Biomechanical effects of flow and coculture on human aortic and cord blood-derived endothelial cells

Human endothelial cells derived from umbilical cord blood (hCB-ECs) represent a promising cell source for endothelialization of tissue engineered blood vessels. hCB-ECs cultured directly above human aortic smooth muscle cells (SMCs), which model native and tissue engineered blood vessels, produce a confluent endothelium that responds to flow like normal human aortic endothelial cells (HAECs). The objective of this study was to quantify the elastic modulus of hCB-ECs cocultured with SMCs under static and flow conditions using atomic force microscopy (AFM). Cytoskeleton structures were assessed by AFM cell surface imaging and immunofluorescence of F-actin. The elastic moduli of hCB-ECs and HAECs were similar and significantly smaller than the value for SMCs in monoculture under static conditions (p<0.05). In coculture, hCB-ECs and HAECs became significantly stiffer with moduli 160-180% larger than their corresponding values in monoculture. While the moduli of hCB-ECs and HAECs almost doubled in monoculture and flow condition, their corresponding values in coculture declined after exposure to flow. Both the number and diameter of cortical stress fiber per cell width increased in coculture and/or flow conditions, whereas the subcortical stress fiber density throughout the cell interior increased by a smaller amount. These findings indicate that changes to biomechanical properties in coculture and/or exposure to flow are correlated with changes in the cortical stress fiber density. For ECs, fluid shear stress appeared to have greater effect on the elastic modulus than the presence of SMCs and changes to the elastic modulus in coculture may be due to EC-SMC communication.