Porcine Temporomandibular Joint Discs Research Articles

Microstructural, Micromechanical Atlas of the Temporomandibular Joint Disc

The temporomandibular joint (TMJ) disc is mainly composed of collagen, with its arrangement responding to efficient stress distribution. However, microstructural and micromechanical transformations of the TMJ disc under resting, functional, and pathological conditions remain unclear. To address this, our study presents a high-resolution microstructural and mechanical atlas of the porcine TMJ disc. First, the naive microstructure and mechanical properties were investigated in porcine TMJ discs (resting and functional conditions). Subsequently, the perforation and tear models (pathological conditions) were compared. Following this, a rabbit model of anterior disc displacement (abnormal stress) was studied. Results show diverse microstructures and mechanical properties at the nanometer to micrometer scale. In the functional state, gradual unfolding of the crimping cycle in secondary and tertiary structures leads to D-cycle prolongation in the primary structure, causing tissue failure. Pathological conditions lead to stress concentration near the injury site due to collagen interfibrillar traffic patterns, resulting in earlier damage manifestation. Additionally, the abnormal stress model shows collagen damage initiating at the primary structure and extending to the superstructure over time. These findings highlight collagen’s various roles in different pathophysiological states. Our study offers valuable insights into TMJ disc function and dysfunction, aiding the development of diagnostic and therapeutic strategies for TMJ disorders, as well as providing guidance for the design of structural biomimetic materials.

Experimental study of eigenstrains in temporomandibular joint discs using digital image analysis

The temporomandibular joint is one of the most frequently used joints of the human body. Its malfunction can severely influence patient’s well-being. Since the temporomandibular joint disc plays a major role in its functioning, especially in load distribution within the joint, it appears to be a crucial element to understand. This paper aims to improve understanding of the tissues within close in vivo conditions (i.e. hydrated at 37 ° C) by (i) comforting literature by revealing the presence of residual stresses within the temporomandibular joint disc, (ii) quantifying eigenstrains through a relaxation process and finally (iii) evaluating the internal mechanical state in intact temporomandibular joint discs central part, considering the tissue as a thin layer. Both global specimen size measurements and local digital image correlation were used to quantify 6 samples’ deformation through a detailed analysis of approximately 30 images, recorded for approximately one hour, per disc. Thanks to a backward time approach combined to an analytical model, eigenstrains were assessed on discs. For the first time, the presence of complex initial strain fields within cylindrical specimens of porcine temporomandibular joint discs was quantified, confirming indications from literature. Digital image analysis revealed the partial internal stress release through specimen self-deformation. Close to zero in central part, it reached approximately 13% radial strain in the outer ring within a characteristic relaxation time close to 530s. The principal strains’ distribution agrees with the alignment of the collagen fibers in the central part of the discs revealed in many works. It led to deduce that, in the central area of the discs, the matrix undergoes a radial compression within physiological conditions to compensate the daily loading stresses. Therefore, this work improves understanding of the tissues in vivo conditions highlighting extraction cut effect on temporomandibular joint disc’s tissues mechanical state.

Identification of Biomechanical Properties of Temporomandibular Discs.

Material Experimental and model tests were conducted on ten fresh porcine temporomandibular joint discs. The average thickness of disc tissue was, accordingly, 2.77 mm for the anterior zone, 3.98 mm for the posterior, and 1.54 mm for the intermediate. The selection of research material in the form of porcine discs was due to the similarity to human discs. Methods Discs were loaded in cycles, a temporary course with the amplitude 3 N and frequency 0.07 Hz, and growth in the load was 1 N/s. The selection of load frequency was due to real conditions of temporomandibular joint functioning during mastication. The necessary experimental research was conducted on a testing machine with a measurement range of 2.5 kN. Results The obtained numeric calculation results indicate that the number of load cycles has a decisive impact on the limitation of energy dispersion capacity through disc tissue. This phenomenon was observed in all the studies on the disc areas. Along with the growth in load cycles, discs are stiffened, and the most significant stiffness was observed in the intermediate area. Conclusions Based on the conducted research, it should be concluded that excessive load affecting temporomandibular joints caused by the act of mastication and occlusal forces generated during parafunction and in people with defined long-term bruxism has crucial importance on biomechanical disc properties and hence the course of temporomandibular joint conditions.

