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See detailOsteoblasts from the sclerotic subchondral bone downregulate aggrecan but upregulate metalloproteinases expression by chondrocytes. This effect is mimicked by interleukin-6, -1 beta and oncostatin M pre-treated non-sclerotic osteoblasts
Sanchez, Christelle ULg; Deberg, Michelle ULg; Piccardi, Nathalie et al

in Osteoarthritis and Cartilage (2005), 13(11), 979-987

OBJECTIVE: To determine the effects of osteoarthritic (OA) subchondral osteoblasts on the metabolism of human OA chondrocytes in alginate beads. METHODS: Human chondrocytes were isolated from OA cartilage ... [more ▼]

OBJECTIVE: To determine the effects of osteoarthritic (OA) subchondral osteoblasts on the metabolism of human OA chondrocytes in alginate beads. METHODS: Human chondrocytes were isolated from OA cartilage and cultured in alginate beads for 4 days in the absence or in the presence of osteoblasts isolated from non-sclerotic (N) or sclerotic (SC) zones of human OA subchondral bone in monolayer (co-culture system). Before co-culture, osteoblasts were incubated for 72 h with or without 1.7ng/ml interleukin (IL)-1beta, 100 ng/ml IL-6 with its soluble receptor (50 ng/ml) or 10 ng/ml oncostatin M (OSM). Aggrecan (AGG) and matrix metalloproteases (MMP)-3 and -13 mRNA levels in chondrocytes were quantified by real-time polymerase chain reaction. AGG production was assayed by a specific enzyme amplified sensitivity immunoassay. RESULTS: SC, but not N, osteoblasts induced a significant inhibition of AGG production and AGG gene expression by human OA chondrocytes in alginate beads, and significantly increased MMP-3 and MMP-13 gene expression by chondrocytes. When they were pre-incubated with IL-1beta, IL-6 or OSM, N osteoblasts inhibited AGG synthesis and increased MMP-3 and -13 gene expression by chondrocytes in alginate beads in a same order of magnitude as SC osteoblasts. CONCLUSIONS: These results demonstrate that SC OA subchondral osteoblasts could contribute to cartilage degradation by stimulating chondrocytes to produce more MMP and also by inhibiting AGG synthesis. [less ▲]

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See detailSubchondral bone osteoblasts induce phenotypic changes in human osteoarthritic chondrocytes
Sanchez, Christelle ULg; Deberg, Michelle ULg; Piccardi, Nathalie et al

in Osteoarthritis and Cartilage (2005), 13(11), 988-997

Objective: To determine the influence of osteoarthritic (OA) phenotype of subchondral osteoblasts on the phenotype of human chondrocytes Methods: Human chondrocytes were isolated from CA cartilage and ... [more ▼]

Objective: To determine the influence of osteoarthritic (OA) phenotype of subchondral osteoblasts on the phenotype of human chondrocytes Methods: Human chondrocytes were isolated from CA cartilage and cultured in alginate beads for 4 or 10 days in the absence or in the presence of osteoblasts in monolayer. The osteoblasts were either isolated from non-sclerotic (N) or sclerotic (SC) zones of human subchondral bone. Before co-culture, osteoblasts were incubated for 72 h with or without 1.7 ng/ml interleukin (IL)-1 beta, 100 ng/ml IL-6 with its soluble receptor (50 ng/ml) or 10 ng/ml oncostatin M. SOX9, type I, II and X collagen (COL1, COL2, COL10), osteoblasts-stimulating factor (OSF)-1, bone alkaline phosphatase (ALP), parathyroid hormone related peptide (PTHrP) and its receptor (PTH-R) messenger RNA (mRNA) levels in chondrocytes were quantified by real-time polymerase chain reaction. Results: In comparison with chondrocytes cultured alone in alginate beads, chondrocytes after 4 days in co-culture with N or SC osteoblasts expressed significantly less SOX9 and COL2 mRNA. The decrease of SOX9 and COL2 gene expression was significantly more pronounced in the presence of SC than in the presence of N osteoblasts (P < 0.001). OSF-1 mRNA level in chondrocyte was increased by both N and SC osteoblasts, but to a larger extent by SC osteoblasts (P < 0.001). PTHrP expression in chondrocytes was 21 -fold increased by N osteoblasts but four-fold inhibited by SC osteoblasts. PTHrP secretion was also increased by N but reduced by SC osteoblasts. SC, but not N osteoblasts, induced a significant decrease of PTH-R gene expression in chondrocyte. In our experimental conditions, chondrocytes did not express COL1, COL10 or ALP, even after 10 days of co-culture with osteoblasts. Conclusions: In co-culture, SC subchondral osteoblasts decrease SOX9, COL2, PTHrP and PTH-R gene expression by chondrocytes but increase that of OSF-1. These findings suggest that SC osteoblasts could initiate chondrocyte phenotype shift towards hypertrophic differentiation and subsequently further matrix mineralization. (c) 2005 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved. [less ▲]

