Alumina on alumina versus metal on conventional polyethylene: a randomized clinical trial with 9 to 15 years follow-up.

We report the long-term results of a randomized clinical trial that compares, in total hip arthroplasty in a young population, metal-on-conventional polyethylene and alumina-on-alumina ceramic bearings. One hundred and forty hips in 116 patients were randomized. Re-operation, revision rate, clinical scores, and radiological signs of osteolysis and loosening were compared at average follow-up of 123 (9-15) years. At final FU, 107 hips were available for clinical evaluation. Eight (11.6%) revisions were performed in the polyethylene group versus 1 (1.4%) in the ceramic group (p = 0.017). All revisions in the polyethylene group were related to bearing wear: 4 for aseptic loosening with severe osteolysis, 1 for polyethylene induced compressive granulomatous tumor, and 3 for severe liner wear. The only revised case from the ceramic group was secondary to mechanical stem loosening. Mean annual polyethylene wear was 0.19mm/year; wear was not measurable in the ceramic group. Our study confirms, in the long-term, the superiority of ceramic-on-ceramic pairing in comparison to metal-on-conventional polyethylene and supports their use in young, active patients.

Estrogen receptor-related receptor α regulation by interleukin-1ß in prostaglandin E(2)- and cAMP-dependent pathways in osteoarthritic chondrocytes.

OBJECTIVE: We reported previously that the orphan nuclear receptor, estrogen receptor-related receptor α (ERRα), is expressed in articular chondrocytes and is dysregulated in a mouse model of inflammatory arthritis. The aim of this study, therefore, was to determine whether ERRα is also dysregulated in patients with osteoarthritis (OA). METHODS: ERRα messenger RNA (mRNA) and protein were quantified in normal and OA cartilage samples and in OA chondrocytes in vitro, with and without short-term treatment with a variety of OA-associated factors and signaling pathway agonists and inhibitors. RESULTS: ERRα expression was lower in OA than in normal articular cartilage. Interleukin-1ß (IL-1ß) markedly up-regulated ERRα expression in OA chondrocytes in vitro, and agonist or inhibitor treatment indicated that the up-regulation was dependent on cyclooxygenase 2 (COX-2; NS398), prostaglandin E(2), cAMP (8-bromo-cAMP), and protein kinase A (PKA; KT5720). Treatment with the ERRα inverse agonist XCT790 decreased the expression of SOX9 and the up-regulation of ERRα by IL-1ß, suggesting autoregulation of ERRα in the IL-1ß pathway. Matrix metalloproteinase 13 (MMP-13) expression was also decreased by treatment with XCT790 plus IL-1ß versus IL-1ß alone, and the down-regulation of MMP-13 mRNA and protein observed with XCT790 alone suggests that the up-regulation of MMP-13 by IL-1ß is ERRα-dependent. CONCLUSION: We report the first evidence that ERRα expression is regulated by IL-1ß in COX-2-, cAMP-, and PKA-dependent pathways in OA chondrocytes. We confirmed that SOX9 is an ERRα target gene in human, as in rodent, chondrocytes and identified MMP-13 as a potential new target gene, which suggests that ERRα may both respond to the healing signal and contribute to extracellular degradation in OA cartilage.

Contribution of H3K4 methylation by SET-1A to interleukin-1-induced cyclooxygenase 2 and inducible nitric oxide synthase expression in human osteoarthritis chondrocytes.

OBJECTIVE: To investigate the role of histone H3 lysine 4 (H3K4) methylation in interleukin-1ß (IL-1ß)-induced cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) expression in human osteoarthritic (OA) chondrocytes. METHODS: Chondrocytes were stimulated with IL-1, and the expression of iNOS and COX-2 messenger RNA and proteins was evaluated by real-time reverse transcriptase-polymerase chain reaction analysis and Western blotting, respectively. H3K4 methylation and the recruitment of the histone methyltransferases SET-1A and MLL-1 to the iNOS and COX-2 promoters were evaluated using chromatin immunoprecipitation assays. The role of SET-1A was further evaluated using the methyltransferase inhibitor 5'-deoxy-5'-(methylthio)adenosine (MTA) and gene silencing experiments. SET-1A level in cartilage was determined using immunohistochemistry. RESULTS: The induction of iNOS and COX-2 expression by IL-1 was associated with H3K4 di- and trimethylation at the iNOS and COX-2 promoters. These changes were temporally correlated with the recruitment of the histone methyltransferase SET-1A, suggesting an implication of SET-1A in these modifications. Treatment with MTA inhibited IL-1-induced H3K4 methylation as well as IL-1-induced iNOS and COX-2 expression. Similarly, SET-1A gene silencing with small interfering RNA prevented IL-1-induced H3K4 methylation at the iNOS and COX-2 promoters as well as iNOS and COX-2 expression. Finally, we showed that the level of SET-1A expression was elevated in OA cartilage as compared with normal cartilage. CONCLUSION: These results indicate that H3K4 methylation by SET-1A contributes to IL-1-induced iNOS and COX-2 expression and suggest that this pathway could be a potential target for pharmacologic intervention in the treatment of OA and possibly other arthritic diseases.

