganismo sobreviva después del proceso. Para un determinado proceso, la<br />
probabilidad de sobrevivencia es determinada por el número y resistencia de los microorganismos y por el ambiente en el<br />
cual se encuentran. La esterilidad es definida en términos de la probabilidad de que un microorganismo viable esté presente<br />
en una unidad de producto. La especificación de esta probabilidad es definida por la autoridad regulatoria de los países,<br />
siendo este valor, en la mayoría de ellos, de 1:10 6 .<br />
El proceso de esterilización consiste en exponer el producto a la radiación durante un tiempo predeterminado, el necesario<br />
para que absorba la energía suficiente para alcanzar el objetivo deseado. Dado que el proceso de irradiación es parte
PONENCIAS<br />
del proceso de fabricación del tejido para injerto, se deben aplicar Buenas Prácticas de Irradiación, así como también el<br />
tejido debe haber sido producido de acuerdo a las Buenas Prácticas de Tejidos. En una etapa inicial se debe realizar la<br />
validación del proceso, de modo de verificar con un alto grado de seguridad que el proceso producirá en forma consistente<br />
un producto que cumpla con los atributos de calidad y especificaciones requeridas. Una vez cumplimentada esta etapa, el<br />
producto podrá ser procesado y controlado en forma rutinaria en las mismas condiciones establecidas en la etapa anterior.<br />
Es necesario realizar verificaciones periódicas del mantenimiento de la validación.<br />
Una validación apropiada y un proceso de esterilización controlados con precisión no garantizan que los tejidos sean<br />
estériles y aptos para su uso clínico. Se deben tener en cuenta una serie consideraciones, tales como el estado microbiano<br />
del tejido ablacionado; la validación y control de rutina de los procedimientos de limpieza y desinfección de los<br />
componentes y materiales que entran en contacto con el tejido; el control del ambiente donde se procesa y empaca;<br />
control de los equipos y la higiene del personal; la forma y los materiales en que se envasan los tejidos; las condiciones<br />
de almacenamiento.<br />
Existen normas internacionales para la validación del proceso de esterilización de productos para el cuidado de la salud<br />
por radiación. El Organismo Internacional de Energía Atómica (OIEA) ha elaborado un código de prácticas para la<br />
esterilización de tejidos para injertos; en el año 2007 fue publicada la versión actualizada del borrador finalizado en<br />
2004 del Radiation Sterilization of Tissue Allografts: Requirements for Validation and Routine Control. A Code of Practice.<br />
Esta versión en inglés fue revisada, actualizada y traducida al español, en el marco de un proyecto regional de América<br />
Latina, ARCAL CVIII RLA 6062, habiéndose finalizado el borrador final en agosto de 2011.<br />
El Código de Prácticas para la Esterilización por Irradiación de Tejidos Humanos para Uso Clínico: Requisitos para la<br />
Validación y Control de Rutina, está basado en los requisitos especificados en la norma ISO 11137 para el desarrollo,<br />
validación y control de rutina de un proceso de esterilización para dispositivos médicos, que fue adaptado para tejidos<br />
humanos para injerto y para un bajo número de unidad de producto por lote de producción.<br />
El objetivo de este Código de Prácticas es proporcionar una guía para el uso de la radiación ionizante como método de<br />
esterilización de tejidos humanos para uso clínico seguro, en donde se especifican los requerimientos para la validación<br />
del proceso de esterilización final con radiaciones ionizantes y para el control de rutina en el procesamiento de tejidos<br />
humanos para uso clínico, producidos bajo Buenas Prácticas de Producción de Tejidos.<br />
Este documento no es aplicable a tejidos que contengan virus, priones o endotoxinas bacterianas porque no garantiza su<br />
inactivación, por lo que deberán aplicarse con rigor los criterios de selección y de exclusión de los donantes.<br />
El Anexo I describe los métodos para seleccionar la dosis de esterilización. El Anexo II proporciona ejemplos aplicando estos<br />
métodos extraídos de experiencias de la región. El Anexo III presenta una serie de tablas de dosis de radiación requeridas<br />
para alcanzar un determinado Nivel de Aseguramiento de Esterilidad para diferentes cargas microbianas que tienen una<br />
Distribución Estándar de Resistencias. En el Anexo IV, en la bibliografía se proporcionan referencias claves para la<br />
esterilización de los tejidos con radiación ionizante.<br />
El desarrollo, validación y control de rutina de un proceso de esterilización comprende un número de actividades<br />
interrelacionadas, tales como calibración, mantenimiento, definición del producto y del proceso, calificación de la instalación<br />
tanto en lo operacional como de desempeño.<br />
Las etapas del Plan de Validación son:<br />
– Calificación del producto y envase, para establecer la dosis mínima de esterilización (Dmin) y determinar la dosis máxima<br />
(Dmax) tolerada por el producto (tejido en su envase) sin deterioro de sus funcionalidades, estableciendo así los requisitos<br />
del proceso,<br />
– Calificación de la instalación, para demostrar que el irradiador y sistemas auxiliares han sido instalados y operan de<br />
acuerdo a sus especificaciones,<br />
– Calificación de la operación, debe demostrar que el irradiador, tal como está instalado, es capaz de operar de acuerdo<br />
