Mitochondrial protein-enriched artificial nanovesicles: mitochondrial recovery and antioxidation for diabetic wound treatment

Oxidative stress and mitochondrial dysfunction are major barriers to the healing of diabetic wounds (DW). Eliminating reactive oxygen species (ROS) and restoring mitochondrial function are considered effective strategies to accelerate DW healing. Although extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) have shown therapeutic potential, the quality and yield of mitochondrial components in naturally secreted EVs are limited. Thus, we employed a top-down approach, using the self-assembly properties of membrane components to develop artificial nanovesicles enriched with mitochondria-associated proteins derived from human umbilical cord MSCs. These cell-derived nanovesicles (CNVs) selectively encapsulate mitochondrial proteins, effectively reducing intracellular ROS levels and specifically restoring mitochondrial membrane potential (∆Ψm) and morphology. Furthermore, the CNVs demonstrate remarkable antioxidant and mitochondrial functional restoration capacity, involving the restoration of mitochondrial complexes I, Ⅲ, V and the uncoupling process, as well as multiple mitochondrial function-associated pathways, such as the ALDH2/HADHA/HADHB axis, the IDH2/GSR/GSH axis, and the Ca²⁺/VDAC1 axis. In vivo experiments further validated the therapeutic potential of CNVs, which significantly promoted wound healing in diabetic mice. In conclusion, our study emphasizes the potential of artificial nanovesicles containing organelle-associated proteins in DW therapy, providing a novel and promising strategy for organelle-based disease treatment.