Optimizing Endothelialization of Prevascularized Tissue Constructs.
Luks, Valerie Louise.
- Background: Due largely to the exponential increase in demand for replacement organs, tissue engineering has become an area of increasing technological focus. One significant challenge for the successful development of large-scale engineered tissue constructs is to ensure efficient nutrient delivery throughout the bulk of the tissue. Therefore, it is necessary to develop a viable method for ... read morevascularizing scaffolds so that engineered constructs can be immediately perfused upon implantation. Methods: Soft lithography was used to fabricate micro-scale channels into a porous silk scaffold. Endothelial cells were seeded within these microchannels and cultured under static and shear conditions for up to two weeks. Chemical and physical parameters were manipulated and quantitative and qualitative measurements were gathered to determine optimal conditions for channel endothelialization. Quantitative RT-PCR measured the expression of three genes involved in cell remodeling and lumen formation. Results: Cell growth was observed in all channel sizes using histological and immunohistochemical analysis. After two weeks of culture, endothelial cells still expressed proliferation markers predominantly in the shear culture flasks. Conclusion: The results of this study demonstrate the ability of microchannels to prevascularize a porous, biodegradable scaffold. After 14 days of dynamic culture conditions, the microchannels supported endothelial cell proliferation and stable luminal structures.read less