BMSC-derived extracellular vesicles intervened the pathogenic changes of scleroderma in mice through miRNAs

Abstract

Background Systemic sclerosis (SSc) is a disease that features severe fibrosis of the skin and lacks effective therapy. Bone marrow mesenchymal stem cell (BMSC)-derived extracellular vesicles (EVs) are potential stem cell-based tools for the treatment of SSc. Methods BMSCs were isolated from the bone marrow of mice and identified with surface markers according to multilineage differentiation. EVs were isolated from the BMSC culture medium by ultracentrifugation and identified with a Nanosight NS300 particle size analyzer, transmission electron microscopy (TEM), and western blot. The microRNAs (miRNAs) of BMSC-derived EVs (BMSC-EVs) were studied via miRNA sequencing (miRNA-seq) and bioinformatic analysis. An SSc mouse model was established via subcutaneous bleomycin (BLM) injection, and the mice were treated with BMSCs or BMSC-derived EVs. Skin tissues were dissociated and analyzed with H&E staining, RNA sequencing (RNA-seq), western blot, and immunohistochemical staining. Results Evident pathological changes, like fibrosis and inflammation, were induced in the skin of BLM-treated mice. BMSCs and BMSC-EVs effectively intervened such pathological manifestations and disease processes in a very similar way. The effects of the BMSC-EVs were found to be caused by the miRNAs they carried, which were proven to be involved in regulating the proliferation and differentiation of multiple cell types and in multiple EV-related biological processes. Furthermore, TGF-β1-positive cells and α-SMA-positive myofibroblasts were significantly increased in the scleroderma skin of BLM-treated mice but evidently reduced in the scleroderma skin of the EV-treated SSc group. In addition, the numbers of mast cells and infiltrating macrophages and lymphocytes were evidently increased in the skin of BLM-treated mice but significantly reduced by EV treatment. In line with these observations, there were significantly higher mRNA levels of the inflammatory cytokines Il6, Il10, and Tnf-α in SSc mice than in control mice, but the levels decreased following EV treatment. Through bioinformatics analysis, the TGFβ and WNT signaling pathways were revealed to be closely involved in the pathogenic changes seen in mouse SSc, and these pathways could be therapeutic targets for treating the disease. Conclusions BMSC-derived EVs could be developed as a potential therapy for treating skin dysfunction in SSc, especially considering that they show similar efficacy to BMSCs but have fewer developmental regulatory requirements than cell therapy. The effects of EVs are generated by the miRNAs they carry, which alleviate SSc pathogenic changes by regulating the WNT and TGFβ signaling pathways.

The Therapeutic Effects of Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells on Scleroderma

“…Studies have shown that bone marrow mesenchymal stem cells or adipose mesenchymal stem cells [ 21 ] or their exosomes [ 22 ] achieve good therapeutic effects in the treatment of sclerodermatous mice, but there is no report on HUMSCs-Ex. Almost all cells can release exosomes, but umbilical cord mesenchymal stem cells are cost-effective, productive and acceptable to gain [ 23 ].…”

Human amniotic epithelial cell-derived extracellular vesicles provide an extracellular matrix-based microenvironment for corneal injury repair

“…Raw reads of the library were produced using the Illumina HiSeq 2500 SE50 platform and subjected to an in-house program, ACGT101-miR (LC Sciences, USA), as described previously. 34 High-level miRNAs were filtered according to a higher number of reads than the average copy number of the dataset.…”

“…Hematoxylin and eosin (H&E) staining was used to analyze the pathological morphology of corneas as described in our previous study. 34 …”

Bone Marrow Mesenchymal Stem Cell-Derived Extracellular Vesicles Carrying circ_0050205 Attenuate Intervertebral Disc Degeneration

“…BMSCs were maintained in serum-free DMEM for 2 days and then centrifuged at 800 × g and 4°C for 10 min and 10000 × g for 10 min and filtered through a 40 μ m membrane to remove cell debris and dead cells. The supernatant was subjected to ultracentrifugation at 100000 × g and 4°C for 1 h using a SW70Ti rotor (Beckman Coulter Inc., Chaska, MN) [ 26 ]. The extracted pellets were stored at −80°C.…”