Restoring cellular homeostasis via vesicle-mediated mitochondrial delivery and regulation of mitochondrial turnover

Mitochondrial dysfunction is a fundamental driver of tissue injury across a wide range of pathological conditions. Disruption of mitochondrial pool homeostasis, characterized by the accumulation of dysfunctional mitochondria and insufficient clearance, compromises cellular bioenergetics and regenerative capacity. Therefore, therapeutic strategies that simultaneously replenish functional mitochondria while promoting the removal of damaged mitochondria represent a promising approach to restoring mitochondrial homeostasis. Here, we report the development of a bioinspired nanotherapeutic platform for efficient mitochondrial delivery, termed Urolithin A–preconditioned mitochondria–loaded hybrid extracellular vesicles (UA-Mito@HEVs). This system is engineered to facilitate endothelial delivery of functional mitochondria while concomitantly enhancing mitophagy-associated markers, thereby re-establishing a balanced and functional mitochondrial pool. The application of UA-Mito@HEVs in a methylglyoxal (MGO)-induced oxidative stress injury model in HUVECs significantly enhanced mitochondrial uptake and promoted the mitophagic clearance of damaged mitochondria, thereby reducing oxidative burden and improving cellular function. In vivo, periwound subcutaneous injection of UA-Mito@HEVs significantly accelerated wound healing in a streptozotocin (STZ)-induced diabetic mouse model of full-thickness dorsal wounds (10 mm), while tracheal instillation of UA-Mito@HEVs alleviated pulmonary injury in a lipopolysaccharide (LPS)-induced acute respiratory distress syndrome model. These observed therapeutic outcomes were consistent with the restoration of mitochondrial pool balance and subsequent tissue repair, which was accompanied by reduced expression of endoplasmic reticulum stress‑associated markers, indicating improved inter‑organelle coordination secondary to mitochondrial recovery. Overall, UA-Mito@HEVs represent a biocompatible and versatile mitochondrial therapeutic that integrates organelle delivery with mitophagy activation, providing a generalizable strategy for treating diseases driven by mitochondrial dysfunction.