Effects of low-intensity pulsed ultrasound on the expression of cyclooxygenase-2 in mandibular condylar chondrocytes.

AIMS: To determine the effect of low-intensity pulsed ultrasound (LIPUS) on cyclooxygenase-2 (COX-2) expression and related mechanisms by using cultured articular chondrocytes derived from porcine mandibular condyles after treatment with interleukin-1 beta (IL-1ß). METHODS: Chondrocytes were derived from porcine mandibular condylar cartilage and cultured. The cells were treated with or without 10 ng/mL IL-1ß. At the same time, the cells were exposed to LIPUS for 20 minutes. After LIPUS exposure, the conditioned medium was changed to a fresh one without IL-1ß, and the cells were incubated for 0 to 24 hours. The effects of LIPUS on IL-1ß-treated chondrocytes were examined in terms of the expression of p-integrin ß1, COX-2, and phosphorylated extracellular signal-related kinase (p-ERK) 1/2 by real-time polymerase chain reaction (PCR) and Western blot analyses. Differences in the means among multiple groups were examined by one-way analysis of variance (ANOVA) for all groups at each time point, followed by a Scheffé multiple comparison test as a post-hoc test; Student t test was also used. RESULTS: COX-2 mRNA level was upregulated by the treatment with IL-1ß and was suppressed significantly (P < .01) by LIPUS exposure. Furthermore, LIPUS enhanced gene expression and phosphorylation of integrin ß, and it inhibited the expression of p-ERK 1/2. CONCLUSION: LIPUS exposure inhibited IL-1ß-induced COX-2 expression through the integrin ß1 receptor followed by the phosphorylation of ERK 1/2. Despite the restricted duration of its effect, LIPUS is suggested to be a potential candidate as a preventive and auxiliary treatment to suppress the degradation of articular chondrocytes in temporomandibular joint osteoarthritis.

Interleukin-1 beta affects cyclooxygenase-2 expression and cartilage metabolism in mandibular condyle.

Extracellular matrix degradation in mandibular condylar cartilage is mediated by various cytokines in the temporomandibular joint (TMJ). Interleukin-1 beta (IL-1ß) is detected in joint structures with pathologic status, and participates in catabolic action in the extracellular matrix. The purpose of this study was to investigate the effects of IL-1ß on cyclooxygenase-2 (COX-2) expression and cartilage metabolism using cultured chondrocytes from mandibular condyle. Articular chondrocytes from the porcine mandibular condylar cartilage around the surface were cultured and treated with 0-10 ng/ml IL-1ß or 0-1000 ng/ml prostaglandin (PGE(2)) for 0-24h. The mRNA levels of COX-2, MMP-1, -3, and -13 were evaluated by real-time PCR analysis. The protein levels of PGE(2) and MMPs were examined by ELISA and Western blot analysis, respectively. The expression levels of COX-2 and PGE(2) were enhanced by exogenous IL-1ß in chondrocytes. The mRNA levels of MMP-1, -3, and -13 were up-regulated by PGE(2) treatment dose-dependently. It is shown that the expression of COX-2/PGE(2) was enhanced by IL-1ß in articular chondrocytes from mandibular condyle, and that MMP-1, -3, and -13 were induced by PGE(2), suggesting that IL-1ß-induced COX-2/PGE(2) play a crucial role in catabolic processes of mandibular condylar cartilage under inflammatory conditions.

Applying an excessive mechanical stress alters the effect of subchondral osteoblasts on chondrocytes in a co-culture system.

