Here - EWMA 2013

POSTER: BASIC SCIENCE
Poster: Basic Science
P 208
COLLECTION OF WOUND FLUIDS FROM HORSES USING MICRODIALYSIS
Mette Aamand Sørensen 1 , Louise Bundgaard 1 , Stine Jacobsen 1 ,
Lars Jelstrup Petersen 2
1 University of Copenhagen (Copenhagen, Denmark);
2 Aalborg University Hospital (Aalborg, Denmark).
Aim: To develop a microdialysis method for collection of fluid from horse wounds.
Background: Collection of sample material for wound healing research in experimental
animals is commonly obtained through biopsies. Though, biopsy collection is an invasive
procedure and consequently triggers an inflammatory response. Therefore, wounds
should only be biopsied once to display the undisturbed, natural healing process. This
necessitates the creation of a wound for every required collection time-point. To limit the
number of wounds created on each experimental animal, a new method that allows
repeated collection from wounds was sought.
Methods: Microdialysis is a minimally invasive method for sampling of compounds from
the extracellular fluid, where a small probe is inserted into the target tissue and flux of
solutes into the probe occurs by simple diffusion. The recovered dialysate reflects
changes in the composition of the extracellular water phase. Sample collection can be
continued for several hours.
Results: Microdialysis was well tolerated by the experimental animal subjects with no
signs of discomfort related to the microdialysis procedure. Collection of sample material
lasted three hours and was carried out with 11 samplings during a 28 day period. It was
safely performed with reliable yield when collecting from vertically positioned
experimental wounds in a standing, sedated horse.
P 209
Poster: Basic Science
CHRONIC WOUNDS: WHAT IS THE ROLE OF THE EXTRACELLULAR MATRIX
(ECM)
Eleri M Jones 1 , Christine A Cochrane 1 , Peter D Clegg 1 , Steven L Percival 2 , John Hunt 1
1 Institute of Ageing and Chronic Disease, University of Liverpool (Leahurst, South Wirral,
United Kingdom);
2 Scapa Healthcare (Manchester, United Kingdom).
Introduction: Extracellular matrix (ECM) molecules play a fundamental role in the
process of wound healing. They are synthesised from fibroblast and keratinocyte cells as
they migrate into the wound space to facilitate healing. However, when a wound fails to
heal becoming chronic the synthesis of ECM molecules becomes impaired. Currently
there is limited knowledge of the presence and composition of ECM molecules within
chronic wounds.
Aim: The aim of this study was to determine any differences in the release of ECM
molecules (collagen, fibronectin, glycosaminoglycans) from fibroblast derived from
normal skin and chronic wound fibroblasts using biochemical and biomolecular
techniques.
Method: Normal skin and chronic wound fibroblast cells (1x105 per well) were seeded
into 12 well plates and incubated for 24, 48, 72 and 96 hours in serum free DMEM
media. After incubation, conditioned media was collected and analysed for the release of
ECM molecules. The ECM present within the cells was also analysed to see any
differences between the two cells types. Dimethylmethylene blue (DMMB) assays were
used to determine levels of glycosaminoglycans, hydroxyproline assays for collagen and
fastin assays for elastin.
Results: Results show that the presence and release of the different ECM molecules
from chronic wound fibroblasts varies significantly compared to normal fibroblasts.
Conclusion: This study has provided simple assays in which the synthesis of ECM
molecules can be established. Future work aims to manipulate the culture conditions to
imitate a chronic wound environment.
Conclusions: Microdialysis allowed the collection of wound fluid samples for research
purposes in a less invasive way than biopsies. This method also allowed repeated
sampling from every wound and the obtained sample material was sufficient in amount
for metabolite and protein analysis.
EWMA 2013
COPENHAGEN
15-17 May · 2013
Danish Wound
Healing Society
133