con sus procedimientos operativos y de entregar las dosis apropiadas dentro de criterios de aceptación definidos
PONENCIAS<br />
– Calificación de desempeño, se debe demostrar que el resultado del proceso aplicado al tejido procesado en su empaque<br />
final cumple con los requisitos especificados. Para realizar este paso en un tejido procesado es necesario conocer<br />
previamente sus características, tipo de tejido y método de conservación, y el rango de dosis (Dmax y Dmin) especificado<br />
por el cliente (banco de tejidos) de acuerdo a los resultados de la calificación del producto. El mapeo de dosis se debe<br />
llevar a cabo con el producto cargado en el contenedor de irradiación de acuerdo al patrón de carga especificado, con<br />
el fin de localizar y conocer las magnitudes de la dosis mínima y máxima, así como, determinar la relación entre ellas y<br />
sus ubicaciones.<br />
– Revisión y Aprobación de los elementos anteriores, se deben verificar, registrar y aprobar los documentos generados a<br />
través de todas las etapas de la validación para su aplicación en el proceso rutinario de irradiación del tejido procesado.<br />
– Mantenimiento de la Validación, mediante revalidaciones periódicas y toda vez que se produzcan modificaciones en la<br />
instalación, producto, sistemas o proceso.
CONTROL MICROBIOLÓGICO EN EL BANCO DE TEJIDOS<br />
¿CÓMO? ¿CUÁNDO? ¿DÓNDE?<br />
Dr. Vicente Mirabet, Banco de Tejidos y Células de Valencia. España.<br />
PONENCIAS<br />
Se ha estimado que, por cada célula presente en el ser humano, se pueden contabilizar hasta 10 microorganismos. Además,<br />
hay una notable diversidad, habiéndose identificado bacterias de más de 200 géneros distintos, sólo sobre la piel. Miles<br />
de años de evolución han propiciado una relación de simbiosis entre el ser humano y su microflora que, en ocasiones,<br />
trasciende el comensalismo para situarse más próxima al mutualismo. Sin embargo, en determinadas circunstancias, este<br />
equilibrio puede romperse y generar un proceso infeccioso.<br />
El número de homoinjertos trasplantados anualmente en el mundo, se cifra en cientos de miles. De acuerdo con ello, se<br />
pueden considerar anecdóticas (en cuanto a proporción) las referencias acerca de efectos adversos de tipo infeccioso, en<br />
las que se haya identificado el homoinjerto como origen del mismo. De hecho, el uso clínico de homoinjertos no se asocia<br />
con un criterio de reversibilidad; es decir, cuando, en lugar de homoinjertos se utiliza material protésico estéril, las tasas<br />
de infección son análogas. No obstante, lo cierto es que algunos de esos eventos, aunque escasos, han tenido consecuencias<br />
muy graves, incluso fatales.<br />
Es un imperativo en la actividad de los bancos de tejidos proporcionar productos biológicamente seguros y clínicamente<br />
eficaces.<br />
Por todo ello, el trasplante de tejidos requiere un exhaustivo control en todas las actividades para evitar el riesgo de<br />
infección. Con este fin, hay que identificar los factores que pueden influir en dicho riesgo y analizar la relación causal de<br />
los mismos. Sólo entonces será posible implantar medidas eficaces dirigidas a los dos principales focos de atención:<br />
– El propio tejido, evitando la extracción de aquéllos que sean considerados como comprometidos.<br />
– El entorno medioambiental, monitorizando los parámetros, especialmente en los procesos críticos (cuando el tejido se<br />
expone a dicho entorno).<br />
La experiencia acumulada en este campo ofrece la posibilidad de abordar el análisis con carácter proactivo, es decir,<br />
tratando de evitar los incidentes, en lugar de actuar en consecuencia una vez que ya se han producido (carácter reactivo).<br />
Seguidamente, repasaremos algunos de los aspectos más representativos a tener en cuenta en cada proceso. Como es<br />
lógico, si el tejido va a ser sometido a algún procedimiento de esterilización secundaria (debidamente validado para este<br />
fin) se podría aplicar, en algunos apartados, otro tipo de consideraciones.<br />
Selección de donantes<br />
El propio donante es la primera fuente a considerar a la hora de evaluar el riesgo de contaminación. El historial médicosocial,<br />
la causa del fallecimiento o el hemocultivo, por ejemplo, van a aportar datos de interés en la toma de decisiones.<br />
La disponibilidad de profesionales especializados en este tipo de actuaciones representa una inestimable ayuda, siendo ésta<br />
una de las funciones asumidas por la figura del coordinador de trasplantes.<br />
La transmisión de enfermedades víricas es uno de los principales focos de atención en este nivel, habiéndose alcanzado un<br />
notable grado de desarrollo en los sistemas de detección de marcadores infecciosos, con una significativa reducción de los<br />
períodos ventana.<br />
Por otro lado, la posible diseminación de flora digestiva hacia otros tejidos, puede condicionar la seguridad de aquéllos.<br />
Éste es un aspecto a valorar, por ejemplo, cuando el potencial donante de tejidos lo ha sido previamente de órganos. En<br />
general, la existencia de traumatismos, con heridas extensas, eleva el riesgo de contaminación.<br />
Obtención<br />
El tiempo de isquemia y la temperatura durante el mismo, el entorno en el que se verifica la extracción de los tejidos y la<br />
composición y experiencia del equipo quirúrgico, son factores determinantes en esta actividad.
