Nano-stenciled RGD-gold patterns that inhibit focal contact maturation induce lamellipodia formation in fibroblasts.
Nano-stenciled RGD-gold patterns that inhibit focal contact maturation induce lamellipodia formation in fibroblasts.
复制标题
抑制焦点接触成熟的纳米模印 RGD-金图案可诱导成纤维细胞中板状伪足的形成。
DOI：
10.1371/journal.pone.0025459
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Chiquet M
中科院分区：
文献类型：
--
作者：
Lutz R;Pataky K;Gadhari N;Marelli M;Brugger J;Chiquet M
培养成纤维细胞通过细胞-基质黏附以分层方式组装，从而获得蛋白质的复杂性和大小。在这里，我们询问限制细胞-基质粘附的大小如何影响细胞形态和行为。利用纳米模板技术，在培养皿上刻上宽度和间距在250 nm到2 μ m之间的金方块。用RGD肽功能化金作为细胞整合素的配体，并将其镀于小鼠胚胎成纤维细胞。将细胞-基质粘附的长度限制在500 nm或更短的时间内，会干扰vinculin阳性的局灶复合物成熟为局灶接触和纤维状粘附，这表明α5-整合素募集不足。我们发现在亚微米模式下，成纤维细胞广泛分布，但不极化。相反，它们形成了大量的板足和一个没有应力纤维的精细肌动蛋白网。此外，这些细胞表现出异常的纤维连接蛋白纤维形成，与2 μ m平方模式的成纤维细胞相比，它们的定向迁移速度明显降低。RhoA/ROCK信号的干扰消除了细胞形态的模式依赖性差异。我们的研究结果表明，通过纳米图案表面操纵细胞基质粘附的成熟可以影响粘附成纤维细胞的形态、肌动蛋白动力学、迁移和ECM组装。
Cultured fibroblasts adhere to extracellular substrates by means of cell-matrix adhesions that are assembled in a hierarchical way, thereby gaining in protein complexity and size. Here we asked how restricting the size of cell-matrix adhesions affects cell morphology and behavior. Using a nanostencil technique, culture substrates were patterned with gold squares of a width and spacing between 250 nm and 2 µm. The gold was functionalized with RGD peptide as ligand for cellular integrins, and mouse embryo fibroblasts were plated. Limiting the length of cell-matrix adhesions to 500 nm or less disturbed the maturation of vinculin-positive focal complexes into focal contacts and fibrillar adhesions, as indicated by poor recruitment of α5-integrin. We found that on sub-micrometer patterns, fibroblasts spread extensively, but did not polarize. Instead, they formed excessive numbers of lamellipodia and a fine actin meshwork without stress fibers. Moreover, these cells showed aberrant fibronectin fibrillogenesis, and their speed of directed migration was reduced significantly compared to fibroblasts on 2 µm square patterns. Interference with RhoA/ROCK signaling eliminated the pattern-dependent differences in cell morphology. Our results indicate that manipulating the maturation of cell-matrix adhesions by nanopatterned surfaces allows to influence morphology, actin dynamics, migration and ECM assembly of adhering fibroblasts.
登录
查看更多内容
影响因子：
2.9
作者：
Arnold, M;Cavalcanti-Adam, EA;Spatz, JP
通讯作者：
Spatz, JP
影响因子：
3.7
作者：
Sarasa-Renedo, A;Tunç-Civelek, V;Chiquet, M
通讯作者：
Chiquet, M
影响因子：
4.1
作者：
May, T;Mueller, PP;Hauser, H
通讯作者：
Hauser, H
影响因子：
64.5
作者：
NOBES, CD;HALL, A
通讯作者：
HALL, A
影响因子：
56.9
作者：
Chen, CS;Mrksich, M;Ingber, DE
通讯作者：
Ingber, DE