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%, w/v) 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. In vitro 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 CTELNs 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.