Procesamiento<br />
La utilización de cabinas de flujo laminar, dentro de salas limpias, garantiza un óptimo control sobre el aire medioambiental.<br />
Existe una estandarización en cuanto a la tolerancia de partículas presentes y la existencia de unidades formadoras de<br />
colonias en cultivo, que se traduce en una gradación cualitativa.<br />
La introducción del material de trabajo en las salas limpias debe realizarse en condiciones que no afecte significativamente<br />
al riesgo de contaminación medioambiental. Además, las pautas de trabajo del personal y su indumentaria se ajustarán<br />
a las recomendaciones en esta materia. Hay que evitar la difusión de partículas.<br />
Por otro lado, es necesaria una adecuada protocolización de la tarea de limpieza y desinfección de las distintas superficies<br />
presentes en la estancia, alternando soluciones con diferentes principios activos.<br />
Almacenamiento<br />
Aunque en esta fase es raro que el tejido pueda verse expuesto directamente a las condiciones medioambientales, hay que<br />
aplicar los medios para evitar que esto suceda. Utilizar recipientes debidamente validados para asegurar su integridad en<br />
las condiciones de almacenamiento no siempre garantiza el aislamiento. Siendo la congelación uno de los métodos más<br />
frecuentemente utilizados para el almacenamiento de tejidos, con la consiguiente rigidez en los materiales que esto conlleva,<br />
se puede dañar el producto durante la manipulación dentro de los contenedores de almacenamiento (generalmente<br />
congeladores eléctricos y tanques con nitrógeno).<br />
Cuando el entorno en la fase de almacenamiento es gaseoso, una eventual fractura de un recipiente (generalmente bolsas<br />
y frascos), difícilmente podrá comprometer más tejidos que el contenido en el propio recipiente. Sin embargo, cuando se<br />
trata de un medio líquido (como el nitrógeno en dicho estado), éste puede ser utilizado por elementos infecciosos como<br />
vehículo de difusión. Un eventual sellado defectuoso en el recipiente que contiene el tejido y la condensación de la atmósfera<br />
en su interior (como consecuencia de la baja temperatura, -196ºC), puede generar vacío (con la consiguiente penetración<br />
de líquido) y, posteriormente, al manipular dicho recipiente, si se evapora el nitrógeno y se expande, puede liberar el<br />
contenido del recipiente al líquido.<br />
En un tanque de nitrógeno líquido, en contra de lo que se pudiera pensar, se pueden encontrar diversidad de<br />
microorganismos ambientales (funcionalmente activos cuando se ponen en las condiciones adecuadas) que son atrapados<br />
en los cristales de hielo microscópicos que se generan al abrir el tanque y se depositan en el interior del mismo. Lógicamente,<br />
estas partículas potencialmente infecciosas se van a encontrar también en las superficies de los recipientes que contiene el<br />
tanque.<br />
El empleo de un sistema de doble embalaje durante esta etapa va a minimizar el riesgo de contaminación.<br />
PONENCIAS<br />
Trasplante<br />
Obviamente, los tejidos congelados tienen que ser necesariamente descongelados antes de su utilización clínica. Éste es otro<br />
procedimiento en el que suele estar presente el medio líquido (agua del baño termostático o batea con solución salina a<br />
37ºC). Por lo expuesto en el apartado anterior, el producto se manipulará considerando no estéril la superficie que haya<br />
estado en contacto con el medio ambiente durante la fase de almacenamiento.<br />
El equipo quirúrgico y el paciente receptor pueden también ser origen de una eventual contaminación del tejido a<br />
trasplantar.<br />
Metodología<br />
Hay distintas opciones, algunas de las cuales dependen de las características físicas del material objeto de análisis. Así,<br />
por ejemplo, para obtener muestras de aire, utilizaremos aparatos especialmente diseñados para muestrear entornos<br />
gaseosos; unos están calibrados para el recuento de partículas y otros se han diseñado para la siembra en placa de Petri,<br />
