Carrot extracellular nanovesicles as carotenoid carriers in an in vitro macular degeneration model

Plant-derived exosome-like nanovesicles (PENs) offer a promising biocompatible platform for the stable and targeted delivery of bioactive compounds, including conditions associated with oxidative stress, such as age-related macular degeneration (AMD). In this study, PENs were isolated from Daucus carota (carrot) juice and callus cultures, characterized as spherical vesicles, with PEN-like morphology and negative zeta potentials. Juice-derived PENs contained at least 12 carotenoids, including three xanthophylls and five carotenes, whereas no endogenous carotenoids were detected in callus-derived PENs. Lutein and zeaxanthin were exogenously encapsulated in both types of PENs using electroporation and passive diffusion, with electroporation at 200 mV and 50 µF achieving up to encapsulation efficiency of up to 80%. These xanthophyll-loaded PENs were delivered to ARPE-19 cells to assess their capacity to maintain cell viability under oxidative stress conditions. In vitro assays demonstrated that xanthophyll-encapsulated callus-derived PENs conferred superior protection maintaining over 95% cell viability, outperforming both juice-derived PENs and free carotenoid extracts. Despite their broad polydispersity and low zeta potential, PENs retained functional bioactivity and enabled efficient intercellular delivery of carotenoids. These findings suggest that carrot-derived PENs can be efficiently encapsulated with bioactive molecules and hold potential as highly biocompatible nanocarriers for targeted drug delivery.