Acellular matrix hydrogel for repair of the temporomandibular joint disc.

Application of tissue-derived extracellular matrix (ECM) biomaterials in the repair of the temporomandibular joint (TMJ) disc is a promising approach for the treatment of disc abrasion and perforation, particularly for the young patient population. Although decellularized ECM (dECM) scaffolds preserve tissue-specific structures as well as biological and biomechanical properties, they require surgical implantation. To address this issue, we prepared porcine TMJ discs into decellularized ECM with serial detergent and enzyme treatments, and the TMJ disc-derived ECM was then processed into hydrogels via pepsin digestion. The decellularization efficiency was assessed by quantification of the DNA and matrix component contents. The fibrous ultrastructure of the hydrogel was observed by scanning electron microscopy (SEM). Rheological characterization and mechanical properties were measured. in vitro experiments with costal chondrocytes ensured the cellular proliferative capacity and compatibility in the injectable disc-derived ECM hydrogel. The results showed that a large amount of DNA (>95%) was removed after decellularization; but, the collagen was retained. SEM of the hydrogels demonstrated a multiaperture fiber ultrastructure. Rheological studies revealed a rapid gelation temperature (37°C) and injectable properties. The mechanical properties of the hydrogels were adjusted by changing the ECM concentration. The in vitro studies revealed that the hydrogels are not cytotoxic, but instead showed good cytocompatibility. The hydrogel also showed good injectability and degradability through an in vivo study. Overall, these results suggest the great potential of injectable disc-derived hydrogels for TMJ disc repair and regeneration applications.

Alteration of structural and mechanical properties of the temporomandibular joint disc following elastase digestion.

The temporomandibular joint disc is a fibrocartilaginous structure, composed of collagen fibers, elastin fibers, and proteoglycans. Despite the crucial role of elastin fibers in load‐bearing properties of connective tissues, its contribution in temporomandibular joint disc biomechanics has been disregarded. This study attempts to characterize the structural–functional contribution of elastin in the temporomandibular joint disc. Using elastase, we selectively perturbed the elastin fiber network in porcine temporomandibular joint discs and investigated the structural, compositional, and mechanical regional changes through: (a) analysis of collagen and elastin fibers by immunolabeling and transmission electron microscopy; (b) quantitative analysis of collagen tortuosity, cell shape, and disc volume; (c) biochemical quantification of collagen, glycosaminoglycan and elastin content; and (d) cyclic compression test. Following elastase treatment, microscopic examination revealed fragmentation of elastin fibers across the temporomandibular joint disc, with a more pronounced effect in the intermediate regions. Also, biochemical analyses of the intermediate regions showed significant depletion of elastin (50%), and substantial decrease in collagen (20%) and glycosaminoglycan (49%) content, likely due to non‐specific activity of elastase. Degradation of elastin fibers affected the homeostatic configuration of the disc, reflected in its significant volume enlargement accompanied by remarkable reduction of collagen tortuosity and cell elongation. Mechanically, elastase treatment nearly doubled the maximal energy dissipation across the intermediate regions while the instantaneous modulus was not significantly affected. We conclude that elastin fibers contribute to the restoration and maintenance of the disc resting shape and actively interact with collagen fibers to provide mechanical resilience to the temporomandibular joint disc.

The dynamic mechanical viscoelastic properties of the temporomandibular joint disc: The role of collagen and elastin fibers from a perspective of polymer dynamics

The temporomandibular joint disc is a structure, characterized as heterogeneous fibrocartilage, and is composed of macromolecular biopolymers. Despite a large body of characterization studies, the contribution of matrix biopolymers on the dynamic viscoelastic behavior of the disc is poorly understood. Given the high permeability and low concentration of glycosaminoglycans in the disc, it has been suggested that poro-elastic behavior can be neglected and that the intrinsic viscoelastic nature of solid matrix plays a dominant role in governing its time-dependent behavior. This study attempts to quantify the contribution of collagen and elastin fibers to the viscoelastic properties of the disc. Using collagenase and elastase, we perturbed the collagen and elastin fibrillar network in porcine temporomandibular joint discs and investigated the changes of dynamic viscoelastic properties in five different regions of the disc. Following both treatments, the storage and loss moduli of these regions were reduced dramatically up to the point that the tissue was no longer mechanically heterogeneous. However, the proportion of changes in storage and loss moduli were different for each treatment, reflected in the decrease and increase of the loss tangent for collagenase and elastase treated discs, respectively. The reduction of storage and loss moduli of the disc correlated with a decrease of biopolymer length. The present study indicates that the compositional and structural changes of collagen and elastin fibers alter the viscoelastic properties of the disc consistent with polymer dynamics.

