Nano- and microscale holes modulate cell-substrate adhesion, cytoskeletal organization, and -β1 integrin localization in SV40 human corneal epithelial cells
Nano- and microscale holes modulate cell-substrate adhesion, cytoskeletal organization, and -β1 integrin localization in SV40 human corneal epithelial cells
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DOI：
10.1109/tnb.2006.886570
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发表时间：
2006-12-01
影响因子：
3.9
通讯作者：
Nealey, Paul F.
中科院分区：
文献类型：
--
作者：
Karuri, Nancy W.;Porri, Teresa J.;Nealey, Paul F.
人角膜上皮细胞(HCECs)在体内与基底膜接触，具有复杂的纳米级形貌特征。我们报道了带有纳米和微米尺度孔图案的合成底物作为特征尺寸的函数来不同地调节SV40人角膜上皮细胞(SV40-HCECs)的增殖、形状和黏附：1)在相同的培养条件下，纳米尺度特征(特征尺寸小于800 nm)与微尺度特征或平面底物相比，细胞增殖受到抑制。2)纳米孔上的细胞具有星状形态，而微米级孔上的细胞分布更均匀。3)当细胞在层流室中受到剪应力时，细胞更多地附着在纳米尺度的特征上，而不是微米尺度的特征上。透射电子显微镜显示，与1600 nm螺距培养的细胞相比，400 nm螺距培养的细胞有更长更多的丝状基孔和回缩纤维。-β(1)整合素的免疫金标记法显示，这些受体定位于细胞外周和上述细胞骨架元件中。我们的发现表明，表面不连续和机械力化学细胞信号机制的激活可能有助于观察到在纳米和微尺度地形上培养的SV40-HCECs的反应。
Human corneal epithelial cells (HCECs) interface with a basement membrane in vivo that possesses complex nanoscale topographic features. We report that synthetic substrates patterned with nano- and microscale holes differentially modulate the proliferation, shape and adhesion of SV40 human corneal epithelial cells (SV40-HCECs) as a function of feature size: 1) Cell proliferation was inhibited on nanoscale features (features size less than 800 nm in pitch) compared to microscale features or planar substrates in identical culture conditions. 2) Cells on nanoscale holes had a stellate morphology compared to those on microscale features that were more evenly spread. 3) Cells adhered more to nanoscale features than to microscale features when exposed to shear stress in a laminar flow chamber. Transmission electron microscopy showed that cells cultured on the 400 nm pitch patterns had longer and more numerous filopodia and retraction fibers than cells cultured on the 1600 nm pitch patterns. Immunogold labeling of -beta(1) integrins revealed that these receptors were localized at the cell periphery and in the aforementioned cytoskeletal elements. Our findings indicate that surface discontinuities and the activation of mechanochemical cell signaling mechanisms may contribute to the observed responses exhibited by SV40-HCECs cultured on nano- and microscale topography.