Osteoarthritis (OA) sometimes occurs as a consequence of repeated microtrauma involved in parafunction, which may lead to microfracture in the subchondral bone. The aim of this in vitro study was to evaluate the effects of subchondral osteoblasts in loading with repeated excessive mechanical stress on the metabolism of overlying chondrocytes. A high-magnitude cyclic tensile stress of 15 kPa (30 cycles min(-1)) was applied to the cultured osteoblasts obtained from porcine mandibular condyles. The chondrocytes in alginate beads were then co-cultured with mechanically stressed or unstressed osteoblasts. Chondrocytes co-cultured with unstressed osteoblasts showed a phenotypic shift to hypertrophic chondrocytes, characterized by decreased expression of type II collagen, aggrecan, Sry-related HMG box (SOX-9), and cartilage oligomeric matrix protein (COMP) genes and increased expression of type X collagen and bone sialoprotein (BSP) genes, suggesting that the co-culture may change the chondrocyte differentiation to some extent. These changes were more distinct in chondrocytes co-cultured with excessively mechanically stressed osteoblasts. After co-culture with stressed osteoblasts, the expressions of matrix metalloproteinase (MMP)1, MMP3 and MMP13 genes were also enhanced and the synthesis of DNA, proteoglycan and collagen were significantly decreased in chondrocytes. These results demonstrate that alterations in cartilage metabolism can be induced by stressed osteoblasts, indicating a possible explanation for the onset and progression of OA.

Dynamic shear behavior of mandibular condylar cartilage is dependent on testing direction.

Little information is available on the direction-dependency of shear behavior in mandibular condylar cartilage. Therefore, we tested the hypothesis that such a dependency of the dynamic shear properties is present in mandibular condylar cartilage. From each of 17 condyles, two cartilage-bone plugs were dissected and tested in a simple shear sandwich configuration under a compressive strain of 10%. Sinusoidal shear strain (frequency range: 0.01-10 Hz) was applied in the medio-lateral or antero-posterior direction with an amplitude of 1.0%, 2.0%, and 3.0%. The magnitudes of the dynamic shear moduli, as calculated from the resulting shear stress, were found to increase with applied frequency and the shear strain amplitude. The values |G*|, G' and G'' for a medio-laterally applied shear were about 20-33% of those in the antero-posterior shear, although the loss tangent (elasticity/viscosity ratio) was almost the same. In conclusion, the present results clearly show the direction-dependent characteristic of the mandibular condylar cartilage in dynamic shear.

Modeling of the effect of friction in the temporomandibular joint on displacement of its disc during prolonged clenching.

PURPOSE: The effect of the frictional coefficient in the temporomandibular joint on the disc during prolonged clenching was examined. MATERIALS AND METHODS: For this purpose, a finite element model of the temporomandibular joint based on magnetic resonance images from a volunteer subject was used. Muscle forces applied for clenching were used as a loading condition for stress analysis during 10 minutes. With respect to the frictional coefficient between articular surfaces, 3 different values ranging from micro = 0.001 to micro = 0.1 were established. RESULTS: At the onset of clenching, large stresses were found in the central and lateral part of the intermediate zone in the disc, and its stress distribution was not markedly changed during 10 minutes of clenching. In the retrodiscal tissue, stress relaxation occurred during the first 2 minutes of clenching. When the frictional coefficient between articular surfaces increased, the anterior, lateral, and central points in the disc moved further forward. At the end of 10 minutes of clenching, the disc showed a more anterior position as the frictional coefficient increased. CONCLUSION: This result indicates that increase of the frictional coefficient between the articular surfaces may be a major cause for the onset of the disc displacement.

Biomechanical response of condylar cartilage-on-bone to dynamic shear.

Shear stress can result in fatigue, damage, and irreversible deformation of the mandibular condylar cartilage. However, little information is available on its dynamic properties in shear. We tested the hypothesis that the dynamic shear properties of the condylar cartilage depend on the frequency and amplitude of shear strain. Ten porcine mandibular condyles were used for dynamic shear tests. Two cartilage-bone plugs were dissected from each condyle and tested in a simple shear sandwich configuration under a compressive strain of 10%. Sinusoidal shear strain was applied with an amplitude of 1.0, 2.0, and 3.0% and a frequency range between 0.01 and 10 Hz. The magnitudes of the shear dynamic moduli were found to be dependent on the frequency and the shear strain amplitude. They increased with shear strain. tan delta ranged from 0.2 to 0.4, which means that the cartilage is primarily elastic in nature and has a small but not negligible viscosity. In conclusion, the present results show that the shear behavior of the mandibular condylar cartilage is dependent on the frequency and amplitude of the applied shear strain. The observed shear characteristics suggest a significant role of shear strain on the interstitial fluid flow within the cartilage.