DEXTRAN INCREASES SURVIVAL OF SUBCONFLUENT ENDOTHELIAL-CELLS EXPOSED TO SHEAR-STRESS

In vitro devices in combination with cultured cells have been used to study the relationship between shear stress and endothelial injury. Almost exclusively, these investigations have used confluent monolayers and conventional culture media as perfusates and reported little cell loss over a wide range of shear stress conditions. In this investigation when subconfluent endothelial cells were exposed to 22 and 88 dyn/cm2 for 2, 8, and 24 h in a perfusate of medium and 5% serum, a progressive cell loss was observed. Lower cell densities were a product of decreased cell proliferation as measured by bromodeoxyuridine (BrdU) incorporation and loss of the initial cell population. Video recordings indicated that cells characteristically detached by proximal cell peeling from the substrate and an aneurysmal rupture of the cell membrane. Cell retention was increased by including 250 and 475 muM neutral dextran (70 kDa) in perfusates. Experimental evidence suggests dextran does not directly stimulate proliferation or correct an osmotic imbalance. This investigation has substantiated that fluid-generated shear stress can cause endothelial denudation and that conditions (subconfluency, time, and perfusate supplementation) under which shear stress is applied are as important for cell survival as shear stress magnitude.