Effect of Sustained Joint Loading on TMJ Disc Nutrient Environment.

The temporomandibular joint (TMJ) disc nutrient environment profoundly affects cell energy metabolism, proliferation, and biosynthesis. Due to technical challenges of in vivo measurements, the human TMJ disc extracellular nutrient environment under load, which depends on metabolic rates, solute diffusion, and disc morphometry, remains unknown. Therefore, the study objective was to predict the TMJ disc nutrient environment under loading conditions using combined experimental and computational modeling approaches. Specifically, glucose consumption and lactate production rates of porcine TMJ discs were measured under varying tissue culture conditions (n = 40 discs), and mechanical strain-dependent glucose and lactate diffusivities were measured using a custom diffusion chamber (n = 6 discs). TMJ anatomy and loading area were obtained from magnetic resonance imaging of healthy human volunteers (n = 11, male, 30 ± 9 y). Using experimentally determined nutrient metabolic rates, solute diffusivities, TMJ anatomy, and loading areas, subject-specific finite element (FE) models were developed to predict the 3-dimensional nutrient profiles in unloaded and loaded TMJ discs (unloaded, 0% strain, 20% strain). From the FE models, glucose, lactate, and oxygen concentration ranges for unloaded healthy human TMJ discs were 0.6 to 4.0 mM, 0.9 to 5.0 mM, and 0% to 6%, respectively, with steep gradients in the anterior and posterior bands. Sustained mechanical loading significantly reduced nutrient levels (P < 0.001), with a critical zone in which cells may die representing approximately 13.5% of the total disc volume. In conclusion, this study experimentally determined TMJ disc metabolic rates, solute diffusivities, and disc morphometry, and through subject-specific FE modeling, revealed critical interactions between mechanical loading and nutrient supply and metabolism for the in vivo human TMJ disc. The results suggest that TMJ disc homeostasis may be vulnerable to pathological loading (e.g., clenching, bruxism), which impedes nutrient supply. Given difficulties associated with direct in vivo measurements, this study provides a new approach to systematically investigate homeostatic and degenerative mechanisms associated with the TMJ disc.

The Zonal Architecture of the Mandibular Condyle Requires ADAMTS5.

Temporomandibular joint (TMJ) osteoarthritis (TMJOA) disrupts extracellular matrix (ECM) homeostasis, leading to cartilage degradation. Upregulated a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-5 leads to cleavage of its substrate aggrecan (Acan) and is considered a hallmark of TMJOA. However, most research on ADAMTS5-Acan turnover has focused on hyaline cartilage, not fibrocartilage, which comprises the TMJ. The mandibular condylar cartilage (MCC) of the TMJ is organized in zones, and chondrocytes are arranged in axial rows, yet the molecular mechanisms required to generate the MCC zonal architecture have not been elucidated. Here, we test the hypothesis that ADAMTS5 is required for development of the TMJ MCC. Adamts5+/+ and Adamts5-/- murine TMJs were harvested at postnatal day 7 (P7), P21, 2 mo, and 6 mo of age; histomorphometrics indicated increased ECM. Immunohistochemistry and Western blots demonstrated the expanded ECM correlated with increased Acan localization in Adamts5-/- compared to Adamts5+/+. Cell volume was also decreased in the MCC of Adamts5-/- due to both a reduction in cell size and less mature hypertrophic chondrocytes. Analysis of chondrogenic maturation markers by quantitative real-time polymerase chain reaction indicated Col2a1, Col10a1, and Sox9 were significantly reduced in Adamts5-/- MCC compared to that of Adamts5+/+. The older (6 mo) Adamts5-/- MCC exhibited changes in chondrogenic cell arrangements, including clustering and chondrogenic atrophy, that correlated with early stages of TMJOA using modified Mankin scoring. These data indicate a potentially novel and critical role of ADAMTS5 for maturation of hypertrophic chondrocytes and establishment of the zonal architecture that, when disrupted, may lead to early onset of TMJOA.

Fluid pressurization and tractional forces during TMJ disc loading: A biphasic finite element analysis.

