Extracellular matrix remodelling during cell adhesion monitored by the quartz crystal microbalance
Extracellular matrix remodelling during cell adhesion monitored by the quartz crystal microbalance
复制标题
DOI：
10.1016/j.biomaterials.2008.03.002
复制
发表时间：
2008-06-01
期刊：
影响因子：
14
通讯作者：
Milthorpe, Bruce K.
中科院分区：
文献类型：
--
作者：
Lord, Megan S.;Modin, Charlotte;Milthorpe, Bruce K.
细胞重塑表面吸附蛋白质层的能力受到蛋白质层本身性质的影响。通常需要重塑来实现细胞粘附和细胞外基质形成，其形成细胞铺展、增加粘附和不同表型表达的基础。用石英晶体微天平（QCM-D）和荧光显微镜检测NIH 3 T3（EGFP）成纤维细胞与血清蛋白（白蛋白或纤连蛋白）预涂钽（Ta）和氧化聚苯乙烯（PS（ox））表面的粘附。细胞未经处理或用放线菌酮处理，以检查在细胞粘附期间内源性蛋白质产生对2小时内QCM-D响应的贡献。在将白蛋白吸附到Ta和PS上之后，在接种未处理的细胞和放线菌酮处理的细胞的反应之间没有检测到差异。QCM-D能够检测到未处理的细胞对纤连蛋白与血清预涂Ta和PSox底物的反应的差异，而放线菌酮处理的细胞对纤连蛋白和血清预涂在每种材料上产生相同的QCM-D反应。这证实了细胞的基质重塑过程取决于底层基质和预吸附蛋白质，并且QCM-D响应主要由底层蛋白质层的变化决定。消散的变化对应于肌动蛋白细胞骨架的发展，如通过肌动蛋白染色所观察到的。(c)2008爱思唯尔有限公司版权所有。
A cell's ability to remodel adsorbed protein layers on surfaces is influenced by the nature of the protein layer itself. Remodelling is often required to accomplish cellular adhesion and extracellular matrix formation which forms the basis for cell spreading, increased adhesion and expression of different phenotypes. The adhesion of NIH3T3 (EGFP) fibroblasts to serum protein (albumin or fibronectin) precoated tantalum (Ta) and oxidised polystyrene (PS(ox)) surfaces was examined using the quartz crystal microbalance with dissipation (QCM-D) monitoring and fluorescence microscopy. The cells were either untreated or treated with cycloheximide to examine the contribution of endogenous protein production during cell adhesion to the QCM-D response over a period of 2 h. Following adsorption of albumin onto Ta and PS., there was no difference detected between the response to seeding untreated and cycloheximide treated cells. The QCM-D was able to detect differences in the untreated cellular responses to fibronectin versus serum precoated Ta and PSox substrates, while cycloheximide treatment of the cells produced the same QCM-D response for fibronectin and serum precoatings on each of the materials. This confirmed that the process of matrix remodelling by the cells is dependent on the underlying substrate and the preadsorbed proteins and that the QCM-D response is dominated by changes in the underlying protein layer. Changes in dissipation correspond to the development of the actin cytoskeleton as visualised by actin staining. (c) 2008 Elsevier Ltd. All rights reserved.