Endothelial actin and cell stiffness is modulated by substrate stiffness in 2D and 3D.
Endothelial actin and cell stiffness is modulated by substrate stiffness in 2D and 3D.
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DOI：
10.1016/j.jbiomech.2009.02.012
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发表时间：
2009-05-29
影响因子：
2.4
通讯作者：
Gooch KJ
中科院分区：
文献类型：
--
作者：
Byfield FJ;Reen RK;Shentu TP;Levitan I;Gooch KJ
人们越来越认识到被动的机械性能，特别是局部环境的刚性，可以对细胞功能产生深远的影响。许多实验都是在2D几何结构(即生长在不同硬度的底物上的细胞)中进行的，这是体内细胞经常经历的3D环境的简化。为了确定基质的维度如何调节基质硬度对肌动蛋白和细胞硬度的影响，在不同硬度的基质上和基质内培养内皮细胞。内皮细胞在顺应性(1.0~1.5 mg/ml，124±8~202±27 pA)和刚性(3.0 mg/ml，502±48 pA)I型胶原凝胶中培养。细胞在这两组凝胶中拉长并形成微血管样网络，这在以前的研究中可以看到。较硬凝胶中的细胞显示出更明显的应力纤维和~1.5倍的肌动蛋白染色。由于肌动蛋白是细胞机械性能的主要决定因素，我们假设硬凝胶中的细胞本身会更坚硬。为了验证这一假设，从凝胶中分离细胞，并使用微吸管吸入法评估它们的硬度。从相对顺应性凝胶中分离的细胞比从相对硬的凝胶中分离的细胞顺应性高1.9倍(p<；0.05)。类似地，在1700帕的聚丙烯酰胺凝胶上培养的细胞比在9000帕上培养的细胞顺应性高2.0倍(p<；0.05)。这些数据表明，细胞外基质硬度在三维和二维环境中调节内皮硬度，尽管细胞对硬度的反应范围在不同的环境中显著不同。
There is a growing appreciation of the profound effects that passive mechanical properties, especially the stiffness of the local environment, can have on cellular functions. Many experiments are conducted in a 2D geometry (i.e., cells grown on top of substrates of varying stiffness), which is a simplification of the 3D environment often experienced by cells in vivo. To determine how matrix dimensionality might modulate the effect of matrix stiffness on actin and cell stiffness, endothelial cells were cultured on top of and within substrates of various stiffnesses. Endothelial cells were cultured within compliant (1.0–1.5 mg/ml, 124±8 to 202±27 Pa) and stiff (3.0 mg/ml, 502±48 Pa) type-I collagen gels. Cells elongated and formed microvascular-like networks in both sets of gels as seen in previous studies. Cells in stiffer gels exhibited more pronounced stress fibers and ~1.5-fold greater staining for actin. As actin is a major determinant of a cell’s mechanical properties, we hypothesized that cells in stiff gels will themselves be stiffer. To test this hypothesis, cells were isolated from the gels and their stiffness was assessed using micropipette aspiration. Cells isolated from relatively compliant gels were 1.9-fold more compliant than cells isolated from relatively stiff gels (p<0.05). Similarly, cells cultured on top of 1700 Pa polyacrylamide gels were 2.0-fold more compliant that those cultured on 9000 Pa (p<0.05). These data demonstrate that extracellular substrate stiffness regulates endothelial stiffness in both three- and two-dimensional environments, though the range of stiffnesses that cells respond to vary significantly in different environments.
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影响因子：
56.9
作者：
Cukierman, E;Pankov, R;Yamada, KM
通讯作者：
Yamada, KM
影响因子：
3.4
作者：
Engler, A;Bacakova, L;Discher, D
通讯作者：
Discher, D
影响因子：
3.7
作者：
Sieminski, AL;Hebbel, RP;Gooch, KJ
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影响因子：
3.3
作者：
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通讯作者：
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影响因子：
64.8
作者：
Storm, C;Pastore, JJ;Janmey, PA
通讯作者：
Janmey, PA