Mechanosensitivity of fibroblast cell shape and movement to anisotropic substratum topography gradients.
Mechanosensitivity of fibroblast cell shape and movement to anisotropic substratum topography gradients.
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成纤维细胞形状和各向异性基质地形梯度的机械敏感性。
DOI：
10.1016/j.biomaterials.2009.06.042
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发表时间：
2009-10
期刊：
影响因子：
14
通讯作者：
Levchenko, Andre
中科院分区：
文献类型：
--
作者：
Kim, Deok-Ho;Han, Karam;Gupta, Kshitiz;Kwon, Keon W.;Suh, Kahp-Yang;Levchenko, Andre
在本报告中，我们描述了使用紫外线（UV）辅助毛细管力光刻（CFL）来创建具有可变局部密度的各向异性微米和纳米形貌图案阵列的模型基质，用于分析细胞-基质相互作用。具有恒定脊宽度 (1 µm) 和深度 (400 nm) 以及可变凹槽宽度（1 µm 至 9.1 µm）的单细胞粘附基质使我们能够表征细胞反应（包括细胞形状、方向和迁移）对可变微米和纳米形貌图案的各向异性和局部密度的依赖性。我们发现，与稀疏区域上的成纤维细胞相比，粘附在较密集图案区域的成纤维细胞沿着脊的方向排列和伸长更强烈，表现出迁移速度对图案密度的双相依赖性。此外，细胞通过改变形态并沿着图案取向（短期）和图案密度（长期）方向偏置的凹槽方向迁移，对形貌的局部变化做出反应。分子动态活细胞成像和粘着斑和肌动蛋白细胞骨架的免疫细胞化学分析表明，可变的基质地形可以导致不同类型的细胞骨架重组。我们还证明，在同一培养基上单层培养的成纤维细胞保留了单细胞显示的大部分特性。这一结果除了证明了一种更复杂的方法来研究伤口愈合过程的各个方面之外，还强烈表明，即使存在相当大的细胞间相互作用，底层地形提供的线索仍然对细胞行为产生重大影响。所描述的实验平台不仅可以进一步加深我们对细胞-基质相互作用的生物力学调节的理解，而且还有助于具有细胞-材料界面的最佳结构设计的设备的生物工程。
In this report, we describe using ultraviolet (UV)-assisted capillary force lithography (CFL) to create a model substratum of anisotropic micro- and nanotopographic pattern arrays with variable local density for the analysis of cell-substratum interactions. A single cell adhesion substratum with the constant ridge width (1 µm), and depth (400 nm) and variable groove widths (1 µm to 9.1 µm) allowed us to characterize the dependence of cellular responses, including cell shape, orientation, and migration, on the anisotropy and local density of the variable micro- and nanotopographic pattern. We found that fibroblasts adhering to the denser pattern areas aligned and elongated more strongly along the direction of ridges, vs. those on the sparser areas, exhibiting a biphasic dependence of the migration speed on the pattern density. In addition, cells responded to local variations in topography by altering morphology and migrating along the direction of grooves biased by the direction of pattern orientation (short term) and pattern density (long term). Molecular dynamic live cell imaging and immunocytochemical analysis of focal adhesions and actin cytoskeleton suggest that variable substratum topography can result in distinct types of cytoskeleton reorganization. We also demonstrate that fibroblasts cultured as monolayers on the same substratum retain most of the properties displayed by single cells. This result, in addition to demonstrating a more sophisticated method to study aspects of wound healing processes, strongly suggests that even in the presence of considerable cell-cell interactions, the cues provided by the underlying substratum topography continue to exercise substantial influence on cell behavior. The described experimental platform might not only further our understanding of biomechanical regulation of cell-matrix interactions, but also contribute to bioengineering of devices with the optimally structured design of cell-material interface.
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