| Tissue engineering of large diameter blood vessels can offer a promising long-term solution to the large population suffering from congenital vascular defects and other vascular disease. In this report design, assembly, in vitro maturation and evaluation of a large diameter, chitosan-based prosthesis is described. To facilitate cell adhesion and proliferation, collagen was included as a scaffold component to a chitosan scaffold. In vitro studies evaluated the role of collagen content, crosslinker type and crosslinking density on degradation kinetics, mechanical properties and cellular interactions. Finally, the vessel scaffold (ID = 12 mm, OD = 15 mm) was fabricated from a moderately cross-linked, 90%/10% chitosan/collagen material. A tubular scaffold with gradient porosity and interconnected pores was generated by controlled freezing and lyophilization of the polymer.;For graft culture laminar and pulsatile flow systems were designed and porous scaffolds were seeded with vascular cells under static conditions. Laminar system grafts were seeded and cultured/analyzed over an 8 week period (15ml/min). For the pulsatile system SMC were seeded and after 2 weeks of pulsatile flow culture (360ml/min, 82 beats/min) microvascular EC were seeded lumenally to initiate a microvascular network followed by aortic EC seeding at 3 weeks. For both systems, cell viability at different culture periods showed the formation of high density of cell within few weeks of graft culture. However, the pulsatile flow system graft showed a significant increase in mechanical properties and ECM protein (collagen and elastin) deposition overtime. This novel chitosan based tissue engineered vascular graft shows promising results for large vessel replacements. |
