Effects of Pulsatile Flow on Cultured Vascular Endothelial Cell Morphology

Abstract

Endothelial cells (EC) appear to adapt their morphology and function to the in vivo hemodynamic environment in which they reside. In vitro experiments indicate that similar alterations occur for cultured EC exposed to a laminar steady-state flow-induced shear stress. However, in vivo EC are exposed to a pulsatile flow environment; thus, in this investigation, the influence of pulsatile flow on cell shape and orientation and on actin microfilament localization in confluent bovine aortic endothelial cell (BAEC) monolayers was studied using a 1-Hz nonreversing sinusoidal shear stress of 40 ± 20 dynes/cm2 (type I), 1-Hz reversing sinusoidal shear stresses of 20 ± 40 and 10 ± 15 dynes/cm2 (type II), and 1-Hz oscillatory shear stresses of 0 ± 20 and 0 ± 40 dynes/cm2 (type III). The results show that in a type I nonreversing flow, cell shape changed less rapidly, but cells took on a more elongated shape than their steady flow controls long-term. For low-amplitude type II reversing flow, BAECs changed less rapidly in shape and were always less elongated than their steady controls; however, for high amplitude reversal, BAECs did not stay attached for more than 24 hours. For type III oscillatory flows, BAEC cell shape remained polygonal as in static culture and did not exhibit actin stress fibers, such as occurred in all other flows. These results demonstrate that EC can discriminate between different types of pulsatile flow environments. Furthermore, these experiments indicate the importance of engineering the cell culture environment so as to include pulsatile flow in investigations of vascular endothelial cell biology, whether these studies are designed to study vascular biology and the role of the endothelial cell in disease processes, or are ones leading to the development of hybrid, endothelial cell-preseeded vascular grafts.