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Bone tissue engineered product based on human bone derived cells and polyurethane scaffold
|Piotr Woźniak 1, Monika Bil 2, Joanna Ryszkowska 2, Piotr Wychowański 3, Grażyna A. Hoser 4, Małgorzata Lewandowska-Szumieł 1, Krzysztof J. Kurzydlowski 2|
1. Medical University of Warsaw, Department of Biophysics and Human Physiology, Chałubińskiego 5, Warszawa 02-004, Poland
Biodegradable polyurethanes are widely investigated as candidate material for scaffolds in bone tissue engineering (1-2). There are, however, only a few papers describing the formation of Tissue Engineered Product (TEP) based on polyurethane support and primary human osteoblasts. The aim of the present study was to fabricate TEP consisting of Human Bone Derived Cells (HBDCs) seeded on 3-D biodegradable polyurethane scaffolds and to evaluate its biocompatibility in vivo.
HBDCs were seeded (3,0x105 cells/sample) and cultured on specially developed polyurethane scaffolds in spinner flask type bioreactor for 14 days. The experiment was performed twice. Each time 6 samples were used for the evaluation of TEP in vitro and additional 22 scaffolds were used for the in vivo experiment. TEP samples and control samples (scaffolds without cells) were implanted subcutaneously into the SCID mice for the 4 and 13 weeks periods.
TEP examination in vitro was performed by means of: cell number (XTT assay), cell phenotype (RT-PCR) and cell distribution within the scaffold (fluorescence microscopy). Explants harvested from the animals were examined using histological (HE) and immunohistochemical (Collagen I and Osteopontin) methods.
It was found that dynamic cell seeding and cell culture conditions enable homogenous distribution, maintaining the proliferative potential and osteogenic phenotype of the HBDCs cultured on the selected three dimensional polyurethane scaffold – fabrication of TEP.
It was also found, that Human Bone Derived Cells implanted into the experimental animals as a component of TEP keep their ability to synthesize and secrete the specific components of the bone tissue Extracellular Matrix.
Acknowledgment: This work was supported by the Ministry of Science and Higher Education, grant R13 01901 and by the MUW (NZME/W2/08).
1 Guelcher, S.A. (2005) In An Introduction to Biomaterials (Eds, Guelcher, S.A. and Hollinger, J.O.) pp. 576.
2 Bil et all. J Mater. Sci. in press.
Presentation: Oral at E-MRS Fall Meeting 2008, Symposium L, by Piotr Woźniak
See On-line Journal of E-MRS Fall Meeting 2008
Submitted: 2008-06-10 15:36 Revised: 2009-06-07 00:48