NMN-loaded hucMSC-derived small extracellular vesicles (NMN-sEVs) may improve skin aging in mice.
NMN-sEVs treatment is associated with increased intracellular NAD levels and SIRT3 expression.
The treatment may delay cellular and restore mitochondrial dysfunction.
NMN-sEVs could influence mitochondrial function by promoting mitochondrial autophagy.
Inhibition of SIRT3 with 3-TYP suppressed the beneficial effects of NMN-sEVs on cellular senescence and mitochondrial function.
Simplified
BACKGROUND: Recently, the beneficial effects of human umbilical cord mesenchymal stromal cell (hucMSC)-derived small extracellular vesicles (sEVs) in mitigating skin aging through multiple mechanisms have been widely reported. β-Nicotinamide mononucleotide (NMN) is an iconic anti-aging drug that increases NADlevels in the body to slow down, ameliorate, and prevent various phenotypes associated with aging, but its high water solubility, low permeability, and instability limit its clinical application. Based on this, we applied electroporation to construct NMN-loaded hucMSC-sEVs (NMN-sEVs) to improve their stability and efficacy and to enhance their potential for translational application in medical aesthetics and anti-aging. +
METHODS: D-galactose was applied to construct a mouse skin aging model, based on which comparative analyses of topical and nano-microneedle administration were performed to determine the optimal delivery method of sEVs in vivo experiments. After constructing NMN-sEVs by electroporation, high-performance liquid chromatography was applied to detect the loading efficiency, and the effects of NMN-sEVs on delaying skin aging were assessed by histological analysis. In addition, the defense effects of NMN-sEVs against cellular were verified by reactive oxygen species assay, β-galactosidase staining, qRT-PCR, Western blot, and cellular immunofluorescence. Finally, the roles of NMN-sEVs in remodeling mitochondrial function and delaying cellular senescence through mitochondrial autophagy were assessed by mitochondrial mass, function, and autophagy level assays.
RESULTS: Our data suggested that NMN-sEVs could improve skin aging in mice, delay cellular senescence, and restore cellular mitochondrial dysfunction. Notably, NMN-sEVs treatment increased intracellular NADlevels and SIRT3 expression, as well as rescued the inhibition of senescence-induced mitochondrial autophagy, suggesting a role for NMN-sEVs in the remodeling of mitochondrial function through mitochondrial autophagy. Additionally, the use of the SIRT3 inhibitor 3-TYP suppressed the positive effects of NMN-sEVs on cellular senescence, mitochondrial function, and mitochondrial autophagy while restoring senescence-associated characteristics. +
CONCLUSION: Overall, our findings revealed a mechanism by which NMN-sEVs attenuated mitochondrial dysfunction and rescued cellular senescence by promoting NAD/SIRT3 pathway-mediated and might provide a promising strategy for anti-aging pharmaceuticals. +
Key numbers
44.13%
Encapsulation Efficiency
Encapsulation efficiency of NMN in 1×10 particles/mL sEVs.
4424.57 µg/10 sEV
Loading Capacity
Loading capacity of NMN in 1×10 particles/mL sEVs.
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Declarations. Ethical approval and consent to participate: The Ethics Committee of the Fourth Affiliated Hospital of Jiangsu University approved the experimental protocol for the study of umbilical cord mesenchymal stem cells and exosomes for delaying skin aging (Project title: Study of the roles of umbilical cord mesenchymal stem cells and exosomes in the repair of tissue injuries; Approval number: 202301; Approval date: January 4, 2023). The Fourth Affiliated Hospital of Jiangsu University confirmed that there was initial ethical approval for collection of hucMSCs, and the donors had signed informed consent. All experiments involving animals were approved by the Institutional Animal Care and Use Committee of Jiangsu University and were conducted in accordance with the ARRIVE guideline 2.0 (Project title: NMN-engineered hucMSC-sEV delays D-gal-induced skin aging in mice; Approval number: UJS-IACUC-2024070902; Approval date: July 9, 2024). Consent for publication: Not applicable. Competing interests: The authors confirm that there are no conflicts of interest regarding the publication of this article.
