Elevation of a collagenase generated type II collagen neoepitope and proteoglycan epitopes in synovial fluid following induction of joint instability in the dog

Chu, Qili; Lopez, Mandi J.; Hayashi, Kei; Ionescu, Mirela; Billinghurst, R. Clark; Johnson, Kenneth A.; Poole, A. Robin; Markel, Mark D.

doi:10.1053/joca.2002.0812

Cited by 72 publications

(87 citation statements)

References 29 publications

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“…The assays for aggrecan 846 epitope and Col2-3/ 4C long mono in canine body fluids and cartilage tissue extracts have been recently described (24). Similarly, the assays for CPII and KS in canine samples demonstrated close parallelism compared with the human or bovine standards used with the assays (Poole AR, Ionescu M: unpublished observations).…”

Section: Methodsmentioning

confidence: 78%

“…KS, an index of proteoglycan catabolism, was quantified by inhibition RIA using the monoclonal antibody AN9P1 Fab fragment, 125 Ilabeled adult human aggrecan as a tracer, and unlabeled aggrecan as the standard (2,21). The amino-terminal (TC A ) fragment cleaved by collagenase from triple-helical type II collagen includes (at its carboxy terminus) the neoepitope Col2-3/4C long mono , which was measured using a competitive inhibition monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) (7,8,23,24). All of these epitopes are present in canine articular cartilage and their extracts and in canine body fluids, as determined by immunohistochemistry (see below) and Western blotting.…”

Section: Methodsmentioning

confidence: 99%

“…Similarly, the assays for CPII and KS in canine samples demonstrated close parallelism compared with the human or bovine standards used with the assays (Poole AR, Ionescu M: unpublished observations). To reduce viscosity and improve pipetting accuracy, joint fluid samples were digested overnight at 37°C with 0.4 turbidityreducing units/ml of Streptomyces hyaluronidase prior to assay for aggrecan 846, KS, CPII, and Col2-3/4C long mono (24). Serum samples were analyzed undiluted, urine samples were diluted 2-fold to 4-fold as needed to ensure that values fell within the linear part of the standard curve, and joint fluid samples were diluted ϳ5% during hyaluronidase digestion.…”

Section: Methodsmentioning

confidence: 99%

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Analysis of cartilage biomarkers in the early phases of canine experimental osteoarthritis

Matyas

Atley

Ionescu

et al. 2004

Arthritis & Rheumatism

141

124

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Objective. To study 3 body fluids for changes in the levels of 5 biomarkers of cartilage metabolism during the early phases of experimental osteoarthritis (OA).Methods. Twenty skeletally mature mixed-breed canines underwent unilateral surgical transection of the anterior cruciate ligament. Samples of joint fluid, serum, and urine were obtained preoperatively and just before necropsy (3 weeks or 12 weeks postoperatively). Biomarkers included 2 markers of cartilage matrix synthesis/turnover (aggrecan 846 epitope and C-propeptide of type II collagen) and 3 markers of cartilage degradation (keratan sulfate proteoglycan epitope, the collagenase-generated cleavage epitope of type II collagen [Col2-3/4C long mono , or CIIC], and crosslinked peptides from the C-telopeptide domain of type II collagen [Col2CTx]). Significant changes in the levels of these biomarkers were determined by paired analyses.Results. Joint pathology was more severe in the 12-week group compared with the 3-week group. In joint fluid, due to limited volume, only Col2-3/4C long mono and Col2CTx were measured. Significant elevations in the levels of both of these markers were observed in experimental joints in both the 3-week group and the 12-week group. In serum, the level of aggrecan 846 epitope was elevated at both 3 weeks and 12 weeks, the level of Col2-3/4C long mono was elevated at 12 weeks, and the level of Col2CTx was elevated at both 3 weeks and 12 weeks. In urine, the level of Col2-3/4C long mono was elevated at 12 weeks after surgery.Conclusion. Levels of biomarkers of intact aggrecan proteoglycan (aggrecan 846 epitope) and type II collagen degradation (Col2-3/4C long mono and Col2CTx) were elevated early after unilateral stifle joint injury, suggesting that these markers are sensitive and specific for early cartilage changes associated with isolated joint injury in this established model of experimental OA.The early stages of osteoarthritis (OA) are difficult to diagnose. Joint structure and function are typically altered substantially before symptoms cause patients to seek medical care, that is, the osteoarthritic process begins long before OA presents as a clinical disease. The insidious onset and "silent" progression of primary OA not only obscure an early diagnosis, but also delay treatment that may help prevent further cartilage destruction and joint failure. Hence, diagnostic tests that can detect and monitor molecular events early in the pathogenesis of OA would be potentially very useful.For nearly a century, synovial fluid analysis has been important in the differential diagnosis of arthritis. Although synovial fluid analysis is useful in distinguishing between inflammatory, septic, and crystal-induced arthritis, its value in OA is limited. Similarly, routine blood and urine analyses can identify certain inflammatory arthritides and metabolic defects that cause joint pain and pathology but, as yet, reveal little about the OA process. No specific molecular markers of the OA process are currently available for clinical use, and the...

