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Objective: Rupture of the anterior cruciate ligament (ACL) increases the risk of developing osteoarthritis (OA). Delayed Gadolinium enhanced magnetic resonance imaging (MRI) of cartilage (dGEMRIC) investigates cartilage integrity through T1-analysis after intravenous contrast injection. A high dGEMRIC index represents good cartilage quality. The main purpose of this prospective cohort study was to investigate the prognostic value of the dGEMRIC index regarding future knee OA., Method: 31 patients with ACL injury (mean age 27 ± 6.7 (±SD) years, 19 males) were examined after 2 years with 1.5T dGEMRIC of femoral cartilage. Re-examination 14 years post-injury included weight-bearing knee radiographs, Lysholm and Knee Osteoarthritis Outcome Score (KOOS)., Results: At the 14-year follow up radiographic OA (ROA) was present in 68% and OA symptoms (SOA) in 42% of the injured knees. The dGEMRIC index of the medial compartment was lower in knees that developed medial ROA, 325 ± 68 (ms±SD) vs 376 ± 47 (51 (7-94)) (difference of means (95% confidence interval (CI))), in patients that developed symptomatic OA (SOA), 327 ± 61 vs 399 ± 42 (52 (11-93)), and poor knee function 337 ± 54 vs 381 ± 52 (48 (7-89)) compared to those that did not develop ROA, SOA or poor function. The dGEMRIC index correlated negatively with the OARSI osteophyte score in medial (r = -0.44, P = 0.01) and lateral (r = -0.38, P = 0.03) compartments., Conclusion: The associations between a low dGEMRIC index and future ROA, as well as SOA, are in agreement with previous studies and indicate that dGEMRIC has a prognostic value for future knee OA., (Copyright © 2019 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

Objective: To determine if type III collagen is concentrated in the chymotrypsin-extractable collagen pool from osteoarthritic articular cartilage to assess its potential as a biomarker of Osteoarthritis (OA) pathogenic mechanisms., Methods: Full thickness articular cartilage from grossly normal surfaces was analyzed from femoral heads, obtained at hip replacement surgery, from OA (n = 10) and fracture (n = 10) patients. Collagen, extracted by α-chymotrypsin, was characterized by SDS-PAGE/Western blot analysis, ELISA and immunohistochemistry using monoclonal antibodies specific to collagens types II and III., Results: α-Chymotrypsin extracted more collagen from OA than control cartilage. The extractable pool included collagen types II and III from both OA and control hips. Importantly, OA cartilage contained 6-fold more collagen type III than control cartilage, based on ELISA. The estimated total tissue ratio of collagen III/II was in the 1-10% range for individual OA cartilage samples, based on pepsin-solubilized collagen using SDS-PAGE densitometry. Collagen type III N-propeptide trimers were the main molecular fragments seen on Western blot analysis of OA and control extracts. The chymotrypsin-extracted type II collagen gave primarily full-length α1(II) chains and chain fragments of α1(II) on Western blot analysis from both OA and control tissues. Immunohistochemistry showed that type III collagen was more concentrated in the upper half of OA cartilage and in the territorial matrix around individual chondrocytes and chondrocyte clusters., Conclusions: The findings confirm that collagen type III deposition occurs in adult articular cartilage but significantly more pronounced in osteoarthritic joints, presenting a potential marker of matrix repair or pathobiology., (Copyright © 2016 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

Background: Previously, a positive effect of exercise on cartilage structure was indicated with delayed gadolinium-enhanced magnetic resonance imaging (MRI) of cartilage (dGEMRIC). However, in that study only one full-thickness region of interest (ROI) in the medial femoral condyle was analyzed., Purpose: To improve the knowledge about exercise effects on cartilage structure by re-analyzing previous images with regional dGEMRIC analysis., Material and Methods: Thirty patients (age range, 38-50 years) with a previous medial meniscus resection were divided into three groups according to self-reported change in physical activity (PA) level in a 4-month exercise intervention study: Group I (n = 11), increased PA level; Group II (n = 13), no change in PA level; and Group III (n = 6), reduced PA level. dGEMRIC index was analyzed at inclusion and after 4 months. Anterior (less load) and posterior (more load) ROIs of medial and lateral femoral condyles were analyzed, as well as superficial and deep cartilage regions., Results: Group I increased the dGEMRIC index in the posterior cartilage (P = 0.004). The increase was larger in the lateral (P = 0.005) than the medial compartment in both superficial and deep cartilage regions. The dGEMRIC index did not change in Group II. In Group III, the dGEMRIC index decreased in the medial posterior cartilage (P = 0.03)., Conclusion: In patients with a previous medial meniscectomy, the beneficial effect of exercise varies between different locations within the joint, the largest improvement being observed in lateral posterior cartilage, i.e. the load-bearing cartilage in the compartment without a meniscus lesion. The effects of exercise do not seem to vary with cartilage depth., (© The Foundation Acta Radiologica 2015.)

