Characterization of Rosa damascena Callus-derived Exo-some-like Vesicles and Their Multifunctional Activities in Skin-related Cellular Models

Plant-derived extracellular vesicles (PDEVs) are emerging as promising bioactive materials for biomedical and dermatological applications. In this study, we isolated and characterized exosome-like vesicles derived from Rosa damascena callus culture medium (RSC-EXO) and evaluated their molecular features and biological activities in skin-related cellular models. Nanoparticle tracking analysis and cryo-electron microscopy showed that RSC-EXO exhibited a nanoscale size distribution and spherical morphology. Western blotting confirmed enrichment of the plant EV-associated markers PEN1 and TET8. RSC-EXO were efficiently internalized by human dermal fibroblasts and showed markedly improved biocompatibility compared with crude conditioned medium (RSC-CM). Functionally, RSC-EXO significantly increased collagen synthesis and showed a trend toward enhanced wound closure in fibroblasts. In addition, RSC-EXO reduced melanin production in -MSH-stimulated B16F10 melanoma cells and suppressed the secretion of pro-inflammatory cytokines, including IL-1, IL-6, and TNF-, in LPS-stimulated RAW 264.7 macrophages. Proteomic analysis revealed a distinct cargo enriched in stress-, defense-, and metabolism-related proteins, while small RNA sequencing identified a heterogeneous small RNA population containing a limited fraction of miRNA-sized reads. Collectively, these findings suggest that RSC-EXO represents a biologically active plant-derived vesicle population with regenerative and anti-inflammatory activity observed in vitro in skin-related cellular models and support its potential as a promising platform for future cosmeceutical and dermatological applications.