Effect of substrate topography on the regulation of human corneal stromal cells
Effect of substrate topography on the regulation of human corneal stromal cells
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DOI：
10.1016/j.colsurfb.2020.110971
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发表时间：
2020-06-01
影响因子：
5.8
通讯作者：
Ahearne, Mark
中科院分区：
文献类型：
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作者：
Bhattacharjee, Promita;Cavanagh, Brenton L.;Ahearne, Mark
天然角膜基质的最佳功能取决于细胞外基质（ECM）的有序排列。为了开发体外角膜模型，需要复制角膜体内微环境。在这项研究中，地形线索对角膜细胞表型的影响。采用光刻和聚合物模压法在聚二甲基硅氧烷（PDMS）上制作了2-2.5 μ m深、5 μ m、10 μ m和20 μ m宽的微槽。微槽使细胞体受到限制，细胞核受到挤压，细胞骨架发生重组。它还影响肌动蛋白丝的浓度，染色质的凝聚和细胞增殖。随着微槽宽度的增加，细胞变得更加分散，肌动蛋白丝浓度降低。微沟宽度和细胞过程如粘附、迁移和基因表达之间的关系也被证明。免疫细胞化学和基因表达（RT-PCR）分析表明，微沟宽度上调角膜细胞特异性基因。与其他微槽宽度或平面基板相比，具有5 μ m宽度的微槽导致细胞沿微通道边缘沿着显著对齐，并且更好地支持细胞极化和迁移。这些发现提供了重要的基础知识，可以作为更好地控制组织生长和细胞工程应用的基础，通过地形图案的角膜基质再生。
Optimal functionality of native corneal stroma depends on a well-ordered arrangement of extracellular matrix (ECM). To develop an in vitro corneal model, replication of the corneal in vivo microenvironment is needed. In this study, the impact of topographic cues on keratocyte phenotype is reported. Photolithography and polymer moulding were used to fabricate microgrooves on polydimethylsiloxane (PDMS) 2-2.5 mu m deep and 5 mu m, 10 mu m or 20 mu m in width. Microgrooves constrained the cells body, compressed nuclei and led to cytoskeletal reorganization. It also influenced the concentration of actin filaments, condensation of chromatin and cell proliferation. Cells became more spread and actin filament concentration decreased as the microgroove width increased. Relationships were also demonstrated between microgroove width and cellular processes such as adhesion, migration and gene expression. Immunocytochemistry and gene expression (RT-PCR) analysis showed that microgroove width upregulated keratocyte specific genes. A microgroove with 5 mu m width led to a pronounced alignment of cells along the edges of the microchannels and better supported cell polarization and migration compared with other microgroove widths or planar substrates. These findings provide important fundamental knowledge that could serve as a basis for better-controlled tissue growth and cell-engineering applications for corneal stroma regeneration through topographical patterns.