Interaction with hyaluronan matrix and miRNA cargo as contributors for in vitro potential of mesenchymal stem cell-derived extracellular vesicles in a model of human osteoarthritic synoviocytes

Abstract

Background Osteoarthritis (OA) is the most prevalent joint disease, and to date, no options for effective tissue repair and restoration are available. With the aim of developing new therapies, the impact of mesenchymal stem cells (MSCs) has been explored, and the efficacy of MSCs started to be deciphered. A strong paracrine capacity relying on both secreted and vesicle-embedded (EVs) protein or nucleic acid-based factors has been proposed as the principal mechanism that contributes to tissue repair. This work investigated the mechanism of internalization of extracellular vesicles (EVs) released by adipose-derived MSCs (ASCs) and the role of shuttled miRNAs in the restoration of homeostasis in an in vitro model of human fibroblast-like synoviocytes (FLSs) from OA patients. Methods ASC-EVs were isolated by differential centrifugation and validated by flow cytometry and nanoparticle tracking analysis. ASC-EVs with increased hyaluronan (HA) receptor CD44 levels were obtained culturing ASCs on HA-coated plastic surfaces. OA FLSs with intact or digested HA matrix were co-cultured with fluorescent ASC-EVs, and incorporation scored by flow cytometry and ELISA. ASC-EV complete miRNome was deciphered by high-throughput screening. In inflamed OA FLSs, genes and pathways potentially regulated by ASC-EV miRNA were predicted by bioinformatics. OA FLSs stimulated with IL-1β at physiological levels (25 pg/mL) were treated with ASC-EVs, and expression of inflammation and OA-related genes was measured by qRT-PCR over a 10-day time frame with modulated candidates verified by ELISA. Results The data showed that HA is involved in ASC-EV internalization in FLSs. Indeed, both removal of HA matrix presence on FLSs and modulation of CD44 levels on EVs affected their recruitment. Bioinformatics analysis of EV-embedded miRNAs showed their ability to potentially regulate the main pathways strictly associated with synovial inflammation in OA. In this frame, ASC-EVs reduced the expression of pro-inflammatory cytokines and chemokines in a chronic model of FLS inflammation. Conclusions Given their ability to affect FLS behavior in a model of chronic inflammation through direct interaction with HA matrix and miRNA release, ASC-EVs confirm their role as a novel therapeutic option for osteoarthritic joints. Electronic supplementary material The online version of this article (10.1186/s13287-019-1215-z) contains supplementary material, which is available to authorized users.

microRNAs in Ex Vivo Human Adipose Tissue Derived Mesenchymal Stromal Cells (ASC) Undergo Rapid Culture-Induced Changes in Expression, Including miR-378 which Promotes Adipogenesis

“…Shuttling of select-miRNAs in microvesicles and delivery to target cells has been observed in human bone-marrow MSCs [7]. A similar phenomenon was also recently reported in ASC, where exosome-shuttled miRNAs were found to reduce the expression of pro-inflammatory cytokines and chemokines in an in vitro model of chronic inflammation [8]. Therefore, the therapeutic potency of ASC is likely to be linked in part to microRNA expression [5].…”

Extracellular vesicles derived from different sources of mesenchymal stem cells: therapeutic effects and translational potential

“…CXCL8 showed significant contractions, suggesting that EVs regulates inflammation through the CREB pathway. In summary, AD-MSCs EVs decreased the expression of pro-inflammatory cytokines and chemokines in the chronic FLS inflammatory model [86]. Moreover, AD-MSCs EVs down-regulated β-galactosidase activity and the accumulation of γH2AX foci which was associated with senescence.…”

MiRNA Profiles of Extracellular Vesicles Secreted by Mesenchymal Stromal Cells—Can They Predict Potential Off-Target Effects?

“…Our literature search revealed that most studies focused on the therapeutic aspect of miRNAs of MSC-derived EVs. Only a few studies made the data of their miRNA arrays publicly available [32][33][34]. Additionally, the role of MSC-derived miRNAs in cancer biology has been discussed and investigated by other groups.…”

“…Even here, however, only few groups made all collected data available for secondary analysis. In the case of AT-MSCs, one group has investigated the role of AT-MSC-derived EVs in the development and treatment of osteoarthritis [32,33]. In both publications, the raw data of the miRNA array were made available by the authors.…”