Enhanced tenogenic differentiation and tendon-like tissue formation by tenomodulin overexpression in murine mesenchymal stem cells.
As a highly specific marker of tenocytes, tenomodulin (Tnmd) functions remain largely unexplored. We investigated the effect of Tnmd overexpression on tenogenic differentiation of murine mesenchymal stem cells (mMSCs) via plasmid-mediated overexpression in the C3H10T1/2 cell line. The results showed that overexpressed Tnmd could significantly enhance cell proliferation (p < 0.05) and the gene expressions of tenogenic-related molecules, including Tnmd, Scleraxis (Scx), collagens I, III and VI and decorin (p < 0.05), and significantly inhibit mMSCs differentiation towards the adipogenic, chondrogenic and osteogenic lineages (p < 0.05). Upon in vivo implantation with rat tail collagen gel subcutaneously in nude mice, Tnmd-overexpressed C3H10T1/2 cells formed neotendon-like tissue, which revealed a histological feature of wave-like dense collagen fibres and cells aligned in parallel. By contrast, a disorganized connective tissue structure with randomly distributed cells was observed in the control group. To further confirm this finding, a conditional Tnmd-overexpressing mouse model was established and the derived primary mMSCs could be induced to overexpress Tnmd with > two-fold upregulated gene expression (p < 0.05) by the treatment of doxycycline (Dox). Similarly, conditional overexpression of Tnmd in primary mMSCs also led to faster proliferation (p < 0.05), enhanced gene expression of tenogenic markers (p < 0.05) and the inhibited expressions of adipogenic and osteogenic markers (p < 0.05). The results of enhanced tenogenic differentiation and neotendon formation indicated that Tnmd may serve not only as a tenogenic marker but also as a positive regulator of MSCs tenogenic differentiation, which might be applied to MSCs-mediated tendon regeneration. Copyright © 2016 John Wiley & Sons, Ltd.
Department of Plastic and Reconstructive Surgery shanghai 9th People's Hospital, People's Republic of China.
Yongkang
Y
Jiang
Department of Plastic and Reconstructive Surgery shanghai 9th People's Hospital, People's Republic of China.
Yuan
Y
Shi
Department of Anatomy and Neurobiology, Tongji University School of Medicine, Shanghai, People's Republic of China.
Jing
J
He
Department of Plastic and Reconstructive Surgery shanghai 9th People's Hospital, People's Republic of China.|National Tissue Engineering Centre of China, Shanghai, People's Republic of China.
Zhiyong
Z
Zhang
Department of Plastic and Reconstructive Surgery shanghai 9th People's Hospital, People's Republic of China.|National Tissue Engineering Centre of China, Shanghai, People's Republic of China.
Guangdong
G
Zhou
Department of Plastic and Reconstructive Surgery shanghai 9th People's Hospital, People's Republic of China.|National Tissue Engineering Centre of China, Shanghai, People's Republic of China.
Wenjie
W
Zhang
Department of Plastic and Reconstructive Surgery shanghai 9th People's Hospital, People's Republic of China.|National Tissue Engineering Centre of China, Shanghai, People's Republic of China.
Yilin
Y
Cao
Department of Plastic and Reconstructive Surgery shanghai 9th People's Hospital, People's Republic of China.|National Tissue Engineering Centre of China, Shanghai, People's Republic of China.
Wei
W
Liu
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D005786:Gene Expression Regulation; D008565:Membrane Proteins / Q000096:biosynthesis*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D051381:Rats; D013710:Tendons / Q000166:cytology / Q000378:metabolism*
D008565:Membrane Proteins; C486026:Tnmd protein, mouse
MSCs; neotendon formation; overexpression; tenogenic differentiation; tenomodulin