Optimal H2O2 preconditioning to improve bone marrow mesenchymal stem cells' engraftment in wound healing.

Background: The transplantation of bone marrow mesenchymal stem cells (BMSCs) is a promising therapeutic strategy for wound healing. However, the poor migration capacity and low survival rate of transplanted BMSCs in wounds weaken their potential application. Objective: To identify the optimal protocol for BMSCs preconditioned with H₂O₂ and improve the therapeutic efficacy using H₂O₂-preconditioned BMSCs in wound healing. Methods: Mouse BMSCs were exposed to various concentrations of H₂O₂, and the key cellular functional properties were assessed to determine the optimal precondition with H₂O₂. The H₂O₂-preconditioned BMSCs were transplanted into mice with full-thickness excisional wounds to evaluate their healing capacity and tissue engraftment. Results: Treatment BMSCs with 50 μM H₂O₂ for 12 h could significantly enhance their proliferation, migration, and survival by maximizing the upregulation of cyclin D1, SDF-1, and its receptors CXCR4/7 expressions, and activating the PI3K/Akt/mTOR pathway, but inhibiting the expression of p16 and GSK-3β. Meanwhile, oxidative stress-induced BMSC apoptosis was also significantly attenuated by the same protocol pretreatment with a decreased ratio of Bax/Bcl-2 and cleaved caspase-9/3 expression. Moreover, after the identification of the optimal protocol of H₂O₂ precondition in vitro, the migration and tissue engraftment of transfused BMSCs with H₂O₂ preconditioning were dramatically increased into the wound site as compared to the un-preconditioned BMSCs. The increased microvessel density and the speedy closure of the wounds were observed after the transfusion of H₂O₂-preconditioned BMSCs. Conclusions: The findings suggested that 50 μM H₂O₂ pretreated for 12 h is the optimal precondition for the transplantation of BMSCs, which gives a considerable insight that this protocol may be served as a promising candidate for improving the therapeutic potential of BMSCs for wound healing. [ABSTRACT FROM AUTHOR]