Viscoelastic properties of the central region of porcine temporomandibular joint disc in shear stress-relaxation

In this study, shear relaxation properties of the porcine temporomandibular joint (TMJ) disc are investigated. Previous studies have shown that, in fatigue failure and damage of cartilage and fibrocartilage, shear loads could be one of the biggest contributors to the failure. The aim of the present study is to develop an evaluation method to study shear properties of the disc and to do a mathematical characterization of it. For the experiments, twelve porcine discs were used. Each disc was dissected from the TMJ and, then, static strain control tests were carried out to obtain the shear relaxation modulus for the central region of the discs. From the results, it was found that the disc presents a viscoelastic behavior under shear loads. Relaxation modulus decreased with time. Shear relaxation was 10% of the instantaneous stress, which implies that the viscous properties of the disc cannot be neglected. The present results lead to a better understanding of the discs mechanical behavior under realistic TMJ working conditions.

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.

Effect of freezing storage time on the elastic and viscous properties of the porcine TMJ disc

The correct characterisation of the articular disc of the temporomandibular joint (TMJ) is key to study the masticatory biomechanics. For the interval from extraction until testing, freezing is the most used preservation technique for biological tissues, but its influence on their behaviour is still unclear. An important error can be committed in the characterisation of such tissues if freezing has any effect on their mechanical properties. Thus, the aim of this study was to determine whether the freezing storage time causes any change in the mechanical properties of the TMJ discs. To check that, the specimens were stored in a −20°C freezer during different time intervals: 1 day, 1 week, 1 month and 3 months. Fresh specimens, tested right after extraction, were used as the control group. Compressive stress relaxation tests were carried out on the specimens and a quasi-linear viscoelastic (QLV) model was used to fit the experimental curves. A statistical analysis detected significant differences among the groups. Post-hoc tests determined that freezing the specimens more than 30 days may lead to changes in the viscoelastic properties of the tissue.

The contribution of collagen fibers to the mechanical compressive properties of the temporomandibular joint disc

The Temporomandibular Joint (TMJ) disc is a fibrocartilaginous structure located between the mandibular condyle and the temporal bone, facilitating smooth movements of the jaw. The load-bearing properties of its anisotropic collagenous network have been well characterized under tensile loading conditions. However, recently it has also been speculated that the collagen fibers may contribute dominantly in reinforcing the disc under compression. Therefore, in this study, the structural-functional role of collagen fibers in mechanical compressive properties of TMJ disc was investigated. Intact porcine TMJ discs were enzymatically digested with collagenase to disrupt the collagenous network of the cartilage. The digested and non-digested articular discs were analyzed mechanically, biochemically and histologically in five various regions. These tests included: (1) cyclic compression tests, (2) biochemical quantification of collagen and glycosaminoglycan (GAG) content and (3) visualization of collagen fibers' alignment by polarized light microscopy (PLM). The instantaneous compressive moduli of the articular discs were reduced by as much as 50-90% depending on the region after the collagenase treatment. The energy dissipation properties of the digested discs showed a similar tendency. Biochemical analysis of the digested samples demonstrated an average of 14% and 35% loss in collagen and GAG, respectively. Despite the low reduction of collagen content the PLM images showed considerable perturbation of the collagenous network of the TMJ disc. The results indicated that even mild disruption of collagen fibers can lead to substantial mechanical softening of TMJ disc undermining its reinforcement and mechanical stability under compression.