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See detailNew serum biochemical markers (Coll 2-1 and Coll 2-1 NO2) for studying oxidative-related type II collagen network degradation in patients with osteoarthritis and rheumatoid arthritis
Deberg, Michelle ULg; Labasse, Alain ULg; Christgau, S. et al

in Osteoarthritis and Cartilage (2005), 13(3), 258-265

Objective: Protein nitration is a prominent feature of inflammatory processes in the joint. We have developed immunoassays specific for a peptide of the alpha-helical region of type II collagen (108 ... [more ▼]

Objective: Protein nitration is a prominent feature of inflammatory processes in the joint. We have developed immunoassays specific for a peptide of the alpha-helical region of type II collagen (108)HRGYPGLDG(116) (Coll 2-1) and its nitrated form (108)HRGY(NO2)PGLDG(116) (Coll 2-1 NO2) in biological fluids. Design: Coll 2-1 and Coll 2-1 NO2 peptides were injected into rabbits. Two antisera (D3 and D37) were selected for their specificity and affinity and used to develop specific immunoassays. Coll 2-1 and Coll 2-1 NO2 were measured in sera of 242 healthy subjects (N), 67 patients with primary knee osteoarthritis (OA) and 19 patients with rheumatoid arthritis (RA). Results: In healthy subjects, Coll 2-1 and Coll 2-1 NO2 concentrations were 125.13 +/- 3.71 nM and 0.16 +/- 0.08 nM, respectively. In OA and RA, Coll 2-1 and Coll 2-1 NO2 serum levels were found to be significantly increased compared to controls of the same range of age (Coll 2-1: OA: 200.80 +/- 8.98 nM, RA: 172.30 +/- 19.05 nM, normal: 126.60 +/- 6.70 nM and Coll 2-1 NO2: OA: 0.26 +/- 0.02, RA: 0.38 +/- 0.05, normal: 0.12 +/- 0.01 nM). Coll 2-1 NO2 levels were significantly more elevated in RA than in OA patients (P < 0.05). As a consequence, the ratio Coll 2-1 NO2/Coll 2-1 was 1.6 times higher in RA than in OA subjects. No relationship was found between the radiological OA severity and the levels of Coll 2-1 and Coll 2-1 NO2 in serum. Coll 2-1 NO2, but not Coll 2-1, was correlated with C-reactive protein in the sera of OA and RA patients. Conclusions: The determination of both Coll 2-1 and Coll 2-1 NO2 in serum of arthritic patients seems to be a promising useful tool for the detection of oxidative-related cartilage degradation episode. Further, these markers could be helpful for monitoring the effects of anti-inflammatory or antioxidant drugs on cartilage degradation. (c) 2004 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved. [less ▲]