Ceramic-on-ceramic total hip arthroplasty is superior to metal-on-conventional polyethylene at 20-year follow-up: A randomised clinical trial.

BACKGROUND: Metal-on-conventional polyethylene (MoPc) bearing wear-related biological reactions in total hip arthroplasty (THA) continue to raise concerns among young, active patients. Ceramic-on-ceramic (CoC) bearings may offer improved outcomes in this patient population. QUESTIONS/PURPOSES: The aim of this study was to determine if, more than 20years postoperatively, there is a difference between MoPc and CoC THA in terms of (1) survivorship, (2) related complications, (3) radiographic signs of wear, and (4) functional scores. HYPOTHESIS: CoC bearing THAs have superior clinical results compared to MoPc THAs. PATIENTS AND METHODS: A total of 140 hips in 116 patients with a mean age of 42years were randomised to receive CoC or MoPc THA between 1996 and 2001. Sixty-nine hips in 58 patients received MoP and 71 hips in 68 patients received CoC. Revision rate, WOMAC score, and radiological signs of osteolysis and loosening were compared at last follow-up. RESULTS: After a mean follow-up of 21years (19-23), 40 patients (48 THAs; 34%) had died and 6 patients (6 THAs; 4%) were lost to follow-up. Aseptic revision rate was significantly higher in the MoPc group (17/69; 24.6%) versus CoC (2/71; 2.8%; p<0.001). Kaplan-Meier survivorship estimator with revision for aseptic reasons was 73.6% (95% CI: 63.3-84.9%) for MoPc and 96.9% (95% CI: 92.8-100%) for CoC (p<0.001). On radiographic evaluation, 13% (3/23) MoPc were considered loose versus no CoC, and 61% (14/23) MoPc versus 6% (2/33) CoC showed osteolytic signs (p<0.001). CoC had better mean WOMAC scores than MoPc (11.0 vs. 19.4; p=0.048). No ceramic fracture was observed. CONCLUSION: In this RCT, CoC bearings provided excellent results and were safer than MoPc bearings at more than 20-year follow-up. The long-term in vivo behaviour of CoC bearing makes it a great THA option for middle-aged patients and should be compared to newer polyethylene bearings. LEVEL OF EVIDENCE: I.

Modulation of OPG, RANK and RANKL by human chondrocytes and their implication during osteoarthritis.

OBJECTIVES: Earlier studies suggest the involvement of osteoprotegerin (OPG), RANK and RANK ligand (RANKL) in OA subchondral bone metabolism; however, few studies have looked at their functional consequences on chondrocytes. We compared the expression/production of OPG, RANK and RANKL on human normal and OA chondrocytes, and evaluated, on OA chondrocytes, their modulation by some catabolic factors. Furthermore, the role of OPG and RANKL on the production of catabolic/anabolic factors was assessed. METHODS: Expression was determined using real-time PCR, production of RANK and RANKL by flow cytometry and that of OPG by ELISA. Modulation of these factors was determined upon treatment with IL-1beta, TNF-alpha and PGE(2). The functional consequences were examined following treatment with soluble RANKL or OPG-Fc (OPG without the heparin-binding domain). RESULTS: OPG, RANK and RANKL were expressed and produced by human chondrocytes. Membranous RANK was produced only by an OA chondrocyte subpopulation (29%) localized throughout the cartilage. The OPG/RANKL ratio was significantly (P = 0.05) reduced on the OA chondrocytes, whereas the RANK/RANKL ratio was significantly (P < 0.03) increased. OPG and membranous RANKL levels were significantly enhanced by IL-1beta, TNF-alpha and PGE(2), whereas membranous RANK was significantly increased only with IL-1beta. Administration of soluble RANKL had no effect on the OA chondrocytes. However, addition of OPG-Fc significantly stimulated MMP-13 (P = 0.05) and protease-activated receptor-2 (PAR-2) (P < 0.04) production. CONCLUSIONS: Our findings showed that human chondrocytes express and produce OPG, RANK and RANKL. OA chondrocyte treatment with catabolic factors pointed towards an increased biological effect of OPG. Interestingly, OPG appears to be involved in OA progression by increasing two catabolic factors involved in cartilage pathophysiology.

