The Prevention of Collagen Breakdown in Bovine Nasal Cartilage by TIMP, TIMP-2 and a Low Molecular Weight Synthetic Inhibitor

Ellis, A.J.; Curry, Valerie; Powell, Eleanor A.; Cawston, TE

doi:10.1006/bbrc.1994.1673

Cited by 85 publications

(79 citation statements)

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“…However, given the correlation between the release of sGAG and G1-NITEGE 373 in response to ADAMTS-4 or ADAMTS-5 treatment observed in the current study, it is unlikely that G1-VDIPES 341 fragments were generated under the conditions tested. Furthermore, while the concentrations of ADAMTS-4 and ADAMTS-5 (and MMP-13) used to treat the cartilage discs in this study may appear to be supraphysiologic, poor penetration and/or poor transport of exogenously added proteins into the highly anionic cartilage matrix has been described previously (17,38,39). In addition, the concentration and localization of ADAMTS-4 or ADAMTS-5 in the cartilage matrix are also likely to be dependent on the enzyme isoform(s) that is present.…”

mentioning

confidence: 70%

Release of hyaluronan and hyaladherins (aggrecan G1 domain and link proteins) from articular cartilage exposed to ADAMTS‐4 (aggrecanase 1) or ADAMTS‐5 (aggrecanase 2)

Chockalingam¹,

Zeng²,

Morris³

et al. 2004

Arthritis & Rheumatism

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Objective. To determine whether aggrecanase (ADAMTS) activities in articular cartilage can directly lead to the release of hyaluronan (HA) and hyaladherins (aggrecan G1 domain and link proteins), as may occur ex vivo during stimulation of cartilage explants with interleukin-1 (IL-1) or retinoic acid or in vivo in synovial joints during aging and joint pathology.Methods. Bovine articular cartilage discs (live or freeze-killed) were cultured in the presence of IL-1 or were incubated in digestion buffer containing recombinant human ADAMTS-4 (rHuADAMTS-4; aggrecanase 1) or rHuADAMTS-5 (aggrecanase 2). Culture media, digestion supernatants, and tissue extracts were assayed for sulfated glycosaminoglycan (sGAG) content and analyzed by Western blotting to detect aggrecanasegenerated G1 domain (using neoepitope monoclonal antibody AGG-C1/anti-NITEGE 373 ) and link proteins (using monoclonal antibody 8-A-4), as well as by quantitative enzyme-linked immunosorbent assays to detect aggrecanase-generated G1 domain (G1-NITEGE 373 ) and HA.Results. IL-1 treatment of live cartilage explants induced a time-dependent release of sGAG, aggrecanase-generated G1 domain (G1-NITEGE 373 ), and HA into the culture media. Exposure of live or freeze-killed articular cartilage discs to rHuADAMTS-4 or rHuADAMTS-5 resulted in a dose-and timedependent release of sGAG and hyaluronan from the tissue, accompanied by a concomitant release of functionally intact hyaladherins (aggrecan G1-NITEGE 373 and link proteins).Conclusion. Coincident with aggrecanolysis, aggrecanase activities in articular cartilage may actuate the release of HA and associated hyaladherins, thereby further compromising the integrity of the cartilage matrix during degenerative joint diseases such as osteoarthritis.

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mentioning

confidence: 70%

Release of hyaluronan and hyaladherins (aggrecan G1 domain and link proteins) from articular cartilage exposed to ADAMTS‐4 (aggrecanase 1) or ADAMTS‐5 (aggrecanase 2)

Chockalingam¹,

Zeng²,

Morris³

et al. 2004

Arthritis & Rheumatism

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“…MT1-MMP can cleave type II collagen (13), which is the major type of collagen in articular cartilage. However, in the bovine nasal cartilage assay, we have shown that the addition of TIMP-1 to resorbing cartilage completely blocks collagen release (17,45). Since TIMP-1 is a poor inhibitor of MT1-MMP (56), these data suggest that MT1-MMP is not directly involved in degrading cartilage collagen.…”

