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Human mesenchymal stem cells-conditioned medium improves diabetic wound healing mainly through modulating fibroblast behaviors

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Abstract

Fibroblast plays a key role in wound healing, and the advantages of mesenchymal stem cells (MSC) secretome in wound healing have previously been reported. In the present study, we investigated the impact of human bone marrow MSC-conditioned media (CM) on skin wound healing in diabetic rats and found that some improvements occurred mainly through fibroblast functions. Then, we scrutinized the impact of MSC-CM treatment on fibroblast cellular behavior by culturing human dermal fibroblasts (HDFs) in a high-glucose (HG) medium, as an in vitro diabetic model. In vivo findings revealed significant improvements in some healing kinetics of diabetic wound which received MSC-CM. Particularly, MSC-CM-treated diabetic wounds reached considerably higher percentages of wound closure. Also, the granulation tissue of these wound had less pronounced inflammatory response, better tissue remodeling, and more vascularization compared with non-treated diabetic ones. Gene expression analyses indicated that MSC-CM treatment leads to upregulation of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) genes. In addition, a significantly higher cell viability/proliferation, migration, and bFGF gene expression were observed when MSC-CM was used to treat HDFs in HG culture media. Based on these findings, it is suggested that MSC-CM could promote wound repair and skin regeneration, in some major processes, via improvement of cellular behaviors of fibroblasts in the diabetic microenvironment. The beneficial advantages of mesenchymal stem cells-conditioned media on fibroblast cellular behaviors and wound healing may lead to establish a novel approach as an alternative therapeutic procedure to cure chronic wounds in diabetic conditions.

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Acknowledgements

This study was funded by financial grants from Shahid Beheshti University of Medical Sciences, Tehran, Iran (Grant No: 9720082) and Iran National Science Foundation (INSF, Grant No: 97009568).

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Authors and Affiliations

  1. Department of Anatomical Sciences, School of Medicine, Afzalipour University of Medical Sciences, Kerman, Iran

    Mona Saheli

  2. Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, 1985717443, Tehran, Iran

    Mohammad Bayat, Rasoul Ganji, Farzane Hendudari, Raziyeh Kheirjou, Baran Najar & Abbas Piryaei

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

    Mohammad Bayat

  4. Department of Radiology, Ayatollah Khansari Hospital, Arak University of Medical Sciences, Arak, Iran

    Farzane Hendudari

  5. Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran

    Mohammad Pakzad & Abbas Piryaei

  6. Urogenital Stem Cell Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Abbas Piryaei

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  1. Mona Saheli
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Supplementary data 1. Characterization of human bone marrow mesenchymal stem cells.

Passage 4 MSCs in the cell culture flask grew as a fibroblastic, homogeneous monolayer cell population (A). Immunophenotypes of the MSCs done, using flowcytometry, and the results showed that the cells expressed mesenchymal markers, CD73, CD90, and CD105, while they did not express hematopoietic markers CD34 and CD45 (B). Mesenchymal stem cells multi-lineage potency was evaluated by inducing adipogenic and osteogenic differentiation and demonstrated with alizarin-red staining to show mineral deposits in extracellular precipitation (C), or oil-red staining to identify accumulation of intracellular lipid droplets (D) (TIF 447 kb)

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Saheli, M., Bayat, M., Ganji, R. et al. Human mesenchymal stem cells-conditioned medium improves diabetic wound healing mainly through modulating fibroblast behaviors. Arch Dermatol Res 312, 325–336 (2020). https://doi.org/10.1007/s00403-019-02016-6

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  • DOI: https://doi.org/10.1007/s00403-019-02016-6

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