Regulation of the IGFBP-5 and MMP-13 genes by the microRNAs miR-140 and miR-27a in human osteoarthritic chondrocytes.

BACKGROUND: MMP-13 and IGFBP-5 are important factors involved in osteoarthritis (OA). We investigated whether two highly predicted microRNAs (miRNAs), miR-140 and miR-27a, regulate these two genes in human OA chondrocytes. METHODS: Gene expression was determined by real-time PCR. The effect of each miRNA on IGFBP-5 and MMP-13 expression/production was evaluated by transiently transfecting their precursors (pre-miRNAs) and inhibitors (anti-miRNAs) into human OA chondrocytes. Modulation of IGFBP-5, miR-140 and miR-27a expression was determined upon treatment of OA chondrocytes with cytokines and growth factors. RESULTS: IGFBP-5 was expressed in human chondrocytes with its level significantly lower (p < 0.04) in OA. Five computational algorithms identified miR-140 and miR-27a as possible regulators of MMP-13 and IGFBP-5 expression. Data showed that both miRNAs were expressed in chondrocytes. There was a significant reduction (77%, p < 0.01) in miR-140 expression in OA compared to the normal chondrocytes, whereas miR-27a expression was only slightly decreased (23%). Transfection with pre-miR-140 significantly decreased (p = 0.0002) and with anti-miR-140 significantly increased (p = 0.05) IGFBP-5 expression at 24 hours, while pre-miR-27a did not affect either MMP-13 or IGFBP-5. Treatment with anti-miR-27a, but not with anti-miR-140, significantly increased the expression of both MMP-13 (p < 0.05) and IGFBP-5 (p < 0.01) after 72 hours of incubation. MMP-13 and IGFBP-5 protein production followed the same pattern as their expression profile. These data suggest that IGFBP-5 is a direct target of miR-140, whereas miR-27a down-regulates, likely indirectly, both MMP-13 and IGFBP-5. CONCLUSION: This study is the first to show the regulation of these miRNAs in human OA chondrocytes. Their effect on two genes involved in OA pathophysiology adds another level of complexity to gene regulation, which could open up novel avenues in OA therapeutic strategies.

Differential modulation of RANKL isoforms by human osteoarthritic subchondral bone osteoblasts: influence of osteotropic factors.

BACKGROUND: Osteoarthritis (OA) is the most common human joint disease. Recent studies suggest that an abnormal subchondral bone metabolism is intimately involved in the genesis of this disease. Bone remodelling is tightly regulated by a molecular triad composed of OPG/RANK/RANKL. RANKL exists as 3 isoforms: RANKL1, 2, and 3. RANKL1 and 2 enhance osteoclastogenesis whereas RANKL3 inhibits this phenomenon. We previously reported that human OA subchondral bone osteoblasts can be discriminated into two subgroups according to their level of PGE2 [low (L) or high (H)]. Moreover, we also showed that L-OA osteoblasts express higher levels of total RANKL compared to H-OA osteoblasts. In this study, we investigated the level of membranous RANKL, comparing L- and H-OA subchondral bone osteoblasts, as well as its modulation by osteotropic factors. The impact of the modulation of RANKL1 and 3 on the membranous RANKL level was also studied. METHODS: Gene expression was determined using real-time PCR for RANKL1 and semi-quantitative PCR for RANKL3. Membranous RANKL was measured by flow cytometry. The modulation of membranous RANKL and RANKL isoforms was monitored on the L- and H-OA osteoblasts and also following treatment with osteotropic factors, including vitamin D3 (50 nM), IL-1beta (100 pg/ml), TNF-alpha (5 ng/ml), PGE2 (500 nM), PTH (100 nM), IL-6 (10 ng/ml) and IL-17 (10 ng/ml). RESULTS: Membranous RANKL levels were significantly increased in L-OA osteoblasts compared to normal (p<0.01) and H-OA (p<0.05). The gene expression level of the RANKL1 profile was reminiscent of the membranous RANKL level. Although RANKL3 gene expression was lower on the H-OA osteoblasts than on normal and L-OA osteoblasts (p<0.03), the overall outcome favoured RANKL1. Treatment with the tested factors showed a significant increase in membranous RANKL on the L-OA osteoblasts, with the exception of PTH and IL-17. Interestingly in this subpopulation, the RANKL3 gene expression level was significantly increased upon PTH and IL-17 treatment. No effect of the tested osteotropic factors was found on the H-OA. CONCLUSION: Our findings showed that the normal, L- and H-OA subchondral bone osteoblasts differentially express membranous RANKL and RANKL isoforms, and that treatment with osteotropic factors generally favours increased membranous localization of RANKL on L-OA compared to H-OA osteoblasts. This phenomenon appears to take place through differential modulation of each RANKL isoform.