Objective: To examine the association between the relaxation time (T1Gd) of delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) and grade of tibiofemoral joint space narrowing (JSN) and osteophytosis 11 years later, in a cohort of meniscectomized patients., Design: Patients (n = 45) aged 35-50 who had undergone an arthroscopic partial medial meniscectomy 1-6 years earlier, due to degenerative meniscal tear, were examined using dGEMRIC. These patients had no cartilage changes defined as deep clefts or visible bone at the time of arthroscopy. Eleven years later (12-16 years after surgery) 34 of these subjects (76%) were evaluated by weight-bearing knee radiography, and tibiofemoral joint changes were graded according to the Osteoarthritis Research Society International Atlas., Results: Lower T1Gd in the medial compartment was associated with higher grade of medial JSN (grade 0, 351 ms; grade 1, 386 ms; grade 2, 342 ms; grade 3, 259 ms [P for trend < 0.001]) and more osteophytosis (score 0, 371 ms; score 1, 389 ms; score 2, 354 ms; score 3, 289 ms; score 4, 265 ms; score 5, 275 ms [P for trend = 0.001]). Lower T1Gd in the lateral compartment was associated with higher grade of lateral JSN (grade 0, 436 ms; grade 1, 346 ms [P for trend = 0.026])., Conclusion: The current study suggests that lower T1Gd measured with dGEMRIC of medial and lateral femoral cartilage is associated with higher grade of JSN 11 years later, and medially, also with more osteophytosis., (Copyright © 2014 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

Objective: Ankle fracture patients were used as a model to study the long-term effect of the removal of joint loading on knee cartilage quality in human subjects., Design: The knees of 10 patients with ipsilateral ankle fractures were investigated using delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) at the time of ankle injury. After 6 weeks' prescribed unloading of the affected leg, but no restrictions regarding knee movement, the cast was removed from the ankle and the patient underwent a second dGEMRIC examination. Physiotherapy was then initiated. A third dGEMRIC examination was performed 4 months after remobilization, and a final examination 1 year after the injury., Results: Baseline T1Gd values for the 10 patients were within a narrow range. No significant change in mean T1Gd was observed after 6 weeks' prescribed unloading, but the T1Gd range had increased significantly. Four months after remobilization, the mean T1Gd was significantly lower than in the previous examinations, and the range remained significantly broader than at baseline. At the 1-year follow-up, the mean T1Gd was almost identical to the value after remobilization, and the T1Gd range still showed a significant increase compared to the baseline investigation., Conclusions: Removal of knee cartilage loading for 6 weeks resulted in a measurable effect on the cartilage matrix, as evidenced by a broader T1Gd range. A decrease in mean T1Gd was observed 4 months after remobilization. These differences persisted a year after injury compared to baseline., (Copyright © 2014 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

Objective: To assess knee cartilage quality and subjective knee function, 20 years after injury in anterior cruciate ligament (ACL) injured copers., Method: We examined 32 knees using delayed gadolinium-enhanced magnetic resonance imaging (MRI) of cartilage (dGEMRIC), 20 years after a complete ACL tear. Only subjects who had coped with the ACL injury without ACL reconstruction (ACLR), and who presented without radiographic signs of osteoarthritis (OA) at an earlier 16-year follow-up, were included in this study. The quality of the central weight-bearing parts of the medial and lateral femoral cartilage was estimated with dGEMRIC (T1Gd). These results were compared with corresponding results in 24 healthy individuals, and with the subjects' self-reported subjective knee function using the Knee Injury and Osteoarthritis Outcome Score (KOOS) questionnaire., Results: The values of T1Gd in the medial and lateral femoral cartilage of the study group (mean (95% CI)), were 404 (385-423) and 427 (399-455) ms, not statistically different from those of the healthy reference group (P = 0.065 and 0.31). The subjective knee function 20 years after the injury, according to the five domains of the KOOS score, was good, with a mean score of 90 ± 11. Values of T1Gd for the medial femoral cartilage were correlated with the KOOS subgroup QOL (P = 0.021, Pearson correlation)., Conclusions: Subjects who have managed to cope with their ACL injury for 20 years with sustained good subjective knee function also seem to have knee cartilage of good quality, with T1Gd values not very different from a healthy reference group., (Copyright © 2013 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

