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Basic science of articular cartilage and osteoarthritis.

Articular cartilage is a specialized tissue uniquely suited for load distribution with a low-friction articulating surface. Its compressive and tensile properties are determined by its matrix and fluid composition, and are maintained by chondrocytes in the homeostatic joint. Osteoarthritis (OA) is increasingly understood as a family of disorders in which the biomechanical properties of cartilage are altered and ultimately fail as the tissue is degraded by local proteases. Mechanically mediated and cytokine-mediated pathways of cartilage degeneration have been identified in the pathogenesis of OA. Further insight into the basic science of cartilage and OA is necessary to develop diagnostic and treatment strategies for this pervasive disease.

Biomechanical Phenomena↗

A role for the interleukin-1 receptor in the pathway linking static mechanical compression to decreased proteoglycan synthesis in surface articular cartilage.

Loading of articular cartilage during weight bearing is essential for the maintenance of cartilage function. Although certain cyclic loading protocols stimulate extracellular matrix synthesis, constant or static compression decreases proteoglycan and collagen synthesis in cartilage explants. The goal of this study was to determine whether the compression-induced decrease in proteoglycan synthesis involves an interleukin-1 (IL-1) signaling pathway. Cartilage explants were compressed 50% in the presence of IL-1 receptor antagonist (IL-1ra), and the incorporation of [35S]sulfate into macromolecules was measured. IL-1ra increased sulfate incorporation in compressed cartilage but not in cartilage maintained at the in situ thickness (0% compression). IL-1alpha and IL-1beta mRNAs were detected in cartilage compressed 50% for at least 3h, while nitric oxide synthase II mRNA was only detected in cartilage compressed 50% for 6h. The data support a role for the IL-1 receptor in the pathway linking static compression to reduced proteoglycan synthesis.

Animals↗

Matrix loss and synthesis following a single impact load on articular cartilage in vitro.

Articular cartilage biopsies were subjected to a single impact load and the metabolic response of the chondrocytes investigated using radiolabelled precursors for protein ([3H]leucine) and glycosaminoglycan ([35S]sulfate). The severity of the impact was controlled by using different masses and drop heights in a purpose built drop tower. Loss of matrix components was studied by prelabelling prior to loading, the possible repair response by pulse labelling at defined intervals after loading. There was an increase in the loss of both labels from the tissue with increasing severity of impact though the patterns of loss were different. Only 25%-40% of the sulfate was lost over a two week period and the loss increased with the severity of impact. This contrasted with 60% of the leucine being lost over the same period independently of loading. In addition to the loss of synthetic activity caused by cell death, there was a suppression of incorporation immediately following loading. This eventually recovered and increased above control values but the recovery time appeared to depend on the severity of the impact. These results provide preliminary evidence for a repair response.

Animals↗

Identification of a hyaluronic acid-binding protein that interferes with the preparation of high-buoyant-density proteoglycan aggregates from adult human articular cartilage.

Adult human articular cartilage contains a hyaluronic acid-binding protein of Mr 60 000-75 000, which contains disulphide bonds essential for this interaction. The molecule can compete with proteoglycan subunits for binding sites on hyaluronic acid, and can also displace proteoglycan subunits from hyaluronic acid if their interaction is not stabilized by the presence of link proteins. The abundance of this protein in the adult accounts for the reported inability to prepare high-buoyant-density proteoglycan aggregates from extracts of adult human cartilage [Roughley, White, Poole & Mort (1984) Biochem. J. 221, 637-644], whereas the deficiency of the protein in newborn human cartilage allows the normal recovery of proteoglycan aggregates from this tissue. The protein shares many common features with a hyaluronic acid-binding region derived by proteolytic treatment of a proteoglycan aggregate preparation, and this may also represent its origin in the cartilage, with its production increasing during tissue maturation.

Adult↗

A triphasic theory for the swelling and deformation behaviors of articular cartilage.

