Objective To explore the effect of grapefruit-derived exosome-like vesicles on melanin synthesis of mouse skin melanoma cells(B16F10)and zebrafish larvae,in order to obtain a safe and effective natural skin whitening agent.Methods Grapefruit-derived exosome-like vesicles(GFEVs)were isolated by differential speed centrifugation and ultracentrifugation,and then were characterized in terms of the particle size,concentration,and zeta potential by nanoparticle tracking analyzer.The morphological integrity of the GFEVs was detected by transmission electron microscope.Their cytotoxicity was assessed according to the viability of human fibroblasts(HFF-1)and human keratinocytes(HaCaT)by CCK8 assay.B16F10 cells and zebrafish larvae were used to study the whitening efficacy of grapefruit-derived exosome-like vesicles.The oxidative damage models of hydrogen peroxide-induced HFF-1 and HaCaT cells were used to explore the antioxidant effect of the GFEVs.Finally,the cargoes of grapefruit-derived exosome-like vesicles were analyzed by HPLC/MS method.Results Isolation and characterization of GFEVs showed they had a small average size,high concentration,high yield and purity,and complete vesicle morphology.Cytotoxicity assessment demonstrated little cytotoxicity of the GFEVs to HFF-1 and HaCaT cells.The efficacy data showed that the GFEVs significantly inhibited melanogenesis in B16F10 cells and zebrafish larvae.Preliminary mechanistic analysis showed that these vesicles are rich in a variety of flavonoids and have obvious antioxidant effects in both human fibroblasts and human keratinocytes.Conclusion GFEVs are rich in a variety of antioxidant flavonoids,with little cytotoxicity to human skin-derived cells and significant whitening effect.Therefore,the products of GFEVs have a good application potential in the field of whitening and freckle skin care.
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