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PubMed2002

Serum concentrations of cartilage oligomeric matrix protein, fibrinogen and hyaluronan distinguish inflammation and cartilage destruction in experimental arthritis in rats.

OBJECTIVES: We investigated if changes in serum/plasma fibrinogen (FIB), hyaluronan (HA) and cartilage oligomeric matrix protein (COMP) levels can be used to differentiate between inflammation and cartilage involvement during arthritis. METHODS: Collagen-induced arthritis (CIA), oil-induced arthritis (OIA) and for comparison, experimental autoimmune encephalitis (EAE) induced in DA rats were investigated. RESULTS: Elevations of FIB concentrations were apparent at days 4-7 post-immunization in both arthritis models reaching a maximum on day 20-21, i.e. before peak arthritis. Elevations of HA in both models were seen shortly before macroscopically apparent arthritis, and peaked at or just before maximal arthritis, i.e. later in CIA than in OIA. COMP levels increased only after onset of arthritis and peaked late in disease (days 34-37), being significantly higher in the more destructive CIA compared with the less destructive OIA. During EAE flares, only FIB levels increased. CONCLUSIONS: FIB is a general inflammation marker, HA appears to be a marker for synovitis and changes in COMP levels appear to reflect the cartilage destruction process.

Animals↗
PubMed1994

Cartilage oligomeric matrix protein and thrombospondin 1. Purification from articular cartilage, electron microscopic structure, and chondrocyte binding.

Cartilage oligomeric matrix protein (COMP) and thrombospondin 1 (TSP1) were purified in a native form from normal bovine articular cartilage. The key step in the purification scheme was selective extraction with EDTA-containing buffer. Final separation of these two molecules was achieved by heparin affinity chromatography. Particles viewed by electron microscopy after rotary shadowing and negative staining revealed structures similar to their prototype molecules; from the Swarm rat chondrosarcoma for COMP, or from platelets for TSP1. Attachment of primary bovine chondrocytes to purified matrix proteins was investigated. Cells attached to COMP but not to the structurally related TSP1 indicating separate functions for these proteins in cartilage.

Amino Acid Sequence↗
PubMed2006

Morphogenesis of the second pharyngeal arch cartilage (Reichert's cartilage) in human embryos.

This study was performed on 50 human embryos and fetuses between 7 and 17 weeks of development. Reichert's cartilage is formed in the second pharyngeal arch in two segments. The longer cranial or styloid segment is continuous with the otic capsule; its inferior end is angulated and is situated very close to the oropharynx. The smaller caudal segment is in contact with the body and greater horn of the hyoid cartilaginous structure. No cartilage forms between these segments. The persistent angulation of the inferior end of the cranial or styloid segment of Reichert's cartilage and its important neurovascular relationships may help explain the symptomatology of Eagle's syndrome.

Branchial Region↗
PubMed1982

Characterisation of human articular cartilage link proteins from normal and osteoarthritic cartilage.

Proteoglycan link proteins were isolated from human articular cartilage obtained from normal and osteoarthritic femoral heads and purified to homogeneity employing a method previously described by this laboratory. The link proteins were analysed for amino acid composition, molecular weight on sodium dodecyl sulphate polyacrylamide gels, and ability to stabilise proteoglycan aggregates. The results of these studies were compared with those obtained with bovine link proteins. Two link proteins were identified in the purified fraction from normal and osteoarthritic human cartilage with apparent molecular weights of 54 000 (link 1) and 48 000 (link 2). Functionally the link proteins, isolated from osteoarthritic and normal cartilage, were indistinguishable as measured by their ability to stabilise aggregate. The amino acid compositions of normal and osteoarthritic link proteins were also found to be similar to each other but significantly different from the amino acid composition reported for the bovine link proteins. The quantities of these proteins in extracts from normal and diseased tissue were similar, as was the ratio of link protein 1 to link protein 2.

Aged↗
PubMed1994

Changes in cartilage metabolism in arthritis are reflected by altered serum and synovial fluid levels of the cartilage proteoglycan aggrecan. Implications for pathogenesis.

The metabolism of the cartilage proteoglycan aggrecan was studied in patients with osteoarthritis (OA, n = 83), rheumatoid arthritis (RA, n = 127), and in controls (n = 117) using monoclonal antibody-based radioimmunoassays for glycosaminoglycans in the serum and synovial fluid (SF) to detect epitope 846 on chondroitin sulfate (probably only on recently synthesized molecules) and a keratan sulfate (KS) epitope AN9PI, present on intact and degraded molecules. Epitope 846 levels were always elevated in SF over serum (mean 38-fold in OA and 8.6-fold in RA) being highest in OA patients with the longest disease duration and greatest loss of cartilage, and lowest in RA joints with high leucocyte counts. Serum levels were more often elevated in RA (56%) than in OA (19%) and probably reflect increased aggrecan synthesis in diseased joints. KS levels were higher in SF than in serum in 69% of patients (up to 2.3-fold); levels were inversely (OA) and directly (RA) related to SF leucocyte counts. Serum KS was reduced in both diseases and in RA was inversely related to both systemic and joint inflammation markers. SF 846 levels were inversely related to SF KS in both diseases. These epitopes may provide a measure of the balance between cartilage synthesis and degradation in these diseases.

