Endothelial cell monolayer formation: Effect of substrate and fluid shear stress

The formation of an endothelial cell (EC) monolayer is one critical factor in the development of a tissue engineered vascular graft. One potential method of endothelialization is the migration of native ECs from the surrounding blood vessel onto the newly implanted graft. In the present study, in vitro experiments were performed to investigate the potential of ECs to migrate on a tissue-engineered blood vessel wall model (TEWM) and form a new monolayer. The TEWM was composed of a three-dimensional, type I collagen matrix seeded with smooth muscle cells. The potential of ECs to form new monolayer was evaluated in the absence and presence of fluid shear stress (10 dynes/cm(2)). The monolayer formation on the TEWM was compared to a control, two-dimensional substrate of glass coated with type I collagen. Results from these studies showed that EC monolayer formation was inhibited on the TEWM in comparison to glass coated with collagen. This correlated with an inhibition of proliferation on the TEWM. The application of shear stress to the EC on the glass coated with collagen also caused an inhibition of monolayer formation, with a corresponding inhibition of proliferation. Furthermore, blocking proliferation by incubation with mitomycin C caused a dose-response inhibition of monolayer formation. In contrast, stimulating proliferation with basic fibroblast growth factor (bFGF) did not further increase monolayer formation on glass coated with collagen. These results suggest that proliferation is one necessary factor for monolayer formation, although not the only factor, where EC proliferation is regulated by its environment, including both substrate and the local shear stress. Continued investigation into the mechanism and stimulation of EC proliferation on the TEWM may lead to developing new strategies for the endothelialization of a tissue engineered vascular graft.