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Further insights into the structural principles governing the function of articular cartilage.

A new experimental technique involving the observation of an artificial notch propagating through articular cartilage has been used to examine the biomechanical properties of this tissue. By predetermining both the orientation of the notch and its location with respect to the primary functional zones a more rigorous description of the structure/function relationships in cartilage has been achieved. The principal findings are: A primary 'strain-locking' role for the superficial zone has been demonstrated experimentally in articular cartilage. Comparison of the behaviour of radial and transverse notches has revealed a primary structural anisotropy in the general matrix. This is strong evidence in support of the morphological model proposed in a recent paper by the present author. A range of mechanical responses is shown to be reflected consistently in structural features considered to arise principally from variations in the degree of crosslinking between the overall radial configuration of collagen fibres. It is possible to separate mechanically the collagen fibres from the general matrix and the bonding relationship between them is time-dependent. Measurement of loads required to propagate a radial notch suggest (a) that the strength of the fibres and/or that of the crosslinks between fibres increases with depth through the cartilage thickness, and (b) that the radial columns of chondrocytes typical of the deep zone do not represent planes of significantly reduced strength relative to the adjacent matrix. A major structural discontinuity exists in normal articular cartilage in a plane parallel to and below the articular surface. It is argued that this plane represents a major change in overall orientation of the collagen fibres. Finally, by applying the experimental techniques described in this paper both to degenerative articular cartilage and to healthy articular cartilage in which the primary components have been selectively degraded enzymatically it should be possible to gain a more precise picture of the structural origin of malfunction in this tissue.

Animals↗

Influence of stress rate on water loss, matrix deformation and chondrocyte viability in impacted articular cartilage.

The biomechanical response of articular cartilage to a wide range of impact loading rates was investigated for stress magnitudes that exist during joint trauma. Viable, intact bovine cartilage explants were impacted in confined compression with stress rates of 25, 50, 130 and 1000 MPa/s and stress magnitudes of 10, 20, 30 and 40 MPa. Water loss, cell viability, dynamic impact modulus (DIM) and matrix deformation were measured. Under all loading conditions the water loss was small (approximately 15%); water loss increased linearly with increasing peak stress and decreased exponentially with increasing stress rate. Cell death was localized within the superficial zone (< or =12% of total tissue thickness); the depth of cell death from the articular surface increased with peak stress and decreased with increasing stress rate. The DIM increased (200-700 MPa) and matrix deformation decreased with increasing stress rate. Initial water and proteoglycan (PG) content had a weak, yet significant influence on water loss, cell death and DIM. However, the significance of the inhomogeneous structure and composition of the cartilage matrix was accentuated when explants impacted on the deep zone had less water loss and matrix deformation, higher DIM, and no cell death compared to explants impacted on the articular surface. The mechano-biological response of articular cartilage depended on magnitude and rate of impact loading.

Animals↗

Magic-angle effect in magnetic resonance imaging of articular cartilage: a review.

RATIONALE AND OBJECTIVES: The laminar appearance of articular cartilage in magnetic resonance (MR) images has been a source of confusion, especially concerning the number, intensity, thickness, and origin of the layers. The laminar appearance is associated with the magic-angle effect in the MR imaging (MRI) of articular cartilage. METHODS: This article introduces the topic with background information about cartilage and the magic-angle effect and then reviews the literature about the magic-angle effect. The review concludes with a brief discussion of the future directions of study and the potential clinical relevance of the laminae in MR images of articular cartilage. CONCLUSIONS: The magic-angle effect is commonly seen in MR images of several tissues. The direct cause of the laminar appearance of articular cartilage is the T2 relaxation anisotropy in the tissue, which is closely linked to the structure of the collagen fibers, their orientation in the magnetic field, and the water-proteoglycan interaction that amplifies the prevailing orientation of the collagen fiber network. The laminar appearance of cartilage has an intrinsic spatial heterogeneity over the two-dimensional joint surface, which leads to inconsistencies in the reported total number of cartilage laminae and the laminar patterns observable in MRI, depending on where the sample was taken. Two additional thin, low-intensity laminae may also be visible at the boundaries of the cartilage with fluid and with bone; whether these boundary laminae are identified and counted with the others may introduce inconsistency in the results reported by various researchers.

Cartilage, Articular↗

The effects of indirect blunt trauma on adult canine articular cartilage.