Adult↗
PubMed1987

The effect of parathyroid hormone (1-34) on cyclic AMP level, ornithine decarboxylase activity, and glycosaminoglycan synthesis of chondrocytes from mandibular condylar cartilage, nasal septal cartilage, and spheno-occipital synchondrosis in culture.

Previously, we reported methods for isolating chondrocytes from the craniofacial complex and their culture in vitro. The response of these chondrocyte cultures to bovine parathyroid hormone (1-34) (PTH) has now been investigated. PTH stimulated glycosaminoglycan (GAG) synthesis, a characteristic of the cartilage phenotype in cultured chondrocytes isolated from mandibular condylar cartilage (MCC), nasal septal cartilage (NSC), and spheno-occipital synchondrosis (SOS). These stimulations of GAG synthesis by PTH were dose-dependent. PTH also increased accumulation of cyclic AMP (cAMP) and the activity of ornithine decarboxylase (ODC), a rate-limiting enzyme in polyamine biosynthesis. However, PTH did not stimulate DNA synthesis. The increases in the cAMP level, ODC activity, and GAG synthesis after addition of PTH (10(-7) mol/L) were greatest in MCC-chondrocytes and least in NSC-chondrocytes. The difference in the responses to PTH of these three types of chondrocytes may reflect differences of the characteristics of these cells in vivo.

Animals↗
PubMed2001

Articular cartilage and changes in arthritis: noncollagenous proteins and proteoglycans in the extracellular matrix of cartilage.

Cartilage contains numerous noncollagenous proteins in its extracellular matrix, including proteoglycans. At least 40 such molecules have been identified, differing greatly in structure, distribution, and function. Some are present in only selected cartilages or cartilage zones, some vary in their presence with a person's development and age, and others are more universal in their expression. Some may not even be made by the chondrocytes, but may arise by absorption from the synovial fluid. In many cases, the molecules' function is unclear, but the importance of others is illustrated by their involvement in genetic disorders. This review provides a selective survey of these molecules and discusses their structure, function, and involvement in inherited and arthritic disorders.

Arthritis↗
PubMed2002

Transplantation of cartilage-like tissue made by tissue engineering in the treatment of cartilage defects of the knee.

We investigated the clinical, arthroscopic and biomechanical outcome of transplanting autologous chondrocytes, cultured in atelocollagen gel, for the treatment of full-thickness defects of cartilage in 28 knees (26 patients) over a minimum period of 25 months. Transplantation eliminated locking of the knee and reduced pain and swelling in all patients. The mean Lysholm score improved significantly. Arthroscopic assessment indicated that 26 knees (93%) had a good or excellent outcome. There were few adverse features, except for marked hypertrophy of the graft in three knees, partial detachment of the periosteum in three and partial ossification of the graft in one. Biomechanical tests revealed that the transplants had acquired a hardness similar to that of the surrounding cartilage. We conclude that transplanting chondrocytes in a newly-formed matrix of atelocollagen gel can promote restoration of the articular cartilage of the knee.

Adolescent↗
PubMed2000

Equine carpal articular cartilage fibronectin distribution associated with training, joint location and cartilage deterioration.

Processes involved in equine carpal osteochondral injury have not been established. In other species, fibronectin appears important in chondrocyte-matrix interactions, and levels are increased in osteoarthritis. This investigation aimed to (a) describe fibronectin immunoreactivity in the middle carpal joint of 2-year-old Thoroughbreds, (b) determine topographical variations, (c) compare strenuously trained (Group 1) or gently exercised horses (Group 2) and (d) describe sites with early osteoarthritis. Group 1 (n = 6) underwent a 19 week high intensity treadmill training programme. Group 2 (n = 6) underwent 40 min walking until euthanasia. Dorsal and palmar sites on radial, intermediate and third carpal articular surfaces were prepared. Immunohistochemistry was performed using a biotin-streptavidin/peroxidase method. Cross-reactivity of rabbit antihuman fibronectin antiserum with equine fibronectin was confirmed using Western blotting. Results showed: (a) fibronectin was present primarily in pericellular and interterritorial matrix locations, (b) dorsal sites had zonal immunoreactivity compared to palmar sites, (c) Group 1 dorsal radial carpal cartilage had increased superficial staining compared to Group 2 and (d) fibrillated cartilage showed increased intracellular and local matrical immunoreactivity (superficial zone). These findings suggest topographical and exercise-related variations in fibronectin distribution, and indicate equine fibronectin is localised at sites of cartilage degeneration and released into the matrix by chondrocytes in the local area.

