3D TGF-β1-expressing mesenchymal stem cells enhance wound healing in a diabetic mouse model

Human mesenchymal stem cells (MSCs) are a promising stem cell source; however, their therapeutic efficacy in chronic wound healing remains limited. This study evaluates the therapeutic potential of transforming growth factor (TGF)-β1-modified, three-dimensionally cultured MSCs (A/T-3D) for enhancing wound healing. Methods: The TGF-β1 gene was inserted into a safe genomic locus in adipose-derived MSCs (ASCs) using transcription activator-like effector nucleases. Quantitative polymerase chain reaction (qPCR) analysis showed that A/T-3D upregulated key factors related to wound healing, including epidermal growth factor (EGF), TGF-β1, fibroblast growth factor (FGF), and vascular endothelial growth factor (VEGF)-A, compared to unmodified ASCs. Results: In vitro scratch wound assays indicated that co-culture with A/T-3D-conditioned medium significantly accelerated wound closure in fibroblasts. In vivo, skin excision in nude mice treated with A/T-3D injections resulted in rapid wound closure, enhanced cellularity, and increased reepithelialization. High engraftment rates of A/T-3D and elevated expression of angiogenic factors were observed in the wound bed, highlighting their direct contribution to tissue repair. Conclusions: Collectively, these findings suggest that A/T-3D MSCs have significant therapeutic potential for wound healing through the secretion of epithelialization and angiogenic factors, along with enhanced engraftment capacity.