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

• Published in: Shanghai jiao tong da xue xue bao 2023-08, Vol.28 (4), p.495-506
• Main Authors: Liu, Yang, Wang, Yajing, Wen, Dawei, Zhang, Quanyou, Wang, Li, An, Meiwen, Liu, Yong
• Format: Article
• Language: English
• Subjects: Actin; Architecture; Computer Science; Cytology; Electrical Engineering; Engineering; Fabrication; Fibroblasts; Fluorescence; Grooves; Immunofluorescence; Life Sciences; Materials Science; Microenvironments; Microfilaments; Morphology; Original Paper; Psychological stress; Stiffness; Substrates; Topography; Topology
• ISSN: 1007-1172; 1674-8115; 1995-8188
• DOI: 10.1007/s12204-022-2542-2

Hyperplastic scar is a common fibrotic disease that may ultimately lead to severe dysfunction and deformity, causing physical and psychological distress. Therefore, we aim to evaluate the effect of the mechanical microenvironment 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 with the stiffness of 19.3 kPa and 90.1 kPa coupled with parallel groove and cross groove, respectively, to simulate the mechanical microenvironment of skin wounds and scar tissues. The morphological changes in HFbs in different substrates were observed, and the changes in the cell-long axis length, area, and the angle between cell-long axis and 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 cells were elongated in parallel grooves as well as in the area where cross grooves restricted groove length, the cell length was restricted, and the angle between the long axis and the groove was increased. The topography exerted no significant effect on the cell area, but the cell area increased with increasing the stiffness. The parallel groove promoted the expression of the F-actin to a certain extent, and the fluorescence intensity of F-actin decreased with increasing the stiffness. Studying the effect of the mechanical microenvironment of substrates on HFb morphology is of great importance for understanding the mechanisms of scar formation and prevention.