Effects of Extracellular Vesicles Secreted by TGFβ-Stimulated Umbilical Cord Mesenchymal Stem Cells on Skin Fibroblasts by Promoting Fibroblast Migration and ECM Protein Production

Abstract

Umbilical cord-derived mesenchymal stem cells (UCMSCs) have been illustrated for their roles in immunological modulation and tissue regeneration through the secretome. Additionally, culture conditions can trigger the secretion of extracellular vesicles (EVs) into extracellular environments with significant bioactivities. This study aims to investigate the roles of three EV sub-populations released by UCMSCs primed with transforming growth factor β (TGFβ) and their capacity to alter dermal fibroblast functions for skin aging. Results show that three EV sub-populations, including apoptotic bodies (ABs), microvesicles (MVs), and exosomes (EXs), were separated from conditioned media. These three EVs carried growth factors, such as FGF-2, HGF, and VEGF-A, and did not express noticeable effects on fibroblast proliferation and migration. Only EX from TGFβ-stimulated UCMSCs exhibited a better capacity to promote fibroblasts migrating to close scratched wounds than EX from UCMSCs cultured in the normal condition from 24 h to 52 h. Additionally, mRNA levels of ECM genes (COL I, COL III, Elastin, HAS II, and HAS III) were detected with lower levels in fibroblasts treated with EVs from normal UCMSCs or TGFβ-stimulated UCMSCs compared to EV-depleted condition. On the contrary, the protein levels of total collagen and elastin released by fibroblasts were greater in the cell groups treated with EVs compared to EV-depleted conditions; particularly elastin associated with TGFβ-stimulated UCMSCs. These data indicate the potential roles of EVs from UCMSCs in protecting skin from aging by promoting ECM protein production.

Bio-Pulsed Stimulation Effectively Improves the Production of Avian Mesenchymal Stem Cell-Derived Extracellular Vesicles That Enhance the Bioactivity of Skin Fibroblasts and Hair Follicle Cells

“…Thus, collagen is among the most vital proteins of the ECM that imparts a fuller and younger appearance to tissues, and elastin is essential for skin elasticity [ 54 ]. Our findings revealed that the effects of bio-pulsed AMSC-sEVs on skin fibroblasts are consistent with previous results that EVs secreted by TGFβ-stimulated umbilical cord mesenchymal stem cells on skin fibroblasts promoted fibroblast migration and ECM protein production [ 55 ].…”

Cytokine-primed umbilical cord mesenchymal stem cells enhanced therapeutic effects of extracellular vesicles on osteoarthritic chondrocytes

“…(C1) An AB representative with a diameter ranging from 500 nm to 2000 nm (scale bar 9 μm); (C2) A MV representative with irregular shapes and diameters from 200 nm to 400 nm (scale bar 500 nm); (C3) An EX representative that has cup-shaped morphology and sizes ranging from 40 to 200 nm (scale bar 2 µm). (D) A representative of EV markers ( 22 ). 15 μg total EV protein was loaded in each lane.…”

Absolute Quantification of Dynamic Cellular Uptake of Small Extracellular Vesicles via Lanthanide Element Labeling and ICP–MS

“…Once the confluency of HeLa cells reached 80%, the original culture medium was replaced with EV-depleted culture medium (DMEM supplemented with 10% EV-depleted FBS and antibiotics), and the cells were cultured for another 24 h. Then, all of the supernatant was collected for sEV isolation. EV-depleted FBS was obtained by ultracentrifuging FBS at 120,000 × g for 18 h at 4 °C, − followed by careful collection of the supernatant.…”