Swelling of articular cartilage depends on its fixed charge density and distribution, the stiffness of its collagen-proteoglycan matrix, and the ion concentrations in the interstitium. A theory for a tertiary mixture has been developed, including the two fluid-solid phases (biphasic), and an ion phase, representing cation and anion of a single salt, to describe the deformation and stress fields for cartilage under chemical and/or mechanical loads. This triphasic theory combines the physico-chemical theory for ionic and polyionic (proteoglycan) solutions with the biphasic theory for cartilage. The present model assumes the fixed charge groups to remain unchanged, and that the counter-ions are the cations of a single-salt of the bathing solution. The momentum equation for the neutral salt and for the intersitial water are expressed in terms of their chemical potentials whose gradients are the driving forces for their movements. These chemical potentials depend on fluid pressure p, salt concentration c, solid matrix dilatation e and fixed charge density cF. For a uni-uni valent salt such as NaCl, they are given by mu i = mu io + (RT/Mi)ln[gamma 2 +/- c(c + cF)] and mu w = mu wo + [p-RT phi (2c + cF) + Bwe]/pwT, where R, T, Mi, gamma +/-, phi, pwT and Bw are universal gas constant, absolute temperature, molecular weight, mean activity coefficient of salt, osmotic coefficient, true density of water, and a coupling material coefficient, respectively. For infinitesimal strains and material isotropy, the stress-strain relationship for the total mixture stress is sigma = - pI-TcI + lambda s(trE)I + 2 musE, where E is the strain tensor and (lambda s, mu s) are the Lamé constants of the elastic solid matrix. The chemical-expansion stress (-Tc) derives from the charge-to-charge repulsive forces within the solid matrix. This theory can be applied to both equilibrium and non-equilibrium problems. For equilibrium free swelling problems, the theory yields the well known Donnan equilibrium ion distribution and osmotic pressure equations, along with an analytical expression for the "pre-stress" in the solid matrix. For the confined-compression swelling problem, it predicts that the applied compressive stress is shared by three load support mechanisms: 1) the Donnan osmotic pressure; 2) the chemical-expansion stress; and 3) the solid matrix elastic stress. Numerical calculations have been made, based on a set of equilibrium free-swelling and confined-compression data, to assess the relative contribution of each mechanism to load support. Our results show that all three mechanisms are important in determining the overall compressive stiffness of cartilage.

Biomechanical Phenomena↗

Differences in collagen metabolism between normal and osteoarthritic human articular cartilage.

Normal human articular cartilage synthesizes only one type of a chain, which exhibits the chromatographic behavior of the alpha(l)(II) chains described for chick and bovine cartilage. Osteoarthritic cartilage, on the other hand, synthesizes in addition a collagen containing alpha(2) chains and beta components. The different structural features of the two types of collagen may account for some of the functional defects of osteoarthritic cartilage.

Cartilage, Articular↗

Growth factors and articular cartilage.

Studies of articular cartilage over the decades have demonstrated a surprisingly brisk rate of synthesis of the matrix proteins which appears to vary considerably with metabolic, physicochemical and pathological state of the tissue. It has become evident that much of this activity is directed by low molecular weight protein mediators which act at specific receptor sites. Platelet derived growth factor (PDGF) is of limited action in normal cartilage, but insulin and its analogues, insulin growth factor-I and II are powerful stimulants of DNA synthesis. Basic fibroblast growth factor stimulates both DNA and protein synthesis and works synergistically with other factors. Transforming growth factor beta potentiates the action of the mitogens and enhances and regulates proteoglycan synthesis. These actions may be of special importance in osteoarthritis and lacerative injury to cartilage.

Cartilage, Articular↗

The preservation of articular cartilage.

Slivers of articular cartilage were stored in Ham's medium, plasma, polyvinyl pyrrolidone, and dimethyl sulphoxide at 0, -20, 4, and 38C. Survival was monitored by potassium determinations and autoradiography using S-35 and by matrix staining. Survival of 48 days was obtained in Ham's medium at 4C.

Animals↗

[The early development of the articular cartilage. IV. The metamorphosing cartilage].

The definite articular cartilage originate from 2 anlagen, the primordial tangential layer and the greater part including the joint bone plate from the metamorphosing cartilage. The tangential layer grow by apposition from the perichondrium. Additional the layer becomes also dilatated as a result of the growing volume of the ossification center. In this way the Lamina splendens with residues of cells may be formed. The chondrocytes resemble partly fibroblasts, in older animals possibly even tendocytes. Moreover the cells exhibit a varying different shape. Today it is impossible to interpret the polymorphism of the cells. In the primordial state, the chondrocyts are embedded in a network from thin cartilage fibrils. Later on collagen fibrils from varied thickness (up to 900 A) are formed. The fibrils run only partly parallel to each other, in general they form a network, in which they cross with a low angle. There are great local differences in the fibrillar structure by the same animal.

Animals↗

Interaction of polymorphonuclear leukocytes with immune complexes trapped in rheumatoid articular cartilage.

When rheumatoid articular cartilage samples were incubated with normal polymorphonuclear leukocytes (PMN) in vitro, large numbers of PMN were seen attached to the articular surface. As observed by electron microscopy, significant numbers of these cells invaded the cartilage tissue and phagocytosed amorphous material which presumably contained immune complexes. In control cartilage from osteoarthritic, pyogenic, and normal subjects, only a few PMN were attached to the articular surface after incubation with PMN. These results demonstrate that immune complexes trapped in the superficial region of the rheumatoid articular cartilage may play an important role in the destruction of cartilage by the release of lysosomal enzymes from PMN.

Antigen-Antibody Complex↗

Early changes in the composition of rabbit articular cartilage following experimentally produced intra-articular fractures. A histological and thermoanalytical study.

