CL-EVLPs Promote Corneal Repair in a BAC-Induced Corneal Injury Model via Ocular Microenvironment Reconstruction.

Abstract

PURPOSE: To investigate the therapeutic potential of Curcuma longa-derived extracellular vesicle-like particles (CL-EVLPs) in a benzalkonium chloride (BAC)-induced corneal injury model. METHODS: CL-EVLPs were isolated via ultracentrifugation and characterized. A murine dry eye model was induced by topical application of BAC. Corneal injury and repair outcomes were assessed using clinical scoring, histopathology, and proteomic analysis. Western blotting and real-time quantitative polymerase chain reaction were conducted to quantify protein or mRNA levels. Bioinformatics analysis was performed using STRING and Cytoscape software. RESULTS: CL-EVLPs, with a typical saucer- or cup-shaped structure with a peak diameter of 104 nm and an average diameter of 138.4 ± 64.3 nm, were efficiently taken up by corneal epithelial cells. Treatment with CL-EVLPs significantly accelerated corneal epithelial repair, reduced pathologic neovascularization, promoted corneal nerve regeneration, and suppressed inflammatory responses in a dry eye mouse model. Proteomic analysis revealed that CL-EVLPs are involved mainly in biological processes such as inflammation, oxidative stress, and immune regulation. Mechanistically, CL-EVLPs upregulated the cornea-specific protein keratin 12 but downregulated the hyperproliferation-associated protein Keratin 10. Additionally, we confirmed that key factors such as proangiogenic VEGFA, proteolytic MMP9, the inflammatory mediators IL-1β and S100A8/A9, and the related transcription factor NF-κB were significantly suppressed, whereas the expression levels of the transcription activator Nrf2 increased. CONCLUSIONS: CL-EVLPs represent a novel natural nanobiologic that restores ocular surface homeostasis, effectively enhancing corneal epithelial regeneration and neural repair in the ocular surface microenvironment. This study provides a foundation for the development of plant-derived EVLPs as a safer and more economical therapeutic strategy for ocular surface disorders.