Characterization and Bioactivity of Nanovesicles Recovered From Industrial Cheesemaking Whey Wastewater

Food-derived nanovesicles (NVs), and particularly, those derived from milk, are considered promising nutraceutical candidates and potential carriers for drug delivery. Here, as an alternative to milk, we used a dairy byproduct as a source of NVs. Specifically, we isolated and characterized NVs from cheese whey wastewater (WWW) generated during the Grana Padano cheese production. The results of this study demonstrated that WWW-NVs shared key features with milk-derived vesicles in terms of biophysical properties (mean size <200 nm and negative zeta potential) and membrane lipid composition, characterized by the high content of phosphatidylcholine and sphingomyelin. WWW-NVs were efficiently internalized by monocytic THP-1 cells. In H₂O₂-treated cells, WWW-NVs significantly increased cell viability and reduced intracellular ROS levels to those of controls. Moreover, WWW-NVs abrogated lipopolysaccharide and ATP-induced inflammasome activation, as demonstrated by the suppression of caspase-1 activation. Overall, WWW-NVs exhibited biophysical and functional properties comparable to those reported for milk-derived vesicles, while presenting advantages in terms of sustainability and waste valorization. The valorization of NVs from this industrial byproduct may bypass the need for pretreatment steps and avoid competition with the food chain. Therefore, WWW might represent a sustainable and alternative source of bioactive NVs, with promising applications in human health and biotechnology. PRACTICAL APPLICATIONS: This study demonstrated that nanovesicles isolated from cheese whey wastewater, an industrial dairy by-product, share morphological and biochemical properties with milk-derived vesicles. The whey wastewater-derived nanovesicles exerted antioxidant and anti-inflammatory effects on mammalian cells. These findings support the potential use of dairy by-products as an abundant, alternative, and sustainable source of bioactive nanovesicles avoiding competition with the food chain.