Transfersomal gel loading of amniotic mesenchymal stem cell metabolite product for diabetic wound healing in alloxan-induced diabetic mice

Amniotic mesenchymal stem cell metabolite product (AMSC-MP), a metabolite derived from cell cultures, contains cytokines and growth factors that support skin regeneration, making it a promising therapeutic candidate for chronic wounds, such as diabetic foot ulcers. However, because of their large molecular size, AMSC-MPs cannot effectively penetrate the skin’s primary barrier, the stratum corneum. To address this issue, transfersomes have been developed as flexible vesicular carriers capable of delivering active compounds through intercellular gaps. This study aimed to evaluate the physicochemical properties of AMSC-MP transfersomes and their wound healing efficacy in a mouse model of diabetes. Transfersomes (Ts), composed of L-alpha phosphatidylcholine and sodium cholate, were prepared in different AMSC-MP concentrations, 5% (T-5) and 25% (T-25), using the thin-film hydration method, and hyaluronic acid (HA) was added. The results showed that the addition of HA increased particle size, polydispersity index, and pH, while reducing zeta potential, and maintaining high entrapment efficiency. In vivo wound healing evaluation showed that the T-25 HA gel formulation achieved the highest wound contraction percentage, followed by T-5 HA, THA, Blank gel, free AMSC-MPs (25% and 5%), and transfersomes without HA. Histopathological observations and collagen density analyses confirmed that HA improved healing by enhancing tissue regeneration, epithelialization, and collagen deposition. In conclusion, the AMSC-MP transfersomal gel, particularly the 25% formulation with HA, demonstrated the most effective wound-healing performance and showed strong potential as a therapeutic strategy for chronic diabetic wounds.