Objective: This study investigates sulphated glycosaminoglycans (sGAG) content changes in early osteoarthritis (OA), and whether contrast-enhanced magnetic resonance imaging (MRI) of cartilage in vitro may identify early event of OA pathology., Method: Osteochondral plugs from patients with hip OA or femoral neck fracture (reference group) were collected and analysed by 1.5 T MRI with ΔR1 as a measure of cartilage contrast concentration. Cartilage hydration, contents of sGAG, cartilage oligomeric matrix protein (COMP), hydroxyproline, denatured collagen, and aggrecan TEGE(392) neoepitope were determined and histological grading was performed., Results: sGAG content correlated to ΔR1, although no difference in either of these parameters was detectable between OA and reference cartilage at 4 h of contrast equilibration. In contrast, biochemical analysis of other cartilage matrix constituents showed distinct alterations typical for early cartilage degradation in OA cartilage and with clear evidence for increased aggrecan turnover., Conclusion: In the present in vitro study, cartilage sGAG content could not distinguish between early OA cartilage and reference cartilage. Given, that delayed gadolinium enhanced MRI of cartilage (dGEMRIC) indicates early events in the pathogenesis of OA in vivo, our results from the in vitro studies imply other, additional factors than cartilage sGAG content, e.g., alterations in diffusion or increased supply of contrast agent in the diseased joint. Alternatively, an altered dGEMRIC reflects later stages of OA, when sGAG content decreases. Further investigations are warranted, to understand variations in sGAG content in pathology, an essential background for interpreting dGEMRIC measurements., (Copyright © 2012 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

Objective: In this observational longitudinal study we estimate knee joint cartilage glycosaminoglycan (GAG) content, in patients with an acute anterior cruciate ligament (ACL) injury, with or without a concomitant meniscus injury., Methods: 29 knees (19 men/10 women) were prospectively examined by repeat delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC), approximately 3 weeks and 2.3±1.3 (range 4.5) years after the injury. We estimated the GAG content (T1Gd) in the central weight-bearing parts of the medial and lateral femoral cartilage and compared results with a reference cohort (n=24) with normal knees and no history of injury examined by dGEMRIC at one occasion previously., Results: The healthy reference group had longer T1Gd values compared with the ACL-injured patients at follow-up both medially: 428±38 vs 363±61ms (P<0.0001) and laterally: 445±41 vs 396±48ms (P=0.0002). At follow-up T1Gd was lower in meniscectomized patients compared to those without a meniscectomy, both medially (-84ms, P=0.002) and laterally (-38ms, P=0.05). In the injured group, the medial femoral cartilage showed similar T1Gd at the two dGEMRIC investigations: 357±50 vs 363±61ms (P=0.57), whereas the lateral femoral cartilage T1Gd increased: 374±48 vs 396±48ms (P=0.04)., Conclusions: The general decrease in cartilage T1Gd in ACL-injured patients compared with references provide evidence for structural matrix GAG changes that seem more pronounced if a concomitant meniscal injury is present. The fact that post-traumatic OA commonly develops in ACL-injured patients, in particularly those with meniscectomy, suggests that shorter T1Gd may be an early biomarker for OA., (Copyright © 2011 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.)

Purpose: To present an evaluation method for three-dimensional Look-Locker (3D-LL) based T1 quantification, calculating correct T1 values independent of local flip angle (FA) variations. The method was evaluated both in phantoms and in vivo in a delayed Gadolinium Enhanced MRI of Cartilage (dGEMRIC) study with 33 subjects., Materials and Methods: T1 was measured with 3D-LL, using both local FA correction and a precalculated FA slice profile, and compared with standard constant FA correction, for all slices in phantoms and in both femur condyles in vivo. T1 measured using two-dimensional Inversion Recovery (2D-IR) was used as gold standard., Results: Due to the FA being slice dependent, the standard constant FA correction results in erroneous T1 (systematic error = 109.1 ms in vivo), especially in the outer slices. With local FA correction, the calculated T1 is excellent for all slices in phantoms (<5% deviation from 2D-IR). In vivo the performance is lower (systematic error = -57.5 ms), probably due to imperfect inversion. With precalculated FA correction the performance is very good also in vivo (systematic error = 13.3 ms)., Conclusion: With the precalculated FA correction method, the 3D-LL sequence is robust enough for in vivo dGEMRIC, even outside the centermost slices., ((c) 2009 Wiley-Liss, Inc.)