Comparison of alumina-alumina to metal-polyethylene bearing surfaces in THA: a randomized study with 4- to 9-years follow-up.

We report the clinical and radiological results of 140 primary THAs, randomized to receive metal-polyethylene or alumina-alumina bearing surfaces. At last follow-up (average 79 months), no significant difference was found on clinical scores (WOMAC and Merle D'Aubigné) between the two groups. However, linear wear of 1 mm or more of the liner was observed in 89% (50/56) of polyethylene cases, whereas no measurable wear was noted in the alumima-on-alumina group. Calcar resorption was noted in 34% (19/56) of cases in the polyethylene group versus 6% (3/51) in the alumina group. Although no aseptic loosening was present in either group, 2 hips in the polyethylene group underwent revision for severe liner wear, and 2 more are pending. No specific complication was associated with alumina components. This study is in line with other reports indicating that alumina-alumina bearing surfaces have better wear properties than metal-on standard polyethylene and should be considered for THA in young and active patients.

Gesture standardization increases the reproducibility of 3D kinematic measurements of the knee joint.

BACKGROUND: The literature on the 3D kinematics of the knee suggests that the gesture accomplished during kinematic assessment might play a significant role in the values measured. The purpose of this study is to demonstrate that a standardized gesture leads to an increased reproducibility in 3D kinematic measurements of the knee. METHODS: Seventeen healthy male subjects performed series of knee-bends in standardized and unconstrained conditions while their left knee's 3D kinematics were recorded using an optical motion-recording system. Standardized knee-bends were performed in a specially designed structure stabilizing the shoulders, pelvis and feet. Coefficient of multiple correlation were calculated for the ascent and the descent phases of the knee-bends for the tibial rotation and abduction/adduction components of the knee movement. FINDINGS: Comparisons between coefficient of multiple correlation of the different gesture conditions showed a statistically significant increase in reproducibility for the tibial rotation during the standardized knee-bends. INTERPRETATION: It appears that gesture standardization is an interesting option to consider for precise kinematic assessment of the living human knee.

A reproducible method for studying three-dimensional knee kinematics.

The methods used in movement analysis often rely on the definition of joint coordinate systems permitting three-dimensional (3D) kinematics. The first aim of this research project was to present a functional and postural method (FP method) to define a bone-embedded anatomical frame (BAF) on the femur and tibia, and, subsequently, a knee joint coordinate system. The repeatability of the proposed method was also assessed. Using FP method to define the BAFs, 4 kinematic parameters (flexion/extension, abduction/adduction, tibial internal/external rotation, and antero-posterior translation) were computed for 15 subjects walking on a treadmill. The repeatability for all four kinematic parameters was then assessed, using intra- and inter-observer settings. After pooling the results for all observers, the mean repeatability value ranged between 0.4 degrees and 0.8 degrees for rotation angles and between 0.8 and 2.2 mm for translation.

Comparison of 2 surgical techniques of posterolateral corner reconstruction of the knee.

