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.

Approaches to improve integration and regeneration of an ex vivo derived temporomandibular joint disc scaffold with variable matrix composition.

Due to their natural biochemical and biomechanical characteristics, using ex vivo tissues as platforms for guided tissue regeneration has become widely accepted, however subsequent attachment and integration of these constructs in vivo is often overlooked. A decellularized porcine temporomandibular joint (TMJ) disc has shown promise as a scaffold to guide disc regeneration and preliminary work has shown the efficacy of surfactant (SDS) treatment within the fibrocartilaginous disc to remove cellular components. The majority of studies focus on the intermediate region of the disc (or disc proper). Using this approach, inherent attachment tissues can be maintained to improve construct stability and integration within the joint. Unlike human disc attachment tissue, the porcine attachment tissues have high lipid content which would require a different processing approach to remove immunogenic components. In order to examine the effect of delipidation on the attachment tissue properties, SDS and two organic solvent mixtures (acetone/ethanol and chloroform/methanol) were compared. Lipid and cellular solubilization, ECM alteration, and seeded human mesenchymal stem cell (MSC) morphology and viability were assessed. Quantitative analysis showed SDS treatments did not effectively delipidate the attachment tissues and cytotoxicity was noted toward MSC in these regions. Acetone/ethanol removed cellular material but not all lipids, while chloroform/methanol removed all visible lipid deposits but residual porcine cells were observed in histological sections. When a combination of approaches was used, no residual lipid or cytotoxicity was noted. Preparing a whole TMJ graft with a combined approach has the potential to improve disc integration within the native joint environment.

Determination of VEGF, collagen type 1 and versican in the discus articularis of the temporomandibular joint in relation to dental status.

The aim of this study was the investigation and comparison of the presence of vascular endothelial growth factor (VEGF), collagen type 1 and the proteoglycan versican in the discus articularis in relation to dental status (full dentition [1], partial dentition [2] and edentulous [3]). The right disci articulares were removed from 17 donated bodies (6 with full dentition, 5 with partial dentition and 6 edentulous). The specimens were immunohistochemically stained for VEGF, collagen type 1 and versican. Semiquantitative analysis of the disci was conducted within the groups based on the intensity of immunoreactivity of VEGF, collagen type 1 and versican. In addition, a pairwise comparison was carried out between the three experimental groups. The results revealed significantly higher immunoreactivity for VEGF and versican in groups 2 and 3 than in group 1. Conversely, determination of immunoreactivity was significantly higher in group 1 for collagen type 1 than in the other two groups. These results indicate an elevated presence of the proteoglycan versican and the neoangiogenesis factor VEGF when the occlusal supporting zone has been lost. By contrast, detection of collagen type 1 is reduced. The loss of collagen type 1 and rise in versican and VEGF suggest increasing degeneration when the supporting zone is lost due to the loss of teeth.

Proteoglycan expression is influenced by mechanical load in TMJ discs.

The expression and assembly of the extracellular matrix are profoundly associated with adaptive and pathological responses of the temporomandibular joint (TMJ). To better understand the adaptive responses of the TMJ disc to mechanical loading, we examined the expression of 2 modular proteoglycans and 10 small leucine-rich proteoglycans (SLRPs) at the mRNA and protein levels and determined the contents of proteoglycan-related glycosaminoglycans (GAGs) in rat TMJ discs in response to altered mechanical loading caused by an incisal bite plane. One hundred thirty 7-week-old male Wistar rats were assigned to control and bite plane groups. TMJ disc thickness and the intensity of toluidine blue staining of metachromasia increased in the posterior band after 2 weeks of wearing the bite plane. GAG content increased significantly in the bite plane group after 2 weeks. Quantitative real-time RT-PCR (reverse transcription polymerase chain reaction) analysis indicated that biglycan and chondroadherin mRNA levels increased after 2 weeks and that the level of decorin mRNA increased at 4 weeks. Versican mRNA levels increased after 3 weeks, particularly for the V0 and V1 versican isoforms, which carry more GAG attachment sites than do the V2 and V3 isoforms. Western analysis demonstrated a corresponding increase in the levels of versican, biglycan, and decorin core proteins at 4 weeks in the bite plane group. These results indicate that mechanical loading differentially influences proteoglycan mRNA expression and protein accumulation in the TMJ disc. The change in proteoglycan mRNA and protein levels may lead to the modulation of matrix-matrix and cell-matrix interactions and has important biological significance for adaptation to complicated biomechanical requirements and for tissue maintenance in the TMJ disc.

