The arterial intima is continuously under pulsatile wall shear stresses (WSS) imposed by the circulating blood. The knowledge of the contribution of smooth muscle cells (SMC) to the response of endothelial cell (EC) to WSS is still incomplete. We developed a co-culture model of EC on top of SMC that mimics the inner in vivo structure of the arterial intima of large arteries. The co-cultured model, as well as a monolayer model of EC, were developed in custom-designed wells that allowed for mechanobiology experiments. Both the monolayer and co-culture models were exposed to steady flow induced WSS of up to 24 dyne/cm² and for lengths of 60 min. Quantification of WSS induced alterations in the cytoskeletal actin filaments (F-actin) and vascular endothelial cadherin (VE-cadherin) junctions were utilized from confocal images and flow cytometry. High confluency of both models was observed even after exposure to the high WSS. The quantitive analysis revealed larger post WSS amounts of EC F-actin polymerization in the monolayer, which may be explained by the relative help of the SMC to resist the external load of WSS. The VE-cadherin demonstrated morphological alterations in the monolayer model, but without significant changes in their content. The SMC in the co-culture maintained their contractile phenotype post high WSS which is more physiological, but not post low WSS. Generally, the results of this work demonstrate the active role of SMC in the intima performance to resist flow induced WSS.

动脉内膜持续受到循环血液施加的脉动壁切应力（WSS）的作用。关于平滑肌细胞（SMC）对内皮细胞（EC）对WSS反应的贡献的知识仍然不完整。我们在SMC之上开发了EC的共培养模型，该模型模仿了大动脉的动脉内膜的体内体内结构。共培养模型以及EC的单层模型是在定制设计的孔中开发的，该孔允许进行机械生物学实验。单层和共培养模型均暴露于高达24达因/ cm ^2的稳定流诱导的WSS长度为60分钟。从共聚焦图像和流式细胞仪中，利用WSS诱导的细胞骨架肌动蛋白丝（F-肌动蛋白）和血管内皮钙粘蛋白（VE-钙粘蛋白）连接的改变。甚至在暴露于高WSS后也观察到两个模型的高度融合。定量分析显示，WSS后单层中的EC F-肌动蛋白聚合量更大，这可以通过SMC抵抗WSS外部负载的相对帮助来解释。VE-钙粘着蛋白在单层模型中表现出形态学改变，但其含量没有明显变化。共培养物中的SMC在高WSS后保持其收缩表型，这更生理，但在低WSS后不保持。通常，