BACKGROUND: Various surgical techniques to treat posterolateral knee instability have been described. To date, the recommended treatment is an anatomical form of reconstruction, in which the 3 key structures of the posterolateral corner are addressed: the lateral collateral ligament, the popliteofibular ligament, and the popliteus tendon. HYPOTHESIS: Two methods of surgical reconstruction will restore posterolateral knee instability, in terms of static laxity as well as dynamic 6 degrees of freedom kinematics, to statistically significant levels compared with the intact state. STUDY DESIGN: Controlled laboratory study. METHODS: Two surgical techniques (A and B) were used to reconstruct the posterolateral structures in 10 cadaveric knees. Static tests were performed on the intact, sectioned, and reconstructed knees at 30 degrees and 90 degrees of flexion for anterior-posterior laxity and external rotational laxity, as well as at 0 degrees and 30 degrees of flexion for varus laxity; dynamic 6 degrees of freedom kinematic testing, through a path of motion from 90 degrees of flexion to full extension, was also performed. RESULTS: For the static varus tests, external rotation and varus laxity were significantly increased after the posterolateral structures were cut. Both reconstruction techniques restored external rotation and varus laxity to levels not significantly different from the intact state. For technique B, dynamic testing did not show any significant difference for all degrees of freedom kinematics compared with the intact state. However, for technique A, a significant internal tibial rotation was observed throughout the entire path of motion from 0 degrees to 90 degrees of knee flexion. CONCLUSIONS: Both surgical techniques for anatomical posterolateral corner reconstruction showed good results in the static laxity tests. The anatomical reconstruction of all structures, including the popliteus tendon, resulted in an abnormal internal tibial rotation during dynamic testing.

Quantitative comparison of three different types of anterior cruciate ligament reconstruction methods: laxity and 3-D kinematic measurements.

In this project, we compared knee laxity and 3-D knee kinematics after ACL reconstruction on cadaver knees using (1) bone-patellar tendon-bone two-tunnel; (2) synthetic ligament two-tunnel; and (3) synthetic ligament "over-the-top" technique. We used a computer assisted system, based on the acquisition of the knee's movement with magnetic sensors (Polhemus, Vermont, USA). The use of personalised three-dimensional (3D) models of the bones enabled us to ensure a reproducible measurement of three-dimensional kinematic and laxity parameters. Our results showed that even when knee laxity was restored to normal, 3D kinematic measurements revealed that the reconstructions tended to under or over-constrain the knee's movement. This study shows that 3D kinematics is a complementary measurement that can be useful to get a better comprehension of the knee's function after ligament reconstruction.

Egr-1 contributes to IL-1-mediated down-regulation of peroxisome proliferator-activated receptor γ expression in human osteoarthritic chondrocytes.

INTRODUCTION: Peroxisome proliferator-activated receptor (PPAR)γ has been shown to exhibit anti-inflammatory and anti-catabolic properties and to be protective in animal models of osteoarthritis (OA). We have previously shown that interleukin-1ß (IL-1) down-regulates PPARγ expression in human OA chondrocytes. However, the mechanisms underlying this effect have not been well characterized. The PPARγ promoter harbors an overlapping Egr-1/specificity protein 1 (Sp1) binding site. In this study, our objective was to define the roles of Egr-1 and Sp1 in IL-1-mediated down-regulation of PPARγ expression. METHODS: Chondrocytes were stimulated with IL-1 and the expression levels of Egr-1 and Sp1 mRNAs and proteins were evaluated using real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting, respectively. The role of de novo protein synthesis was evaluated using the protein synthesis inhibitor cycloheximide (CHX). The recruitment of Sp1 and Egr-1 to the PPARγ promoter was evaluated using chromatin immunoprecipitation (ChIP) assays. The PPARγ promoter activity was analyzed in transient transfection experiments. The roles of Egr-1 and Sp1 were further evaluated using small interfering RNA (siRNA) approaches. The level of Egr-1 in cartilage was determined using immunohistochemistry. RESULTS: Down-regulation of PPARγ expression by IL-1 requires de novo protein synthesis and was concomitant with the induction of the transcription factor Egr-1. Treatment with IL-1 induced Egr-1 recruitment and reduced Sp1 occupancy at the PPARγ promoter. Overexpression of Egr-1 potentiated, whereas overexpression of Sp1 alleviated, the suppressive effect of IL-1 on the PPARγ promoter, suggesting that Egr-1 may mediate the suppressive effect of IL-1. Consistently, Egr-1 silencing prevented IL-1-mediated down-regulation of PPARγ expression. We also showed that the level of Egr-1 expression was elevated in OA cartilage compared to normal cartilage. CONCLUSIONS: Our results indicate that induction and recruitment of Egr-1 contributed to the suppressive effect of IL-1 on PPARγ expression. They also suggest that modulation of Egr-1 levels in the joint may have therapeutic potential in OA.

