A simple multi-well stretching device to induce inflammatory responses of vascular endothelial cells
A simple multi-well stretching device to induce inflammatory responses of vascular endothelial cells
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一种诱导血管内皮细胞炎症反应的简单多孔拉伸装置
DOI：
10.1039/c5lc01416f
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发表时间：
2016-01-01
期刊：
影响因子：
6.1
通讯作者：
Ding, Yongsheng
中科院分区：
文献类型：
--
作者：
Wang, Jiasheng;Fan, Beiyuan;Ding, Yongsheng
本文介绍了一种新型的多孔拉伸装置，该装置由三层聚二甲基硅氧烷层制成，包括顶部穿孔层、中间薄膜和底部图案化层。这是第一次，这样一个简单的装置已被用来提供轴对称和不均匀的应变培养的细胞在井底是可拉伸的。这些机械刺激可以在一定程度上模拟血管弯曲部位的拉伸，其中应变很复杂。在该装置中，在将一定体积的水注入到中间膜和底层之间的腔室中期间，通过膜从平坦表面变形为球形帽，向细胞施加不均匀的应变。EA.将Hy 926细胞（人脐静脉内皮细胞系）接种在孔底上，并在5、10、15和20%的膜变形度下暴露于轴对称应变。根据细胞形态学、细胞活力以及炎性mRNA和蛋白质的表达来表征细胞应答。随着变形程度的增加，细胞有脱落和凋亡的趋势，MCP-1、IL-8、IL-6和ICAM-1等炎性mRNA和蛋白的表达也明显增加。实验结果表明，电针可明显抑制炎症反应. hy 926细胞可以通过增加应变的大小来诱导。这种简单的装置为血管疾病相关炎症机制的体外研究提供了有用的工具。
We herein introduce a novel multi-well stretching device that is made of three polydimethylsiloxane layers, consisting of a top hole-punched layer, middle thin membrane, and bottom patterned layer. It is the first time that such a simple device has been used to supply axisymmetric and nonuniform strains to cells cultured on well bottoms that are stretchable. These mechanical stimuli can somewhat mimic the stretching at the bending sites of blood vessels, where the strains are complicated. In this device, nonuniform strain is given to cells through the deformation of a membrane from a flat surface to a spherical cap during the injection of a certain volume of water into the chamber between the middle membrane and bottom layer. EA. hy926 cells (a human umbilical vein endothelial cell line) were seeded on the well bottoms and exposed to axisymmetric strain under a 5, 10, 15, and 20% degree of deformation of the membrane. The cellular responses were characterized in terms of cell morphology, cell viability, and expression of inflammatory mRNAs and proteins. With increasing the degree of deformation, the cells exhibited an inclination toward detachment and apoptosis; meanwhile the expression of inflammatory mRNAs and proteins, such as MCP-1, IL-8, IL-6 and ICAM-1, showed a significant increment. The obtained results demonstrate that the inflammatory responses of EA. hy926 cells can be induced by increasing the magnitude of the strain. This simple device provides a useful tool for in vitro investigation of the inflammatory mechanisms related to vascular diseases.