title: Induction of predominant tenogenic phenotype in human dermal fibroblasts via synergistic effect of TGF-β and elongated cell shape.
abstract: Micropattern topography is widely investigated for its role in mediating stem cell differentiation, but remains unexplored for phenotype switch between mature cell types. This study investigated the potential of inducing tenogenic phenotype in human dermal fibroblasts (hDFs) by artificial elongation of cultured cells. Our results showed that a parallel microgrooved topography could convert spread hDFs into an elongated shape and induce a predominant tenogenic phenotype as the expression of biomarkers was significantly enhanced, such as scleraxis, tenomodulin, collagens I, III, VI, and decorin. It also enhanced the expression of transforming growth factor (TGF)-β1, but not α-smooth muscle actin. Elongated hDFs failed to induce other phenotypes, such as adiopogenic, chondrogenic, neurogenic, and myogenic lineages. By contrast, no tenogenic phenotype could be induced in elongated human chondrocytes, although chondrogenic phenotype was inhibited. Exogenous TGF-β1 could enhance the tenogenic phenotype in elongated hDFs at low dose (2 ng/ml), but promoted myofibroblast transdifferentiation of hDFs at high dose (10 ng/ml), regardless of cell shape. Elongated shape also resulted in decreased RhoA activity and increased Rho-associated protein kinase (ROCK) activity. Antagonizing TGF-β or inhibiting ROCK activity with Y27632 or depolymerizing actin with cytochalasin D could all significantly inhibit tenogenic phenotype induction, particularly in elongated hDFs. In conclusion, elongation of cultured dermal fibroblasts can induce a predominant tenogenic phenotype likely via synergistic effect of TGF-β and cytoskeletal signaling.
authors.affiliation: Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China;
authors.forename: Wenbo
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China;
authors.forename: Jie
authors.initials: J
authors.lastname: Li
authors.affiliation: National Chromatography R&A Centre, CAS Key Lab of Separation for Analytical Chemistry, Dalian Institute of Chemical Physics, CAS, Dalian, China; and University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Keyun
authors.initials: K
authors.lastname: Wang
authors.affiliation: Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Tissue Engineering Center of China, Shanghai, China;
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Tissue Engineering Center of China, Shanghai, China;
authors.forename: Wenjie
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Tissue Engineering Center of China, Shanghai, China;
authors.forename: Guangdong
authors.initials: G
authors.lastname: Zhou
authors.affiliation: Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Tissue Engineering Center of China, Shanghai, China;
authors.forename: Yilin
authors.initials: Y
authors.lastname: Cao
authors.affiliation: National Chromatography R&A Centre, CAS Key Lab of Separation for Analytical Chemistry, Dalian Institute of Chemical Physics, CAS, Dalian, China; and University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Mingliang
authors.initials: M
authors.lastname: Ye
authors.affiliation: National Chromatography R&A Centre, CAS Key Lab of Separation for Analytical Chemistry, Dalian Institute of Chemical Physics, CAS, Dalian, China; and University of Chinese Academy of Sciences, Beijing, China.
authors.forename: Hanfa
authors.initials: H
authors.lastname: Zou
authors.affiliation: Department of Plastic and Reconstructive Surgery, Shanghai Key Laboratory of Tissue Engineering, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Tissue Engineering Center of China, Shanghai, China; liuwei_2000@yahoo.com mingliang@dicp.ac.cn.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
mesh_terms: D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D048430:Cell Shape / Q000502:physiology*; D054338:Cell Transdifferentiation; D002478:Cells, Cultured / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D058628:Myofibroblasts / Q000166:cytology*; D010641:Phenotype; D015398:Signal Transduction / Q000187:drug effects; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: D016212:Transforming Growth Factor beta
keywords: ROCK activity; TGF-β; cell elongation; dermal fibroblasts; tenogenic phenotype