Biomechanical and biochemical outcomes of porcine temporomandibular joint disc deformation

ObjectiveThe structure–function relationship in the healthy temporomandibular joint (TMJ) disc has been well established, however the changes in dysfunctional joints has yet to be systematically evaluated. Due to the poor understanding of the etiology of temporomandibular disorders (TMDs) this study evaluated naturally occurring degenerative remodeling in aged female porcine temporomandibular joint (TMJ) discs in order to gain insight into the progression and effects on possible treatment strategies of TMDs. DesignSurface and regional biomechanical and biochemical properties of discal tissues were determined in grossly deformed (≥Wilkes Stage 3) and morphologically normal (≤Wilkes Stage 2) TMJ discs. ResultsCompared to normal disc structure the deformed discs lacked a smooth biconcave shape and characteristic ECM organization. Reduction in tensile biomechanical integrity and increased compressive stiffness and cellularity was found in deformed discs. Regionally, the posterior and intermediate zones of the disc were most frequently affected along with the inferior surface. ConclusionsThe frequency of degeneration observed on the inferior surface of the disc (predominantly posterior), suggests that a disruption in the disc-condyle relationship likely contributes to the progression of joint dysfunction more than the temporodiscal relationship. As such, the inferior joint space may be an important consideration in early clinical diagnosis and treatment of TMDs, as it is overlooked in techniques performed in the upper joint space, including arthroscopy and arthrocentesis. Furthermore, permanent damage to the disc mechanical properties would limit the ability to successfully reposition deformed discs, highlighting the importance of emerging therapies such as tissue engineering.

Dynamic and stress relaxation properties of the whole porcine temporomandibular joint disc under compression

In this study, the dynamic and static compressive properties of the whole porcine temporomandibular joint (TMJ) disc were investigated. The aim of the study was to develop a new simple method for the evaluation of joint viscoelasticity, enabling examination of the load-bearing capacity and joint flexibility of the entire disc. For the experiments, a novel testing fixture that reproduces the condylar and fossa surfaces of the TMJ was developed to replicate TMJ disc geometry. Ten porcine discs were used in the experiments. Each disc was dissected from the TMJ and sinusoidal compressive strain was applied to obtain the storage and loss moduli. Static strain control tests were carried out to obtain the relaxation modulus. The result of static and dynamic tests indicated that the whole disc presented viscoelastic behavior under compression. Storage and loss moduli increased with frequency and the relaxation modulus decreased over time. The loss tangent showed less frequency dependence, with values ranging from 0.2 to 0.3, suggesting that the viscous properties of the disc cannot be neglected. These results provide a better understanding of whole disc mechanical compression behavior under realistic TMJ working conditions.

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

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. 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. 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). 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.

Analysis of Compressive Properties of Porcine Temporomandibular Joint Disc

Since the temporomandibular joint (TMJ) disc material exhibits a non-homogenous and viscoelastic structure, the compressive properties in five different regions of eleven porcine TMJ discs were investigated over a wide range of loading frequencies. The results obtained suggest that the dynamic viscoelastic compressive modulus is region-specific and depends on the loading frequency, thus having important implications for the transmission of load in the TMJ. The dynamic storage and loss moduli increase with frequency, the highest values being attained at the posterior region, followed by the central and anterior regions. Loss tangent, tan δ, ranged from 0.20 to 0.35, which means that the disc is primarily elastic in nature and has a small but not negligible viscosity.

Relationship between anisotropic diffusion properties and tissue morphology in porcine TMJ disc

To investigate the relationship between anisotropic solute diffusion properties and tissue morphology in porcine temporomandibular joint (TMJ) discs. TMJ discs from eleven pigs aged 6-8 months were divided into five regions: anterior, intermediate, posterior, lateral, and medial. The transport properties and tissue morphology were investigated in three orthogonal orientations: anteroposterior (AP), mediolateral (ML), and superoinferior (SI). The anisotropic diffusivity of fluorescein (332Da) in the right discs was determined by the fluorescence recovery after photobleaching (FRAP) protocols. The tissue morphology in the left discs was quantified by scanning electron microscopy. The diffusivities of fluorescein in the TMJ disc were significantly anisotropic, except for the anterior region. In the medial, intermediate, and lateral regions, the diffusion along the fiber orientation (i.e., AP direction) was significantly faster than the diffusion in ML and SI directions. In the posterior region, the diffusion along the fiber orientation (i.e., ML direction) was significantly faster than the diffusion in AP and SI directions. The diffusion in the anterior region was mostly isotropic with the lowest degree of diffusion anisotropy, as well as collagen fiber alignment, likely due to the multi-directional fiber arrangement. The anterior region had the highest mean diffusivity [65.6 (49.3-81.8)μm(2)/s] in the disc, likely due to its high water content. The overall average diffusivity of fluorescein across the TMJ disc was 57.0 (43.0-71.0)μm(2)/s. The solute diffusion in porcine TMJ discs was strongly anisotropic and inhomogeneous, which associated with tissue structure (i.e., collagen fiber alignment) and composition (e.g., water content).

