# Plant Extracellular Vesicles: Investigating Their Utilization as Beneficial Nutrients in Diet
## 4 P-EV Biological Properties and Therapeutic Applications  
In the last decade, plant extracellular vesicles have been exploited as excellent candidates for nanomedicine and nutraceutical applications due to their unique biological functions; thus, the research in this field has increased exponentially [2]. As already mentioned, because P-EVs can establish cell-to-cell communication between different species, they can be internalized and modulate host cell processes [7]. As such, the intrinsic bioactive cargos are believed to account for the beneficial effects of these vesicles, including anti-inflammatory, anticancer, antioxidant, and regenerative properties [18]. In addition to the promising application as a new class of therapeutics, P-EVs can also be developed as novel biologically derived drug vehicles with further advantages over current synthetic carriers, exhibiting in vitro and in vivo high biocompatibility, as most of them are isolated from edible plants, and stability in biological fluids even when administered orally [51]. Their great potential as oral drug carriers stems from their ability to withstand the harsh gastrointestinal environments and cross-biological barriers [51]. In vitro studies have demonstrated that plant vesicles can remain stable at different temperatures and pH values, mainly due to the protective functions against degradation exerted by the lipid bilayer on the bioactive cargos [51]. In particular, aquaporins were reported to play a key role in preserving the integrity of plasma membrane in broccoli-derived vesicles under high salt stress, which induced a significant accumulation of these proteins to ensure permeability as an adaptive mechanism [52]. In a previous study, the same authors demonstrated that changes in the lipid/protein ratio of Brassica species confer different physical properties to the lipid bilayer, leading to a degree of salt tolerance related to the presence of membrane proteins [53]. Because of their stability in the gastrointestinal tract, P-EVs can be administered orally and reach the target site efficiently as intact structures [54]. For example, [PERSON] et al. showed that ginger-derived EVs were taken up by intestinal stem cells and macrophages, and after oral administration for the treatment of inflammatory bowel disease, they promoted intestinal wound healing [55]. These authors also used ginger-nanoparticles to deliver siRNA-CD98 into colon tissue, and they found that administration of these vesicles decreased CD98 expression and prevented inflammation [55]. In addition, orally given vesicles, isolated from broccoli extracts, can regulate intestinal immune homeostasis in mice by targeting dendritic cells [41]. Inflammation is part of the innate immune response that, under damaging stimuli, may evolve toward acute or chronic inflammatory diseases, such as colitis [11]. In particular, plant vesicles have shown immunomodulatory effects on gastrointestinal homeostasis [56]. For example, [PERSON] et al. [57] demonstrated that grapefruit-derived vesicles could alleviate dextran sulfate sodium (DSS)-induced colitis in mice without exerting toxic effects. It is likely that these vesicles enhance the anti-inflammatory response of intestinal macrophages by upregulating the expression of anti-inflammatory cytokines such as IL-10, which in turn decreases the expression of proinflammatory cytokines and chemokines. In addition, the same study reported that miRNAs from ginger and grapefruit vesicles can target genes of the probiotic _Lactobacillus rhamnus_ in mice gut, thus promoting antimicrobial immunity [57]. Similar studies have also been reported on broccoli, grapes, and carrots [41,58]. Several studies have demonstrated the anti-proliferative and pro-apoptotic effects of P-EVs, which account for their anticancer properties [59]. Vesicles isolated from lemon juice were found to inhibit cancer cells growth in vitro, without affecting healthy cells, by selective activation of the Trail-mediated apoptotic pathway.