Fluid flow-induced shear stress results in a variety of morphological and metabolic changes in cultured bovine auricular chondrocytes (BAC). However, the mechanism by which the flow signal is transduced into a biological response is unknown. Therefore, we investigated the effects of fluid flow on intracellular Ca2+ concentration ([Ca2+]i) in BAC. Cells loaded with fura 2 were exposed to steady and pulsatile (0.5 Hz) flow at 9, 18, and 34 ml/min in a parallel-plate flow chamber. In response to flow, there was a significant and flow rate-dependent increase in the percentage of cells showing a rise in [Ca2+]i, but no effect on the [Ca2+]i response amplitude. There was no significant difference between the [Ca2+]i responses to steady and pulsatile flow. Mean intracellular Ca2+ response values ranged between 26.2 ( 1.6 (9 ml/min) and 38.0 ( 6.8 nM (34ml/min) above basal [Ca2+]i (81.3 ( 24.1 nM, n=90). Removal of extracellular Ca2+ or addition of Gd3+ significantly reduced the percentage of cells responding, suggesting that influx of Ca2+, possibly through mechanosensitive channels, contributes to the rise in intracellular Ca2+.
Our data suggest fluid flow-induced mobilization of intracellular Ca2+ may contribute to the mechanism by which mechanical loads are transduced by chondrocytes.