UNLABELLED: In order to determine the effect of subfracture loads on articular cartilage, we impacted twelve adult canine patellofemoral joints utilizing a drop-tower with two different force-levels. The joints were examined with light and electron microscopy at two, four, and six weeks after impaction. In ten additional animals a single knee was impacted and they were analyzed biochemically at similar time-periods, using the contralateral joint as a control. In all impacted specimens changes were observed in the zone of calcified cartilage, represented by an increase in cellular clones, vascular invasion, and proteoglycan content of the matrix. Ultrastructural evaluation of the superficial and deep radial zones of the articular cartilage revealed loss of the cellular processes and territorial matrices of chondrocytes in both layers. Ruthenium-red staining of impacted samples revealed a 40 per cent decrease in proteoglycan associated with collagen fibers in the extraterritorial matrix. An increase in collagen-fiber width was observed in the four and six-week groups. The earliest changes in articular cartilage included activation of the zone of calcified cartilage as well as ultrastructural alterations in the superficial and radial zones. Biochemical analysis revealed an increase in water content and hexuronic acid at two weeks. These changes occurred at a subfracture level in the absence of surface disruption. CLINICAL RELEVANCE: These animal experiments indicate that adult articular cartilage may show significant alterations in its histological, biochemical, and ultrastructural characteristics without disruption of the articular surface. This model of articular cartilage "contusion" may represent a corollary to the joint damage that is observed following direct blunt trauma transmitted across articular surfaces without radiographic evidence of fracture. The possibility that this form of injury may be the precursor of chondromalacic changes in patellar or femoral cartilage merits further study.

Animals↗

Mechanical properties of articular cartilage covered by the meniscus.

OBJECTIVE: To investigate the mechanical properties and morphological characteristics of articular cartilage on the tibial plateau of human knees, including the region covered by the meniscus. DESIGN: Using a 1-mm diameter flat-ended cylindrical probe to apply a constant load (0.6 MPa) at specific sites on the tibial plateau, the mechanical properties of articular cartilage were studied using seven cadaver knees. Comparison was made between data obtained by the cartilage covered by the meniscus and that not covered. This was done for both the medial and lateral plateaus. Histological sections of the articular cartilage were also performed to study differences between cartilage from these regions of the tibial plateau. RESULTS: Compared to cartilage that was not covered by the meniscus, the articular cartilage beneath the meniscus showed a significantly (P<0.05) larger modulus by as much as 70%, and was less thick by about 40%. Also, the subchondral bone quantity and calcified layer thickness were observed to be significantly lesser in the regions covered by the meniscus. CONCLUSIONS: Our findings revealed a significant difference between the mechanical properties and associated structures of articular cartilage in the region covered by the meniscus compared with the articular cartilage not covered by the meniscus.

Aged↗

Biomechanical and histological evaluation of hydrogel implants in articular cartilage.

We evaluated the mechanical behavior of the repaired surfaces of defective articular cartilage in the intercondylar region of the rat femur after a hydrogel graft implant. The results were compared to those for the adjacent normal articular cartilage and for control surfaces where the defects remained empty. Hydrogel synthesized by blending poly(2-hydroxyethyl methacrylate) and poly(methyl methacrylate-co-acrylic acid) was implanted in male Wistar rats. The animals were divided into five groups with postoperative follow-up periods of 3, 5, 8, 12 and 16 weeks. Indentation tests were performed on the neoformed surfaces in the knee joint (with or without a hydrogel implant) and on adjacent articular cartilage in order to assess the mechanical properties of the newly formed surface. Kruskal-Wallis analysis indicated that the mechanical behavior of the neoformed surfaces was significantly different from that of normal cartilage. Histological analysis of the repaired defects showed that the hydrogel implant filled the defect with no signs of inflammation as it was well anchored to the surrounding tissues, resulting in a newly formed articular surface. In the case of empty control defects, osseous tissue grew inside the defects and fibrous tissue formed on the articular surface of the defects. The repaired surface of the hydrogel implant was more compliant than normal articular cartilage throughout the 16 weeks following the operation, whereas the fibrous tissue that formed postoperatively over the empty defect was stiffer than normal articular cartilage after 5 weeks. This stiffness started to decrease 16 weeks after the operation, probably due to tissue degeneration. Thus, from the biomechanical and histological point of view, the hydrogel implant improved the articular surface repair.

Animals↗

Expression of clusterin in the superficial zone of bovine articular cartilage.

