Biomedical Potential of Keratin-Biphalin Wound Dressing in Diabetic Mice: In Vitro and In Vivo Studies

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ABSTRACTStrategies for successful healing of diabetic wounds are lacking. Natural-derived dressings have gained much attention due to their promising properties. We hypothesized that keratin-biphalin fiber-dressing accelerates skin wound healing in a mouse model of diabetes. Keratin fibers were obtained from mouse fur and soaked in a biphalin solution. Studies on NIH/3T3 cells showed that keratin-biphalin fibers increase cell viability. In vivo studies were made on C57BL/6J mice with iatrogenically induced diabetes. Two full-thickness wounds were created on the back of the mice, one untreated and the other treated with an experimental dressing. Western blot analysis, histopathological and immunohistochemical staining were performed. Biphalin was slowly released from keratin fibers. The obtained keratin-biphalin fibers were biocompatible and supported cell growth. Western blot analysis showed that cells treated with our dressing had an increasing expression level of mTOR, and p-AKT 72 h post-treatment compared to 24 h and 48 h. Animal studies showed that the keratin-biphalin dressing accelerated wound healing on days 5 and 15. Treated wounds showed faster reepithelization and developed thicker epidermis. Histopathological and immunohistochemical studies confirmed that keratin-biphalin fibers stimulate macrophage infiltration, which promotes tissue remodeling and regeneration. Exogenous keratin-biphalin fibers p-AKT/mTOR expression protein in vitro and accelerate skin wound healing in diabetic mice.