mediante aspiración de un volumen predeterminado.<br />
Más compleja puede resultar la decisión para el control microbiológico del propio tejido. Generalmente, se emplea uno o<br />
más de los siguientes métodos de recogida de muestras para cultivo microbiológico:
PONENCIAS<br />
– Fragmento de tejido: puede estimarse como poco representativo del total de la pieza y conlleva cierta pérdida de material,<br />
pero aporta información de las regiones tisulares profundas.<br />
– Frotado de la superficie con una torunda: más representativo que el anterior, pero el análisis se suele limitar a la cara<br />
externa.<br />
– Lavado con solución salina: requiere más tiempo de dedicación que los anteriores y la manipulación es más compleja (lo<br />
que podría elevar el riesgo de contaminación), pero ofrece la opción de incorporar un procedimiento de concentración,<br />
ya sea mediante centrifugación o filtrado.<br />
Independientemente del método utilizado, es importante obtener al menos una muestra relativa al tejido antes de una<br />
eventual exposición del mismo a una solución desinfectante, con el fin de evitar el enmascaramiento de resultados por este<br />
motivo.<br />
Los productos de consistencia fluida, como suspensiones de células precursoras hematopoyéticas o sobrenadantes de cultivo<br />
utilizados durante la expansión «ex vivo» para terapia celular, suelen ser evaluados mediante cultivo microbiológico de una<br />
alícuota. En el segundo de los ejemplos citados conviene, además, disponer una fracción de las células cultivadas para<br />
pruebas adicionales, por ejemplo, para detección de micoplasmas. También los diferentes lotes de las soluciones que van<br />
a estar en contacto directo con el tejido, por ejemplo las soluciones crioprotectoras, deben ser testadas mediante el cultivo<br />
de una alícuota.<br />
En todos los casos, se utilizarán sistemas de detección para el crecimiento de bacterias aerobias y anaerobias, y hongos.<br />
En relación con la fase de almacenamiento, si se utiliza nitrógeno líquido y hay muestras procedentes de pacientes con<br />
marcadores virales positivos (en el caso de tejidos para autotrasplante, por ejemplo), no hay que descartar la opción de<br />
extraer muestras del propio nitrógeno y de los detritus congelados que se depositan en el fondo del recipiente para<br />
analizarlos, por ejemplo, con técnicas NAT (nucleic acid testing).<br />
Los resultados de los cultivos microbiológicos de las muestras obtenidas en diferentes fases de la manipulación y el historial<br />
médico-social del donante aportarán un conjunto de datos sobre los que aplicar los algoritmos de decisión del banco.<br />
Los distintos procedimientos a los que es sometido un tejido, iniciándose con la detección del potencial donante y finalizando<br />
con el trasplante, se deben desarrollar en una cadena de asepsia que no debe romperse en ningún momento.
PROCESAMIENTO DE TEJIDOS Y TERAPIAS AVANZADAS EN AMBIENTES<br />
CONTROLADOS<br />
Fernández Pérez, M. Auditorio Centre Salut Mental. Sant Boi de Llobregat. Barcelona<br />
PONENCIAS<br />
1) Visión general de la normativa aplicable. Se hablará sobre las diferencias que las autoridades sanitarias realizan sobre<br />
los diferentes tipos de manipulaciones, tejidos o terapias celulares: Directiva 2003/63/EC, Directiva 2004/23/EC y<br />
Normas de Correcta Fabricación de Medicamentos.<br />
2) Consideraciones de diseño de instalaciones bajo perspectiva GMP. Se realizará un repaso de las consideraciones claves<br />
en el diseño de las instalaciones: salas, acabados, sistemas de tratamiento de aire y flujos de materiales y personal.<br />
3) Cómo evitar la contaminación cruzada. ¿Qué es la contaminación cruzada? Se tratarán aspectos sobre cómo evitarla<br />
en el diseño de flujos, adaptando filosofías de vestuario, enclavamientos, cómo afecta el tratamiento de aire y la<br />
minimización de intervenciones en abierto, procesamiento por lotes y cómo afecta la limpieza y diferentes tipos de<br />
limpieza y descontaminaciones.
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