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Section: Methodsmentioning

confidence: 78%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Analysis of cartilage biomarkers in the early phases of canine experimental osteoarthritis

Matyas

Atley

Ionescu

et al. 2004

Arthritis & Rheumatism

141

124

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“…25 Surgery was carried out under general anesthesia with halothane and oxygen inhalation via endotracheal intubation. Using aseptic techniques, arthroscopy was performed in one randomly selected ovine stifle.…”

Section: Methodsmentioning

confidence: 99%

Development of partial thickness articular cartilage injury in an ovine model

Markel

Swain

et al. 2006

Journal Orthopaedic Research

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The purpose of this study was to create a controlled partial thickness cartilage lesion in a sheep model, and to provide a foundation to study the natural history of the progression of this lesion. Twenty-eight sheep divided into four groups (1, 12, 24, and 52 weeks, n ¼ 7/group) were used in this study. In one stifle, a mechanical tool was used to create a 200 mm partial thickness lesion (1.5 Â 1.5 cm 2 ) on the medial femoral condyle via arthroscopy. Joint fluid was drawn presurgery and after euthanasia for analysis of collage II 3/4 C long (C2C). After euthanasia, the condyle was analyzed by gross appearance, confocal laser microscopy (CLM) for cell viability, scanning electronic microscopy (SEM) for surface roughness, Artscan for cartilage stiffness, and histology for cartilage morphology. The gross appearance of the treated area appeared rough, soft, and swollen compared to untreated control over time. CLM demonstrated that the depth of cell death increased to 590 mm at 52 weeks after surgery. SEM demonstrated that the treated area became more irregular over time. Stiffness of the treated area was significantly less than control by 12 weeks after surgery. Histologic analysis demonstrated that the 12, 24, and 52 week groups had significantly poorer histologic scores than the 1 week group. Joint fluid analysis demonstrated that the treatment group at 1 week had significant higher levels of C2C than the pretreatment baseline data. The results of this study demonstrated that partial thickness injury of cartilage continued to propagate and degenerate over time in this sheep model. Options for the prevention or treatment of this lesion may be tested using this model in the future. ß

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“…Measuring CII cleavage products in SF has the potential advantage of assessing CII turnover in a single joint. Such an assay indicated increased degradation of CII in a canine model of joint instability (31). However, interpretation of results from such a generic CII marker may be limited if the assay cannot distinguish between structural and newly synthesized molecules, since collagen synthesis was reported to increase after joint injury (2,10).…”

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confidence: 99%

The release of crosslinked peptides from type II collagen into human synovial fluid is increased soon after joint injury and in osteoarthritis

Lohmander¹,

Atley²,

Pietka³

et al. 2003

Arthritis & Rheumatism

234

165

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Objective. To determine concentrations of crosslinked C-telopeptide fragments of type II collagen (CTX-II) in synovial fluid (SF) from patients with joint injury, osteoarthritis (OA), or other knee arthritides.Methods. Two study groups were used: a crosssectional group, which included healthy-knee volunteers (reference group [REF]) and patients with pseudogout (PPA), an anterior cruciate ligament tear with or without a meniscus tear (INJ), or primary knee OA (POA); and a longitudinal group, which included patients with arthroscopic cartilage changes or septic arthritis. CTX-II was quantified by competition enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody that recognized the C-terminus of the peptide EKGPDP as a proteolytic neoepitope. Aggrecan fragments, matrix metalloproteinases 1 and 3, and tissue inhibitor of metalloproteinases 1 were determined by ELISAs.Results. Concentrations of CTX-II in SF were higher in patients with PPA, INJ, and POA than in the REF group (P < 0.001). After joint injury, mean levels of CTX-II in SF were increased above REF levels at all time intervals (P < 0.001), and were highest within hours after trauma. In those in the longitudinal study group with joint cartilage damage, variation coefficients for CTX-II were 81% (between patients) and 64% (within patient), monitored over 1 year. In a patient with septic arthritis, SF CTX-II increased at the onset of symptoms, and peaked 30-fold higher than the baseline. Concentrations of all biomarkers decreased with successful treatment.Conclusion. This is the first report to describe the release into SF of soluble molecular fragments specific for the degradation of mature, crosslinked, type II collagen (CII) in human OA and joint injury. The results provide strong evidence that the integrity of the CII network of cartilage is compromised soon after joint injury and in arthritis. This early degradation of CII may represent an important treatment target.Osteoarthritis (OA) and joint injury are characterized by remodeling and degradation of cartilage, bone, and other joint tissues. The normally very slow turnover rate of cartilage matrix (1) is increased, as observed by several techniques in both animal OA models and human OA. Phasic changes in both synthesis and degradation of collagen and other matrix molecules (2-6) are associated with altered tissue structure and material properties (7-9).Increased joint tissue turnover after injury and in OA can be detected by a release of molecules and molecular fragments into synovial fluid (SF), blood, and urine. For example, the stimulated synthesis of type II collagen (CII) results in higher levels of CII C-propeptide in SF (10,11). Stimulated aggrecan synthesis results in more aggrecan fragments carrying the 846 epitope (12). Higher synovial concentrations of matrix metalloproteinases (MMPs), such as MMP-1 and MMP-3, and proteinase activity after joint injury and in OA are consistent with the observed expression of these

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Elevation of a collagenase generated type II collagen neoepitope and proteoglycan epitopes in synovial fluid following induction of joint instability in the dog

Cited by 72 publications

References 29 publications

Analysis of cartilage biomarkers in the early phases of canine experimental osteoarthritis

Analysis of cartilage biomarkers in the early phases of canine experimental osteoarthritis

Development of partial thickness articular cartilage injury in an ovine model

The release of crosslinked peptides from type II collagen into human synovial fluid is increased soon after joint injury and in osteoarthritis

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