Lola Dechêne1,2, Margaux Colin3, Catherine Demazy1,4, Maude Fransolet1,4, Ariane Niesten5, Thierry Arnould1, Didier Serteyn2, Marc Dieu1,4, Patricia Renard6,7. 1. Laboratory of Biochemistry and Cell Biology (URBC), NARILIS (Namur Research Institute for Life Sciences), University of Namur (UNamur), Rue de Bruxelles 61, 5000, Namur, Belgium. 2. Department of Clinical Sciences, Anesthesiology and Equine Surgery, Faculty of Veterinary Medicine, B41, University of Liege, Sart Tilman, 4000, Liège, Belgium. 3. Department of Pharmacotherapy and Pharmaceuticals, Faculty of Pharmacy, Université Libre de Bruxelles (ULB), 1050, Brussels, Belgium. 4. Mass Spectrometry Platform (MaSUN) - Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000, Namur, Belgium. 5. Centre of Oxygen, Research and Development (CORD), Institute of Chemistry B6a, University of Liege (ULiège), Sart Tilman, 4000, Liège, Belgium. 6. Laboratory of Biochemistry and Cell Biology (URBC), NARILIS (Namur Research Institute for Life Sciences), University of Namur (UNamur), Rue de Bruxelles 61, 5000, Namur, Belgium. patsy.renard@unamur.be. 7. Mass Spectrometry Platform (MaSUN) - Namur Research Institute for Life Sciences (Narilis), University of Namur (UNamur), 5000, Namur, Belgium. patsy.renard@unamur.be.
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. 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.
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. 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.
Authors: C Wayne McIlwraith; Christopher E Kawcak; David D Frisbie; Christopher B Little; Peter D Clegg; Mandy J Peffers; Morten A Karsdal; Stina Ekman; Sheila Laverty; Richard A Slayden; Linda J Sandell; L S Lohmander; Virginia B Kraus Journal: J Orthop Res Date: 2018-01-24 Impact factor: 3.494
Authors: Inês Esteves Dias; Pedro Olivério Pinto; Luís Carlos Barros; Carlos Antunes Viegas; Isabel Ribeiro Dias; Pedro Pires Carvalho Journal: BMC Vet Res Date: 2019-10-22 Impact factor: 2.741