Effects of SW033291 on the myogenesis of muscle-derived stem cells and muscle regeneration

Yuanqiang Dong,Lijia Liu,Chuan Zhang,Haibin Chen,Simeng Chen,Yuan Li

doi:10.1186/s13287-020-1574-5

Yuanqiang Dong, Lijia Liu + Show 4 more

Open Access

https://doi.org/10.1186/s13287-020-1574-5

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Abstract

BackgroundThe unmet medical needs in repairing large muscle defects promote the development of tissue regeneration strategy. The use of bioactive molecules in combination with biomaterial scaffold has become an area of great interest. SW033291, a small-molecule inhibitor targeting 15-hydroxyprostaglandin dehydrogenase (15-PDGH) and subsequently elevating the production of prostaglandin E2 (PGE2), has been proved to accelerate the recovery and potentiate the regeneration of multiple tissues including the bone, liver, and colon. The limited understanding of the potential therapeutic effects on myogenesis motivated us to investigate the role of SW033291 in regulating muscle-derived stem cell (MDSC) myogenic differentiation and MDSC-mediated muscle regeneration.MethodsThe characteristics of rat MDSCs, including cell-specific markers and myogenic differentiation potential, were determined. MDSCs were incubated with SW033291 to evaluate PGE2 production and cytotoxicity. The effects of SW033291 on MDSC myogenic differentiation were assessed by quantitative real-time polymerase chain reaction (qPCR), western blot, and immunocytochemistry. The fibrin gel containing MDSCs and SW033291 was used for muscle regeneration in a tibialis anterior muscle defect model.ResultsOur data demonstrated that MDSCs were well-tolerated to SW033291 and treatment with SW033291 significantly promoted the production of PGE2 by MDSCs. In vitro analysis showed that SW033291 enhanced the myogenic differentiation and myotube formation by upregulating a series of myogenic markers. Additionally, the activation of PI3K/Akt pathway was involved in the mechanism underlying these promotive effects. Then, in situ casting of fibrin gel containing MDSCs and SW033291 was used to repair the tibialis anterior muscle defect; the addition of SW033291 significantly promoted myofiber formation within the defect region with mild immune response, less fibrosis, and sufficient vascularization.ConclusionSW033291 acted as a positive regulator of MDSC myogenic differentiation, and incorporating the compound with MDSCs in fibrin gel could serve as an effective method to repair large skeletal muscle defects.

Highlights

Skeletal muscle consists of myofibers, connective tissues, and an organized network of vascular and nerves, which is responsible for body movement and locomotion [1]
To investigate the myogenic differentiation potential, muscle-derived stem cell (MDSC) were cultured in differentiation medium containing 2% horse serum; quantitative real-time polymerase chain reaction was performed to detect myogenic-specific gene expression, along with immunocytochemistry for the detection of myosin heavy chain (MHC) expression. qPCR results showed that the expression of myogenesis-related genes such as myoD, myoG, and Myf5 increased under myogenic induction as compared to the control (Fig. 1d)
To test the effects of SW033291 on cell senescence, senescence-associated β-galactosidase (SA β-Gal) staining was performed; our results showed that MDSCs treated with different concentration of SW033291 displayed minimal SA β-Gal staining with a cell positive rate around 5%, and there was no significant difference among each treatment group and between Growth medium (GM) and differentiation medium (DM) (Fig. 2b)

Summary

Introduction

Skeletal muscle consists of myofibers, connective tissues, and an organized network of vascular and nerves, which is responsible for body movement and locomotion [1]. The repair of volumetric muscle loss still remains a great challenge, as the current standard of care in clinic, engraftment of autologous muscle flaps, is usually limited by insufficient supply of muscle tissue and considerable donor site morbidity. Alternatives such as biological acellular scaffolds and minced muscle tissue have been applied in repairing VML; all these techniques suffer from multiple disadvantages [4, 5]. The limited understanding of the potential therapeutic effects on myogenesis motivated us to investigate the role of SW033291 in regulating muscle-derived stem cell (MDSC) myogenic differentiation and MDSC-mediated muscle regeneration

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