Functional recovery in photo-damaged human dermal fibroblasts by human adipose-derived stem cell extracellular vesicles

Abstract

Ultraviolet-B (UVB) irradiation causes imbalance between dermal matrix synthesis and degradation through aberrant upregulation of matrix metalloproteinases (MMPs), which leads to overall skin photoaging. We investigated the effects of extracellular vesicles (EVs) derived from human adipose-derived stem cells (HASCs) on photo-damaged human dermal fibroblasts (HDFs). EVs were isolated from conditioned media of HASCs with tangential flow filtration and characterized using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), western blotting, micro RNA (miRNA) arrays, cytokine arrays and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The effects of EVs on the UVB-irradiated HDFs were evaluated using scratch assay, ELISA and real-time PCR. Microarrays exhibited that EVs are rich in various miRNAs and proteins, and that these EV contents are linked to a broad range of biological functions, including fibroblast proliferation, UV protection, collagen biosynthesis, DNA repair and cell ageing. A scratch assay showed that HASC-EVs enhanced the migration ability of UVB-irradiated HDFs. Real-time RT-PCR and ELISA analyses revealed that the HASC-derived EVs significantly suppressed the overexpression of MMP-1, -2, -3 and -9 induced by UVB irradiation and enhanced the expression of collagen types I, II, III and V and elastin. In particular, tissue inhibitor of metalloproteinase (TIMP)-1 and transforming growth factor (TGF)-beta 1, which are important factors involved in MMP suppression and ECM synthesis, were upregulated in EV-treated HDFs after UVB irradiation. Overall results suggest that diverse components that are enriched in HASC-derived EVs could act as a biochemical cue for recovery from skin photoaging.