Substrate Stiffness and Topography Affect the Morphology of Human Fibroblasts in Mechanical Microenvironment

doi:10.1007/s12204-022-2542-2

Abstract

Abstract: Hyperplastic scar is a common fibrotic disease that may ultimately lead to severe dysfunction anddeformity, causing physical and psychological distress. Therefore, we aim to evaluate the effect of the mechanicalmicroenvironment of scar substrates on the morphology of human fibroblasts (HFbs). The micro-modular fabrication technique was used to design a new cross-groove topology and to construct four elastic substrates withthe stiffness of 19.3 kPa and 90.1 kPa coupled with parallel groove and cross groove, respectively, to simulate themechanical microenvironment of skin wounds and scar tissues. The morphological changes in HFbs in differentsubstrates were observed, and the changes in the cell-long axis length, area, and the angle between cell-long axisand grooves were recorded. Immunofluorescence staining was performed to observe the distribution of microfilaments. The results indicated that substrate stiffness and topography affected the morphology of HFbs. The cellswere elongated in parallel grooves as well as in the area where cross grooves restricted groove length, the celllength was restricted, and the angle between the long axis and the groove was increased. The topography exertedno significant effect on the cell area, but the cell area increased with increasing the stiffness. The parallel groovepromoted the expression of the F-actin to a certain extent, and the fluorescence intensity of F-actin decreased withincreasing the stiffness. Studying the effect of the mechanical microenvironment of substrates on HFb morphologyis of great importance for understanding the mechanisms of scar formation and prevention.

Key words: mechanical micro-environment, matrix stiffness, topological structure, fibroblast

摘要： 增生性瘢痕是一种常见的纤维化疾病，最终可导致严重的功能障碍和畸形，本文旨在研究瘢痕基底力学微环境对人体正常皮肤成纤维细胞（HFbs）形态的影响。采用微模式化制作技术，设计了新型交叉沟槽拓扑结构，构建刚度（19.3 kPa、90.1 kPa）分别耦合平行沟槽、交叉沟槽的4种弹性基底，模拟皮肤创面及瘢痕组织的力学微环境，观察不同组别基底上HFbs的形态变化，统计细胞长轴长度、面积、细胞长轴与沟槽夹角的变化，免疫荧光染色观察微丝分布情况。结果表明，基底刚度和拓扑结构影响HFbs形态：平行沟槽上细胞被拉长，在交叉沟槽限制沟槽长度的区域，细胞长度被限制且细胞长轴与沟槽夹角有所增大；拓扑结构对细胞面积无明显影响，但刚度增加，细胞面积增大；平行沟槽在一定程度上促进F-actin的表达，刚度增加时，F-actin荧光强度下降；研究基底力学微环境对HFbs形态的影响对于瘢痕的形成及预防具有重要意义。

关键词: 力学微环境，基底刚度，拓扑结构，成纤维细胞

CLC Number:

R 318.5

LIU Yang-,2 (刘阳), WANG Yajing- (王雅靖)，WEN Daweil (温大渭)，ZHANG Quanyoul (张全有), WANG Li (王立)，AN Meiwen1* (安美文), LIU Yong3* (刘勇). Substrate Stiffness and Topography Affect the Morphology of Human Fibroblasts in Mechanical Microenvironment[J]. J Shanghai Jiaotong Univ Sci, 2023, 28(4): 495-.

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