Plant-derived extracellular vesicles as a dual-function nanoplatform for synergistic neurovascular repair in ischemic...

Current therapies for ischemic stroke lack the capacity to simultaneously restore metabolic homeostasis, repair the neurovascular unit, and deliver hydrophobic neuroprotectants across the blood-brain barrier. Here, we demonstrate that extracellular vesicles derived from Ligusticum sinense chuanxiong (CXEVs)-nanoscale particles of 167.1 ± 3.3 nm-are naturally enriched in phthalides (∼60%), including ligustilide and butylphthalide derivatives. Following systemic administration, CXEVs efficiently cross the blood-brain barrier, accumulating in ischemic brain tissue with peak concentration at 12 h. In photothrombotic stroke mice, CXEVs dose-dependently improved motor coordination and reduced anxiety-like behaviors. Untargeted metabolomics revealed that CXEVs reprogrammed 30 key metabolites across seven pathways, notably restoring arginine-proline, methionine, purine, and tyrosine metabolism-thereby mitigating ammonia toxicity, oxidative stress, and energy failure. Concurrently, CXEVs activated VEGF signaling by upregulating VEGFA and NOS3 while normalizing KDR and MAPK1 expression, driving endothelial migration, tube formation in vitro, and vascular regeneration in zebrafish. To enhance therapeutic potency, we engineered G3702-loaded CXEVs (G3702@CXEVs) with optimal loading efficiency (1:2 w/w), exceptional stability over 30 days, and sustained release without burst effect. Critically, G3702@CXEVs outperformed either free G3702 or blank CXEVs alone in promoting functional recovery, preserving cortical architecture, and synergistically enhancing both neurogenesis (BrdU⁺/DCX⁺ cells) and angiogenesis (BrdU⁺/CD31⁺ microvessels). CXEVs represent a novel, multifunctional nanoplatform that integrates intrinsic phytochemical-mediated metabolic reprogramming with innate brain-targeting capability. When loaded with G3702, they form a synergistic "therapy-and-delivery" system that concurrently rescues neuronal and vascular injury after stroke. This work establishes plant-derived EVs as a low-cost, scalable, and dual-action nanomedicine platform for complex neurological disorders.