EPOTF-Enabled Scalable Production of Structurally Intact Lemon-Derived Extracellular Vesicles with Enhanced Bioactivity.

Lemon-derived extracellular vesicles (LDEVs) have attracted increasing attention as promising bioactive nanovesicles for cosmetic and skin engineering applications owing to their biocompatibility, bioactive cargo, and skin-friendly lipid bilayer structure. However, the lack of standardized purification methods and limited scalability remain critical bottlenecks for the practical application and industrial translation of LDEVs. In this study, we developed an electrophoretic oscillation-assisted tangential flow filtration (EPOTF) platform using a SiNx membrane nanofilter for the scalable and high-purity purification of structurally intact LDEVs. The EPOTF process effectively suppressed membrane cake formation and membrane fouling, enabling stable size-selective purification under large-volume conditions. EPOTF-purified LDEVs exhibited stable physicochemical characteristics, high recovery yield, and excellent batch-to-batch reproducibility. Cryo-TEM analysis confirmed preservation of a distinct lipid bilayer structure and near-ideal spherical morphology with low spherical deviation, indicating minimal structural damage during purification. Furthermore, EPOTF-purified LDEVs significantly enhanced hydration, antioxidant activity, skin barrier recovery, and anti-inflammatory responses in an ex vivo human skin explant model. Taken together, these findings establish EPOTF as a robust and scalable purification platform for the biomanufacturing of high-quality LDEVs, with strong potential for cosmetic and skin-engineering applications.