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Research Day
AGA2012-219
Einfluss dynamischer Zellkultur auf die Proliferation von stromalen Knochenmarkszellen auf
einem Polyurethan Meniskus Implantat
Effect of dynamic culture on the proliferation of bone marrow stromal cells seeded in
polyurethane meniscus scaffolds
Authors
* Michael Jagodzinski Medizinische Hochschule Hannover Zentrum Chirurgie Klinik für Unfallchirurgie,
Hannover, Germany
* Chaoxu Liu Medizinische Hochschule Hannover Zentrum Chirurgie Klinik für Unfallchirurgie,
Hannover, Germany
Carl Haasper Medizinische Hochschule Hannover Zentrum Chirurgie Klinik für Unfallchirurgie,
Hannover, Germany
Daniel Günther Medizinische Hochschule Hannover Zentrum Chirurgie Klinik für Unfallchirurgie,
Hannover, Germany
Henning Windhagen Medizinische Hochschule Hannover Orthopädische Klinik im Annastift, Hannover,
Germany
Christian Krettek Medizinische Hochschule Hannover Zentrum Chirurgie Klinik für Unfallchirurgie,
Hannover, Germany
Gabriela von Lewinski Medizinische Hochschule Hannover Orthopädische Klinik im Annastift, Hannover,
Germany
Abstract
Fragstellung: The objective of this study was to investigate the effect of dynamic culture on the proliferation of
BMSC seeded in polyurethane meniscus scaffold.
Methodik: The Institutional Ethical Committee had approved all procedures and written informed consent had
been obtained from all subjects. 20-80 ml bone marrow aspirates from the iliac crest were collected from 7 donors
undergoing dorsal instrumentation and fusion because of vertebral fractures after informed consent. Isolation and
cultivation of human BMSC were performed according to a modified protocol as previously described. 6×10 6 cells
of the third passage resuspended in 1ml culture medium were seeded into one ActifitTM scaffold using a 27G
syringe. The cells were allowed to adhere a 4 hours period if incubation. Then the scaffolds were cultures under
three different conditions: static control, perfusion (10ml/min), perfusion and mechanical stimulation (10%
compression, 3x4 hours per day @ 1Hz). The scaffolds were harvested for further analysis after 24 hours, 1 week
and 2 weeks. Proliferation was investigated using the MTS assay. The total protein concentration was measured
with Coomassie Plus Reagent. In addition, light and scanning electron microscope analysis were performed to
observe the proliferation and distribution of cells. The data of all groups were compared using one-way ANOVA at
each time point, a significance level of 0.05 was used (SPSS 15.0).
Ergebnis: The data obtained from MTS assay demonstrated a significant increase in proliferation over time in all
groups. In addition, statistical differences were found between the individual groups. Both perfusion and
mechanical stimulation enhanced the proliferation of BMSC compared with static control. There was no significant
difference between the two dynamic culture groups. Similar results were observed from the total protein
concentration assay. Light microscope and SEM analysis showed the cells distributed throughout the entire
scaffold after incubation for 24 hours. After one and two weeks of static and dynamic culture, cells were observed
growing through the pores within the scaffolds, spreading uniformly and extensively. Compared to static culture,
cell density appeared to be higher in dynamic culture groups after the same interval.
Schlussfolgerung: Perfusion culture system shortened the period of building a cell-laden polyurethane construct
by enhancing the cell proliferation, which could be a further step towards the tissue engineered meniscus.
Keywords
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Vortrag
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