Arugula leaf nanovesicles as sustainable food ingredient: processing, stability, and bioavailability

Plant-derived nanovesicles (PDVs) are emerging as natural nanostructures with potential applications in food and nutraceutical systems. In this study, we report the scalable production of arugula leaf–derived nanovesicles (ALVs) through ultrafiltration-size exclusion purification followed by spray-drying stabilization. This workflow ensured preservation of nanoscale morphology, biochemical cargo, and stability during long-term storage. Comprehensive characterization by cryo-EM, Raman spectroscopy, circular dichroism, and LC-HRMS revealed that ALVs contain amino acids, fatty acids, phenolic acids, flavonoids, and lignans, along with regulatory plant miRNAs. Importantly, ALVs were able to cross differentiated Caco-2 intestinal monolayers without compromising barrier integrity, suggesting safe oral bioavailability. Overall, this work establishes arugula-derived nanovesicles as sustainable, stable, and bioavailable food nano-ingredient, highlighting their potential integration into functional foods and nutraceutical applications.