The success of therapeutic vascularization and tissue engineering will rely on our ability to create vascular networks using clinically friendly biomaterials. The ability to influence the direction and structure in the formation of a vascular system is crucial in engineering tissue. Traditional approaches for three-dimensional scaffold fabrication are not suitable for generating large area and complex structures due to the lack of control of the architecture, microvasculature, and cell-cell interactions. The proposed photolithography platform allows us to design uniform large area, multilayers and complex tissue scaffolds with arbitrary architecture that can mimic the microarchitecture of tissues and to augment regeneration therapies. The photolithography system was developed and optimized to fabricate complex three-dimensional architecture using gelatin methacrylate biomaterial by optimizing a number of process parameters to get good nutrient diffusion and endothelial cell viability. Variation of the methacrylation degrees enabled tailoring mechanical and degradation rate properties of the scaffolds. A dynamic cell seeding method was designed to optimize the coverage of the scaffold. Scaffolds were incubated in a cell suspension and submitted to constant agitation, resulting in enhanced cell adhesion compared to conventional cell seeding. Our results demonstrates we can scale up our scaffolds up to 6 cm in diameter and fabricate multilayers which allowed for uniform endothelial cells distributed and proliferated throughout whole scaffolds, resulting into high cell density and homogeneous distribution at the end of the culture period. Moreover, results obtained at day 1, day 2 and day 4 clearly shows an increase cells in migration, proliferation and spreading throughout the full thickness of the scaffolds as well as a complete three-dimensional cell coverage of the material by endothelial cells.
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