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A Novel Ecm Patch Combines Human Fibroblast-Derived Matrix and Mesenchymal Stem Cells for Advanced Deep Wound Healing

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Document pages: 33 pages

Abstract: Decellularized extracellular matrix (ECM)-based scaffold has been a very useful resource for effective tissue regeneration. In this study, we report a novel ECM patch that physically combines human fibroblast-derived matrix (hFDM) and polyvinyl alcohol (PVA) hydrogel. hFDM was obtained after decellularization of in vitro cultured human fibroblasts. We investigated the basic characteristics of hFDM itself using immunofluorescence (fibronectin, collagen type Ⅰ) and angiogenesis related factors analysis. Successful incorporation of hFDM with PVA produced an hFDM PVA patch, which showed an excellent cytocompatibility with human mesenchymal stem cells (hMSCs), as assessed via cell adhesion, viability, and proliferation. Moreover, in vitro scratch assay using human dermal fibroblasts showed a significant improvement of cell migration when treated with the paracrine factors orignated from the hMSCs-incorporated hFDM. To evaluate therapeutic effect on wound healing, hMSCs were seeded on the hFDM PVA patch and they were then transplanted into mouse full-thickness wound model. Among four experimental groups (control, PVA, hFDM PVA, hMSC hFDM PVA), we found that hMSC hFDM PVA patch accelerated the wound closure with time. More notably, histology and immunofluorescence demonstrated that compared to normal skin tissue, hMSC hFDM PVA patch could lead to significantly advanced tissue regeneration, as confirmed via nearly normal epidermis thickness, skin adnexa regeneration (hair follicle), mature collagen deposition, and neovascularization. Additionally, cell tracking of pre-labelled hMSCs suggegsts the in vivo retention of transplanted cells in the wound region after transplantation of hMSC hFDM PVA patch. Taken together, our engineered ECM patch demonstrates a strong regeneration potential toward advanced wound healing.

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