Nondestructive fluorescence lifetime imaging and time-resolved fluorescence spectroscopy detect cartilage matrix depletion and correlate with mechanical properties.

Tissue engineers utilize a battery of expensive, time-consuming and destructive techniques to assess the composition and function of engineered tissues. A nondestructive solution to monitor tissue maturation would reduce costs and accelerate product development. As a first step toward this goal, two nondestructive, label-free optical techniques, namely multispectral fluorescent lifetime imaging (FLIm) and time-resolved fluorescence spectroscopy (TRFS), were investigated for their potential in evaluating the biochemical and mechanical properties of articular cartilage. Enzymatic treatments were utilized to selectively deplete cartilage of either collagen or proteoglycan, to produce a range of matrix compositions. Samples were assessed for their optical properties using a fiber-coupled optical system combining FLIm and TRFS, their biochemical and mechanical properties and by histological staining. Single and multivariable correlations were performed to evaluate relationships among these properties. FLIm- and TRFS-derived measurements are sensitive to changes in cartilage matrix and correlate with mechanical and biochemical assays. Mean fluorescence lifetime values extracted from FLIm images (375-410 nm spectral band) showed strong, specific correlations with collagen content (R2 = 0.79, p < 0.001) and tensile properties (R2 = 0.45, p = 0.02). TRFS lifetime measurements centered at 520 nm (with a 5 nm bandwidth) possessed strong, specific correlations with proteoglycan content (R2 = 0.59, p = 0.001) and compressive properties (R2 = 0.71, p < 0.001). Nondestructive optical assessment of articular cartilage, using a combination of FLIm- and TRFS-derived parameters, provided a quantitative method for determining tissue biochemical composition and mechanical function. These tools hold great potential for research, industrial and clinical settings.

Tendon and ligament as novel cell sources for engineering the knee meniscus.

OBJECTIVE: The application of cell-based therapies in regenerative medicine is hindered by the difficulty of acquiring adequate numbers of competent cells. For the knee meniscus in particular, this may be solved by harvesting tissue from neighboring tendons and ligaments. In this study, we have investigated the potential of cells from tendon and ligament, as compared to meniscus cells, to engineer scaffold-free self-assembling fibrocartilage. METHOD: Self-assembling meniscus-shaped constructs engineered from a co-culture of articular chondrocytes and either meniscus, tendon, or ligament cells were cultured for 4 weeks with TGF-ß1 in serum-free media. After culture, constructs were assessed for their mechanical properties, histological staining, gross appearance, and biochemical composition including cross-link content. Correlations were performed to evaluate relationships between biochemical content and mechanical properties. RESULTS: In terms of mechanical properties as well as biochemical content, constructs engineered using tenocytes and ligament fibrocytes were found to be equivalent or superior to constructs engineered using meniscus cells. Furthermore, cross-link content was found to be correlated with engineered tissue tensile properties. CONCLUSION: Tenocytes and ligament fibrocytes represent viable cell sources for engineering meniscus fibrocartilage using the self-assembling process. Due to greater cross-link content, fibrocartilage engineered with tenocytes and ligament fibrocytes may maintain greater tensile properties than fibrocartilage engineered with meniscus cells.

Inhibition of CDK9 prevents mechanical injury-induced inflammation, apoptosis and matrix degradation in cartilage explants.

Joint injury often leads to post-traumatic osteoarthritis (PTOA). Acute injury responses to trauma induce production of pro-inflammatory cytokines and catabolic enzymes, which promote chondrocyte apoptosis and degrade cartilage to potentiate PTOA development. Recent studies show that the rate-limiting step for transcriptional activation of injury response genes is controlled by cyclin-dependent kinase 9 (CDK9), and thus it is an attractive target for limiting the injury response. Here, we determined the effects of CDK9 inhibition in suppressing the injury response in mechanically-injured cartilage explants. Bovine cartilage explants were injured by a single compressive load of 30 % strain at 100 %/s, and then treated with the CDK9 inhibitor Flavopiridol. To assess acute injury responses, we measured the mRNA expression of pro-inflammatory cytokines, catabolic enzymes, and apoptotic genes by RT-PCR, and chondrocyte viability and apoptosis by TUNEL staining. For long-term outcome, cartilage matrix degradation was assessed by soluble glycosaminoglycan release, and by determining the mechanical properties with instantaneous and relaxation moduli. Our data showed CDK9 inhibitor markedly reduced injury-induced inflammatory cytokine and catabolic gene expression. CDK9 inhibitor also attenuated chondrocyte apoptosis and reduced cartilage matrix degradation. Lastly, the mechanical properties of the injured explants were preserved by CDK9 inhibitor. Our results provide a temporal profile connecting the chain of events from mechanical impact, acute injury responses, to the subsequent induction of chondrocyte apoptosis and cartilage matrix deterioration. Thus, CDK9 is a potential disease-modifying agent for injury response after knee trauma to prevent or delay PTOA development.

