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Vascular endothelial growth factor (VEGF) has been extensively investigated to promote vascularization at damaged or diseased sites and in tissue implants. Here we are interested in determining if the manner in which VEGF is presented from a scaffold to endothelial cells influences the architecture of the blood vessels formed. We bound VEGF to nanoparticles and placed these nanoparticles inside fibrin hydrogels, which contained human umbilical vein endothelial cells (HUVECs) bound to cytodex beads. Fibroblast cells are plated on top of the fibrin gel to further mimic a physiologic environment. In addition, we used a chorioallantoic membrane (CAM) assay to determine the role of VEGF presentation on angiogenesis in vivo. We tested VEGF bound in high density and low density to study differences between growth factor presentation in heterogeneous nanodomains and homogenous distribution. VEGF covalently bound to nanoparticles at high density led to an increase in HUVEC tube branching, thickness, and total vessel network length compared to soluble VEGF. While VEGF bound electrostatically exhibited no significant difference with covalently bound VEGF in the tube formation assay, this method failed to promote host vessel infiltration into the fibrin implant on the CAM. Together our data suggest that the mode of VEGF presentation to endothelial cells influences the vessel architecture and vascularization of implants in vivo.

Sean M Anderson, Shayne N Siegman, Tatiana Segura

Anderson, SM; Siegman, SN; Segura, T