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See detailSubchondral Bone Osteoblasts Induce Phenotypic Changes in Human Osteoarthritic Chondrocytes
Sanchez, Christelle ULg; Deberg, Michelle ULg; Piccardi, Nathalie et al

in Osteoporosis International (2005), 16(Suppl.3), 55-56

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See detailAvocado/soybean unsaponifiables prevent osteoarthritic subchondral osteoblasts-induced cartilage degradation
Sanchez, Christelle ULg; Deberg, Michelle ULg; Piccardi, Nathalie et al

in Osteoporosis International (2005), 16(Suppl.3), 56

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See detailInterleukin-6 mediates subchondral osteoblasts-induced cartilage degradation
Henrotin, Yves ULg; Kaut, Elisabeth; Deberg, Michelle ULg et al

Poster (2005)

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See detailInterleukin-6 mediates subchondral osteoblasts-induced cartilage degradation
Sanchez, Christelle ULg; Kaut, Elisabeth; Deberg, Michelle ULg et al

in Osteoarthritis and Cartilage (2005), 13(Suppl A), 157

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See detailEtude in vitro de l’axe physiopathologique os-cartilage dans l’arthrose : implication de l’interleukin-6
Henrotin, Yves ULg; Kaut, Elisabeth; Deberg, Michelle ULg et al

in Revue du Rhumatisme (2005), 72(10-11), 958-986

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See detailInterleukin-1, Interleukin-6 and Oncostatin M Stimulate Normal Subchondral Osteoblasts to Induce Cartilage Degradation
Sanchez, Christelle ULg; Deberg, Michelle ULg; Piccardi, Nathalie et al

in Osteoporosis International (2005), 16(Suppl 3), 54-55

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See detailOne-year increase of Coll 2-1, a new marker of type II collagen degradation, in urine is highly predictive of radiological OA progression.
Deberg, Michelle ULg; Labasse, Alain ULg; Collette, Julien ULg et al

in Osteoarthritis and Cartilage (2005), 13(12), 1059-65

OBJECTIVE: To analyse the relationship between the levels of urinary biochemical markers of type II collagen degradation and the clinical and radiological severity and progression of knee osteoarthritis ... [more ▼]

OBJECTIVE: To analyse the relationship between the levels of urinary biochemical markers of type II collagen degradation and the clinical and radiological severity and progression of knee osteoarthritis (OA). METHOD: Seventy-five patients with primary knee OA were included in this 3-year follow-up study. Mean joint space width (JSW) of the medial compartment of the femorotibial joint was measured with a computer assisted method on standardized radiographs taken at baseline and after a 3-year follow-up. Pain, stiffness, and physical function subscales of the Western Ontario and McMaster Universities (WOMAC) were assessed at the same time points. Type II collagen peptides Coll 2-1 and Coll 2-1 NO(2), as well as pyridinoline (Pyr) and deoxypyridinoline (D-Pyr) were measured in urines at baseline, after 1 year and 3 years, with specific immunoassays. RESULTS: At baseline, significant correlations were found between the urinary Coll 2-1 and Coll 2-1 NO(2) levels and the global WOMAC score (Coll 2-1: r=0.28, P=0.01; Coll 2-1 NO(2): r=0.27, P=0.02) and its subscales for pain (Coll 2-1: r=0.27, P=0.01; Coll 2-1 NO(2): r=0.30, P=0.01) and function (Coll 2-1: r=0.29, P=0.01; Coll 2-1 NO(2): r=0.27, P=0.02). Pyr and D-Pyr levels were not significantly correlated with the WOMAC scores. One-year change in Coll 2-1 and Coll 2-1 NO(2) urinary levels were negatively correlated with a 3-year change in JSW (Coll 2-1: r=-0.31, P=0.03; Coll 2-1 NO(2): r=-0.31, P=0.03), indicating that an increase of Coll 2-1 or Coll 2-1 NO(2) over 1 year is predictive of subsequent joint space narrowing. Neither Pyr nor D-Pyr was correlated with radiological OA progression. CONCLUSIONS: At baseline, Coll 2-1 and Coll 2-1 NO(2) urinary levels were indicative of the clinical activity of knee OA and the increase of these peptides over 1 year was predictive of the radiological progression of knee OA. [less ▲]

