Regulation of Keratocyte Phenotype and Cell Behavior by Substrate Stiffness
Regulation of Keratocyte Phenotype and Cell Behavior by Substrate Stiffness
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基质硬度对角膜细胞表型和细胞行为的调节
DOI：
10.1021/acsbiomaterials.0c00510
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发表时间：
2020-09-01
影响因子：
5.8
通讯作者：
Danielson, Patrik
中科院分区：
文献类型：
--
作者：
Chen, Jialin;Backman, Ludvig J.;Danielson, Patrik
角膜组织工程是解决角膜移植中角膜供体组织缺乏的一种替代方法。在组织工程角膜中，具有正常表型和功能的角质细胞对角膜再生至关重要。尽管众所周知，细胞外/底物硬度在许多不同细胞类型中调控细胞表型和细胞行为，但其在角化细胞培养中的作用尚未得到充分研究。本项目研究底物硬度对角化细胞表型标志物表达和典型细胞行为（细胞粘附、增殖和迁移）的影响及其可能的机制。将人原代角质细胞培养在组织培养塑料（TCP，类似于10(6)kPa）或硬度相当于生理性人角膜基质（25 kPa）或玻璃体（1 kPa）的板上。比较角质细胞表型标志物的表达、细胞粘附、增殖和迁移。结果表明，基质材料的硬度调节了培养的角质细胞的表型标记表达和细胞行为。与TCP和1 kPa组相比，生理角膜硬度（25 kPa）更能保留细胞表型。与TCP培养皿相比，25 kPa培养皿培养的角化细胞面积更大。用NSC 23766 （Rac1抑制剂）处理的细胞模拟了从软材料到硬材料转变过程中细胞表型和行为的反应，包括细胞骨架结构、角质细胞表型标记物的表达和细胞行为。总之，本研究表明底物硬度通过rac1介导的细胞骨架重组调节细胞表型标记物的表达和细胞行为。这些知识有助于角膜组织工程的发展。
Corneal tissue engineering is an alternative way to solve the problem of lack of corneal donor tissue in corneal transplantation. Keratocytes with a normal phenotype and function in tissue-engineered cornea would be critical for corneal regeneration. Although the role of extracellular/substrate material stiffness is well-known for the regulation of the cell phenotype and cell behavior in many different cell types, its effects in keratocyte culture have not yet been thoroughly studied. This project studied the effect of substrate stiffness on the keratocyte phenotype marker expression and typical cell behavior (cell adhesion, proliferation, and migration), and the possible mechanisms involved. Human primary keratocytes were cultured on tissue culture plastic (TCP, similar to 10(6) kPa) or on plates with the stiffness equivalent of physiological human corneal stroma (25 kPa) or vitreous body (1 kPa). The expression of keratocyte phenotype markers, cell adhesion, proliferation, and migration were compared. The results showed that the stiffness of the substrate material regulates the phenotype marker expression and cell behavior of cultured keratocytes. Physiological corneal stiffness (25 kPa) superiorly preserved the cell phenotype when compared to the TCP and 1 kPa group. Keratocytes had a larger cell area when cultured on 25 kPa plates as compared to on TCP. Treatment of cells with NSC 23766 (Rac1 inhibitor) mimicked the response in the cell phenotype and behavior seen in the transition from soft materials to stiff materials, including the cytoskeletal structure, expression of keratocyte phenotype markers, and cell behavior. In conclusion, this study shows that substrate stiffness regulates the cell phenotype marker expression and cell behavior of keratocytes by Rac1-mediated cytoskeletal reorganization. This knowledge contributes to the development of corneal tissue engineering.