The region-dependent dynamic properties of porcine temporomandibular joint disc under unconfined compression

In this study, the dynamic compressive properties in five different regions of the porcine temporomandibular joint (TMJ) disc are investigated over a wide range of loading frequencies. The aim was, thus far, to evaluate the regional difference and the frequency-related effect of the applied load on these properties. Eleven porcine TMJ discs were used; each disc was divided into 5 regions, anterior, central, posterior, lateral and medial. Sinusoidal compressive strain was applied with an amplitude of 1.0% and a frequency range between 0.01 and 10Hz. The dynamic storage and loss moduli increase with frequency, the highest values being attained at the posterior region, followed by the central and anterior regions. Loss tangent, tanδ, ranged from 0.20 to 0.35, which means that the disc is primarily elastic in nature and has a small but not negligible viscosity. The present results suggest that the dynamic viscoelastic compressive modulus is region-specific and depends on the loading frequency, thus having important implications for the transmission of load to the TMJ.

Shear Mechanics of the TMJ Disc

The temporomandibular joint (TMJ) is a complex hinge and gliding joint that induces significant shear loads onto the fibrocartilage TMJ disc during jaw motion. The purpose of this study was to assess regional variation in the disc’s shear loading characteristics under physiologically relevant loads and to associate those mechanical findings with common clinical observations of disc fatigue and damage. Porcine TMJ discs were compressed between an axially translating bottom platen and a 2.5-cm-diameter indenter within a hydrated testing chamber. Discs were cyclically sheared at 0.5, 1, or 5 Hz to 1, 3, or 5% shear strain. Within the anterior and intermediate regions of the disc when sheared in the anteroposterior direction, both shear and compressive moduli experienced a significant decrease from instantaneous to steady state, while the posterior region’s compressive modulus decreased approximately 5%, and no significant loss of shear modulus was noted. All regions retained their shear modulus within 0.5% of instantaneous values when shear was applied in the mediolateral direction. The results of the disc’s regional shear mechanics suggest an observable and predictable link with the common clinical observation that the posterior region of the disc is most often the zone in which fatigue occurs, which may lead to disc damage and perforation.

Aquaporin 1 expression in human temporomandibular disc

Aquaporins (AQPs) are a family of hydrophobic membrane channel proteins. The expression of several AQP isoforms has been investigated in different human tissues, including the orofacial region. However, information on the role and localization of AQP1 in joints is limited, and no data are available on aquaporins in the normal temporomandibular joint (TMJ) disc. Sixteen human TMJ discs without degenerative changes were taken from fresh cadavers to investigate the presence and distribution of AQP1 by immunohistochemistry. The aim of the study was to gain additional insights into the biomolecular composition of aquaporins and their role in homeostasis of the TMJ. Porcine TMJ discs were also studied by Western blotting for comparison. Scattered AQP1 immunoexpression was detected in human disc cells, documenting its constitutive expression, but differences amongst the three disc regions were not significant. AQP1 expression was demonstrated in porcine TMJ disc by Western blotting. Our findings suggest that AQP1 is normally expressed in the TMJ disc and confirm a role for it in the maintenance of TMJ homeostasis. Further studies are needed to elucidate expression patterns of aquaporins in diseased TMJ discs.

Effect of Mechanical Loading on Electrical Conductivity in Porcine TMJ Discs

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 region-dependent. 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.

Regional cell density distribution and oxygen consumption rates in porcine TMJ discs: an explant study

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. 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 recorded 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. The overall cell density [mean, 95% confidence interval (CI)] was 51.3 (21.3-81.3) × 10(6) cells/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/h and 28.7 (12.2-45.2)nmol/10(6)cells/h, 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%. 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.