Collagenase and collagenolytic activity in human osteoarthritic cartilage

245

136

Objective. To quantify stromelysin and collagenase in synovial fluid (SF) from patients with rheumatoid arthritis (RA) or traumatic knee injury.Methods. Stromelysin and collagenase were measured in the SF of 33 patients with RA or pttraumatic knee injury, using specific double-antibody sandwich enzyme-linked immunosorbent assays. Stromelysin was fractionated from representative SF, and the molecular form was identilied by immunoblot analysis.Results. The stromelysin concentration was -20-fold higher than the collagenase concentration in the fluids from patients with RA and -8-fold higher in the fluids from patients with traumatic iaury. For both metalloproteinases, there was a higher enzyme concentration in RA SF than in the SF from patients with trauma (stromelysin 40.1 f 26 pg/ml [mean f SDI in RA SF, 8.5 +: 15 d m l in trauma SF; collagenase 2.2 f 3.3 pg/ml in RA SF, 1.1 f 2.3 pg/ml in trauma SF).The majority of the stromelysin within the SF bound to reactive red-agarose and was identified as prostrome-

Section: Resultsmentioning

confidence: 99%

Detection of Stromelysin and Collagenase in Synovial Fluid From Patients with Rheumatoid Arthritis and Posttraumatic Knee Injury

Walakovits

Moore

Bhardwaj

et al. 1992

245

136

“…Also, proteases that are relevant for articular cartilage degradation might primarily diffuse in from the synovial fluid, originating in synovial cells (31,32) rather than articular chondrocytes. It is well known from biochemical studies that OA cartilage shows increased collagenolytic activity (9,33,34) and severe collagen denaturation and degradation (35).…”

Section: Discussionmentioning

confidence: 99%

Relative messenger RNA expression profiling of collagenases and aggrecanases in human articular chondrocytes in vivo and in vitro

Bau¹,

Gebhard²,

Haag³

et al. 2002

370

322

Objective. Osteoarthritic (OA) cartilage destruction depends on collagen-and aggrecan-degrading proteases such as collagenases (MMP-1 and MMP-13), stromelysin (MMP-3), MMP-14, as well as the so-called aggrecanases (ADAM-TS4 and ADAM-TS5). In this study, we tried to clarify whether these proteases are expressed in vivo in human normal and OA cartilage (and whether they are up-regulated or down-regulated during the disease process) and in interleukin-1␤ (IL-1␤)-stimulated chondrocytes in vitro.Methods. Quantitative polymerase chain reaction assays were developed and performed on RNA isolated directly from normal and degenerative cartilage tissue as well as from primary human articular chondrocytes cultured with and without IL-1␤.Results. In vivo, MMP-1 was detectable only at very low levels in any condition. MMP-13 expression was low in normal and early degenerative cartilage but was strongly up-regulated in late-stage OA specimens. MMP-1 and MMP-13 were expressed much higher in vitro than in vivo and were up-regulated by IL-1␤. Among all proteases, MMP-3 was by far the most strongly expressed, although it was strongly downregulated in late-stage OA specimens. Expression of MMP-3 was higher in vitro than in vivo and was up-regulated by IL-1␤. ADAM-TS5 and MMP-14 were expressed in all sample groups. Expression of ADAM-TS4 was very low in vivo and was induced in vitro after stimulation by IL-1␤.Conclusion. Our expression data clearly support MMP-13 as the major collagenase in OA cartilage. The most strongly expressed aggrecanase was ADAM-TS5. ADAM-TS4 was expressed only at a very low level in normal cartilage and was only slightly up-regulated in OA cartilage, casting doubt on this enzyme being the relevant aggrecanase of articular cartilage. Results of our study show that expression of many enzymes is significantly different in vitro and in vivo and suggest that IL-1␤ stimulation of articular chondrocytes might not be a good model for the matrix catabolism in OA cartilage.Osteoarthritic (OA) cartilage degeneration as well as cartilage destruction in rheumatoid arthritis depend, at least to a significant degree, on enzymatic degradation of matrix components. Two types of molecules, collagen fibrils and the proteoglycan aggrecan, represent the major targets in terms of enzymatic activity and functional loss of the cartilage matrix as a result of damage to these molecules. Whereas degradation of collagen fibrils leads to matrix instability with tissue swelling, degradation of proteoglycans leads to cartilage softening and loss of fixed charges (1), both of which are classic features of cartilage destruction.Catabolism of both types of molecules is supposed to involve different types of enzymes, largely from 2 protease families: matrix metalloproteinases (MMPs) and the "A disintegrin and metalloproteinases with thrombospondin type 1 motif" (ADAM-TS). The initial degradation of collagen fibrils (within the triple-helical region) depends on cleavage at the collagenase site, for which there exist 2 major candidate enzymes...

“…In previous studies (4,15,17) we have shown that OA cartilage contains proteases capable of digesting collagen and proteoglycan macromolecules within the matrix at physiologic pH. These enzymes belong to the metalloprotease class.…”

Section: Discussionmentioning

confidence: 99%

Proteoglycan‐degrading acid metalloprotease activity in human osteoarthritic cartilage, and the effect of intraarticular steroid injections

Pelletier¹,

Martel‐Pelletier²,

Cloutier³

et al. 1987

Cartilage samples from both the immediate and remote lesion areas were obtained from the tibia1 plateaus of 21 patients with osteoarthritis, and were subjected to histologic and enzymatic study. There was a frequent loss of pericellular metachromatic staining in the OA cartilage. Seven patients had received intraarticular injections of steroids, and in 21 % of those cartilage samples, a pericellular halo was seen. This halo was seen in 71 % of patients who had not received steroid injections. The total acid metalloprotease activity was increased more than twofold in specimens from OA lesions and in those samples graded moderate, as compared with age-matched control cartilages. These differences were greater when the specimens from patients who had received steroid therapy were excluded from the data. The cartilage specimens from steroid-treated patients were not significantly different from those of controls with respect to the enzyme activity in the lesions or in cartilage with moderate disease. The active form of the protease was suppressed by steroids. In samples from patients who did not receive steroid injections and who had a moderate grade of OA, a significantly elevated level of the active protease was present, as compared with control samples. Those samples graded moderate which came from patients who received steroid treatments showed no difference in the active protease level versus that of controls. Our results are consistent with the hypothesis that acid metalloprotease activity is involved in the degradation of the cartilage matrix in OA. Since the protease retains a significant fraction (40%) of its activity at neutral pH, its physiologic role might occur either at acid pH or at neutral pH. The halo surrounding the chondrocytes (the result of a local loss of proteoglycan) might be related to the action of this enzyme. Preliminary evidence has suggested that steroids reduce cartilage enzyme activity. Address reprint requests to J. p. Pelletier, MD, Unit6 des Maladies Rhumatismales, HGpital Notre-Dame, 1560 rue Sherbrooke Est, Montreal, Quebec, Canada H2L 4K8.vised form November 11, 1986. tent (1). Recent reports by Sachs et a1 (2) and Goldberg et (3) describe types Of changes in metachromatic staining. Type A is characterized by a lack of perkellular staining, so that a halo appearance is present around the lacunae. The remainder Of the intertenitorial matrix has norma] metachromatic staining. The