Functional Neovascularization in Tissue Engineering with Porcine Acellular Dermal Matrix and Human Umbilical Vein Endothelial Cells

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Abstract
Endothelial cells–matrix interactions play an important role in promoting and controlling network formation. In this study, porcine acellular dermal matrix (PADM) was used to guide human umbilical vein endothelial cells (HUVECs) adhesion and proliferation as a potential system for vascularization of engineered tissues. We fabricated PADM using a modified protocol and assessed their composition and ultrastructures. Subsequently, the viability of HUVECs and the formation of capillary-like networks were evaluated by seeding cells directly on PADM scaffolds or PADM digests in vitro. We further investigated the function of the HUVECs seeded on the PADM scaffolds after subcutaneous transplantation in athymic mice. Moreover, the function of the neovessels formed in the PADM scaffolds was assessed by implantation into cutaneous wounds on the backs of mice. The results showed that PADM scaffolds significantly increased proliferation of HUVECs, and the PADM digest induced HUVECs formed many tube-like structures. Moreover, HUVECs seeded on the PADM scaffolds formed numerous capillary-like networks and some perfused vascular structures after implantation into mice. PADM seeded with HUVECs and fibroblasts were also able to form many capillary-like networks in vitro. Further, these neovessels could inosculate with the murine vasculature after implantation into cutaneous wounds in mice. The advantage of this method is that the decellularized matrix not only provides signals to maintain the viability of endothelial cells but also serves as the template structure for regenerated tissue. These findings indicate that PADM seeded with HUVECs may be a potential system for successful engineering of large, thick, and complex tissues.

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