The Effects of Substrate Elasticity on Endothelial Cell Network Formation and Traction Force Generation

While the growth factors and cytokines known to influence angiogenesis and vasculogenesis have garnered widespread attention, less is known about how the mechanical environment affects blood vessel formation and cell assembly. In this study, we investigate the relationship between substrate elasticity, endothelial cell-cell connectivity and traction force generation. We find that on more compliant substrates, endothelial cells self-assemble into network-like structures independently of additional exogenous growth factors or cytokines. These networks form from the assembly of sub-confluent endothelial cells on compliant (E=200-1000Pa) substrates, and results from both the proliferation and migration of endothelial cells. Interestingly, stabilization of these cell-cell connections and networks requires fibronectin polymerization. Traction Force Microscopy measurements indicate that individual endothelial cells on compliant substrates exert forces which create substrate stains that propagate from the cell edge. We speculate that these strains draw the cells together and initiate self-assembly. Notably, endothelial cell network formation on compliant substrates is dynamic and transient; as cell number and substrate strains increase, the networks fill in through collective cell movements from the network edges. Our results indicate that network formation is mediated in part by substrate mechanics and that cellular traction force may promote cell-cell assembly by directing cell migration.