Downregulation of RUNX1-activated osteopontin facilitates burn wound healing by activating the MAPK pathways

Burn wounds require intervention to ensure timely progression to reduce morbidity and mortality. The migrative and proliferative capabilities of keratinocytes are impaired in wounds. Matrix Metalloproteinases (MMPs) can degrade the extracellular matrix (ECM), allowing epithelial cells to migrate. As reported, osteopontin can regulate cell migration, cell adhesion, and ECM invasion in endothelial and epithelial cells, and its expression is significantly increased in chronic wounds. Therefore, this study investigates the biological functions of osteopontin and its related mechanisms involved in burn wounds. We established cellular and animal models of burn injury. Levels of osteopontin, RUNX1, MMPs, collagen I, CK19, PCNA and pathway-associated proteins were measured by RT-qPCR, western blotting and immunofluorescence staining. Cell viability and migration were examined by CCK-8 and wound scratch assays. Histological changes were analyzed by hematoxylin and eosin staining and Masson's trichrome staining. For in vitro analysis, osteopontin silencing facilitated the growth and migration of HaCaT cells and promoted ECM degradation in HaCaT cells. Mechanistically, RUNX1 bound to osteopontin promoter, and RUNX1 upregulation attenuated the promoting efficacy of osteopontin silencing on cell growth and migration and ECM degradation. Additionally, RUNX1-activated osteopontin inactivated the MAPK signaling pathway. For in vivo analysis, osteopontin depletion facilitated burn wound healing by promoting re-epithelialization and ECM degradation. In conclusion, RUNX1 activates the osteopontin expression at the transcriptional level and osteopontin depletion facilitates the recovery of burn wounds by promoting the migration of keratinocytes and re-epithelization and ECM degradation by activating the MAPK pathway.

ECM; MAPK signaling; RUNX1; burn wounds; keratinocytes; migration; osteopontin; re-epithelization.