Animals↗
PubMedDate not supplied

Osteoarthritic patients with high cartilage turnover show increased responsiveness to the cartilage protecting effects of glucosamine sulphate.

OBJECTIVE: Glucosamine sulphate has been shown in a large double-blind, placebo-controlled clinical trial to prevent structural damage and improve clinical symptoms of osteoarthritis (OA). We investigated whether early response in a newly developed biochemical marker of collagen type II degradation (CTX-II, CartiLaps ELISA) could reflect the long-term preservation of hyaline cartilage. METHODS: Study subjects comprised 212 knee OA patients participating in a clinical trial of the effects of glucosamine sulphate. Disease symptoms were assessed quarterly by WOMAC scoring and X-ray analysis was performed at baseline and after 3 years. Urine samples were obtained at baseline and after 1, 2 and 3 years for measurement in the CartiLaps assay. The measurements were corrected for creatinine. RESULTS: At baseline the patients had an average concentration of urinary CTX-II of 222.4 +/- 159.5 ng/mmol creatinine. This was significantly above the CTX-II levels measured in urine samples from 415 healthy controls (169.1 +/- 92.3 ng/mmol, p < 0.0001). There was no significant difference in the CTX-II response in the placebo group and the glucosamine treated group. However, those with high cartilage turnover presented a significant decrease in CTX-II after 12-month glucosamine treatment. Thus, three group with CTX II concentrations above normal average + 1SD decreased 15.5% after 12-month therapy. The 12 months change in CTX-II in OA patients with elevated CTX-II at baseline correlated with the change in average joint space width observed after 36 months (R = 0.43, p < 0.05). Increased baseline levels of CTX-II were associated with a worsening of the WOMAC index (p < 0.01). CONCLUSION: The data indicate that measurement of urinary collagen type II C-telopeptide fragments enables the identification of OA patients with high cartilage turnover who at the same time are most responsive to therapy with structure modifying drugs.

Aged↗
PubMed1976

Cathepsin D activity in bovine articular cartilage, synovial membrane and fluid: degradation of cartilage proteoglycans from same joint.

Cathepsin D type proteases were extracted from articular cartilage, synovial membrane, and synovial fluid from normal, adult bovine knee joints. A sensitive enzyme assay made it possible to measure protease activity in the different tissues from individual joints. Highest activity was found in the synovial membrane, while cell free synovial fluids contained comparatively low activity. The degrading effect on articular cartilage proteoglycans (PGC and PGS), isolated from the same joints, was demonstrated by gelfiltration on Sepharose columns and by viscometry. Gelfiltration profiles of incubation mixtures indicated a proteolytic effect on PGC and on PGS), at pH 3.5, in concentrations of enzyme and proteoglycans found in cartilage tissue. No effect at neutral pH was obtained despite a 100-fold increase of enzyme concentration. These findings were supported by viscometry data. The degrading effect of enzymes from all sources was completely inhibited by pepstatin.

Animals↗
PubMed1990

Immobilization aggravates cartilage damage during antigen-induced arthritis in mice. Attachment of polymorphonuclear leukocytes to articular cartilage.

The early and late effects of short-term immobilization on arthritic joints have been studied. Knee joints of mice in which an antigen-induced unilateral arthritis was elicited were immobilized in extension for 3, 5, and 7 days. After 5 and 7 days' immobilization, arthritis was significantly more severe. More leukocytes infiltrated the periarticular tissues and more cellular exudate was found in the joint space. A striking observation was that large numbers of polymorphonuclear leukocytes (PMN) attached to the surface of the cartilage, a phenomenon not found in mobile arthritic joints. Electron-microscopy confirmed PMN adhesion and showed severe ruffling of the cartilage surface under immobilized conditions. Further examination of factors determining PMN sticking revealed that attachment is rapid when the cartilage surface is already damaged, and that retained immune complexes and complement play a pivotal role. The late effects of immobilization were studied after a remobilization period of 2 weeks. Enhanced matrix depletion and chondrocyte death persisted in arthritic joints that were previously immobilized for 5 and 7 days, and the latter also showed significantly increased osteophyte formation. Although these results are speculative for the human situation, this study indicates that treatment of arthritic joints by complete rest should be applied with caution.

Animals↗
PubMedDate not supplied

[Ultrastructure of reparation collagen and its relations to residual hyaline joint cartilage in inflammatory cartilage destruction].

The development of collagenous fibrils in reparative tissues replacing hyaline cartilage in arthritic joints was investigated by electron microscopy: granulation tissue consists of a loose network of collagenous fibrils, scar tissue is characterized by dense bundles of parallel arranged collagenous fibrils, chondroid tissue that may develop under certain instances is characterized by a network of random orientated fibrils that differ from those of the normal hyaline cartilage by their uniformity. The fibrils of the reparative tissues often exhibited an intense contact with those of the original hyaline cartilage.

