Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5896866 | Cytokine | 2015 | 8 Pages |
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 (1 μg/ml and 10 μg/ml) of canakinumab alone or with TNF-α (10 ng/ml) for 48 h. 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 (1 μg/ml, p < 0.01; 10 μg/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.
Keywords
TGFβIL-1 RaTNFFDADMOADABBEMEAproteoglycansACRiNOSNSAIDNGSPBSMmpsqPCRDMEMBSADMSODulbecco’s modified Eagle mediumMTTQuantitative PCRbovine serum albuminenzyme-linked immune-sorbent assayEuropean Medicines agencyDrug administrationOsteoarthritisTemInterleukin-1βTransforming Growth Factor BetaELISAtumor necrosis factor-αnonsteroidal anti-inflammatory drugDimethyl sulfoxidenormal goat seruminducible nitric oxide synthaseActbPhosphatidylserineMetalloproteinasesPhosphate-buffered salineTransmission electron microscopeNitric oxidePropidium iodideoptical densityAmerican College of RheumatologycanakinumabHuman chondrocytes
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Authors
Sara Cheleschi, Luca Cantarini, Nicola Antonio Pascarelli, Giulia Collodel, Orso Maria Lucherini, Mauro Galeazzi, Antonella Fioravanti,