Gelatin-based perfusable, endothelial carotid artery model for the study of atherosclerosis

Abstract

Background: Carotid artery geometry is important for recapitulating a pathophysiological microenvironment to study wall shear stress (WSS)-induced endothelial dysfunction in atherosclerosis. Endothelial cells (ECs) cultured with hydrogel have been shown to exhibit in vivo-like behaviours. However, to date, studies using hydrogel culture have not fully recapitulated the 3D geometry and blood flow patterns of real-life healthy or diseased carotid arteries. In this study, we developed a gelatin-patterned, endothelialized carotid artery model to study the endothelium response to WSS. Results: Two representative regions were selected based on the computational fluid dynamics on the TF-shaped carotid artery: Region ECA (external carotid artery) and Region CS (carotid sinus). Progressive elongation and alignment of the ECs in the flow direction were observed in Region ECA after 8, 16 and 24 h. However, the F-actin cytoskeleton remained disorganized in Region CS after 24 h. Further investigation revealed that expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) was greatly increased in Region CS relative to that in Region ECA. The physiological WSS in the carotid artery system was found to stimulate nitric oxide (NO) and prostacyclin (PGI2) release and inhibit endothelin-1 (ET-1) release after 24-h perfusion experiments. The effective permeability (E.P) of fluorescein isothiocyanate (FITC)-dextran 40 kDa in Regions ECA and CS was monitored, and it was found that the turbulence WSS value (in Region CS) was less than 0.4 Pa, and there was a significant increase in the E.P relative to that in Region ECA, in which laminar WSS value was 1.56 Pa. The tight junction protein (ZO-1) production was shown that the low WSS in Region CS induced ZO-1-level downregulation compared with that in Region ECA. Conclusions: The results suggested that the gelatin-based perfusable, endothelial carotid artery model can be effective for studying the pathogenesis of atherosclerosis by which flow dynamics control the endothelium layer function in vitro.