Low Shear-induced Fibrillar Fibronectin: comparative analyses of morphologies and cellular effects on Bovine Aortic Endothelial Cell Adhesion and Proliferation

Phys Biol. 2024 Oct 4. doi: 10.1088/1478-3975/ad838c. Online ahead of print.

Abstract

Wall shear stress (WSS) is a critical factor in vascular biology, and both high and low WSS are implicated in atherosclerosis. Fibronectin (FN) is a key extracellular matrix protein that plays an important role in cell activities. Under high shear stress, plasma FN (pFN) undergoes fibrillogenesis; however, its behavior under low shear stress remains unclear. This study aimed to investigate the formation of in vitro cell-free fibrillar FN (FFN) under low shear rate conditions and its effect on bovine aortic endothelial cell behavior. FN (500 µg/ml) was perfused through slide chambers at three flow rates (0.16 ml/h, 0.25 ml/h, and 0.48 ml/h), corresponding to low shear rates of 0.35 s-1, 0.55 s-1, and 1.05 s-1, respectively, for 4 hours at room temperature. The formed FN matrices were observed using fluorescence microscopy and SEM. Under low shear rates, distinct FN matrix structures were observed. FFN0 .48 formed immense fibrils with smooth surfaces, FFN0.25 formed a matrix with a rough surface, and FFN16 exhibited nodular structures. FFN0.25 supported cell activities to a greater extent than native FN and other FFN surfaces. Our study suggests that abnormally low shear conditions impact FN structure and function and enhance the understanding of FN fibrillogenesis in vascular biology, particularly in atherosclerosis.

Keywords: fibronectin; fibronectin fibrillogenesis; hemodynamic; low shear rate.