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A Parallel-Plate Flow Chamber for Mechanical Characterization of Endothelial Cells Exposed to Laminar Shear Stress

机译:平行板流动室用于表征层流剪切应力作用下的内皮细胞

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摘要

Shear stresses induced by laminar fluid flow are essential to properly recapitulate the physiological microenvironment experienced by endothelial cells (ECs). ECs respond to these stresses via mechanotransduction by modulating their phenotype and biomechanical characteristics, which can be characterized by Atomic Force Microscopy (AFM). Parallel Plate Flow Chambers (PPFCs) apply unidirectional laminar fluid flow to EC monolayers in vitro. Since ECs in sealed PPFCs are inaccessible to AFM probes, cone-and-plate viscometers (CPs) are commonly used to apply shear stress. This paper presents a comparison of the efficacies of both methods. Computational Fluid Dynamic simulation and validation testing using EC responses as a metric have indicated limitations in the use of CPs to apply laminar shear stress. Monolayers subjected to laminar fluid flow in a PPFC respond by increasing cortical stiffness, elongating, and aligning filamentous actin in the direction of fluid flow to a greater extent than CP devices. Limitations using CP devices to provide laminar flow across an EC monolayer suggest they are better suited when studying EC response for disturbed flow conditions. PPFC platforms allow for exposure of ECs to laminar fluid flow conditions, recapitulating cellular biomechanical behaviors, whereas CP platforms allow for mechanical characterization of ECs under secondary flow.
机译:层流引起的剪切应力对于正确概括内皮细胞(EC)经历的生理微环境至关重要。 EC通过调节其表型和生物力学特性,通过机械转导来响应这些压力,这可以用原子力显微镜(AFM)来表征。平行板流动室(PPFC)在体外将单向层流应用于EC单层。由于AFM探头无法获得密封PPFC中的EC,因此通常使用锥板粘度计(CP)施加剪切应力。本文介绍了两种方法的效果比较。使用EC响应作为度量标准的计算流体动力学模拟和验证测试表明,使用CP施加层流切应力的局限性。与CP装置相比,在PPFC中经受层流流动的单分子层通过增加皮质的刚度,延长并在流体流动方向上排列丝状肌动蛋白来做出响应。使用CP设备在EC单层上提供层流的局限性表明,它们更适合研究扰动流量条件下的EC响应。 PPFC平台允许EC暴露于层流条件下,概括了细胞的生物力学行为,而CP平台允许在二次流动下对EC进行机械表征。

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