Isolation of Red Beet Plant-Derived Nanovesicles, and Characterization of Their Molecular Content and Biological Activities in Human Cells

Nowadays, growing evidence indicates that plant-derived nanovesicles cross biological barriers between species, including humans, and deliver therapeutic molecules that influence gene expression, affecting various processes such as inflammation, oxidative stress, and cancer. For these reasons, plant-derived nanovesicles are gaining attention as a valuable substitute for mammalian exosomes as they offer benefits such as reduced immunogenicity, enhanced bioavailability, and the inclusion of beneficial plant metabolites. However, the development of affordable plant-derived nanovesicle-based therapies requires a robust characterization of their molecular structure and cargo, which in turn depends on obtaining sufficient quantities of homogeneous nanovesicle populations. In this study, we used an advanced purification platform combining ultrafiltration and anion exchange chromatography to isolate highly pure plant-derived nanovesicles from a new source, <i>Beta vulgaris</i> L. These particles were characterized in terms of size, charge, and morphology, and their molecular content was analyzed by omic technologies, including proteomics, lipidomics, and miRNomics. Their ability to promote wound healing and reduce inflammation was demonstrated in vitro using human cells. Furthermore, bioinformatic analysis linking the microRNA profile with potential human target genes provides insights into the biochemical pathways that underlie the bioactivity of nanovesicles.