Characterization and antioxidant mechanisms of cherry-tomato derived exosome-like nanoparticles: Physicochemical properties, biomolecular cargo, and bioactivity

Edible plant derived exosome-like nanoparticles (ELNs) have been shown to possess numerous bioactive properties with potential health benefits. Cherry tomatoes contain a wide variety of bioactive substances, however, cherry-tomato derived exosome-like nanoparticles (CTELNs) remain unexplored. This study employed a polyethylene glycol (PEG) precipitation method for the efficient isolation of CTELNs, evaluating five PEG concentrations (ranging from 10% to 30%, <em>w</em>/<em>v</em>) to determine the optimal precipitation conditions. Characterization and compositional analysis demonstrated that CTELNs isolated using 20% PEG displayed superior physicochemical properties. The small RNA sequencing analysis revealed a distinct enrichment of specific microRNAs (miRNA) implicated in the regulation of oxidative stress in CTELNs, such as miR156e-3p, miR156e-5p, miR390a-5p and miR9474-5p. Target gene prediction indicated that CTELNs potentially regulated pathways related to binding and catalytic activity. Notably, nine miRNAs were predicted to target the genes related to oxidative stress. <em>In vitro</em> cellular assays conducted on RAW264.7 cells showed that CTELNs exhibited no significant toxicity. Furthermore, functional validation further confirmed that CTELNs ameliorated H₂O₂-induced oxidative stress. The results of this work suggest that PEG precipitation for CTELN<span style="text-decoration: line-through;">s</span> isolation is feasible and the optimal PEG concentration is determined, which facilitates further exploration of edible plant derived exosome-like nanoparticles. Moreover, the remarkable antioxidant properties of CTELNs support their potential application as natural antioxidative agents in functional foods or nutraceuticals.