ORIGINAL RESEARCH article
Front. Cell Dev. Biol.
Sec. Cell Growth and Division
Volume 13 - 2025 | doi: 10.3389/fcell.2025.1636884
The effects of TGF-β receptor I inhibitors on myofibroblast differentiation and myotube formation
Provisionally accepted
- Radboud universitair medisch centrum, Nijmegen, Netherlands
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Fibrosis frequently occurs in muscle wounds, ultimately leading to suboptimal function. This study investigates the effects of TGF-βRI inhibitors AZ12799734, Galunisertib, and SM16, on myofibroblast differentiation and myotube formation. Human gingival fibroblasts were treated with TGF-β1 (0, 1, 5, and 10 ng/mL) to induce myofibroblasts. Then, fibroblasts were incubated with TGF-βRI inhibitors (0, 1, 5, 10, and 20 μM) together with 10 ng/ml TGF-β1. Myofibroblast marker expression was assessed using RT-PCR (day 3), while myofibroblast differentiation was analyzed by immunofluorescence staining for α-SMA (day 6). C2C12 myoblasts were also cultured with TGF-βRI inhibitors, and gene expression (day 3) and myotube formation (day 6) were analyzed. TGF-β1 (10 ng/ml) increased the proportion of myofibroblasts from 9.3±3.5% to 38.1±4.4%, which was reduced by all TGF-βRI inhibitors even at 1 μM (for example, Galunisertib 23.5±2.1% (p<0.05)). All inhibitors reduced ACTA2 and COL1A1 gene expression, while only AZ12799734 and SM16 inhibited Ki-67 expression. In C2C12 cultures, AZ12799734 and SM16 reduced the fusion index, whereas Galunisertib did not. Moreover, only Galunisertib increased myotube size from 0.09±0.01 to 0.13±0.01mm 2 /nucleus (p<0.05). Galunisertib inhibited MyoD gene expression (at 20 µM), but not MyoG nor MyHC. In conclusion, Galunisertib may have potential for improving muscle wound healing following injury.
Keywords: TGF-βRI inhibitors, Myotube, myofibroblast, C2C12, Fibrotic
Received: 28 May 2025; Accepted: 10 Jul 2025.
Copyright: © 2025 Wang, Ongkosuwito, Von den Hoff and Wagener. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Frank Wagener, Radboud universitair medisch centrum, Nijmegen, Netherlands
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