Abstract
Extracellular vesicles (EVs) are nanoscale membrane-bound vesicles released by any cell type under both physiological and pathological conditions. They carry a wide array of bioactive molecules, including microRNAs (miRNAs), lipids, proteins and other small biomolecules, and therefore play a key role in intercellular communication by transferring functional cargo between cells. Plant-derived EVs (PDVs) are secreted by plant cells and, because of their natural abundance, PDVs are a compelling alternative to synthetic nanoparticles, particularly for drug delivery applications, due to their biocompatibility, low immunogenicity and intrinsic ability to encapsulate and transport therapeutic molecules. PDVs can enhance drug delivery by improving efficacy and safety, reducing dosage and toxicity, and enabling environmentally sustainable approaches. They also hold promises in diagnostics and nutraceuticals, where their immunomodulatory effects, disease-specific molecular signatures, and incorporation into functional foods may lead to significant health benefits. Despite these advances, challenges remain in standardizing PDVs isolation and characterization techniques and in ensuring reproducibility for clinical translation. Moreover, the transformative potential of PDVs to drive pharmaceutical and nutraceutical innovation and to promote sustainable healthcare solutions has also been underlined. This review provides a comprehensive analysis of PDVs, integrating research findings from 2014 to 2025 to summarize their sources, biochemical composition, isolation, characterization and numerous uses in the fields of biomedical and biotechnological research. Emphasis has been placed on the use of flow cytometry as an emerging, robust, and rapid technique for PDVs identification and characterization, underlining its relevance in clinical and nutraceutical applications.