Synergistic Anti-Aging Effects of Adipose-Derived Stem Cell Extracellular Vesicles Loaded With Natural Compounds

Background: Small extracellular vesicles from adipose-derived stem cells (ASC-sEVs) are gaining attentions rapidly for inherent therapeutic values in skin care and cosmetics. However, the optimal combinations of ASC-sEVs and certain natural compounds for synergistic anti-aging effects have not been systematically studied. Methods: Human ASC-sEVs were purified from culture supernatant of ASCs and multi-omics datasets of miRNAs, proteins and lipids of ASC-sEVs were analyzed for pathways regulating skin homeostasis. ASC-sEVs were then loaded with nicotinamide riboside (NR), resveratrol (RES), vitamin C (VITC), retinol (RET) and arbutin (ARB) at different concentrations by the sonication-incubation method. Their anti-oxidant, anti-wrinkle and anti-melanogenic effects were tested in vitro using the human keratinocyte HaCaT cells exposed to UVB radiation and human melanocyte B16F10 cells. Results: Multi-omics data analysis of ASC-sEVs identified key bioactive molecules regulating collagen formation, pigmentation, oxidative stress and inflammation. In the in vitro screenings for anti-aging effects, the compound-loaded ASC-sEVs outperformed the sEV- and compound-only treatments. Specifically in UVB-exposed HaCaT cells, 2 mu g/mL sEVs loaded with 20 mu g/mL NR, 2 mu g/mL RES, 5 mu g/mL VITC reduced reactive oxygen species level by 22.0%; while combination of sEVs and 2 mu g/mL RES, 2.8 mu g/mL RET significantly reduced MMP3 and upregulated PLOD1 expressions. B16F10 cells incubated with 2 mu g/mL sEVs loaded with 2 mu g/mL RES, 0.5 mM ARB had intracellular and extracellular melanin content lowered by 21.4% and 22.4% respectively. All the combinations caused no cytotoxicity. Conclusion: Our study demonstrated the superiority of ASC-sEVs to deliver both endogenous biocargos and exogenous compounds to achieve synergistic skin anti-aging effects.