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Zinc and hypoxic preconditioning: a strategy to enhance the functionality and therapeutic potential of bone marrow-derived mesenchymal stem cells

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Abstract

The therapeutic use of bone marrow mesenchymal stem cells (BM-MSCs) requires a large number of cells (1–100 × 106 cells/kg of body weight). Extensive in vitro growth is limited due to the aging of cultured BM-MSCs which leads to abnormal morphology and senescence. Hypoxia increases BM-MSC proliferation, but the question of whether hypoxia preconditioning is safe for clinical application of BM-MSCs remains to be answered. Zinc is essential for cell proliferation and differentiation, especially for the regulation of DNA synthesis and mitosis. It is a structural constituent of numerous proteins on a molecular level, including transcription factors and enzymes of cellular signaling machinery. All the tissues, fluids, and organs of the human body contain zinc. More than 95% of zinc is intracellular, of which 44% is involved in the transcription of DNA. We investigated the effects of ZnCl2 on proliferation, morphology, migration, population doubling time (PDT), and gene expression of BM-MSCs under hypoxic (1% O2) and normoxic (21% O2) environments. BM-MSCs were preconditioned with optimized concentrations of ZnCl2 under normoxic and hypoxic environments and further examined for morphology by the phase-contrast inverted microscope, cell proliferation by MTT assay, PDT, cell migration ability, and gene expression analysis. Zinc significantly enhanced the proliferation of BM-MSCs, and it decreases PDT under hypoxic and normoxic environments as compared to control cells. Migration of BM-MSCs toward the site of injury increased and expression of HIF1-α significantly decreased under hypoxic conditions as compared to non-treated hypoxic cells and control. At late passages (P9), the morphology of normoxic BM-MSCs was transformed into large, wide, and flat cells, and they became polygonal and lost their communication with other cells. Conversely, zinc-preconditioned BM-MSCs retained their spindle-shaped, fibroblast-like morphology at P9. The expression of proliferative genes was found significantly upregulated, while downregulation of genes OCT4 and CCNA2 was observed in zinc-treated BM-MSCs under both normoxic and hypoxic conditions. ZnCl2 treatment can be used for extensive expansion of BM-MSCs in aged populations to obtain a large number of cells required for systemic administration to produce therapeutic efficacy.

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I declare that I have no other data to share. All the information is included in the manuscript.

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Funding

This research received no external funding.

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Authors and Affiliations

  1. Ghulam Muhammad Mahar Medical College Sukkur at Shaheed Mohtarma Benazir Bhutto Medical University Larkana, Larkana, 77150, Pakistan

    Syed Faizan Ali Rizvi

  2. Ziauddin University, Clifton, Karachi, 74700, Pakistan

    Syed Faizan Ali Rizvi, Bushra Wasim, Shumaila Usman & Kevin Joseph Jerome Borges

  3. Dr. Panjwani Center for Molecular Medicine and Drug Research, International Centre for Chemical and Biological Sciences, Karachi, 75270, Pakistan

    Iqra Sahibdad, Asmat Salim & Irfan Khan

  4. Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan

    Irfan Khan

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  1. Syed Faizan Ali Rizvi
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  4. Kevin Joseph Jerome Borges
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  7. Irfan Khan
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Contributions

SFAR performed the experiments and wrote the manuscript. BW evaluated, analyzed, and reviewed the data. SU and KJJB evaluated, analyzed, and reviewed the data, and assisted in manuscript preparation. IS performed experiments. AS analyzed the data and reviewed and edited the manuscript. IK designed the experiments, analyzed the data, and finalized the manuscript.

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Correspondence to Irfan Khan.

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The procedures followed in this study were approved by the local independent ethics committee for human subjects IEC-052-HT-2020 at PCMD, ICCBS, University of Karachi, Karachi.

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Rizvi, S.F.A., Wasim, B., Usman, S. et al. Zinc and hypoxic preconditioning: a strategy to enhance the functionality and therapeutic potential of bone marrow-derived mesenchymal stem cells. Mol Cell Biochem 477, 2735–2749 (2022). https://doi.org/10.1007/s11010-022-04468-3

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