PDMS Elastic Micropost Arrays for Studying Vascular Smooth Muscle Cells.
PDMS Elastic Micropost Arrays for Studying Vascular Smooth Muscle Cells.
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DOI：
10.1016/j.snb.2013.08.018
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发表时间：
2013-11-01
期刊：
影响因子：
--
通讯作者：
Almasri M
中科院分区：
文献类型：
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作者：
Cheng Q;Sun Z;Meininger G;Almasri M
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、制备和表征，用于定量测量离体血管平滑肌细胞（VSMCs）的细胞牵引力和收缩事件。微柱阵列的直径范围为3 ~ 10 μm，边间距为5、7和10 μm，高径比为10。VSMCs施加更大的基底牵引力时，他们生长在更硬的微柱阵列。这些基础牵引力在对照VSMC中比在整合素连接激酶（ILK）使用shRNA敲低的VSMC中大80%。血管紧张素II（ANGII）的加入导致VSMC收缩，如通过施加在细胞下的微柱上的增加的牵引力所证明的。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。在用细胞松弛素D处理VSMC以使肌动蛋白细胞骨架脱粘后，VSMC表现出松弛，这明显表现为施加在细胞下的微柱上的测量牵引力的显著降低。总体而言，这项研究表明微柱阵列的有用性的研究的收缩反应的VSMC和结果表明，ILK起着关键作用的信号通路，导致产生的基板牵引力在VSMC。
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.