Engineered microporosity: enhancing the early regenerative potential of decellularized temporomandibular joint discs.

The temporomandibular joint (TMJ) disc is susceptible to numerous pathologies that may lead to structural degradation and jaw dysfunction. The limited treatment options and debilitating nature of severe temporomandibular disorders has been the primary driving force for the introduction and development of TMJ disc tissue engineering as an approach to alleviate this important clinical issue. This study aimed to evaluate the efficacy of laser micropatterning (LMP) ex vivo-derived TMJ disc scaffolds to enhance cellular integration, a major limitation to the development of whole tissue implant technology. LMP was incorporated into the decellularized extracellular matrix scaffold structure using a 40 W CO2 laser ablation system to drill an 8×16 pattern with a bore diameter of 120 µm through the scaffold thickness. Disc scaffolds were seeded with human neonatal-derived umbilical cord mesenchymal stem cells differentiated into chondrocytes at a density of 900 cells per mm(2) and then assessed on days 1, 7, 14, and 21 of culture. Results derived from histology, PicoGreen DNA quantification, and cellular metabolism assays indicate that the LMP scaffolds improve cellular remodeling compared to the unworked scaffold over the 21-day culture period. Mechanical analysis further supports the use of the LMP showing the compressive properties of the LMP constructs closely represent native disc mechanics. The addition of an artificial path of infiltration by LMP culminated in improved chondrocyte adhesion, dispersion, and migration after extended culture aiding in recapitulating the native TMJ disc characteristics.

FasL expression in articular discs of human temporomandibular joint and association with osteoarthrosis.

BACKGROUND: Apoptosis is a programme of cell death which does not induce an inflammatory response. Recent previous research has suggested a correlation between temporomandibular internal derangement and apoptosis. Fas ligand (FasL) is an apoptosis-inducing factor, known to trigger apoptosis through distinct signal pathways. This study aims to examine, by immunohistochemistry, the expression of FasL in temporomandibular joint (TMJ) articular discs of patients with anterior disc displacement with reduction (ADDwR) and without reduction (ADDwoR) in patients with and without osteoarthrosis (OA). METHODS: Forty-two (n = 42) TMJ articular discs were divided into two cut-offs: (i) 8 control, 17 ADDwR, 17 ADDwoR, and (ii) without OA (n = 25) and with OA (n = 17). The area of immunostaining was compared statistically between groups (P < 0.05). RESULTS: Statistically significant differences were found in the expression of FasL in TMJ discs between the three groups (P = 0.001). ADDwR presented significant higher FasL expression when compared with ADDwoR (P < 0.001). Significant higher FasL expression was observed in the group without OA (P = 0.001). All patients without OA presented ADDwR, while all the patients with OA presented ADDwoR. CONCLUSION: A higher area of in situ immunostaining of FasL was found in temporomandibular discs with reduction, which is the less severe condition. Moreover, a reduced expression of FasL in the discs of patients with osteoarthrosis was found, suggesting that some aspects of apoptosis might underlie the progression of TMJ disorders.

An interspecies comparison of the temporomandibular joint disc.