Section: Discussionmentioning

confidence: 95%

“…Hydroxyproline release was assayed (as a measure of collagen degradation) using a microtiter modification of the method as described previously (45,46).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of furin‐like enzymes blocks interleukin‐1α/oncostatin M–stimulated cartilage degradation

Milner¹,

Rowan²,

Sf³

et al. 2003

Arthritis & Rheumatism

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Objective. To investigate the role of furin-like enzymes in the proteolytic cascades leading to cartilage breakdown and to examine which collagenase(s) contribute to collagen degradation.Methods. Bovine nasal cartilage was stimulated to resorb with the addition of interleukin-1␣ (IL-1␣)/ oncostatin M (OSM) in the presence or absence of a furin inhibitor, Dec-RVKR-CH 2 Cl, or selective matrix metalloproteinase 1 (MMP-1) inhibitors. Collagen and proteoglycan levels were determined by assay of hydroxyproline and sulfated glycosaminoglycan, respectively. Collagenase and gelatinase activity were measured using 3 H-acetylated collagen and gelatin zymography, respectively.Results. The addition of Dec-RVKR-CH 2 Cl to stimulated cartilage reduced the release of collagen fragments and the levels of active collagenase and MMP-2, suggesting that furin-like enzymes are involved in the cascades leading to activation of procollagenases. At MMP inhibitor concentrations that selectively inhibit MMP-1, no inhibition of collagen release was observed, but increasing the concentration to the 50% inhibition concentration for MMP-13 resulted in a 50% blockage of collagen release. The addition of Dec-RVKR-CH 2 Cl to resorbing cartilage also partially blocked proteoglycan release, thus demonstrating a role for furin-activated enzymes in the pathways leading to proteoglycan degradation. Conclusion.Furin-like enzymes are involved in cascades leading to activation of procollagenases and degradation of collagen. MMP-13, which can be activated by furin-processed membrane-type 1 MMP-1, appears to be a major collagenase involved in collagen degradation induced by IL-1␣/OSM. Furin-like enzymes also appear to play a role in the pathways leading to proteoglycan degradation. These findings are of importance when considering proteinase inhibition as a target for therapeutic intervention in arthritic diseases.

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“…Pellets were washed with 0.5 M acetic acid before digestion with pepsin (1:10 wet weight) in 0.5 M acetic acid for 24 h at 48C. Samples were centrifuged at 30,000 g for 15 min at 48C, and the supernatant containing pepsin-soluble collagen was analyzed by hydroxyproline assay 25 before Western blot analysis. An equal amount of collagen from control (untreated) and ZVF-treated samples were diluted 1:1 in sample buffer (0. )…”

Section: Western Blottingmentioning

confidence: 99%

Enhanced Tissue Integration During Cartilage Repair In Vitro Can Be Achieved by Inhibiting Chondrocyte Death at the Wound Edge

Gilbert

Singhrao

Khan

et al. 2009

Tissue Engineering Part A

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The Prevention of Collagen Breakdown in Bovine Nasal Cartilage by TIMP, TIMP-2 and a Low Molecular Weight Synthetic Inhibitor

Cited by 85 publications

References 0 publications

Release of hyaluronan and hyaladherins (aggrecan G1 domain and link proteins) from articular cartilage exposed to ADAMTS‐4 (aggrecanase 1) or ADAMTS‐5 (aggrecanase 2)

Release of hyaluronan and hyaladherins (aggrecan G1 domain and link proteins) from articular cartilage exposed to ADAMTS‐4 (aggrecanase 1) or ADAMTS‐5 (aggrecanase 2)

Inhibition of furin‐like enzymes blocks interleukin‐1α/oncostatin M–stimulated cartilage degradation

Enhanced Tissue Integration During Cartilage Repair In Vitro Can Be Achieved by Inhibiting Chondrocyte Death at the Wound Edge

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