Elevated Dickkopf-2 levels contribute to the abnormal phenotype of human osteoarthritic osteoblasts.

The Wnt signaling pathway is crucial for osteogenesis and regulates terminal osteoblast differentiation. Although osteoarthritic (OA) osteoblasts show an abnormal phenotype and poor in vitro mineralization, the mechanism leading to this situation still remains unknow. Recent evidence indicates that Wnt signaling may be altered in OA osteoblasts. In this study we determined whether an alteration of the Wnt/ß-catenin signaling pathway is responsible for the abnormal phenotype of OA osteoblasts. Expression of the Wnt signaling antagonist Dickkopf-1 (DKK1) was similar in normal and OA osteoblasts, whereas DKK2 expression was higher in OA osteoblasts than in normal osteoblasts. OA osteoblasts showed a decrease of Wnt3a-dependent Wnt/ß-catenin signaling, measured by the TOPflash reporter assay and by Western blot analysis, compared with normal osteoblasts. Correcting DKK2 levels in OA osteoblasts by siRNA techniques enhanced Wnt/ß-catenin signaling. Elevated DKK2 levels could be explained by elevated transforming growth factor ß1 (TGF-ß1) in OA osteoblasts, and exogenous TGF-ß1 increased DKK2 expression in normal osteoblasts, whereas ablating TGF-ß1 expression in OA osteoblasts reduced DKK2 expression. Inhibiting TGF-ß1 or DKK2 expression corrected the abnormal phenotype of OA osteoblasts. In vitro mineralization of OA osteoblasts also was increased by DKK2 siRNA. We conclude that elevated TGF-ß1 levels in OA osteoblasts can stimulate DKK2 expression, which, in turn, is responsible, at least in part, for their abnormal phenotype.

Valproic acid suppresses interleukin-1ß-induced microsomal prostaglandin E2 synthase-1 expression in chondrocytes through upregulation of NAB1.

OBJECTIVE: Microsomal prostaglandin E(2) synthase-1 (mPGES-1) catalyzes the terminal step in the biosynthesis of PGE(2). Early growth response factor-1 (Egr-1) is a key transcription factor in the regulation of mPGES-1, and its activity is negatively regulated by the corepressor NGF1-A-binding protein-1 (NAB1). We examined the effects of valproic acid (VA), a histone deacetylase inhibitor, on interleukin 1ß (IL-1ß)-induced mPGES-1 expression in human chondrocytes, and evaluated the roles of Egr-1 and NAB1 in these effects. METHODS: Chondrocytes were stimulated with IL-1 in the absence or presence of VA, and the level of mPGES-1 protein and mRNA expression were evaluated using Western blotting and real-time reverse-transcription polymerase chain reaction (PCR), respectively. mPGES-1 promoter activity was analyzed in transient transfection experiments. Egr-1 and NAB1 recruitment to the mPGES-1 promoter was evaluated using chromatin immunoprecipitation assays. Small interfering RNA (siRNA) approaches were used to silence NAB1 expression. RESULTS: VA dose-dependently suppressed IL-1-induced mPGES-1 protein and mRNA expression as well as its promoter activation. Treatment with VA did not alter IL-1-induced Egr-1 expression, or its recruitment to the mPGES-1 promoter, but prevented its transcriptional activity. The suppressive effect of VA requires de novo protein synthesis. VA induced the expression of NAB1, and its recruitment to the mPGES-1 promoter, suggesting that NAB1 may mediate the suppressive effect of VA. Indeed, NAB1 silencing with siRNA blocked VA-mediated suppression of IL-1-induced mPGES-1 expression. CONCLUSION: VA inhibited IL-1-induced mPGES-1 expression in chondrocytes. The suppressive effect of VA was not due to reduced expression or recruitment of Egr-1 to the mPGES-1 promoter and involved upregulation of NAB1.

Variable effects of 3 different chondroitin sulfate compounds on human osteoarthritic cartilage/chondrocytes: relevance of purity and production process.

