Plant-Derived Nanovesicles: Resolving Conceptual Confusion, Overcoming Isolation Challenges, and Advancing Translational Potential

Zhaoyu Zhang,1,2,* Guangyang Chen,1,3,* Kaiyuan Zheng,4,* Meifang Lin,1,2 Dali Zheng,1 Youguang Lu,1,2 Li Huang,1,3 Xiaohang Chen,1,2 Ruihuan Gan1,2 1Fujian Key Laboratory of Oral Diseases, School and Hospital of Stomatology, Fujian Medical University, Fuzhou, People’s Republic of China; 2Department of Preventive Dentistry, School and Hospital of Stomatology, Fujian Medical University, Fuzhou, People’s Republic of China; 3Department of Dentistry, The First Affiliated Hospital of Fujian Medical University, Fuzhou, People’s Republic of China; 4School of Stomatology Kunming Medical University Haiyuan College, Kunming, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ruihuan Gan, Email ruihuan_gan@fjmu.edu.cn Li Huang, Email lihuang@fjmu.edu.cnAbstract: 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