Arthritis, Rheumatoid↗
PubMed1988

Reimplantation of autologous septal cartilage in the growing nasal septum. II. The influence of reimplantation of rotated or crushed autologous septal cartilage on nasal growth: an experimental study in growing rabbits.

A series of experiments were carried out on growing rabbits to investigate the effect of rotation (through 90 degrees and 180 degrees) or crushing of autologous cartilage implants on growth of the nose. Reconstruction of the nasal septum with rotated or crushed implants did not restore the normal growth of the nose. This is in accordance with the results of a previously described experiment in which the continuity was repaired with autologous cartilage in the same position (Nolst Trenité et al., 1987). The crushed cartilage implants were as effective as the noncrushed implants in preventing septal perforations. The implants rotated through 90 degrees, showed less intrinsic growth in dorso-ventral direction.

Animals↗
PubMed1987

Reimplantation of autologous septal cartilage in the growing nasal septum. I. The influence of resection and reimplantation of septal cartilage upon nasal growth: an experimental study in rabbits.

The effects of restoration of the dorso-ventral continuity of the nasal septum on the growth of the nose was investigated in growing rabbits. Submucous resection of the middle third of the septal cartilage resulted in characteristic growth disturbances of nose and maxilla. Reimplantation of the resected strip of autologous cartilage did not restore the normal development of the nose. The implants however did prevent septal perforations as seen after resection of cartilage only and showed a considerable growth in dorso-ventral direction.

Animals↗
PubMed1987

In vitro repair potential of articular cartilage: proteoglycan metabolism in the different areas of the femoral condyles in human cartilage explants.

We tested the hypothesis that metabolism of individual cartilage samples, such as those which might be taken at arthroscopy, could reflect the overall intrinsic reparative response of the cartilaginous tissue of that joint. Full thickness samples of articular cartilage, removed from the anteromedial, anterolateral and posterolateral areas of the femoral condyles of 2 normal knees, were placed in tissue culture separately. The incorporation of 14C-glucosamine into matrix molecules, especially proteoglycan, was measured. Three slices of each area were removed after one, 3 or 6 weeks of in vitro culture. The extracted matrix molecules and the media were analyzed and the relative amount of radioactive proteoglycan and proteoglycan aggregates, as well as the total amount of radiolabelled proteoglycan/mg dry weight, were calculated. A progressive increase in these variables during the culture period was demonstrated, suggesting an attempt at repair of the articular cartilage in this in vitro condition. No differences in the incorporation dynamics were found between the anteromedial, anterolateral and posterolateral areas of both subjects.

Cartilage, Articular↗
PubMed1986

Studies on hormonal regulation of the growth of the craniofacial skeleton: IV. Specific binding sites for glucocorticoids in condylar cartilage and their involvement in the biological effects of glucocorticoids on cartilage cell growth.

Using a whole-tissue binding assay and cell-free binding measurements indicated the presence of a specific steroid receptor for triamcinolone in cartilage cells of neonatal mouse mandibular condyle. Analysis of receptor levels showed that whole-tissue preparations bound 1360 fmol triamcinolone/mg protein. Affinity measurements revealed a dissociation constant of 7.6 X 10(-9) M. There was a close correlation between triamcinolone inhibition of DNA synthesis and steroid occupancy of whole-tissue receptors. The inhibitory effect of triamcinolone upon DNA synthesis could be significantly reduced by "blocking" the respective receptors with cortexolone. All the cartilage cells in the condyle revealed distinct intracellular labeling using [3H] dexamethasone autoradiography. Hence, neonatal condylar cartilage, an active site of endochondral bone formation in the craniofacial skeleton, can be regarded as a genuine target tissue for the biological effects of glucocorticoids.

Animals↗
PubMed1980

[Cartilage vessels and the ossification center. Metamorphosing cartilage: 2nd communication].

In this paper we compared the vessels and the surrounding cells in the growing (cattle fetus 520 mm SSL) and metamorphosing cartilage of rats. In the growing cartilage the vessels and sinusoid capillares are accompanied by some morphologic uncharacteristical perivasculare cells which may swell oedematatically. Chondorolytic symptoms appear sporadically but not multinucleated chondroclasts are found. In the metamorphosing cartilage the perivasculare cells grow up to large cells exhibiting an extensive granular reticulum. Firstly the intercellularsubstance seems to be destructed, later on degenerate, they pine away, the chondrocytes as well as the chondrocytes with cyst-like cisternae as such with lacunar cisternae. Also a disintegration of cells occurs. The foci of mineralisation in the neighborhood of the vessels consist of needle-shaped cristals in chains arranged along the fibrils.

Animals↗