Hypoxia-induced collagen crosslinking as a mechanism for enhancing mechanical properties of engineered articular cartilage.

OBJECTIVE: The focus of tissue engineering of neocartilage has traditionally been on enhancing extracellular matrix and thus biomechanical properties. Emphasis has been placed on the enhancement of collagen type and quantity, and, concomitantly, tensile properties. The objective of this study was to improve crosslinking of the collagen network by testing the hypothesis that hypoxia could promote pyridinoline (PYR) crosslinks and, thus, improve neocartilage's tensile properties. METHODS: Chondrocyte expression of lysyl oxidase (LOX), an enzyme responsible for the formation of collagen PYR crosslinks, was first assessed pre- and post- hypoxia application. Then, the mechanical properties of self-assembled neocartilage constructs were measured, after 4 weeks of culture, for groups exposed to 4% O2 at different initiation times and durations, i.e., during the 1st and 3rd weeks, 3rd and 4th weeks, 4th week only, continuously after cell seeding, or never. RESULTS: Results showed that LOX gene expression was upregulated ∼20-fold in chondrocytes in response to hypoxia. Hypoxia applied during the 3rd and 4th weeks significantly increased PYR crosslinks without affecting collagen content. Excitingly, neocartilage tensile properties were increased ∼2-fold. It should be noted that these properties exhibited a distinct temporal dependence to hypoxia exposure, since upregulation of these properties was due to hypoxia applied only during the 3rd and 4th weeks. CONCLUSION: These data elucidate the role of hypoxia-mediated upregulation of LOX and subsequent increases in PYR crosslinks in engineered cartilage. These results hold promise toward applying hypoxia at precise time points to promote tensile integrity and direct construct maturation.

Nondestructive testing of native and tissue-engineered medical products: adding numbers to pictures.

Traditional destructive tests are used for quality assurance and control within manufacturing workflows. Their applicability to biomanufacturing is limited due to inherent constraints of the biomanufacturing process. To address this, photo- and acoustic-based nondestructive testing has risen in prominence to interrogate not only structure and function, but also to integrate quantitative measurements of biochemical composition to cross-correlate structural, compositional, and functional variances. We survey relevant literature related to single-mode and multimodal nondestructive testing of soft tissues, which adds numbers (quantitative measurements) to pictures (qualitative data). Native and tissue-engineered articular cartilage is highlighted because active biomanufacturing processes are being developed. Included are recent efforts and prominent trends focused on technologies for clinical and in-process biomanufacturing applications.

Public knowledge, beliefs and practices in Greece about cancer etiology and prevention.

This questionnaire survey of the parents of elementary schoolchildren in Greece assessed their self-reported knowledge, attitudes and practices towards smoking, diet and exposure to X-radiation. A random sample of 403 household units (379 fathers and 391 mothers) was selected from urban areas of Thessaloniki. Half of the parents who smoked (50.1%) did not ask for permission to smoke from other people and 66.0% regularly smoked in front of their children. On the other hand, 82.6% of smokers recognized the existence of a health risk to children from passive smoking. Parents overestimated the role of nuclear tests and accidents as factors in carcinogenesis. Two-thirds of parents did not know the beneficial role of the Mediterranean diet to health, and dietary intake analysis showed some departure from the traditional Mediterranean diet. A reconsideration of the policy regarding health education programmes concerning cancer prevention in Greece is needed.

Shear stress induced by fluid flow produces improvements in tissue-engineered cartilage.