OBJECTIVES: To investigate the ploughing mechanism associated with tractional force formation on the temporomandibular joint (TMJ) disc surface. SETTING AND SAMPLE POPULATION: Ten left TMJ discs were harvested from 6- to 8-month-old male Yorkshire pigs. MATERIALS AND METHODS: Confined compression tests characterized mechanical TMJ disc properties, which were incorporated into a biphasic finite element model (FEM). The FEM was established to investigate load carriage within the extracellular matrix (ECM) and the ploughing mechanism during tractional force formation by simulating previous in vitro plough experiments. RESULTS: Biphasic mechanical properties were determined in five TMJ disc regions (average±standard deviation for aggregate modulus: 0.077±0.040 MPa; hydraulic permeability: 0.88±0.37×10-3 mm4 /Ns). FE simulation results demonstrated that interstitial fluid pressurization is a dominant loading support mechanism in the TMJ disc. Increased contact load and duration led to increased solid ECM strain and stress within, and increased ploughing force on the surface of the disc. CONCLUSION: Sustained mechanical loading may play a role in load carriage within the ECM and ploughing force formation during stress-field translation at the condyle-disc interface. This study further elucidated the mechanism of ploughing on tractional force formation and provided a baseline for future analysis of TMJ mechanics, cartilage fatigue and early TMJ degeneration.

The effects of oxygen level and glucose concentration on the metabolism of porcine TMJ disc cells.

OBJECTIVE: To determine the combined effect of oxygen level and glucose concentration on cell viability, ATP production, and matrix synthesis of temporomandibular joint (TMJ) disc cells. DESIGN: TMJ disc cells were isolated from pigs aged 6-8 months and cultured in a monolayer. Cell cultures were preconditioned for 48 h with 0, 1.5, 5, or 25 mM glucose DMEM under 1%, 5%, 10%, or 21% O2 level, respectively. The cell viability was measured using the WST-1 assay. ATP production was determined using the Luciferin-Luciferase assay. Collagen and proteoglycan synthesis were determined by measuring the incorporation of [2, 3-(3)H] proline and [(35)S] sulfate into the cells, respectively. RESULTS: TMJ disc cell viability significantly decreased (P < 0.0001) without glucose. With glucose present, decreased oxygen levels significantly increased viability (P < 0.0001), while a decrease in glucose concentration significantly decreased viability (P < 0.0001). With glucose present, decreasing oxygen levels significantly reduced ATP production (P < 0.0001) and matrix synthesis (P < 0.0001). A decreased glucose concentration significantly decreased collagen synthesis (P < 0.0001). The interaction between glucose and oxygen was significant in regards to cell viability (P < 0.0001), ATP production (P = 0.00015), and collagen (P = 0.0002) and proteoglycan synthesis (P < 0.0001). CONCLUSIONS: Although both glucose and oxygen are important, glucose is the limiting nutrient for TMJ disc cell survival. At low oxygen levels, the production of ATP, collagen, and proteoglycan are severely inhibited. These results suggest that steeper nutrient gradients may exist in the TMJ disc and it may be vulnerable to pathological events that impede nutrient supply.

Viscoelastic shear properties of porcine temporomandibular joint disc.

OBJECTIVES: To investigate the intrinsic viscoelastic shear properties in porcine TMJ discs. MATERIALS AND METHODS: Twelve fresh porcine TMJ discs from young adult pigs (6-8 months) were used. Cylindrical samples (5 mm diameter) with uniform thickness (~1.2 mm) were prepared from five regions of the TMJ disc. Torsional shear tests were performed under 10% compressive strain. Dynamic shear was applied in two methods: 1) a frequency sweep test over the frequency range of 0.1-10 rad/s with a constant shear strain amplitude of 0.05 rad and 2) a strain sweep test over the range of 0.005-0.15 rad at a constant frequency of 10 rad/s. Transient stress relaxation tests were also performed to determine the equilibrium shear properties. RESULTS: As the frequency increased in the frequency sweep test, the dynamic shear complex modulus increased, with values ranging from 7 to 17 kPa. The phase angle, ranging from 11 to 15 degrees, displayed no pattern of regional variation as the frequency increased. The dynamic shear modulus decreased as the shear strain increased. The equilibrium shear modulus had values ranging from 2.6 to 4 kPa. The posterior region had significantly higher values for dynamic shear modulus than those in the anterior region, while no significant regional difference was found for equilibrium shear modulus. CONCLUSION: Our results suggest that the intrinsic region-dependent viscoelastic shear characteristics of TMJ disc may play a crucial role in determining the local strain of the TMJ disc under mechanical loading.