Intra-articular fractures were produced in rabbit knee joints. The articular cartilage of the injured joints was processed histologically and analysed by a complex thermoanalytical method. It was demonstrated that intra-articular fractures in the early stages have essentially no irreversible damaging effects on the composition of articular cartilage of injured joints.

Animals↗

Calcified cartilage zone and its dimensional relationship to the articular cartilage in the human temporomandibular joint of elderly individuals.

The aim was to describe the appearance of the calcified cartilage zone (CCZ) and to determine its dimensional relationship to the articular cartilage thickness in the normal human temporomandibular joint. An autopsy material comprising 21 joints from 12 elderly individuals was examined microscopically. The appearance of the CCZ was examined, and the thickness of the CCZ and of the total articular cartilage was measured in 18 different positions in each joint. The CCZ was outlined by a flat or gently undulating tidemark and an irregular osteochondral junction. The cellularity of the CCZ varied extensively. The cells were numerous in the CCZ when the overlying articular cartilage displayed high cellularity. Statistical analysis of the measurements demonstrated a relationship (p < 0.001) between the thickness of the CCZ and of the articular cartilage. Our findings, both qualitative and quantitative, indicate a close relationship between the physiology of the CCZ and of the overlying articular cartilage.

Aged↗

Alteration and recovery of the spatial orientation of the collagen network of articular cartilage in adolescent rabbits following intra-articular chymopapain injection.

We have used polarized light (POL) to monitor changes in the organization of the articular cartilage collagen network and matrix proteoglycans (PGs) after intra-articular injection of chymopapain (CP). POL viewing of sirius red stained sections revealed a loss of normal birefringence suggesting an apparent collapse of the collagen network following intra-articular CP. After 21 days, knees injected with 2.0 mg CP showed no return of normal birefringence, however, normal birefringence was noted in knees injected with only 0.2 mg CP. POL viewing of toluidine blue stained sections revealed a severe loss of matrix PGs followed by PG restoration in animals injected with 0.2 mg CP. The most important inference from the data is that articular cartilage can recover from enzyme-induced alterations in the spatial collapse of its fibrillar network. This is an important finding since it has often been inferred that damage to the collagen network leads invariably to progressive articular cartilage destruction.

Animals↗

The effects of ascorbic acid on cartilage metabolism in guinea pig articular cartilage explants.

Ascorbic acid has been associated with the slowing of osteoarthritis progression in guinea pig and man. The goal of this study was to evaluate transcriptional and translational regulation of cartilage matrix components by ascorbic acid. Guinea pig articular cartilage explants were grown in the presence of L-ascorbic acid (L-Asc), D-isoascorbic acid (D-Asc), sodium L-ascorbate (Na L-Asc), sodium D-isoascorbate (Na D-Asc), or ascorbyl-2-phosphate (A2P) to isolate and analyze the acidic and nutrient effects of ascorbic acid. Transcription of type II collagen, prolyl 4-hydroxylase (alpha subunit), and aggrecan increased in response to the antiscorbutic forms of ascorbic acid (L-Asc, Na L-Asc, and A2P) and was stereospecific to the L-forms. Collagen and aggrecan synthesis also increased in response to the antiscorbutic forms but only in the absence of acidity. All ascorbic acid forms tended to increase oxidative damage over control. This was especially true for the non-nutrient D-forms and the high dose L-Asc. Finally, we investigated the ability of chondrocytes to express the newly described sodium-dependent vitamin C transporters (SVCTs). We identified transcripts for SVCT2 but not SVCT1 in guinea pig cartilage explants. This represents the first characterization of SVCTs in chondrocytes. This study confirms that ascorbic acid stimulates collagen synthesis and in addition modestly stimulates aggrecan synthesis. These effects are exerted at both transcriptional and post-transcriptional levels. The stereospecificity of these effects is consistent with chondrocyte expression of SVCT2, shown previously to transport L-Asc more efficiently than D-Asc. Therefore, this transporter may be the primary mechanism by which the L-forms of ascorbic acid enter the chondrocyte to control matrix gene activity.

Aggrecans↗

Ultrasonic attenuation in articular cartilage.