Tissue engineering aims to create implantable biomaterials for the repair and regeneration of damaged tissues. In vitro tissue engineering is generally based on static culture, which limits access to nutrients and lacks mechanical signaling. Using shear stress is controversial because in some cases it can lead to cell death while in others it promotes tissue regeneration. To understand how shear stress works and how it may be used to improve neotissue function, a series of studies were performed. First, a tunable device was designed to determine optimal levels of shear stress for neotissue formation. Then, computational fluid dynamics modeling showed the device applies fluid-induced shear (FIS) stress spanning three orders of magnitude on tissue-engineered cartilage (neocartilage). A beneficial window of FIS stress was subsequently identified, resulting in up to 3.6-fold improvements in mechanical properties of neocartilage in vitro. In vivo, neocartilage matured as evidenced by the doubling of collagen content toward native values. Translation of FIS stress to human derived neocartilage was then demonstrated, yielding analogous improvements in mechanical properties, such as 168% increase in tensile modulus. To gain an understanding of the beneficial roles of FIS stress, a mechanistic study was performed revealing a mechanically gated complex on the primary cilia of chondrocytes that is activated by FIS stress. This series of studies places FIS stress into the arena as a meaningful mechanical stimulation strategy for creating robust and translatable neotissues, and demonstrates the ease of incorporating FIS stress in tissue culture.

Systematic assessment of growth factor treatment on biochemical and biomechanical properties of engineered articular cartilage constructs.

OBJECTIVE: To determine the effects of bone morphogenetic protein-2 (BMP-2), insulin-like growth factor (IGF-I), and transforming growth factor-beta1 (TGF-beta1) on the biochemical and biomechanical properties of engineered articular cartilage constructs under serum-free conditions. METHODS: A scaffoldless approach for tissue engineering, the self-assembly process, was employed. The study consisted of two phases. In the first phase, the effects of BMP-2, IGF-I, and TGF-beta1, at two concentrations and two dosage frequencies each were assessed on construct biochemical and biomechanical properties. In phase II, the effects of growth factor combination treatments were determined. Compressive and tensile mechanical properties, glycosaminoglycan (GAG) and collagen content, histology for GAG and collagen, and immunohistochemistry (IHC) for collagen types I and II were assessed. RESULTS: In phase I, BMP-2 and IGF-I treatment resulted in significant, >1-fold increases in aggregate modulus, accompanied by increases in GAG production. Additionally, TGF-beta1 treatment resulted in significant, approximately 1-fold increases in both aggregate modulus and tensile modulus, with corresponding increases in GAG and collagen content. In phase II, combined treatment with BMP-2 and IGF-I increased aggregate modulus and GAG content further than either growth factor alone, while TGF-beta1 treatment alone remained the only treatment to also enhance tensile properties and collagen content. DISCUSSION: This study determined systematically the effects of multiple growth factor treatments under serum-free conditions, and is the first to demonstrate significant increases in both compressive and tensile biomechanical properties as a result of growth factor treatment. These findings are exciting as coupling growth factor application with the self-assembly process resulted in tissue engineered constructs with functional properties approaching native cartilage values.

Biomaterial effects in articular cartilage tissue engineering using polyglycolic acid, a novel marine origin biomaterial, IGF-I, and TGF-beta 1.

Bovine articular chondrocytes were seeded on either polyglycolic acid (PGA) non-woven mesh scaffolds or a biomatrix from the species Porites lutea (POR). These constructs were cultured for 6 weeks in the presence of insulin-like growth factor (IGF)-I (10 ng/ml or 100 ng/ml) or transforming growth factor (TGF)-beta 1 (5 ng/ml or 30 ng/ml) to determine the in-vitro articular cartilage regeneration capacity of each. Histology, deoxyribonucleic acid content, collagen I and II (immunohistochemistry and enzyme-linked immunosorbent assay), and glycosaminoglycan (GAG) contents were measured at 0 weeks, 2 weeks, and 6 weeks to assess the characteristics of chondrogenesis. Both scaffolds supported the maintenance of the chondrocytic phenotype, as evidenced by the predominance of collagen II and the presence of rounded chondrocytes embedded in lacunae. Regardless of growth factor treatment, cells cultured on PGA scaffolds produced more collagen type II than those cultured on POR. Conversely, by 6 weeks, cells cultured on POR scaffolds produced more GAG than those cultured on PGA scaffolds, again regardless of the growth factor used. Across the two groups, 100 ng/ml of IGF-I had the greatest overall effect in GAG content. This work indicates that PGA and the POR scaffolds are both effective growth matrices for articular cartilage, with each scaffold exhibiting different yet desirable profiles of articular cartilage growth.