The temporomandibular joint (TMJ) disc plays a critical role in normal function of the joint, and many disorders of the TMJ are a result of disc dysfunction. Previous quantitative TMJ characterization studies examined either the human or a specific animal model, but no single study has compared different species, in the belief that differences in joint morphology, function, and diet would be reflected in the material properties of the disc. In this study, we examined topographical biochemical (collagen, glycosaminoglycan, and DNA content) and biomechanical (tensile and compressive) properties of the human TMJ disc, and also discs from the cow, goat, pig, and rabbit. Regional and interspecies variations were identified in all parameters measured, and certain disc characteristics were observed across all species, such as a weak intermediate zone under mediolateral tension. While human discs possessed properties distinct from those of the other species, pig discs were most similar to the human, suggesting that the pig may be a suitable animal model for TMJ bioengineering efforts.

Occurrence and regional distribution of TRAIL and DR5 on temporomandibular joint discs: comparison of disc derangement with and without reduction.

BACKGROUND: Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is an apoptosis-inducing member of the TNF gene family which triggers apoptotic signals by interaction with its receptors. It has been suggested to be a major contributing factor to tissue degeneration. OBJECTIVE: The present study investigated, through immunohistochemistry, the regional expression of TRAIL and in temporomandibular joint (TMJ) disc of anterior disc displacement with reduction (ADDwR) and without reduction (ADDwoR) patients, to help determine the relationship between TMJ disc displacement and apoptosis. STUDY DESIGN: We studied 18 TMJ diseased discs affected by disc displacement without or with reduction and 4 normal TMJ discs. Specimens were processed for immunohistochemistry to evaluate TRAIL and its receptor DR5 expression. RESULTS: Disc tissues from internal derangements (both ADDwR and ADDwoR) exhibited a much higher percentage of TRAIL- and DR5-positive cells as well as stain intensity compared with normal tissue though with regional variation according to the portion of the disc. There was a significantly higher percentage of stained cells in the posterior disc attachment compared with the anterior or intermediate bands of both ADDwR and ADDwoR discs for TRAIL and DR5. CONCLUSIONS: TRAIL and DR5 are overexpressed in displaced human TMJ disc, especially in the posterior disc attachment. These results suggest a possible pivotal role of the TRAIL/DR5 system in TMJ disc degeneration.

The region-dependent biphasic viscoelastic properties of human temporomandibular joint discs under confined compression.

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+/-11.47kPa compared to 22.0+/-5.15kPa). Permeability showed the inverse trend with the largest values in the anterior and posterior regions (8.51+/-1.36x10(-15)m(4)/Ns compared with 3.75+/-0.72x10(-15)m(4)/Ns). 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 (0.01-3Hz) with average moduli of 171.8-609.3kPa compared with 454.6-1613.0kPa 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.

Biochemical analysis of the porcine temporomandibular joint disc.

Tissue engineering can be a boon in treating lesions of the disc in the temporomandibular joint (TMJ). Unfortunately, little is known about its biochemical content, so we analysed the discs of six slaughtered pigs. We measured the content and distribution of total DNA, glycosaminoglycan, and collagen. The mean (S.D.) content of DNA was 0.14% (0.08%) of the dry weight, of glycosaminoglycan 0.96% (0.39%), and of collagen 68.2% (14.5%). There were no significant differences from top to bottom, but from front to back the smallest concentration of glycosaminoglycan was in the posterior band, and the highest concentration of collagen was in the intermediate zone. The concentrations of DNA and glycosaminoglycan were higher in the medial than in the lateral area of the disc.

Evaluation of three growth factors in combinations of two for temporomandibular joint disc tissue engineering.

Tissue engineering of the temporomandibular joint disc could be a great value in treatments that require discectomy. Potential benefit has been found in the use of three growth factors: insulin-like growth factor-I, basic fibroblast growth factor and transforming growth factor-beta1 in maintaining disc-like tissue in culture. In the present study, these three growth factors were combined in pairs and tested at two different concentrations over a 6-week period. All combinations of the growth factors appear to be beneficial since only three-quarters of the control constructs (without growth factors) retained mechanical integrity, compared with the majority of constructs exposed to growth factors. Importantly, the concentrations of the presented growth factors had a significant impact on the cellularity of constructs at Week 6. When a high concentration of the two growth factors was used, at least twice as many cells remained in the constructs compared with controls.