OBJECTIVE: To investigate the differences between chondrocytes of the superficial and underlying zones of articular cartilage at the level of gene expression. METHODS: Messenger RNA (mRNA) was isolated from chondrocytes harvested from the superficial and deep zones of immature bovine articular cartilage. This mRNA was reverse transcribed, radiolabeled, and then each complementary DNA (cDNA) sample was used to screen duplicate filters of a bovine chondrocyte cDNA library. By comparing autoradiographic signals on matching filter sets, clones exclusively expressed in the superficial zone of articular cartilage were isolated and characterized further. RESULTS: Of the superficial-specific gene clones isolated, 25% were found to be a single gene product, clusterin. Northern hybridization was used to show that clusterin is expressed specifically in the superficial zone of articular cartilage and that its expression is up-regulated in mature cartilage. In situ hybridization was used to precisely localize clusterin transcripts in articular cartilage, where it was found that clusterin expression was confined to the articular surface in both immature and mature samples. CONCLUSION: The discovery of clusterin expression at the articular cartilage surface extends previous observations that superficial articular chondrocytes are highly specialized cells. Clusterin is a multifunctional, secreted glycoprotein that has been shown to be expressed in diverse locations that have in common a tissue-fluid boundary. Additionally, clusterin has been implicated in regulating complement activation and cell death in injured and degenerating tissues.

Animals↗

Symptomatic articular cartilage degeneration: the impact in the new millennium.

The symptomatic degeneration of articular cartilage and associated arthritis is among the most prevalent chronic conditions in the United States and the population most at risk is increasing. It is the leading cause of limitations in activities of daily living and is second to heart disease in causing work disability. The current and future socioeconomic impact of chronic articular cartilage disease on the healthcare system will be magnified by increasing numbers of patients who will seek relief of their symptoms and their disability to remain active. Because these individuals live longer and remain active, the proportion of their life living with symptoms and disability from articular cartilage degeneration increases. The economic, psychologic, and social impact of degenerative articular cartilage can be enormous for these individuals but it also impacts their family and society. The direct traditional medical costs and indirect economic and wage loss from arthritis in individuals the United States has reached in excess of $65 billion annually and is expected to increase as the population ages. In addition, the expenditures for complementary and alternative professional services and therapies for arthritis is increasing and is also in the billions of dollars annually. Because of these escalating costs, documenting the value of the patient and cost effectiveness to society of prevention and treatment programs for symptomatic articular cartilage degeneration will be required.

Cartilage, Articular↗

Articular cartilage collagen birefringence is altered concurrent with changes in proteoglycan synthesis during dynamic in vitro loading.

INTRODUCTION: The articular cartilage collagen network and proteoglycans are subject to changes in deteriorating joint diseases. In this study, we exposed articular cartilage plugs to cyclic loading and investigated the properties of collagen network and proteoglycans in different zones of the articular cartilage. METHODS: Articular cartilage full-depth plugs were exposed in vitro to 4.1 MPa cyclic (0.5 Hz) loading for 1 to 20 hr and investigated using quantitative microscopic methods (i.e., polarized light microscopy, microspectrophotometry, and autoradiography). RESULTS: The loading caused packing or condensation of the tissue. In histological sections, the height of uncalcified articular cartilage decreased by an average of 12.8% (range, 4 to 19.7%). Loading increased the birefringence of collagen in the superficial cartilage (P < 0.05), with thickening of the zone up to 41.4% at 20 hr. The thickness of the intermediate zone increased also (22% at 1 hr and 434% at 20 hr). Concomitantly, the birefringence (P < 0.05) and the thickness of the deep zone decreased (18.5 to 27.8%). Loading for 4 hr increased the 35S-sulphate incorporation of the cartilage explants by an average of 67% (P < 0.05). The increase was most significant in the deep cartilage. A simultaneous increase was observed in the proteoglycan concentration of the cartilage; the staining intensity with safranin-O increased by 8.8% (P < 0.05). After 8 hr loading, this stimulation decreased; at 20 hr, loading caused a clear inhibitory effect on proteoglycan synthesis in the superficial zone. DISCUSSION: According to these results, the chosen loading regimen increased the thickness and collagen orientation in the superficial zone. In contrast, the thickness and birefringence in the deep cartilage were reduced. The proteoglycan metabolism of chondrocytes was first stimulated deep in the cartilage, but as the loading continued, the effect proved to be inhibitory (especially in the superficial part of uncalcified cartilage).

Animals↗

Changes in the hyaline articular cartilage after air exposure.

UNLABELLED: The changes of hyaline articular cartilage from rabbits after air exposure were evaluated. The knee joints were exposed to air for periods of thirty minutes to two hours. The animals were killed periodically, at three days, one week and three weeks postoperatively. After sacrifice, the cartilage was removed and prepared for study by light microscopy and electron microscopy. Exposure to room air for thirty minutes produced chondrocyte necrosis in the upper third of the cartilage, and exposure for 60 minutes or longer produced chondrocyte necrosis of the entire thickness of articular cartilage at three days after arthrotomy. But, three weeks after arthrotomy, we could not find any chondrocyte necrosis in any rabbits at varying periods of air exposure. There was no significant change in proteoglycan content between the aired and control cartilage. CLINICAL RELEVANCE: Exposing cartilage to air can cause transient and reversible cartilage damage. If these changes are not reversible, the orthopedic surgeon should consider avoiding the prolonged exposure of articular cartilage to air, since complete matrix disintegration is known to occur months after chondrocyte necrosis.

