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5-Azacytidine and trichostatin A enhance the osteogenic differentiation of bone marrow mesenchymal stem cells isolated from steroid-induced avascular necrosis of the femoral head in rabbit

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Abstract

Bone marrow mesenchymal stem cells (BMSCs) play an important role in the process of bone repair. The present study investigated the effect of 5-azacytidine (AZA) and trichostatin A (TSA) on BMSC behaviors in vitro. The role of WNT family member 5A (WNT5A)/WNT family member 5A (WNT7A)/β-catenin signaling was also investigated. BMSCs were isolated from a steroid-induced avascular necrosis of the femoral head (SANFH) rabbit model. The third-generation of BMSCs was used after identification. The results revealed obvious degeneration and necrosis in the SANFH rabbit model. AZA, TSA and TSA + AZA increased BMSC proliferation in a time-dependent fashion. AZA, TSA and TSA + AZA induced the cell cycle release from the G0/G1 phase and inhibited apoptosis in BMSCs. AZA, TSA and TSA + AZA treatment significantly decreased caspase-3 and caspase-9 activities. The treatment obviously increased the activity and relative mRNA expression of alkaline phosphatase. The treatment also significantly up-regulated the proteins associated with osteogenic differentiation, including osteocalcin and runt-related transcription factor 2 (RUNX2), and Wnt/β-catenin signal transduction pathway-related proteins β-catenin, WNT5A and WNT7A. The relative levels of Dickkopf-related protein 1 (an inhibitor of the canonical Wnt pathway) decreased remarkably. Notably, TSA + AZA treatment exhibited a stronger adjustment ability than either single treatment. Collectively, the present studies suggest that AZA, TSA and TSA + AZA promote cell proliferation and osteogenic differentiation in BMSCs, and these effects are potentially achieved via up-regulation of WNT5A/WNT7A/β-catenin signaling.

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Acknowledgements

This study was supported by grants from the Shandong Provincial Natural Science Foundation (Number: ZR2013HM069) and Primary Research & Development Plan of Shandong Province (2017GSF218089).

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Authors and Affiliations

  1. Department of Orthopaedics, Shandong Provincial Hospital Affiliated to Shandong University, 324 Jingwu Road, Jinan, 250021, Shandong, People’s Republic of China

    Peng Zhang, Fulin Tao, Qinghu Li, Shuai Wu & Baisheng Fu

  2. Department of Pharmacy, Shandong Provincial Hospital Affiliated to Shandong University, 324 Jingwu Road, Jinan, 250021, Shandong, People’s Republic of China

    Ping Liu

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  1. Peng Zhang
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  2. Fulin Tao
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  3. Qinghu Li
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  4. Shuai Wu
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  5. Baisheng Fu
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  6. Ping Liu
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Correspondence to Baisheng Fu or Ping Liu.

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Animal rights statement

All animal protocols were performed strictly in accordance with the international ethical guidelines and the National Institutes of Health Guide Concerning the Care and Use of Laboratory Animals, and the Animal Experimentation Ethics Committee of our hospital approved all protocols.

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Communicated by BJ RAO

Corresponding editor: BJ Rao

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Zhang, P., Tao, F., Li, Q. et al. 5-Azacytidine and trichostatin A enhance the osteogenic differentiation of bone marrow mesenchymal stem cells isolated from steroid-induced avascular necrosis of the femoral head in rabbit. J Biosci 44, 87 (2019). https://doi.org/10.1007/s12038-019-9901-7

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  • DOI: https://doi.org/10.1007/s12038-019-9901-7

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