Distribution of mucopolysaccharides and glycoproteins in the articular discs of temporomandibular joints in human fetuses.

AIMS: To evaluate the distribution of mucopolysaccharides and glycoproteins in the articular discs of temporomandibular joints (TMJs) in human fetuses at different stages of development in order to test the hypothesis that the development and histological maturation of the articular disc has already begun by the 12th week of gestation. METHODS: Eighteen human fetuses at gestational ages 12, 14, and 16 weeks were used (6 fetuses of each age). Sections (6 microm wide) of the articular discs were stained with trichromic stain for collagen fibers, Mayer's mucicarmine for mucopolysaccharides, and Schiff's periodic acid reaction for glycoproteins. The densities of the stained zones were measured by means of Image J software. The nonparametric Kruskal-Wallis test was used to evaluate the differences among stained zones in the 3 fetus groups. RESULTS: TMJ tissues of four of the six 12-week-old fetuses were stained positive for collagen fibers, mucopolysaccharides, and glycoproteins. In these fetuses the stain was localized to the articular posterior area and was denser in that area than in the middle and anterior areas. The stained areas in the 14-week-old fetuses were distributed throughout the articular discs, with isolated clear unstained areas. The stained areas of all the articular discs of the 16-week-old fetuses were more compact than those of the 12-and 14-week-old fetuses. In all the fetuses examined, the collagen fibers along the articular disc had a wavy appearance. The fossa of the temporal bone was observed in all the fetuses as a straight structure that was similar in the 3 fetus groups. The densities (mean +/- SD) of the stained zones were 38.36% +/- 3.39%, 59.5% +/- 1.56%, and 94.04% +/- 2.04% for 12, 14, and 16 weeks of gestation, respectively; these densities were significantly different (chi2 = 15.16; df = 2, P < .001). CONCLUSION: This study indicates that mucopolysaccharides and glycoproteins as well as collagen fibers are present at 12, 14, and 16 weeks of gestation. This suggests that the histological maturation of the articular disc has already begun at the 12th week and is complete by the 16th week of gestation.

Histochemistry of the foetal human temporomandibular joint articular disc.

The human temporomandibular joint (TMJ) develops from mesenchymal cells that form condensations appearing as condylar and temporal blastema which give rise to the respective anterior and posterior regions of the TMJ articular disc. Previous reports have shown the foetal disc to be avascular, with a high content of organized collagen fibres and a lesser content of elastic fibres. In this study, the articular discs from TMJs of a human foetus at age 22 weeks were evaluated. At this stage of intrauterine (i.u.) development, the disc was found to be a highly cellular, biconcave structure with a dense arrangement of collagen fibres. Cell density was not uniform, with increased density in the intermediate band relative to the anterior and posterior bands. In contrast to earlier reports, capillaries containing red blood cells were observed along the inferior surface of the disc. Immunohistochemical staining for proteoglycans and glycosaminoglycans (GAGS) revealed abundant chondroitin sulphate proteoglycan (CSPG) and hyaluronic acid in the disc while relatively little amounts of dermatan sulphate proteoglycan II (DSPGII) were found. No keratin sulphate proteoglycan (KSPG) was detectable. Foetal human TMJ articular discs at this age were found to have morphology and regional characteristics similar to adult discs.

The immunohistochemical distribution of vimentin in human temporomandibular joint samples.

