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Does survivin overexpression enhance the efficiency of fibroblast cell-based wound therapy?

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Abstract

Cell-based wound therapy is faced with some limiting factors that decrease the therapeutic efficacy of transplanted cells. In this study, we aimed to genetically modify fibroblast cells with anti-apoptotic Survivin gene (Birc5) before cell transplantation. In vitro, pIRES2-eGFP-Survivin plasmid was transfected into the fibroblast cells and the growth curve was evaluated for transfected and normal cells performing MTT assay. In vivo, two 6-diameter cutaneous wounds were created at mice dorsal skin. Fibrin clot was used as a delivery vehicle to transfer cells into the wound bed. The effects of four treatment groups including (a) Cell-SVV-Clot (b) Cell-GFP-Clot, (c) Normal cell-Clot and, (d) Clot alone were evaluated. After 1,2,3,7 and 14 days post-transplantation, the wounds were photographed for evaluating the wound closure rate and wound samples were obtained. Angiogenesis and formation of granulated tissue were assessed via H&E staining for wound samples. The expression levels of Survivin, VEGF, and bFGF genes were also determined using qRT-PCR. The MTT assay showed similar proliferation potential of transfected cells with normal cells verifying that Survivin had no detrimental effect. Compared to the Normal cell-Clot group, the Survivin overexpression was seen for 3 days in the Cell-SVV-Clot group verifying the cell survival during the early stage of wound healing. The Survivin further upregulated VEGF and bFGF expressions resulting in more angiogenesis and formation of granulated tissue by day 3 and 14. The treated wounds with Cell-SVV-Clot were regenerated with a higher wound closure rate by day 7 compared to Normal cell-Clot and Clot groups. Survivin enhanced wound healing through induction of VEGF and bFGF at particular times post-wounding that led to a more structured-epidermis with higher angiogenesis and granulation tissue formation rate.

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Acknowledgements

The authors thank the Cellular and Molecular Research Center of Shahrekord University of Medical Sciences for their supports.

Funding

This research was funded by Shahrekord University of Medical Sciences (Grant Number:1396-08-74-516).

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Authors and Affiliations

  1. Department of Medical Biotechnology, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran

    Fereshteh Shojaei-Ghahrizjani & Seyed Abbas Mirzaei

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

    Shima Rahmati

  3. Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran

    Mehdi Banitalebi-Dehkordi

Authors
  1. Fereshteh Shojaei-Ghahrizjani
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  2. Shima Rahmati
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  3. Seyed Abbas Mirzaei
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  4. Mehdi Banitalebi-Dehkordi
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Correspondence to Mehdi Banitalebi-Dehkordi.

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Authors declare that they have no conflict of interest.

Research involving animals

All animal intervention in this study was carried out according to the National Institute of Health Guide for the Care and Use of Laboratory Animals (NIH Publication No. 86-23, Revised 1985). The ethics code was also received from Ethics Committee of Shahrekord University of Medical Sciences for animal experiments in this research. (Ethics code: IR.SKUMS.RE.1396.203).

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Shojaei-Ghahrizjani, F., Rahmati, S., Mirzaei, S.A. et al. Does survivin overexpression enhance the efficiency of fibroblast cell-based wound therapy?. Mol Biol Rep 47, 5851–5864 (2020). https://doi.org/10.1007/s11033-020-05656-4

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