Bone cells respond to fluid shear loading by activating various biochemical pathways, mediating a dynamic process of bone formation and resorption. The whole-cell volume dilatation [1] and regional deformation of intracellular structures [2] may be able to directly activate and modulate relevant biochemical pathways. Therefore, understanding how bone cells deform under fluid flow can help elucidate the fundamental mechanisms by which mechanical stimuli are able to initiate biochemical responses. Most studies on cell deformation have focused only on cell deformation in the plane parallel to the substrate surface. Height-dependent cell deformation has not been well characterized even though it may contribute greatly to mechanotransduction mechanisms. Traditional techniques to obtain this additional height information of a cell-body, such as confocal and deconvolution microscopy, are inherently limited by the timescale under which the deformational information can be visualized. Previous studies have investigated cell adhesion to substrate under flow using a single view side-view imaging technique [3, 4]. In this study, we present a novel technique that is able to image a single cell simultaneously in two orthogonal planes to obtain real-time images of a cell at a millisecond timescale. Thus, the objectives of this study were to: (1) develop an imaging technique to visualize the depth-directional information of a cell simultaneously with the traditional 2D view; (2) map out the strain fields of the cell by image analysis; and (3) investigate the viscoelastic behavior of osteoblasts under steady fluid flow.

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