The aim of this investigation was to evaluate the immunohistochemical distribution of vimentin in the temporomandibular joint (TMJ) and to compare it with the control specimens. Immunohistochemical distribution in the disc and synovial membrane in 30 human TMJ (internal derangement of TMJ, n = 20; and control, n = 10) was studied immunohistologically using paraffin-embedded tissue and specific anti-human vimentin monoclonal antibody. Vimentin expression was distributed in chondrocyte-like cells, synovial cells and endothelial cells. There was an obvious distinction of vimentin immunoreactivity between the control specimens and internal derangement cases, in the posterior and/or anterior loose connective tissues. In particular, intensive vimentin expression was detected in the hypertrophic synovial membrane of internal derangement cases. The findings of the present study suggest that vimentin might be an important marker of pathological hypertrophy of the synovial membrane and/or connective tissue with internal derangement of TMJ.

Soluble tumour necrosis factor receptors in synovial fluids from temporomandibular joints with painful anterior disc displacement without reduction and osteoarthritis.

The objective of this study was to detect soluble-form tumour necrosis factor receptors (sTNFRs) in temporomandibular joint (TMJ) synovial fluid aspirates, and to compare the sTNFR concentrations between painful anterior disc displacement without reduction and osteoarthritis (ADDwoR/OA) and asymptomatic TMJs. Synovial fluid was sampled from the superior TMJ cavity of 11 painful ADDwoR/OA cases (mean age: 36.9 years) and 10 asymptomatic females (mean age: 24.7 years) by diluted aspiration. The concentrations of sTNFR-I and -II in the synovial fluid were measured using human sTNFR-I and -II enzyme-linked immunosorbent assays. The total protein concentrations in synovial fluids were measured using a bicinchoninic acid protein assay kit. All data were normalised to the total protein concentration of each sample.Two-way factorial analysis of variance and post hoc multiple comparison revealed that: (1). mean normalised sTNFR-I and -II concentrations were higher in TMJ synovial aspirates from ADDwoR/OA patients than from healthy controls; (2). in the ADDwoR/OA patients and the healthy controls, the sTNFR-I concentration in TMJ synovial aspirates was higher than the sTNFR-II concentration; and (3). high TMJ synovial aspirate sTNFR-II seemed to be associated with less TMJ pain and a less restricted range of mouth opening in the ADDwoR/OA patients. The concentrations of sTNFRs in TMJ synovial fluid are higher in the presence of painful ADDwoR/OA, which could modulate intracapsular inflammation.

Immunohistochemical analysis of Bcl-2 and Bax oncoproteins in rabbit craniomandibular joint.

The purpose of this study was to elucidate the expression of proto-oncogene Bcl-2 (anti-apoptotic) and Bax (pro-apoptotic) in fibrocartilage of the disc and hyaline cartilage of the condyle in the rabbit craniomandibular joint (CMJ). Ten New Zealand white rabbit heads were used. Sections were processed by the immunohistochemical techniques using mouse anti-Bcl-2 and anti-Bax antibodies. Intensity levels of immunostaining in condylar cartilage were quantified by a computer-image system. Immunoreactivity for Bcl-2 was mainly observed in the cytoplasm of the reserve cell and chondrocytic cell layers. A mild heterogeneous Bax expression was detected in the cytoplasm of chondrocytes of the upper hypertrophic layer and a few cells of the chondrocytic layer. The cytoplasm of chondrocytes in the disc exhibited a high intensity for Bcl-2, while Bax activity was only sporadically observed. We have shown that Bcl-2 and Bax proteins are present in CMJ cartilage and their expression patterns suggest that these oncoproteins are involved in chondrocyte survival or death via apoptotic pathways.

Biomechanical behavior of the temporomandibular joint disc.

The temporomandibular joint (TMJ) disc consists mainly of collagen fibers and proteoglycans constrained in the interstices of the collagen fiber mesh. This construction results in a viscoelastic response of the disc to loading and enables the disc to play an important role as a stress absorber during function. The viscoelastic properties depend on the direction (tension, compression, and shear) and the type of the applied loading (static and dynamic). The compressive elastic modulus of the disc is smaller than its tensile one because the elasticity of the disc is more dependent on the collagen fibers than on the proteoglycans. When dynamic loading occurs, the disc is likely to behave less stiffly than under static loading because of the difference of fluid flow through and out of the disc during loading. In addition, the mechanical properties change as a result of various intrinsic and extrinsic factors in life such as aging, trauma, and pathology. Information about the viscoelastic behavior of the disc is required for its function to be understood and, for instance, for a suitable TMJ replacement device to be constructed. In this review, the biomechanical behavior of the disc in response to different loading conditions is discussed.

