Intracellular calcium response to directly applied mechanical shearing force in cultured vascular endothelial cells

We studied the responses of cultured endothelial cells to mechanical shearing force directly applied to those cells in vitro to determine changes in the concentration of intracellular calcium ion (Ca++), one of the factors that transfers information within the cell. Cultured bovine fetal aortic endothelial cells containing the Ca++ fluorescence indicator, Fura-2, were rubbed with a latex balloon in a specially designed system, and changes in the fluorescence of Fura-2 caused by this shear stimulation were determined by photometric fluorescence microscopy. Immediately after shear stimulation, the concentration of Ca++ in the cells was increased and reached a peak (511 +/- 165 nM, n = 12) within 15 seconds after stimulation. After the peak, the concentration was gradually restored to the resting level (55 +/- 17 nM, n = 12). The magnitude of the Ca++ response was dependent on the intensity of the shear force applied. Analysis of fluorescence images of Fura-2 revealed that the cells showed this Ca++ reaction without being injured or desquamated, although there were slight differences in the degree and duration of reaction among cells. This reaction appeared even when the cells were placed in the air with no contact with the fluid. This result suggests that neither the fluid flow associated with the balloon movement nor chemical substances in the fluid are involved in the reaction, but that pure physical force alone is responsible for the Ca++ reaction. Further, it suggests that endothelial cells have the ability to perceive such physical stimulation as shear force and to transfer this information to the interior of the cell via changes in the intracellular Ca++ concentration.