Hypoxic chondrogenic differentiation of human embryonic stem cells enhances cartilage protein synthesis and biomechanical functionality.

BACKGROUND: Engineering musculoskeletal cartilages with stem cells remains a challenge because researchers must control many factors, including differentiation and cartilage matrix synthesis, particularly collagen II production. Hypoxia has effects on many cellular processes, though few investigations with hypoxia provide quantitative functional data on engineered cartilage. OBJECTIVE: This study investigated the effects of hypoxia on chondrogenesis with human embryonic stem cells (hESCs). METHODS: The experiment comprised two phases, embryoid body (EB) differentiation for 3 wks followed by a scaffold-less tissue engineering strategy called self-assembly for 4 wks. During each phase, hypoxic conditions (2% O(2)) or normoxic conditions (20% O(2)) were applied, and engineered constructs were analyzed for cellular, morphological, biochemical, and biomechanical properties. RESULTS: Hypoxic conditions significantly altered the chondrogenic differentiation process, whereby cells cultured in these conditions had an enhanced ability to produce collagen II (up to 3.4-times), collagen I (up to 2.9-times), and glycosaminoglycans (GAGs) (up to 1.9-times), resulting in better biomechanical functionality (up to three times in tensile modulus and up to four times in compressive properties). Hypoxic cells had a different expression profile than normoxic cells for cluster of differentiation (CD)44, CD105, and platelet derived growth factor receptor (PDGFR)alpha, further emphasizing that hypoxia altered hESC differentiation and suggesting that these markers may be used to predict an hESC-derived cell population's chondrogenic potential. Also, normoxic self-assembly outperformed hypoxic self-assembly in tensile and compressive biomechanical characteristics. CONCLUSIONS: These results show that oxygen availability has dramatic effects on the differentiation and synthetic potentials of hESCs and may have important implications for the development of strategies to engineer cartilage.

Scaffold and growth factor selection in temporomandibular joint disc engineering.

Temporomandibular joint disc tissue-engineering studies commonly fail to produce significant matrix before construct contraction. We hypothesized that poly-L-lactic acid (PLLA) non-woven meshes would limit contraction, allow for comprehensive mechanical evaluation, and maintain viability relative to polyglycolic acid (PGA) non-woven mesh controls. Additionally, we proposed that growth factor stimulation, while limiting contraction, would increase construct properties relative to previous reports. After 4 wks, cell proliferation and matrix deposition were similar between the two meshes, but PGA constructs had contracted significantly. Furthermore, only PLLA constructs could be tested in tension and compression. Additional PLLA constructs were formed, then treated with insulin-like growth factor-1 (10 ng/mL), transforming growth factor-beta 1 (5 ng/mL), or transforming growth factor-beta 3 (5 ng/mL). Transforming growth factor-beta 1 yielded the most cells, collagen, and glycosaminoglycans at 6 wks; these constructs also demonstrated improved mechanics. Analysis of these data demonstrated significant temporomandibular joint disc-engineering potential for PLLA and transforming growth factor-beta 1.

Clinically relevant cell sources for TMJ disc engineering.

Tissue-engineering of the temporomandibular joint (TMJ) disc aims to provide patients with TMJ disorders an option to replace diseased tissue with autologous, functional tissue. This study examined clinically relevant cell sources by comparing costal chondrocytes, dermal fibroblasts, a mixture of the two, and TMJ disc cells in a scaffoldless tissue-engineering approach. It was hypothesized that all constructs would produce matrix relevant to the TMJ disc, but the mixture constructs were expected to appear most like the TMJ disc constructs. Costal chondrocyte and mixture constructs were morphologically and biochemically superior to the TMJ disc and dermal fibroblast constructs, and their compressive properties were not significantly different. Costal chondrocyte constructs produced almost 40 times more collagen and 800 times more glycosaminoglycans than did TMJ constructs. This study demonstrates the ability of costal chondrocytes to produce extracellular matrix that may function in a TMJ disc replacement.

