Elisa Belluzzi1,2, Eleonora Olivotto3, Giovanna Toso2, Augusto Cigolotti4, Assunta Pozzuoli2, Carlo Biz4, Giovanni Trisolino5,6, Pietro Ruggieri4, Brunella Grigolo3,7, Roberta Ramonda1, Marta Favero1,3. 1. a Rheumatology Unit, Department of Medicine-DIMED , University Hospital of Padova , Padova , Italy. 2. b Musculoskeletal Pathology and Oncology Laboratory, Department of Orthopaedics and Orthopaedic Oncology , University of Padova , Padova , Italy. 3. c RAMSES Laboratory-RIT Department , IRCCS Istituto Ortopedico Rizzoli , Bologna , Italy. 4. d Department of Orthopaedics and Orthopaedic Oncology , University of Padova , Padova , Italy. 5. e Reconstructive Hip and Knee Joint Surgery , IRCCS Istituto Ortopedico Rizzoli , Bologna , Italy. 6. f Pediatric orthopedic and Traumatology , IRCCS Istituto Ortopedico Rizzoli , Bologna , Italy. 7. g Laboratory of Immunorheumatology and Tissue Regeneration , IRCCS Istituto Ortopedico Rizzoli , Bologna , Italy.
Abstract
AIM: Osteoarthritis (OA) is a whole joint pathology involving cartilage, synovial membrane, meniscus, subchondral bone, and infrapatellar fat pad (IFP). Synovitis has been widely documented in OA suggesting its important role in pathogenesis. The aim of this study was to investigate the role of different joint tissues in promoting synovitis. MATERIALS AND METHODS: Conditioned media (CM) from cartilage, synovial membrane, meniscus, and IFP were generated from tissues of five patients undergoing total knee replacement and used to stimulate a human fibroblast-like synoviocytes cell line (K4IM). Cytokines, chemokines, and metalloproteases release was analyzed in all CM by Bio-Plex Assay and sulfated glycosaminoglycan (GAG) content by dimethylmethylene blue assay. Gene expression of several markers was evaluated by real-time PCR in K4IM cells stimulated with the CM obtained from joint tissues. RESULTS: CM from all tissues produced high levels of IL-6, IL-8, and CCL2. CCL21, MMP-3, and -13 levels were detected in all CM except IFP. MMP-10 was present only in CM of cartilage and synovial tissues. IL-1β, IL-15, TNF-α, CCL5, and CCL19 were undetectable. However, only K4IM cells stimulated by the CM from OA synovium showed an increase of IL-6, CXCL-8, CCL21, MMP10, and IL-1β expression. CONCLUSION: Our study showed that K4IM might be a suitable in vitro model for evaluating different cellular pathways in OA studies. Importantly, we demonstrated that in OA, all joint tissues might be involved in the progression of synovitis with a predominant role of synovial membrane itself compared to the other joint tissues.
AIM: Osteoarthritis (OA) is a whole joint pathology involving cartilage, synovial membrane, meniscus, subchondral bone, and infrapatellar fat pad (IFP). Synovitis has been widely documented in OA suggesting its important role in pathogenesis. The aim of this study was to investigate the role of different joint tissues in promoting synovitis. MATERIALS AND METHODS: Conditioned media (CM) from cartilage, synovial membrane, meniscus, and IFP were generated from tissues of five patients undergoing total knee replacement and used to stimulate a human fibroblast-like synoviocytes cell line (K4IM). Cytokines, chemokines, and metalloproteases release was analyzed in all CM by Bio-Plex Assay and sulfated glycosaminoglycan (GAG) content by dimethylmethylene blue assay. Gene expression of several markers was evaluated by real-time PCR in K4IM cells stimulated with the CM obtained from joint tissues. RESULTS: CM from all tissues produced high levels of IL-6, IL-8, and CCL2. CCL21, MMP-3, and -13 levels were detected in all CM except IFP. MMP-10 was present only in CM of cartilage and synovial tissues. IL-1β, IL-15, TNF-α, CCL5, and CCL19 were undetectable. However, only K4IM cells stimulated by the CM from OA synovium showed an increase of IL-6, CXCL-8, CCL21, MMP10, and IL-1β expression. CONCLUSION: Our study showed that K4IM might be a suitable in vitro model for evaluating different cellular pathways in OA studies. Importantly, we demonstrated that in OA, all joint tissues might be involved in the progression of synovitis with a predominant role of synovial membrane itself compared to the other joint tissues.
Authors: Francesca Oliviero; Yessica Zamudio-Cuevas; Elisa Belluzzi; Lisa Andretto; Anna Scanu; Marta Favero; Roberta Ramonda; Giampietro Ravagnan; Alberto López-Reyes; Paolo Spinella; Leonardo Punzi Journal: Foods Date: 2019-11-07
Authors: Elisa Belluzzi; Veronica Macchi; Chiara Giulia Fontanella; Emanuele Luigi Carniel; Eleonora Olivotto; Giuseppe Filardo; Gloria Sarasin; Andrea Porzionato; Marnie Granzotto; Assunta Pozzuoli; Antonio Berizzi; Manuela Scioni; Raffaele De Caro; Pietro Ruggieri; Roberto Vettor; Roberta Ramonda; Marco Rossato; Marta Favero Journal: Int J Mol Sci Date: 2020-08-21 Impact factor: 5.923
Authors: Selene Pérez-García; Valentina Calamia; Tamara Hermida-Gómez; Irene Gutiérrez-Cañas; Mar Carrión; Raúl Villanueva-Romero; David Castro; Carmen Martínez; Yasmina Juarranz; Francisco J Blanco; Rosa P Gomariz Journal: Int J Mol Sci Date: 2021-06-16 Impact factor: 6.208