The covalent attachment of adhesion molecules to silicone membranes for cell stretching applications
The covalent attachment of adhesion molecules to silicone membranes for cell stretching applications
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DOI：
10.1016/j.biomaterials.2008.12.022
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发表时间：
2009-03-01
期刊：
影响因子：
14
通讯作者：
Hinz, Boris
中科院分区：
文献类型：
--
作者：
Wipff, Pierre-Jean;Majd, Hicham;Hinz, Boris
带有可膨胀聚二甲基硅氧烷(PDMS)培养膜的应变装置经常用于在体外拉伸细胞，模拟机械动态的组织环境。为了将细胞黏附分子固定到非黏附的PDMS底物上，已经开发了疏水、静电和共价表面包覆方法。然而，不同的涂层策略将拉伸传输到细胞的效果却很少有文献记载，也没有进行过比较。我们描述了一种新颖而简单的方法，将细胞外基质蛋白共价结合到可伸展的PDMS膜的表面。该方法包括PDMS氧化、硅烷化和共价蛋白质与硅烷的交联。在已建立的涂层程序中，我们展示了成纤维细胞在新涂层上的附着性(类似于2倍)、铺展(类似于2.5倍)和增殖(类似于1.2倍)的改善。此外，我们还比较了不同的PDMS涂层技术对传输拉伸的效率。经过15%的拉伸后，我们的PDMS表面涂层上的最大拉伸细胞数量(15+/-5%)类似于替代涂层方案的7倍。因此，粘合分子的共价键在提供抵抗大变形并将这种拉伸完全传递到培养细胞的涂层方面优于非共价方法。(C)2008爱思唯尔有限公司。保留所有权利。
Strain devices with expandable polydimethylsiloxane (PDMS) culture membranes are frequently used to stretch cells in vitro, mimicking mechanically dynamic tissue environments. To immobilize cell-adhesive molecules to the otherwise non-adhesive PDMS substrate, hydrophobic, electrostatic and covalent surface coating procedures have been developed. The efficacy of different coating strategies to transmit stretches to cells however is poorly documented and has not been compared. We describe a novel and simple procedure to covalently bind extracellular matrix proteins to the surface of stretchable PDMS membranes. The method comprises PDMS oxygenation, silanization, and covalent protein cross-linking to the silane. We demonstrate improved attachment (similar to 2-fold), spreading (similar to 2.5-fold) and proliferation (similar to 1.2-fold) of fibroblasts to our new coating over established coating procedures. Further, we compared the efficiency of different PDMS coating techniques to transmit stretches. After 15% stretch, the number of maximally (15 +/- 5%) stretched cells on our PDMS surface coating was similar to 7-fold higher compared with alternative coating protocols. Hence, covalent linkage of adhesive molecules is superior to non-covalent methods in providing a coating that resists large deformations and that fully transmit this stretch to cultured cells. (C) 2008 Elsevier Ltd. All rights reserved.