Air↗

Chondrocyte allografts for repair of full-thickness defects in the condylar articular cartilage of rabbits.

PURPOSE: This study investigated the feasibility of repairing defects of the condylar articular cartilage by chondrocyte allotransplantation. MATERIAL AND METHODS: A full-thickness defect (2 mm diameter) was made in the condylar articular cartilage of 6-month-old rabbits with a No. 701 dental fissure bur that penetrated both the articular cartilage and the subchondral bone, and entered the marrow cavity. Sixty-four animals were divided into 5 groups. In the cell transplantation group (20 rabbits), the defect was filled with a collagen membrane embedded with chondrocytes of neonate rabbits cultured in vitro for 1 week. In control group 1 (18 rabbits), the defect was left untreated. In control group 2 (18 rabbits), the defect was filled with a collagen membrane without chondrocytes. In sham-operation control group (6 rabbits), the condyle surface was exposed but left unchanged. Two rabbits were added as a normal control group. Mandibular movement was not restricted postoperatively. The macroscopic and microscopic features of the condyles were observed at 1, 2, 4, 8, 12, and 20 weeks following surgery. RESULTS: The defects in the condylar articular cartilage were repaired with cartilage tissue after cell transplantation. CONCLUSION: A defect in the condylar articular cartilage can be repaired with articular cartilage-like tissue by allotransplantation of chondrocytes.

Animals↗

The isolation and characterization of magnesium whitlockite crystals from human articular cartilage.

A number of basic calcium phosphate crystals have been demonstrated in human articular tissues. The exact relationship between crystal deposition and disease remains obscure, although there is evidence supporting a rapid degenerative arthropathy within a specific set of patients. Limited reports of 'cuboid' calcium phosphate microcrystals in articular cartilage have been made over the last 10 years. In this study the occurrence of such crystals, not apparent by light microscopy, in human articular cartilage has been confirmed by transmission electron microscopy and X-ray microanalysis of tissue prepared by aqueous and anhydrous processing techniques. A crystal isolation technique involving collagenase digestion, centrifugation and sodium hypochlorite treatment was developed enabling crystal characterization by electron and X-ray diffraction. Crystals were identified as magnesium whitlockite; the first report of this mineral in articular cartilage. The presence of this mineral phase in normal and osteoarthritic articular cartilage is discussed with consideration given to physical conditions known to favor whitlockite formation and those extant in articular cartilage.

Calcium Phosphates↗

Composition and dynamics of articular cartilage: structure, function, and maintaining healthy state.

Disorders of articular cartilage represent some of the most common and debilitating diseases encountered in orthopaedic practice. Understanding the normal functioning of articular cartilage is a prerequisite to understanding its pathologic processes. The mechanical properties of articular cartilage arise from the complex structure and interactions of its biochemical constituents: mostly water, electrolytes, and a solid matrix composed primarily of collagen and proteoglycan. The viscoelastic properties of cartilage, due primarily to fluid flow through the solid matrix, can explain much of the deformational responses observed under many loading conditions. Degenerative processes can often be explained by a breakdown of the normal load-bearing capacity of cartilage which arises from the mechanics of this fluid flow. Several factors which may lead to such a breakdown include direct trauma to the cartilage, obesity, immobilization, and excessive repetitive loading of the cartilage. Sports activity, without traumatic injury, does not appear to be a risk factor for the development of osteoarthritis in the normal joint; however, such activity may be harmful to an abnormal joint.

Animals↗

Comparative assessment of articular cartilage and synovial membrane in experimental haemarthrosis.