OBJECTIVE: During osteoarthritis (OA), the altered metabolism of cartilage involves proinflammatory factors and matrix metalloprotease (MMP) activity. Studies showed that chondroitin sulfate (CS) may exert a positive effect on the cartilage. Because of differences in CS in terms of purity and the production/purification process, we compared the effects of 3 different types of CS on human OA cartilage. METHODS: Three types of CS were tested: CS1 (porcine, purity 90.4%), CS2 (bovine, purity 96.2%), and CS3 (bovine, purity 99.9%). Treatment with CS at 200 and 1000 microg/ml was performed on human OA cartilage explants in the presence/absence of interleukin 1ss (IL-1ss), and the protein modulations of factors including prostaglandin E(2) (PGE(2)), IL-6, and MMP-1 measured by ELISA. The CS effect on the expression of collagen type II was also investigated on OA chondrocytes using quantitative polymerase chain reaction. RESULTS: In the presence of IL-1ss, CS2 at 1000 microg/ml significantly inhibited IL-6 and PGE(2) production, and CS3 at 200 microg/ml markedly reduced the level of IL-6. CS1 was much less efficient at reducing the catabolic markers and in the absence of IL-1ss, it significantly increased IL-6 and MMP-1. IL-1ss significantly inhibited the gene expression level of collagen type II; only CS3 was able to limit this inhibition. CS1, in the presence or absence of IL-1ss, further markedly decreased collagen type II expression. CONCLUSION: Our data indicate that among the 3 tested CS, CS1 increased production of some catabolic pathways and inhibited the gene expression level of collagen type II. Our study provides new information in the context of prescribing CS for alleviating OA symptoms, as the purity and/or production/purification of the CS compound could orient the current OA disease process toward increased catabolic pathways.

Human articular chondrocytes express 15-lipoxygenase-1 and -2: potential role in osteoarthritis.

INTRODUCTION: 15-Lipoxygenases and their metabolites have been shown to exhibit anti-inflammatory and immunomodulatory properties, but little is known regarding their expression and function in chondrocytes. The objective of this study was to evaluate the expression of 15-lipoxygenase-1 and -2 in human articular chondrocytes, and to investigate the effects of their metabolites 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids on IL-1beta-induced matrix metalloproteinase (MMP)-1 and MMP-13 expression. METHODS: The expression levels of 15-lipoxygenase-1 and -2 were analyzed by reverse transcription PCR and Western blotting in chondrocytes, and by immunohistochemistry in cartilage. Chondrocytes or cartilage explants were stimulated with IL-1beta in the absence or presence of 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids, and the levels of MMP-1 and MMP-13 protein production and type II collagen cleavage were evaluated using immunoassays. The role of peroxisome proliferator-activated receptor (PPAR)gamma was evaluated using transient transfection experiments and the PPARgamma antagonist GW9662. RESULTS: Articular chondrocytes express 15-lipoxygenase-1 and -2 at the mRNA and protein levels. 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids dose dependently decreased IL-1beta-induced MMP-1 and MMP-13 protein and mRNA expression as well as type II collagen cleavage. The effect on MMP-1 and MMP-13 expression does not require de novo protein synthesis. 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids activated endogenous PPARgamma, and GW9662 prevented their suppressive effect on MMP-1 and MMP-13 production, suggesting the involvement of PPARgamma in these effects. CONCLUSIONS: This study is the first to demonstrate the expression of 15-lipoxygenase-1 and -2 in articular chondrocytes. Their respective metabolites, namely 13(S)-hydroxy octadecadienoic and 15(S)-hydroxyeicosatetraenoic acids, suppressed IL-1beta-induced MMP-1 and MMP-13 expression in a PPARgamma-dependent pathway. These data suggest that 15-lipoxygenases may have chondroprotective properties by reducing MMP-1 and MMP-13 expression.

Proteinase-activated receptor (PAR)-2 activation impacts bone resorptive properties of human osteoarthritic subchondral bone osteoblasts.

