Exosome-like nanovesicles isolated from Citrus limon L. exert antioxidative effect.

BACKGROUND

Exosome-like nanovesicles are biological nanostructures mediating cell-to-cell communication and capable to load selected cargos also in the interaction among different species.

OBJECTIVE

We aimed to explore the content of exosome-like nanovesicles derived from Citrus limon L. and to analyze the effects of their uptake on human cells.

METHODS

We isolated exosome-like nanovesicles from Citrus limon L. juice (EXO-CL) by differential centrifugation. EXOCL were analyzed for short RNA content by advanced sequencing technologies, and for ascorbic acid (vitamin C) and citrate content by enzymatic assays. EXO-CL anti-oxidant and pro-differentiative potential was evaluated in vitro on mesenchymal stromal cells (MSC), a common tool for regenerative strategies for several human tissues.

RESULTS

We showed that EXO-CL carry detectable amounts of citrate and vitamin C and, although it was not possible to identify specific miRNAs, we detected short RNA sequences (20-30 bp) with unknown functions and with different distribution size in respect to whole Citrus limon L. juice. In vitro, EXO-CL were uptaken by MSC and had a significant protective effect against oxidative stress. Furthermore, regarding the potential benefit for human bone health, we found that EXO-CL modulate MSC differentiation versus the osteogenic lineage.

CONCLUSIONS

We demonstrated that incubation with EXO-CL exerts antioxidant activity in human cells. This is most likely due to the direct delivery and uptake of micronutrients by human cells that are well preserved inside the nanovesicle membrane, including the unstable vitamin C. Based on our results, we speculate that fruit-derived nanovesicles have the potential to mediate interspecies influence after food intake.