Biochemical and biomechanical characterisation of equine cervical facet joint cartilage.

BACKGROUND: The equine cervical facet joint is a site of significant pathology. Located bilaterally on the dorsal spine, these diarthrodial joints work in conjunction with the intervertebral disc to facilitate appropriate spinal motion. Despite the high prevalence of pathology in this joint, the facet joint is understudied and thus lacking in viable treatment options. OBJECTIVE: The goal of this study was to characterise equine facet joint cartilage and provide a comprehensive database describing the morphological, histological, biochemical and biomechanical properties of this tissue. STUDY DESIGN: Descriptive cadaver studies. METHODS: A total of 132 facet joint surfaces were harvested from the cervical spines of six skeletally mature horses (11 surfaces per animal) for compiling biomechanical and biochemical properties of hyaline cartilage of the equine cervical facet joints. Gross morphometric measurements and histological staining were performed on facet joint cartilage. Creep indentation and uniaxial strain-to-failure testing were used to determine the biomechanical compressive and tensile properties. Biochemical assays included quantification of total collagen, sulfated glycosaminoglycan and DNA content. RESULTS: The facet joint surfaces were ovoid in shape with a flat articular surface. Histological analyses highlighted structures akin to articular cartilage of other synovial joints. In general, biomechanical and biochemical properties did not differ significantly between the inferior and superior joint surfaces as well as among spinal levels. Interestingly, compressive and tensile properties of cervical facet articular cartilage were lower than those of articular cartilage from other previously characterised equine joints. Removal of the superficial zone reduced the tissue's tensile strength, suggesting that this zone is important for the tensile integrity of the tissue. MAIN LIMITATIONS: Facet surfaces were sampled at a single, central location and do not capture the potential topographic variation in cartilage properties. CONCLUSIONS: This is the first study to report the properties of equine cervical facet joint cartilage and may serve as the foundation for the development of future tissue-engineered replacements as well as other treatment strategies.

Improving culture conditions for temporomandibular joint disc tissue engineering.

BACKGROUND: The temporomandibular joint (TMJ) is extremely important for activities like eating and talking, which can become painful and difficult for patients with TMJ dysfunction. Tissue engineering is a potential alternative to current surgical interventions through replacement of diseased or injured tissue with a functional construct. Since research with TMJ disc cells began relatively recently, optimal culturing conditions must be determined. METHODS: Metabolic additives, L-glutamine, L-alanyl-L-glutamine, sodium pyruvate, and insulin, were examined for their effects on TMJ disc cells in monolayer. Effects of L-proline were examined in three-dimensional (3-D) culture at concentrations of 0, 25 and 100 mg/l. RESULTS: The combination of L-glutamine, sodium pyruvate, and insulin improved cell proliferation rates without affecting collagen production or gene expression. No differences were observed in mechanical properties of the engineered constructs; however, collagen and glycosaminoglycan quantities normalized to cell number decreased at the highest concentration of L-proline. CONCLUSION: This work identified supplements for 2-D monolayer expansion. Other supplements or culture conditions still need to be investigated for 3-D tissue production. This work improves upon porcine TMJ disc cell culturing conditions, taking us closer to being able to engineer the TMJ disc.

Design characteristics for temporomandibular joint disc tissue engineering: learning from tendon and articular cartilage.

Tissue engineering of chondrocytic or fibroblastic musculoskeletal tissues has been relatively well studied compared with that of the temporomandibular joint (TMJ) disc. Early attempts at tissue engineering the disc have been misguided owing to a lack of understanding of the composition and function of the TMJ disc. The objective of this review is to compare the TMJ disc with a chondrocytic tissue (hyaline articular cartilage) and a fibroblastic tissue (tendon) to understand better the properties of this fibrocartilaginous tissue. The TMJ disc has 25 times more glycosaminoglycan (GAG) per dry weight than tendon but half that of articular cartilage. The disc's tensile modulus is six times more than cartilage but orders less than tendon. The GAG content and tensile modulus suggest that the TMJ disc is characterized as a tissue between hyaline cartilage and tendon, but the disc appears more tendon like when considering its collagen make-up and cell content. Like tendon, the TMJ disc contains primarily collagen type I at 85 per cent per dry weight, while articular cartilage has 30 per cent less collagen, which is type II. Knowledge of quantitative comparisons between joint tissues can give extensive insight into how to improve tissue engineering of the TMJ 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.

