The objective of this study was to determine the biphasic viscoelastic properties of human temporomandibular joint (TMJ) discs, correlate these properties with disc biochemical composition, and examine the relationship between these properties and disc dynamic behavior in confined compression. The equilibrium aggregate modulus (H A ), hydraulic permeability (k), and dynamic modulus were examined between five disc regions. Biochemical assays were conducted to quantify the amount of water, collagen, and glycosaminoglycan (GAG) content in each region. The creep tests showed that the average equilibrium moduli of the intermediate, lateral, and medial regions were significantly higher than for the anterior and posterior regions (69.75±11.47 kPa compared to 22.0±5.15 kPa). Permeability showed the inverse trend with the largest values in the anterior and posterior regions (8.51±1.36×10 −15 m 4 /N compared with 3.75±0.72×10 −15 m 4 /N). Discs were 74.5% water by wet weight, 62% collagen, and 3.2% GAG by dry weight. Regional variations were only observed for water content which likely results in the regional variation in biphasic mechanical properties. The dynamic modulus of samples during confined compression is related to the aggregate modulus and hydraulic permeability of the tissue. The anterior and posterior regions displayed lower complex moduli over all frequencies (0.01-3 Hz) with average moduli of 171.8-609.3 kPa compared with 454.6-1613.0 kPa for the 3 central regions. The region of the TMJ disc with higher aggregate modulus and lower permeability had higher dynamic modulus. Our results suggested that fluid pressurization plays a significant role in the load support of the TMJ disc under dynamic loading conditions.
A custom-designed three-dimensional additive manufacturing device was developed to fabricate scaffolds for intervertebral disk (IVD) regeneration. This technique integrated a computer with a device capable of 3D movement allowing for precise motion and control over the polymer scaffold resolution. IVD scaffold structures were designed using computer-aided design to resemble the natural IVD structure. Degradable polyurethane (PU) was used as an elastic scaffold construct to mimic the elastic nature of the native IVD tissue and was deposited at a controlled rate using ultra-fine micropipettes connected to a syringe pump. The elastic PU was extruded directly onto a collecting substrate placed on a freezing stage. The three-dimensional movement of the computer-controlled device combined with the freezing stage enabled precise control of polymer deposition using extrusion. The addition of the freezing stage increased the polymer solution viscosity and hardened the polymer solution as it was extruded out of the micropipette tip. This technique created scaffolds with excellent control over macro- and micro-structure to influence cell behavior, specifically for cell adhesion, proliferation, and alignment. Concentric lamellae were printed at a high resolution to mimic the native shape and structure of the IVD. Seeded cells aligned along the concentric lamellae and acquired cell morphology similar to native tissue in the outer portion of the IVD. The fabricated scaffolds exhibited elastic behavior during compressive and shear testing, proving that the scaffolds could support loads with proper fatigue resistance without permanent deformation. Additionally, the mechanical properties of the scaffolds were comparable to those of native IVD tissue.
A new method solely based on spatial Fourier analysis (SFA) was developed to completely determine a two dimensional (2D) anisotropic diffusion tensor in fibrous tissues using fluorescence recovery after photobleaching (FRAP). The accuracy and robustness of this method was validated by using computer simulated FRAP experiments. This method was applied to determine the region-dependent anisotropic diffusion tensor in porcine temporomandibular joint (TMJ) discs. The average characteristic diffusivity of 4kDa FITC-Dextran across the disc was 26.05±4.32μm 2 /s which is about 16% of its diffusivity in water. In the anteroposterior direction, the anterior region (30.99±5.93 μm 2 /s) had significantly higher characteristic diffusivity than the intermediate region (20.49±5.38μm 2 /s) and posterior region (20.97±2.46 μm 2 /s). The ratio of the two principal diffusivities represents the anisotropy of the diffusion and ranged between 0.45~0.51 (1.0=isotropic). Our results indicated that the solute diffusion in TMJ discs is inhomogenous and anisotropic. These findings suggested that diffusive transport in the TMJ disc is dependent upon tissue composition (e.g., water content) and structure (e.g., collagen orientation). This study provides a new method to quantitatively investigate the relationship between solute transport properties and tissue composition and structure.
The objective of this study was to examine the impact of mechanical loading on solute transport in porcine temporomandibular joint (TMJ) discs using the electrical conductivity method. The electrical conductivity, as well as ion diffusivity, of TMJ discs was determined under confined compression with 3 strains in 5 disc regions. The average electrical conductivity over the 5 regions (mean ± SD) at 0% strain was 3.10 ± 0.68 mS/cm, decreased to 2.76 ± 0.58 mS/cm (-11.0%) at 10% strain, and 2.38 ± 0.55 mS/cm (-22.2%) at 20% compressive strain. Correspondingly, the average relative ion diffusivity (mean ± SD) at 0% strain was 0.273 ± 0.055, decreased to 0.253 ± 0.048 (-7.3%) at 10% strain, and 0.231 ± 0.048 (-15.4%) at 20% compressive strain. These results indicated that compressive strain impeded solute transport in the TMJ disc. Furthermore, our results showed that the transport properties of TMJ discs were regiondependent. The electrical conductivity and ion diffusivity in the anterior region were significantly higher than in the posterior region. This regional difference is likely due to the significant differences of tissue hydration between these 2 regions. This study provides important insight into the electrical and solute transport behaviors in TMJ discs under mechanical loading and aids in the understanding of TMJ pathophysiology related to tissue nutrition.
Objective To determine the regional cell density distribution and basal oxygen consumption rates (based on tissue volume and cell number) of temporomandibular joint (TMJ) discs and further examine the impact of oxygen tension on these rates. Design TMJ discs from pigs aged 6–8 months were divided into five regions: anterior, intermediate, posterior, lateral and medial. The cell density was determined using confocal laser scanning microscopy. The change in oxygen tension was recorded while TMJ disc explants were cultured in sealed metabolism chambers. The volume based oxygen consumption rate of explants was determined by theoretical curve fitting of the recoded oxygen tension data with the Michaelis-Menten equation. The rate on a per-cell basis was calculated based on the cell density measurements and volume based rate measured in another group of discs. Results The overall cell density (mean, 95% CI) was 51.3(21.3–81.3)×106cells/mL wet tissue. Along the anteroposterior axis, the anterior band had 25.5% higher cell density than the intermediate zone (p<0.02) and 29.1% higher than the posterior band (p<0.008). Along the mediolateral axes, the medial region had 26.2% higher cell density than the intermediate zone (p<0.04) and 25.4% higher than the lateral region (p<0.045). The overall volume and cell based maximum oxygen consumption rates were 1.44(0.44–2.44) μmol/mL wet tissue/hr and 28.7(12.2–45.2) nmol/106 cells/hr, respectively. The central regions (intermediate, lateral, and medial) had significantly higher volume based (p<0.02) and cell based (p<0.005) oxygen consumption rates than the anterior and posterior bands. At high oxygen tension, the oxygen consumption rate remained constant, but dropped as oxygen tension fell below 5%. Conclusions The TMJ disc had higher cell density and oxygen consumption rates than articular cartilage reported in the literature. These results suggest that a steeper oxygen gradient may exist in the TMJ disc and may be vulnerable to pathological events that impede nutrient supply.
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