Co-culture of Smooth Muscle Cells and Endothelial Cells on Porous 3D Polyurethane Scaffolds for Vascular Tissue Engineering

One of the challenges in the designing of clinically-relevant vascular substitutes is our lack of understanding on how vascular smooth muscle cells (VSMCs) and vascular endothelial cells (VECs) interact in the graft. The aim of this study was to examine the factors that play a role in VSMC and VEC interaction in 3D co-culture. Highly porous 3D poly(carbonate urethane) scaffolds were fabricated using a solvent casting and particulate leaching method. VSMCs and VECs were co-cultured for 48 hours. Immunofluorescence staining showed that VSMCs readily attached to the scaffold and formed dense confluent layers which facilitated the organization of VECs of into a monolayer above the VSMC layer. Western blot analysis showed that co-culture induced an up-regulation of the contractile phenotype in VSMCs. A small interfereing RNA knockdown study of Jagged1 established a direct link between Jagged1 expression in VECs and contractile protein expression in VSMCs