title: Possible chondroprotective effect of canakinumab: an in vitro study on human osteoarthritic chondrocytes.
abstract: Canakinumab is a human IgGκ monoclonal antibody that neutralizes the activity of interleukin (IL)-1β blocking interaction with IL-1β receptors. Our study aimed to evaluate the in vitro effect of canakinumab on human osteoarthritic (OA) chondrocytes cultivated in the presence or absence of tumor necrosis factor (TNF)-α. Articular cartilage was obtained from the femoral heads of patients with osteoarthritis (OA). Chondrocytes were incubated with two concentrations (1μg/ml and 10μg/ml) of canakinumab alone or with TNF-α (10ng/ml) for 48h. We evaluated cell viability, release of proteoglycans (PG) and nitric oxide (NO) in culture medium, inducible nitric oxide synthase (iNOS) and metalloproteinanes (MMP)-1,3,13 gene expression, apoptosis, necrosis and morphological feature by transmission electron microscopy (TEM). Canakinumab alone did not have cytotoxic effect. Cell viability was reduced significantly (p<0.001) by TNF-α and restored by canakinumab at both concentrations used. TNF-α determined a significant decrease of PG (p<0.001) and an increase of NO (p<0.001) and MMP-1,3,13 gene expression. Canakinumab significantly increased the PG levels and decreased (1μg/ml, p<0.01; 10μg/ml, p<0.01) NO levels in cells cultured with TNF-α. The NO data were confirmed by the immunocytochemistry assay for iNOS. A significant reduction of MMP-1,3,13 gene expression was induced by canakinumab. Our experiments confirmed the pro-apoptotic effect of TNF-α and demonstrated a protective role of canakinumab. The results concerning biochemical data were further confirmed by the morphological findings obtained by TEM. We showed that canakinumab counteracts the negative effects of TNF-α on OA chondrocyte cultures and may have a potential chondroprotective role in OA.
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Italy. Electronic address: saracheleschi@hotmail.com.
authors.forename: Sara
authors.initials: S
authors.lastname: Cheleschi
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Italy. Electronic address: cantariniluca@hotmail.com.
authors.forename: Luca
authors.initials: L
authors.lastname: Cantarini
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Italy. Electronic address: pascarelli@unisi.it.
authors.forename: Nicola Antonio
authors.initials: NA
authors.lastname: Pascarelli
authors.affiliation: Department of Molecular and Developmental Medicine, University of Siena, Italy. Electronic address: giulia.collodel@unisi.it.
authors.forename: Giulia
authors.initials: G
authors.lastname: Collodel
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Italy. Electronic address: lucherini9@unisi.it.
authors.forename: Orso Maria
authors.initials: OM
authors.lastname: Lucherini
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Italy. Electronic address: mauro.galeazzi@unisi.it.
authors.forename: Mauro
authors.initials: M
authors.lastname: Galeazzi
authors.affiliation: Department of Medicine, Surgery and Neuroscience, Rheumatology Unit, University of Siena, Italy. Electronic address: fioravanti7@virgilio.it.
authors.forename: Antonella
authors.initials: A
authors.lastname: Fioravanti
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D061067:Antibodies, Monoclonal, Humanized; D017209:Apoptosis / Q000187:drug effects; D002358:Cartilage, Articular / Q000378:metabolism / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D005270:Femur Head / Q000378:metabolism / Q000473:pathology; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D053505:Matrix Metalloproteinases, Secreted / Q000235:genetics / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D009336:Necrosis / Q000209:etiology; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D010003:Osteoarthritis / Q000378:metabolism / Q000473:pathology; D020011:Protective Agents / Q000494:pharmacology*; D011509:Proteoglycans / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D020011:Protective Agents; D011509:Proteoglycans; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; C541220:canakinumab; D052247:Nitric Oxide Synthase Type II; D053505:Matrix Metalloproteinases, Secreted
keywords: Canakinumab; Human chondrocytes; Osteoarthritis; Tumor necrosis factor-α