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Abstract: Plant-derived nanovesicles (PDNVs) are promising bioactive nanoparticles with potential in drug delivery, immune regulation, and tissue repair. However, inconsistent terminology and isolation methods hinder reproducibility and clinical translation. A key confusion lies in their comparison with plant extracellular vesicles (PEVs), which are naturally secreted, whereas PDNVs are typically extracted by breaking plant tissues. This process yields a mix of extracellular and intracellular vesicles, creating both functional diversity and challenges. The heterogeneity complicates standardization, large-scale production, and quality control. Here, we clarify the distinctions between PDNVs and PEVs, and then we explore key factors that affect PDNVs isolation. These include the type of plant used, how the plant is processed, and how the vesicles are purified. We highlight workflow-specific optimizations that boost PDNV recovery (e.g. 4– 5 fold higher yields with optimized PEG precipitation) and enhance purity (e.g. ATPS delivering multi-fold higher recovery while eliminating > 95% of protein contaminants). Finally, we propose strategies to help establish standardized methods for using PDNVs in biomedical applications. Illustration of optimized pretreatment and isolation of plant-derived nanovesicles and extracellular vesicles. Keywords: plant-derived nanovesicles, plant extracellular vesicles, isolation methods, plant pretreatment, clinical translation
Zhang et al. (Wed,) studied this question.