What is the role of the convective current density in the real-time calcium response of cultured bone cells to fluid flow?

ABSTRACT

Cultured cells subjected to fluid flow are exposed to mechanical forces and electrokinetic forces. The convective current establishes an electrokinetic force created by the flow-dependent transport of mobile ions in the media over the charged cell surfaces. This current can be expressed as a current density, the current normalized by the cross-sectional area in which it exists. In this study, we hypothesized that the convective current density has no role in the bone cell real-time intracellular calcium response to fluid flow. Our hypothesis was tested by incorporating electrokinetic measurements and classical electrokinetic double-layer theory to estimate the value of convective current density in a parallel-plate flow chamber and then to apply an external current during the presence of fluid flow that would alter convective current density. There was no difference between the mean peak calcium response of cells exposed to flow with an altered (canceled or doubled) convective current density versus flow with an unmodified convective current density, as was measured with fura-2 fluorescence microscopy. These results suggest that mechanical forces, such as fluid-induced shear stress, rather than concomitant electrokinetic forces are the primary stimuli in eliciting the observed calcium response of bone cells to fluid flow.