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See detailDifferential Regulation of Chondrocyte Metabolism by Oncostatin M and Interleukin-6
Sanchez, Christelle ULg; Deberg, Michelle ULg; Devel, Philippe et al

in Osteoarthritis and Cartilage (2004), 12(10), 801-10

OBJECTIVE: To determine the effects of interleukin (IL)-6 and oncostatin M (OSM) added separately or in combination with IL-1beta on human osteoarthritic (OA) chondrocytes in alginate beads. DESIGN: Human ... [more ▼]

OBJECTIVE: To determine the effects of interleukin (IL)-6 and oncostatin M (OSM) added separately or in combination with IL-1beta on human osteoarthritic (OA) chondrocytes in alginate beads. DESIGN: Human chondrocytes were isolated from OA cartilage and cultured in alginate beads for 12 days, in the absence or in the presence of increasing amounts of IL-6 (20-500ng/ml) with its soluble receptor or OSM (0.1-10ng/ml) and with or without IL-1beta (1.7ng/ml). Aggrecan (AGG), transforming growth factor-beta1 (TGF-beta1), stromelysin-1 [matrix metalloprotease (MMP)-3], tissue inhibitor of metalloproteinases-1 (TIMP-1), macrophage inflammatory protein-1 beta (MIP-1beta), IL-6 and IL-8 productions were assayed by specific enzyme amplified sensitivity immunoassays. Prostaglandin (PG)E(2) was measured by a specific radioimmunoassay and nitrite (NO(2)(-)) by a spectrophotometric method based upon the Griess reaction. RESULTS: OSM, but not IL-6, decreased basal AGG and TGF-beta1 synthesis. Although IL-6 stimulated basal TIMP-1 production, it did not significantly modify MMP-3/TIMP-1 ratio. In contrast, 10ng/ml OSM highly increased TIMP-1 production, and decreased by half the ratio MMP-3/TIMP-1. IL-1beta highly stimulated *NO, IL-8, IL-6, MIP-1beta and PGE(2) synthesis but decreased AGG and TGF-beta1 production. Neither IL-6 nor OSM modulated IL-1beta-inhibitory effect on AGG production. IL-6, but not OSM, reversed IL-1beta-induced TGF-beta1 inhibition. At 1-10ng/ml, OSM significantly decreased IL-1beta-stimulated IL-8, MIP-1beta, PGE(2) and *NO production but amplified IL-1beta stimulating effect on IL-6 production. IL-6 had no effect on these parameters. CONCLUSIONS: OSM and IL-6, two glycoprotein 130 binding cytokines, show different activity profiles on OA chondrocytes, indicating that these cytokines could play different roles in the OA disease process. [less ▲]

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See detailType II collagen peptides for measuring cartilage degradation
Henrotin, Yves ULg; Deberg, Michelle ULg; Dubuc, J. E. et al

in Biorheology (2004), 41(3-4), 543-547

This paper describes two new immunoassays for a peptide of the triple helix of type II collagen (Coll 2-1) and its nitrated form (Coll 2-1 NO2). In healthy subjects aged between 20 and 65 years old, Coll ... [more ▼]

This paper describes two new immunoassays for a peptide of the triple helix of type II collagen (Coll 2-1) and its nitrated form (Coll 2-1 NO2). In healthy subjects aged between 20 and 65 years old, Coll 2-1 and Coll 2-1 NO2 levels in serum were in means 125.13+/-3.71 and 0.16+/-0.08 nmol/l, respectively. These levels did not significantly vary with age. However, up to 45 years of age, Coll 2-1 NO2 levels in women were significantly higher than in men. In patients with knee osteoarthritis (OA), Coll 2-1 in serum was found to be elevated compared to healthy controls (267.45+/-26.42 nmol/l vs 126.78+/-6.61 nmol/l). Further, we have demonstrated that an increase of the urinary levels of Coll 2-1 or Coll 2-1 NO2 over 1 year was predictive of joint space narrowing progression in OA patients. In conclusion, these preliminary results indicate that Coll 2-1 could be a predictive marker of knee OA progression. [less ▲]

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