Characterization of the Proteins Secreted by Equine Muscle-Derived Mesenchymal Stem Cells Exposed to Cartilage Explants in Osteoarthritis Model

Abstract

Background Osteoarthritis (OA) is a highly prevalent joint degenerative disease for which therapeutic treatments are limited or invasive. Cell therapy based on mesenchymal stem/stromal cells (MSCs) is therefore seen as a promising approach for this disease, in both human and horses. As the regenerative potential of MSCs is mainly conferred by paracrine function, the goal of this study was to characterize the secreted proteins of muscle-derived MSCs (mdMSCs) in an in vitro model of OA to evaluate the putative clinical interest of mdMSCs as cell therapy for joint diseases like osteoarthritis. Methods An equine osteoarthritis model composed of cartilage explants exposed to pro-inflammatory cytokines was first developed. Then, the effects of mdMSC co-culture on cartilage explant were studied by measuring the glycosaminoglycan release and the NO2− production. To identify the underlying molecular actors, stable isotope-labeling by amino acids in cell culture based secreted protein analyses were conducted, in the presence of serum. The relative abundance of highly sequenced proteins was finally confirmed by western blot. Results Co-culture with muscle-derived MSCs decreases the cytokine-induced glycosaminoglycan release by cartilage explants, suggesting a protecting effect of mdMSCs. Among the 52 equine proteins sequenced in the co-culture conditioned medium, the abundance of decorin and matrix metalloproteinase 3 was significantly modified, as confirmed by western blot analyses. Conclusions These results suggest that muscle-derived MSCs could reduce the catabolic effect of TNFα and IL-1β on cartilage explant by decreasing the secretion and activity of matrix metalloproteinase 3 and increasing the decorin secretion. Graphical abstract mdMSCs capacity to reduce the catabolic consequences of cartilage exposure to pro-inflammatory cytokines. These effects can be explained by mdMSC-secreted bioactive such as TIMP-1 and decorin, known as an inhibitor of MMP3 and an anti-inflammatory protein, respectively.

Injectable Decorin/Gellan Gum Hydrogel Encapsulating Adipose-Derived Stem Cells Enhances Anti-Inflammatory Effect in Cartilage Injury via Autophagy Signaling

“…Our results showed a synergistic effect of decorin and ADSCs in inflammatory joint disorder; therefore, we assumed that decorin in the matrix could modulate ADSCs immunol phenotype to enhance the anti-inflammatory effect via cytokines paracrine action on chondrocytes. In line with these data, a co-culture-secreted protein study of MSCs and cartilage explants showed a protective effect of MSCs on IL-1β induced cartilage catabolic injury by increasing the decorin secretion 24 .…”

“…Of note, alleviation of antigen-induced arthritis by corticosteroid administration elevated decorin mRNA levels in connective tissues 23 . Furthermore, co-culture MSCs could reduce the pro-inflammatory effect of TNFα and IL-1β on cartilage by increasing decorin secretion 24 , in line with the fact that decorin may interact with extracellular tyrosine kinase receptors for immunol signaling transduction in inflammatory diseases 25 . Given the fact that decorin-overexpressed MSCs significantly attenuate inflammation by regulating immune responses 26 , we therefore wonder whether pericellular decorin would enhance the anti-inflammatory property of ADSCs that facilitate cartilage repair.…”

Menstrual Blood-Derived Mesenchymal Stem Cell Paracrine Factors Stimulate Chondrogenesis in vitro and Possess Protective Effects to Articular Cartilage

“…5), as compared to explants cultured without MenSCs, suggesting a protective effect of MenSCs on cartilage ECM. Similar results were demonstrated with equine derived muscle MSCs cultivated in co-cultures with OA cartilage explants, where signi cantly lower amounts of GAGs was present in the medium of co-cultures, as compared to cartilage explants cultured alone (42). Secretion of growth factors revealed signi cantly higher levels of activin A in co-culture medium with and without IL-1β and signi cantly decreased levels of IGF-1 in co-cultures (Fig.…”

Menstrual Blood-Derived Stem Cell Paracrine Factors Possess Stimulatory Effects on Chondrogenesis In Vitro and Diminish the Degradation of Articular Cartilage during Osteoarthritis

“…GAG analysis also showed a significant decrease of GAG release into the medium after co-culturing explants with MenSCs for 3, 7, and 21 days (Figure 5) as compared to explants cultured without MenSCs, suggesting a protective effect of MenSCs on cartilage ECM. Similar results were demonstrated with equine derived muscle MSCs cultivated in co-cultures with OA cartilage explants, where significantly lower amounts of GAGs were present in the medium of cocultures compared to cartilage explants cultured alone [52]. The secretion of growth factors revealed significantly higher levels of activin A in co-culture medium with and without IL-1β and significantly decreased levels of IGF-1 in co-cultures (Figure 6).…”