The proteoglycan contents of the temporomandibular joint disc influence its dynamic viscoelastic properties.

The collagen fibers and proteoglycans in the disc of temporomandibular joint provide resistance to various loadings. Thus far, however, the role of the proteoglycans in determining the viscoelastic properties of the disc has not been investigated. In the present study the hypothesis was tested that the viscoelastic behavior of the disc decreases by the removal of proteoglycans. In 32 bovine discs, dynamic tensile tests with a wide range of frequencies were performed. Before testing, specimens were treated with different concentrations of alpha-amylase to remove proteoglycans. As the frequency increased from 0.1 to 100 Hz, the disc became more viscoelastic. Increasing the concentration of alpha-amylase significantly decreased its viscoelasticity. It was concluded that proteoglycans play an important role in determining the viscoelastic properties of the disc and, therefore, give the disc a greater capacity for distributing and reducing stresses.

Structure and function of the temporomandibular joint disc: implications for tissue engineering.

The temporomandibular joint (TMJ) disc is a little understood structure that, unfortunately, exhibits a plethora of pathologic disorders. Tissue engineering approaches may be warranted to address TMJ disc pathophysiology, but first a clear understanding of structure-function relationships needs to be developed, especially as they relate to the regenerative potential of the tissue. In this review, we correlate the biochemical content of the TMJ disc to its mechanical behavior and discuss what this correlation infers for tissue engineering studies of the TMJ disc. The disc of the TMJ exhibits a somewhat biconcave shape, being thicker in the anterior and posterior bands and thinner in the intermediate zone. The disc, which is certainly an anisotropic and nonhomogeneous tissue, consists almost entirely of type I collagen with trace amounts of type II and other types. In general, collagen fibers in the intermediate zone appear to run primarily in an anteroposterior direction and in a ringlike fashion around the periphery. Collagen orientation is reflected in higher tensile stiffness and strength in the center anteroposteriorly than mediolaterally and in the anterior and posterior bands than the intermediate zone mediolaterally. Tensile tests have shown the disc is stiffer and stronger in the direction of the collagen fibers. Elastin fibers in general appear along the collagen fibers and most likely function in restoring and retaining disc form after loading. The 2 primary glycosaminoglycans of the disc by far are chondroitin sulfate and dermatan sulfate, although their distribution is not clear. Compression studies are conflicting, but evidence suggests the disc is compressively stiffest in the center. Only a few tissue engineering studies of the TMJ disc have been performed to date. Tissue engineering studies must take advantage of existing information for experimental design and construct validation, and more research is necessary to characterize the disc to create a clearer picture of our goals in tissue engineering the TMJ disc.

Dynamic properties of bovine temporomandibular joint disks change with age.

The temporomandibular joint disk exhibits morphological and biochemical age-related changes. However, the possible age-related changes of the dynamic viscoelasticity in the disk are unclear. We tested the hypothesis that the dynamic viscoelastic properties of the disk change with age. Thirty-six disks from young-adult, adult, and mature-adult cattle were used for dynamic tensile tests. In all disks, the magnitudes of the complex modulus, the storage modulus, and the loss modulus increased as the frequency increased. The mature-adult disks had higher values of these moduli than did the younger disks. The loss tangent ranged from 0.1 to 0.3, which means that the disk has relatively large elasticity and relatively small viscosity. It was concluded that both the elasticity and viscosity of the disk increase with age. This may reflect age-related changes in biochemical composition.