INTRODUCTION: In osteoarthritis (OA), the subchondral bone undergoes a remodelling process involving several factors synthesized by osteoblasts. In this study, we investigated the expression, production, modulation, and role of PAR-2 in human OA subchondral bone osteoblasts. MATERIALS AND METHODS: PAR-2 expression and production were determined by real-time PCR and flow cytometry, respectively. PAR-2 modulation was investigated in OA subchondral bone osteoblasts treated with IL-1 beta (100 pg/ml), TNF-alpha (5 ng/ml), TGF-beta1 (10 ng/ml), PGE(2) (500 nM), IL-6 (10 ng/ml) and IL-17 (10 ng/ml). Membranous RANKL protein was assessed by flow cytometry, and OPG, MMP-1, MMP-9, MMP-13, IL-6 and intracellular signalling pathways by specific ELISAs. Bone resorptive activity was measured by using a co-culture model of human PBMC and OA subchondral bone osteoblasts. RESULTS: PAR-2 expression and production (p<0.05) were markedly increased when human OA subchondral bone osteoblasts were compared to normal. On OA osteoblasts, PAR-2 production was significantly increased by IL-1 beta, TNF-alpha and PGE(2). Activation of PAR-2 with a specific agonist, SLIGKV-NH(2), induced a significant up-regulation of MMP-1, MMP-9, IL-6, and membranous RANKL, but had no effect on MMP-13 or OPG production. Interestingly, bone resorptive activity was also significantly enhanced following PAR-2 activation. The PAR-2 effect was mediated by activation of the MAP kinases Erk1/2 and JNK. CONCLUSION: This study is the first to demonstrate that PAR-2 activation plays a role in OA subchondral bone resorption via an up-regulation of major bone remodelling factors. These results shed new light on the potential of PAR-2 as a therapeutic target in OA.

Gesture as an important factor in 3D kinematic assessment of the knee.

Contradictions exist between studies of the 3D kinematics of the knee. We hypothesize that they are in part due to differences in the gesture performed by the subjects during kinematic assessment. The purpose of this study is to evaluate the impact of gesture variations on knee kinematics. Seventeen healthy male subjects performed 20-s series of knee-bends in a knee-bend standardizing structure. All series differed regarding either foot rotation, knee excursion, or hip rotation. 3D knee kinematics were recorded using optical position sensors mounted on a skin-motion-reducing harness. Kinematic comparisons were made between a gesture of reference (the standard gesture) and every other gesture. Analyses were performed on average differences. Differences of up to 15 degrees of tibial rotation were found for gestures involving different foot rotation. Gestures involving different knee excursion brought on differences of more than 4 degrees of tibial rotation while hip rotation induced more than 5 degrees of tibial rotation. It is hereby demonstrated that gesture differences can have a dramatic impact on measured knee kinematics. Hence gesture performance needs to be carefully monitored during 3D kinematic assessment of the weight-bearing human knee.

Activation of the receptor EphB4 by its specific ligand ephrin B2 in human osteoarthritic subchondral bone osteoblasts.

OBJECTIVE: Abnormal subchondral bone metabolism is involved in osteoarthritis (OA). It has been suggested that ephrin B2 and its specific receptor EphB4 participate in bone homeostasis. We previously reported that human OA subchondral bone osteoblasts could be classified into 2 subpopulations: low (L), having proresorption properties, and high (H), having proformation properties. The purpose of this study was to investigate the importance of the ephrin system in OA subchondral bone osteoblasts. METHODS: The presence of the EphB4 receptor was determined by immunohistochemistry, and its expression level, modulation upon treatment, and consequences of activation by ephrin B2 were determined by quantitative polymerase chain reaction. The effects of ephrin B2 activation of the EphB4 receptor on bone resorption activity were also determined. EphB4 receptor activation signaling pathways were investigated by specific enzyme-linked immunosorbent assay. RESULTS: EphB4 receptors were present in subchondral bone osteoblasts and osteocytes. Compared with normal and H-OA osteoblasts, EphB4 receptor expression levels were significantly increased in L-OA osteoblasts, with no difference between normal and H-OA osteoblasts. EphB4 receptor levels in L-OA osteoblasts were significantly up-regulated by prostaglandin E2 (PGE2) and interleukin-17 (IL-17). Ephrin B2, PGE2, and IL-17 significantly inhibited bone resorption activity in these cells. EphB4 activation by ephrin B2 significantly inhibited the expression of IL-1beta, IL-6, matrix metalloproteinase 1 (MMP-1), MMP-9, MMP-13, and RANKL, but not MMP-2 and osteoprotegerin. EphB4 receptor activation significantly inhibited the phosphatidylinositol 3-kinase/Akt pathway. CONCLUSION: This study is the first to provide evidence that EphB4 receptor activation by ephrin B2 in OA subchondral bone could affect abnormal metabolism in this tissue by inhibiting resorption factors and their activities. Ephrin B2 could be targeted as a specific therapeutic approach in the development of a disease-modifying OA drug.