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Mir-4699 promotes the osteogenic differentiation of mesenchymal stem cells

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Journal of Bone and Mineral Metabolism Aims and scope Submit manuscript

Abstract

Introduction

Mesenchymal stem cells (MSCs) are drawing considerable attention in the field of regenerative medicine due to their differentiation capabilities. The miRNAs are among the most important epigenetic regulators of MSC differentiation. Our previous study identified miR-4699 as a direct suppressor of the DKK1 and TNSF11 gene expression. However, the precise osteogenic-related phenotype or mechanism caused by miR-4699 change has yet to be dealt with in depth.

Material and Methods

In the present study, miR-4699 mimics were transfected into human Adipose tissue-derived mesenchymal stem cells (hAd-MSCs) and osteoblast marker gene expression (RUNX2, ALP, and OCN), was analyzed to investigate whether miR-4699 promotes osteoblast differentiation of hAd-MSCs through targeting the DKK-1 and TNFSF11. We further examined and compared the effects of recombinant human BMP2 with miR-4699 on cell differentiation. In addition to quantitative PCR, analysis of alkaline phosphatase activity, calcium content assay, and Alizarin red staining were used to explore osteogenic differentiation. To evaluate the effect of miR-4699 on its target gene (on protein level) we utilized the western blotting technique.

Results

The overexpression of miR-4699 in hAd-MSCs resulted in the stimulation of alkaline phosphatase activity, osteoblast mineralization, and the expression of RUNX2, ALP, and OCN osteoblast marker genes.

Conclusion

Our findings indicated that miR-4699 supported and synergized the BMP2-induced osteoblast differentiation of mesenchymal stem cells. We suggest, thereof, the utilization of hsa-miR-4699 for further in vivo experimental investigation to reveal the potential therapeutic impact of regenerative medicine for different types of bone defects.

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Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

The author thanks the Cellular and Molecular Biology Research Center and Medical Nanotechnology and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran, for providing technical support.

Funding

This project was funded by Shahid Beheshti University of Medical Sciences, Tehran, Iran (Contact grant No: 32003). The funders had no role in the study design, data collection, analysis, decision to publish, or preparation of the manuscript.

Author information

Authors and Affiliations

  1. Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

    Vahedeh Hosseini

  2. Department of Molecular Medicine and Genetics, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran

    Vahedeh Hosseini

  3. Department of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran

    Mahdi Paryan

  4. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Ameneh Koochaki

  5. Department of Nursing, Saint Joseph’s College, Brooklyn, NY, USA

    Henry Manuel Cesaire

  6. Medical Nanotechnology and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Samira Mohammadi-Yeganeh

  7. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Samira Mohammadi-Yeganeh

Authors
  1. Vahedeh Hosseini
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  2. Mahdi Paryan
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  3. Ameneh Koochaki
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  4. Henry Manuel Cesaire
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  5. Samira Mohammadi-Yeganeh
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Contributions

VH: conceptualization, investigation, formal analysis, methodology, resources, writing—original draft, writing—review, and editing. Ak: data curation, investigation, formal analysis, resources, writing—original draft, writing—review, and editing. HMC: methodology and writing—review and editing. SM-Y and MP: conceptualization, formal analysis, funding acquisition, methodology, writing—original draft and writing—review and editing.

Corresponding authors

Correspondence to Mahdi Paryan or Samira Mohammadi-Yeganeh.

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Conflict of interest

The authors declare no competing conflict of interest concerning the work described.

Ethical approval

This project was approved and supervised by the ethical committee of Shahid Beheshti University of Medical Sciences, (ethics number Iran (IR.SBMU.REC.1401.002)).

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Hosseini, V., Paryan, M., Koochaki, A. et al. Mir-4699 promotes the osteogenic differentiation of mesenchymal stem cells. J Bone Miner Metab 41, 481–491 (2023). https://doi.org/10.1007/s00774-023-01433-y

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  • DOI: https://doi.org/10.1007/s00774-023-01433-y

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