Quantification of Morphological Modulation, F-Actin Remodeling and PECAM-1 (CD-31) Re-distribution in Endothelial Cells in Response to Fluid-Induced Shear Stress under Various Flow Conditions.

Cardiovascular diseases (CVDs) are the number one cause of death globally. Arterial endothelial cell (EC) dysfunction plays a key role in many of these CVDs, such as atherosclerosis. Blood flow-induced wall shear stress (WSS), among many other pathophysiological factors, is known to significantly contribute to EC dysfunction. The present study reports an in vitro investigation of the effect of quantified WSS on ECs, analyzing the EC morphometric parameters as well as cytoskeletal remodeling. The effects of four different flow cases (low steady laminar (LSL), medium steady laminar (MSL), non-zero-mean sinusoidal laminar (NZMSL) and laminar carotid (LCRD) waveforms) on EC area, perimeter, shape index (SI), angle of orientation, F-actin bundle remodeling and PECAM-1 localization were studied. For the first time, a flow facility was fully quantified for the uniformity of flow over ECs as well as for WSS determination (as opposed to relying on analytical equations). The SI and angle of orientation were found to be the most flow-sensitive morphometric parameters. A 2D Fast Fourier Transform based image processing technique was applied to analyze the F-actin directionality and an alignment index (AI) was defined accordingly. Also, a significant peripheral loss of PECAM-1 in ECs subjected to atheroprone cases (LSL and NZMSL) with high cell surface/cytoplasm stain of this protein is reported, which may shed light on of the mechanosensory role of PECAM-1 in mechanotransduction.