Previous studies have utilized articular cartilage from joints as a model to investigate the influence of various constituents in a connective tissue matrix on ultrasonic properties. These studies have assumed a degree of homogeneity of articular cartilage taken from the same joint. However, tactile loads on articular cartilage vary significantly with location in a joint, and the effects of mechanical load on the connective tissue matrix and the resulting effects on ultrasonic properties are not known. This work reports the variations in acoustical properties of bovine articular cartilage from the stifle (knee) joint both among different joints and within each joint. A pulse-echo transmission technique was used to measure acoustic attenuation in the frequency range of 10 to 40 MHz. The attenuation coefficient was characterized by the integrated attenuation (mean value) over the frequency bandwidth considered. Integrated attenuation averaged over each joint varied among joints from 3.2 to 7.5 NP/cm (6.0 +/- 2.0, mean +/- s.d.). Additionally, a linear regression (r = 0.59) of all the data versus location along the patellar groove indicated that within joints integrated attenuation increased from proximal to distal locations by 6% to 60% (32 +/- 25, mean +/- s.d.). The variations observed among joints and along the patellar groove within a given joint suggest that studies utilizing articular cartilage to determine the role of connective tissue constituents on acoustic properties require control for joint and location. An additional outcome of this study was the observation that damage to the load-bearing surface of articular cartilage may be detectable ultrasonically through characteristics of the acoustic reflection from the articular surface.

Animals↗

Repair of articular cartilage injury following intra-articular chymopapain-induced matrix proteoglycan loss.

The intra-articular injection of 0.02, 0.2, or 2.0 mg of chymopapain (CP) into the knee of adolescent rabbits caused the loss of more than 50% of the proteoglycans (PGs) in the cartilaginous tissues within the joint. Sequential measurements of cartilage-derived keratan sulfate epitope in serum and analyses of articular cartilage slices 2 days after the injection revealed that 0.02 mg of CP was nearly as effective as higher doses (0.2 or 2.0 mg of CP) in causing the depletion. The degradation and depletion of PGs in articular cartilage were shown to be localized to the joint and did not affect articular cartilage in the contralateral knee joint (no injection) or other cartilaginous tissues in the body. On day 9, partial replenishment of the articular cartilage PGs had occurred, irrespective of the dose used, and the articular surface within the joint remained intact. However, by day 21, articular cartilage in joints injected with 2.0 mg of CP had begun to show progressive degenerative changes, and these changes became more severe with time. In contrast, joints injected with 0.2 mg of CP continued to repair successfully by the reestablishment of a matrix that retained its integrity and appeared to remain functional for at least 6 months. These results suggest that the model may prove useful for the study of the repair processes that follow matrix injury and severe depletion of PGs from the articular cartilage matrix.

Animals↗

Comparison of age-dependent expression of aggrecan and ADAMTSs in mandibular condylar cartilage, tibial growth plate, and articular cartilage in rats.

A disintegrin and metalloproteinase with thrombospondin motif (adamalysin-thrombospondins, ADAMTS) degrades aggrecan, one of the major extracellular matrix (ECM) components in cartilage. Mandibular condylar cartilage differs from primary cartilage, such as articular and growth plate cartilage, in its metabolism of ECM, proliferation, and differentiation. Mandibular condylar cartilage acts as both articular and growth plate cartilage in the growing period, while it remains as articular cartilage after growth. We hypothesized that functional and ECM differences between condylar and primary cartilages give rise to differences in gene expression patterns and levels of aggrecan and ADAMTS-1, -4, and -5 during growth and aging. We employed in situ hybridization and semiquantitative RT-PCR to identify mRNA expression for these molecules in condylar cartilage and primary cartilages during growth and aging. All of the ADAMTSs presented characteristic, age-dependent expression patterns and levels among the cartilages tested in this study. ADAMTS-5 mainly contributed to ECM metabolism in growth plate and condylar cartilage during growth. ADAMTS-1 and ADAMTS-4 may be involved in ECM turn over in articular cartilage. The results of the present study reveal that ECM metabolism and expression of related proteolytic enzymes in primary and secondary cartilages may be differentially regulated during growth and aging.

ADAM Proteins↗

The role of the surface amorphous layer of articular cartilage in joint lubrication.

Articular cartilage is a complex soft tissue that performs multiple functions in the joint. In particular, the amorphous layer that covers the surface of articular cartilage is thought to play some role in lubrication. This study aimed to characterize the surface amorphous layer (SAL) using a variety of techniques, including environmental scanning electron microscopy, transmission electron microscopy, white light interferometry, and biochemical analysis of its composition. Friction tests were conducted to investigate the role of the SAL in lubrication. A protocol to remove successfully the SAL without damaging the underlying cartilage was developed and the material removed from healthy cartilage was found to contain approximately equal quantities of glycosaminoglycan (GAG), protein, and lipid. Cartilage-on-cartilage friction tests were conducted on fresh, healthy cartilage with and without the SAL, under both dynamic and static operating conditions. Removal of the SAL was not found to change the friction coefficient. However, subsequent staining of specimens indicated that the SAL had replenished during the test following loading. The replenished SAL was characterized and found to contain lipids and sulphated GAGs with undetectable protein. This study revealed experimental evidence of surface layer replenishment in articular cartilage. It was postulated that the surface layer regeneration mechanism was purely mechanical and associated with movement of GAGs and lipids through the cartilage matrix during deformation, since the experimental set-up did not contain any means of biochemical activation.

Animals↗