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Regulation of Keratocyte Phenotype and Cell Behavior by Substrate Stiffness
Umeå University, Faculty of Medicine, Department of Integrative Medical Biology (IMB), Anatomy. Department of Pathogenic Biology and Immunology, School of Medicine and Jiangsu Key Laboratory for Biomaterials and Devices, Southeast University, Nanjing, China.
Umeå University, Faculty of Medicine, Department of Integrative Medical Biology (IMB), Anatomy. Umeå University, Faculty of Medicine, Department of Community Medicine and Rehabilitation, Section of Physiotherapy.ORCID iD: 0000-0002-6091-3982
Umeå University, Faculty of Medicine, Department of Integrative Medical Biology (IMB), Anatomy. Jiangsu Key Laboratory for Biomaterials and Devices and Department of Physiology, School of Medicine, Southeast University, Nanjing, China.ORCID iD: 0000-0003-2700-6739
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2020 (English)In: ACS Biomaterials Science & Engineering, E-ISSN 2373-9878, Vol. 6, no 9, p. 5162-5171Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2020. Vol. 6, no 9, p. 5162-5171
Keywords [en]
keratocytes, stiffness, phenotype, cell behavior, cytoskeletal reorganization, Rac1
National Category
Ophthalmology
Identifiers
URN: urn:nbn:se:umu:diva-176078DOI: 10.1021/acsbiomaterials.0c00510ISI: 000572822300037Scopus ID: 2-s2.0-85092544653OAI: oai:DiVA.org:umu-176078DiVA, id: diva2:1478922
Available from: 2020-10-23 Created: 2020-10-23 Last updated: 2023-03-23Bibliographically approved

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Chen, JialinBackman, Ludvig J.Zhang, WeiDanielson, Patrik

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