Design, validation, and utilization of an articular cartilage impact instrument.

This paper describes the development and use of an instrument mechanically to impact bovine articular cartilage and record the event using a piezoelectric accelerometer, as well as to carry out post-impact characterization of the tissue. Two levels of impact (low: 6 cm drop height, 18.4 N tup; high: 10 cm drop height, 27.8 N tup) were chosen such that the former did not show gross damage upon inspection, while the latter showed substantial gross damage. Peak stress, time to peak stress, and impact duration were taken from data recorded by the instrument. Three cartilage biomechanical properties (aggregate modulus, Poisson's ratio, and permeability) were acquired by creep indentation, and tissue morphology rated on a standardized scale was also determined. When subjected to the high level of impact, articular cartilage showed statistically significant (p < 0.05) differences in all three impact metrics and morphology. This high level of impact also resulted in a 37 per cent decrease in the aggregate modulus of the tissue. Lower drop heights resulted in more consistent impact curves, demonstrated less standard deviation, and did not change the biomechanical properties of the tissues. With the instrument and techniques described in this study, articular cartilage can be subjected to specific levels of impact in order to study injury biomechanics of the tissue at specific levels of mechanical damage.

Effects of ascorbic acid concentration on the tissue engineering of the temporomandibular joint disc.

The temporomandibular joint (TMJ) disc is a specialized fibrocartilaginous tissue. When the disc becomes an obstacle and becomes damaged, surgeons have no choice but to perform a discectomy. Tissue engineering may provide a novel treatment modality for TMJ disorder patients who undergo discectomy. No studies have been conducted on the most favourable media for TMJ disc cells. The objective of the current study was to examine the effects on biochemical and biomechanical properties of varying ascorbic acid concentrations (0, 25, or 50 microg/ml) on TMJ disc cells seeded on non-woven PGA scaffolds. The ascorbic acid concentration of the 25 microg/ml group resulted in more effective cell seeding of the scaffolds, with 1.53 million cells per construct, by comparison with the 0 and 50 microg/ml groups which had 1.20 million and 1.32 million cells per scaffold respectively. At week 4, the 25 microg/ml group had a higher collagen content than the 0 microg/ml group, with 30.4 +/- 2.7 and 24.9 +/- 3.3 microg of collagen per construct respectively. The 25 microg/ml group had a higher aggregate modulus than the 50 microg/ml group, with values of 6.1 +/- 1.3 and 4.0 +/- 0.9 kPa respectively at week 4. The results of this study indicate that the use of 25 microg/ml of ascorbic acid in culture media is effective for the tissue engineering of the TMJ disc, significantly outperforming media without or with 50 microg/ml of ascorbic acid.

Effects of ageing on the biomechanical properties of rat articular cartilage.

Previous studies have demonstrated that male Sprague Dawley (SD) rats experience age-related bone loss with the same characteristics as that in ageing men. As articular cartilage, like bone, is a critical component of the health and function of the musculoskeletal system, the authors hypothesized that articular cartilage in the untreated male SD rats could be a suitable model for studying the age-related deterioration of articular cartilage in men. To test this hypothesis, male SD rats were killed at between 6 and 27 months. The right femur of each rat was removed. The effects of ageing on the structural integrity of the distal femoral articular cartilage were studied by biomechanical testing with a creep indentation apparatus. The aggregate modulus, Poisson's ratio, permeability, thickness, and percentage recovery of articular cartilage were determined using finite element/non-linear optimization modelling. No significant differences were observed in these biomechanical properties of the distal femoral articular cartilage as a function of age. Therefore, untreated male SD rats appear to be unsuitable for studying the age-related changes of articular cartilage as they occur in men. However, and more intriguingly, it is also possible that ageing does not affect the biomechanical properties of articular cartilage in the absence of cartilage pathology.

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.