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用于研究血管平滑肌细胞的聚二甲基硅氧烷弹性微柱阵列

PDMS Elastic Micropost Arrays for Studying Vascular Smooth Muscle Cells.

作者信息

Cheng Qi, Sun Zhe, Meininger Gerald, Almasri Mahmoud

机构信息

Department of Electrical and Computer Engineering, University of Missouri, Columbia, MO 65211 USA.

Dalton Cardiovascular Research Center and Department of Medical Pharmacology and Physiology, University of Missouri, Columbia, MO 65211 USA.

出版信息

Sens Actuators B Chem. 2013 Nov 1;188:1055-1063. doi: 10.1016/j.snb.2013.08.018.

Abstract

This paper describes the design, modeling, fabrication and characterization of a micromachined array of high-density 3-dimensional microposts (100×100) made of flexible material (silicone elastomers) for use to measure quantitatively the cellular traction force and contractile events in isolated vascular smooth muscle cells (VSMCs). The micropost array was fabricated with diameters ranged from 3 to 10 μm, with edge to edge spacing of 5, 7 and 10 μm, and with a height to diameter aspect ratio up to 10. VSMCs exerted larger basal traction forces when they were grown on stiffer micropost arrays. These basal traction forces were 80% larger in control VSMCs than in VSMCs in which integrin linked kinase (ILK) was knocked down using shRNA. The addition of Angiotensin II (ANGII) led to VSMC contraction as evidenced by an increased traction force exerted on the microposts under the cell. This ANGII induced contractile response and change in traction force on the microposts was not observed in VSMCs lacking ILK. Following treatment of VSMCs with Cytochalasin D to depolymerize the actin cytoskeleton, the VSMCs exhibited relaxation that was apparent as a significant reduction in the measured traction force exerted on microposts under the cell. Overall, this study demonstrates the usefulness of micropost arrays for study of the contractile responsiveness of VSMC and the results indicate that ILK plays a critical role in the signaling pathways leading to the generation of substrate traction force in VSMC.

摘要

本文描述了一种由柔性材料(硅橡胶弹性体)制成的高密度三维微柱(100×100)微机械阵列的设计、建模、制造和表征,用于定量测量分离的血管平滑肌细胞(VSMC)中的细胞牵引力和收缩事件。微柱阵列的制造直径范围为3至10μm,边缘到边缘间距为5、7和10μm,高度与直径的长宽比高达10。当VSMC生长在更硬的微柱阵列上时,会施加更大的基础牵引力。对照VSMC中的这些基础牵引力比使用shRNA敲低整合素连接激酶(ILK)的VSMC中的基础牵引力大80%。添加血管紧张素II(ANGII)导致VSMC收缩,这通过细胞下方微柱上施加的牵引力增加得到证明。在缺乏ILK的VSMC中未观察到这种ANGII诱导的收缩反应和微柱上牵引力的变化。用细胞松弛素D处理VSMC以使肌动蛋白细胞骨架解聚后,VSMC表现出松弛,这表现为细胞下方微柱上测量的牵引力显著降低。总体而言,本研究证明了微柱阵列在研究VSMC收缩反应性方面的有用性,结果表明ILK在导致VSMC中产生底物牵引力的信号通路中起关键作用。

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