A systematic investigation of endothelial cell behavior under hydrostatic pressure.

Biomechanical stimuli are critical in regulating cell behavior and phenotype across various tissues and organs, particularly within the cardiovascular system. Endothelial cells, which line blood vessels, are continuously subjected to forces generated by the pulsatile nature of blood flow, including shear stress, strain, and hydrostatic pressure (HP). Among these stimuli, HP remains the least explored, primarily due to the technical challenges of incorporating it into conventional cell culture systems. However, HP significantly influences key biological processes, such as cell differentiation, migration, proliferation, and apoptosis. To facilitate the introduction of HP in vitro, we have previously developed an automated, high-throughput platform compatible with standard 96-well plates capable of delivering up to 12 independent pressure conditions. In this study, we applied this setup to investigate the effects of a wide range of static pressure conditions on the viability, morphology, and cytoskeleton adaptation of Human Umbilical Vein Endothelial Cells (HUVECs).