The changes in articular cartilage and synovial membrane of the knee joints were studied in two groups of rabbits and Wistar rats with experimental haemarthrosis, electron microscopically. Hamarthrosis was produced in group 1 by a single autologous blood injection, in group 2 by intraarticular fracture of the femoral condyles. Samples were taken from the intact articular cartilage, the menisci and the infrapatellar portion of the synovial membrane 12 h to 20 days after intervention. Blood resorption occurs only in the synovial membrane. Fragmentation of erythrocytes and erythrocytophagy by synovial macrophages is documented. The different stages of intracellular digestion of erythrocyte fragments are traced down. Synovial fibroblasts do not participate in erythrocytophagy, although they disclose morphological signs of enhanced functional activity. The findings show changes in the matrix and chondrocytes within the articular cartilage and menisci, and presence of free erythrocytes and lipoprotein complexes amidst the collagen fibres of the matrix. The chondrocytes are poor in cell organelles, while the intracytoplasmic filaments, lipid droplets and glycogen granules are augmented in number. There is no evidence of erythrocytophagy by cartilage cells. On single blood injection in the joint, the ensuing changes are reversible, and the normal synovial membrane structure is restored much quicker than the articular cartilage.

Animals↗

Rapid degradation of articular cartilage proteoglycan by neutrophils: comparison with macrophages and synovial fibroblasts.

OBJECTIVE AND DESIGN: To determine and compare the proteoglycan degradative properties of neutrophils, macrophages and synoviocytes in cultures of articular cartilage. MATERIAL OF SUBJECTS: Bovine articular cartilage was aseptically isolated from metacarpopharyngeal joints. Neutrophils and macrophages were isolated from normal human blood and bovine synovial fibroblasts were isolated from explant cultures before being incubated with the cartilage. TREATMENT: Neutrophils, macrophages or synovial fibroblasts (1 x 10(6)-8 x 10(6)) were incubated with 35SO4 labelled cartilage for 2.5-72 h. METHODS: Cartilage degradation was measured as a loss of 35SO4 into the cartilage medium as a percentage of the total labelled proteoglycan in the cartilage slice. Statistical significances were determined using a 2-tailed unpaired Student's t-test. RESULTS: Neutrophils rapidly degraded articular cartilage. After 2.5 hours of culture, neutrophils degraded cartilage proteoglycan up to 28 times more than either macrophages or synovial fibroblasts. CONCLUSIONS: Neutrophils induce rapid damage to articular cartilage proteoglycan, whereas in comparison, macrophages and synovial fibroblasts degrade articular cartilage proteoglycans poorly. These findings indicate that at least under conditions where the influence of cellular-cellular interactions and soluble mediator action are excluded, adhesion of neutrophils to articular cartilage is sufficient to stimulate rapid and marked cartilage degradation compared to the other two cell types.

Animals↗

Utility of delayed gadolinium-enhanced MRI (dGEMRIC) for qualitative evaluation of articular cartilage of patellofemoral joint.

Delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) was used for the measurement of relative proteoglycan depletion of articular cartilage in the patellofemoral (PF) joint following a proprietary protocol, which was compared with the X-ray images, proton density weighted MR images (PDWI) and arthroscopic findings. The study examined 30 knees. The ages ranged from 16 to 74 (average 40.3) years. The Gd-DTPA(2-)containing contrast medium was used in a single dose. The subjects were made to exercise the knee joint for 10 min; and MR images were taken 2 h after intravenous injection of contrast medium. T1-calculated images were produced and the region of interest (ROI) was set as follows. (1) ROI1: entire articular cartilage in a slice through the center of the patella. (2) ROI2: low signal region in T1-calculated images, which were set in a blind fashion by two observers. (3) ROI3: articular cartilage on one side that includes ROI2 where low signal region were detected (medial or lateral). ROI3 was set to examine the contrast of ROI2 with surrounding articular cartilage. The average T1 values of ROI1 was 393.5+/-33.6 ms for radiographic grade 0 and 361.3+/-11.1 ms for grade I, which showed a significant difference (P=0.036). The T1 value of ROI2 was 351.6+/-28.2 ms for grade I, 361.9+/-38.3 ms for grade II, 362.1+/-67.7 ms for grade III, and 297.8+/-54.1 ms for grade IV according to arthroscopic Outerbridge classification. All cases, that demonstrated decrease of T1 values on dGEMRIC (ROI2), showed abnormal arthroscopic or direct viewing findings. The ratio (ROI3/ROI2) in cases of only slight damage classified as Outerbridge grade I (6 cases) was an average of 1.04+/-0.02 and was 1.0 or greater in all cases, thereby indicating well-defined contrast with the surrounding cartilage. The diagnosis of damage in articular cartilage was possible in all 16 cases with radiographic K-L grade I on dGEMRIC, while the intensity changes were not found in 10 of 16 cases on PDWI. The dGEMRIC with a single-dose would be useful on a diagnosis of the area demonstrating early relative proteoglycan depletion in the articular cartilage of the PF joint prior to any discernible changes in the subchondral bone on X-ray images and exceeds to plain MR images for examining deterioration of articular cartilage.

Adolescent↗