Objective: To study the relationship between cartilage integrity, assessed with [delayed Gadolinium-Enhanced Magnetic Resonance Imaging of Cartilage (dGEMRIC)] and epidemiologic risk factors for knee osteoarthritis (OA) in meniscectomized patients., Methods: Body mass index (BMI) was calculated in 45 patients (16 women), mean age 46, who underwent an arthroscopic medial meniscectomy 1-6 years earlier. The cartilage glycosaminoglycan (GAG) content was estimated by dGEMRIC Index and tests of isokinetic muscle strength and functional performance (one-leg hop test) were conducted., Results: BMI ranged from 20.0 to 34.3 (mean: 26.5). The dGEMRIC Index was 14.4% lower in the medial index compartment (374+/-61 ms, mean+/-SD) than in the lateral reference compartment (437+/-59 ms, mean+/-SD) (P<0.001). The dGEMRIC Index of the medial diseased compartment correlated positively with both knee flexor (r=0.50, P=0.001) and knee extensor strength (r=0.47, P=0.001) relative to body weight and with the one-leg hop test (r=0.42, P=0.004). Furthermore, a negative correlation was found between the dGEMRIC Index of the medial compartment and BMI (r=-0.35, P=0.019). No significant correlations were found in the lateral reference compartment., Conclusion: The lower dGEMRIC Index of the medial compartment suggests decreased cartilage GAG content after medial meniscectomy, indicating an early stage OA. Furthermore, results suggest that overweight is a factor that deteriorates cartilage, whereas strong and co-ordinated thigh muscles may have a protective effect on the cartilage integrity.

Objective: To evaluate the effects of moderate exercise on glycosaminoglycan (GAG) content in knee cartilage in subjects at high risk of knee osteoarthritis (OA)., Methods: Forty-five subjects (16 women, mean age 46 years, mean body mass index 26.6 kg/m(2)) who underwent partial medial meniscus resection 3-5 years previously were randomized to undergo a regimen of supervised exercise 3 times weekly for 4 months or to a nonintervention control group. Cartilage GAG content, an important aspect of the biomechanical properties of cartilage, was estimated by delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC), with results expressed as the change in the T1 relaxation time in the presence of Gd-DTPA (T1[Gd])., Results: Thirty of 45 patients were examined by dGEMRIC at baseline and followup. The exercise group (n = 16) showed an improvement in the T1(Gd) compared with the control group (n = 14) (15 msec versus -15 msec; P = 0.036). To study the dose response, change in the T1(Gd) was assessed for correlation with self-reported change in physical activity level, and a strong correlation was found in the exercise group (n = 16, r(S) = 0.70, 95% confidence interval [95% CI] 0.31-0.89) and in the pooled group of all subjects (n = 30, r(S) = 0.74, 95% CI 0.52-0.87)., Conclusion: This in vivo cartilage monitoring study in patients at risk of knee OA who begin exercising indicates that adult human articular cartilage has a potential to adapt to loading change. Moderate exercise may be a good treatment not only to improve joint symptoms and function, but also to improve the knee cartilage GAG content in patients at high risk of developing OA.

Objective: To examine the glycosaminoglycan (GAG) content in cartilage and that in synovial fluid and determine whether they are associated, in patients with an acute anterior cruciate ligament (ACL) injury., Methods: Twenty-four patients (14 of whom were male) with a mean age of 27 years (range 17-40 years) were assessed with delayed gadolinium-enhanced magnetic resonance imaging (MRI) of cartilage an average of 3 weeks after an ACL rupture and compared with 24 healthy volunteers. Two hours after an intravenous injection of Gd-DTPA(2-) (0.3 mmoles/kg body weight), quantitative measurements of the T1 relaxation time (T1(Gd) [T1 relaxation time in the presence of Gd-DTPA]) were made in lateral and medial femoral weight-bearing cartilage. In the patients, synovial fluid was aspirated immediately before the MRI, and GAG was analyzed using dye precipitation with Alcian blue., Results: Fifteen of the 24 patients had an isolated bone bruise in the lateral femoral condyle, where the cartilage T1(Gd) was shorter than that in the controls (mean +/- SD 385 +/- 83 msec and 445 +/- 41 msec, respectively; P = 0.004), consistent with decreased GAG content. However, the T1(Gd) was also decreased in the medial femoral cartilage, where bone bruises were rare (376 +/- 76 msec in patients versus 428 +/- 38 msec in controls; P = 0.006). The mean +/- SD synovial fluid GAG concentration in patients was 157 +/- 86 mug/ml and showed a positive correlation with the T1(Gd) (r = 0.49, P = 0.02)., Conclusion: This study indicates that an ACL injury causes posttraumatic edema of the lateral femoral cartilage but initializes a generalized biochemical change within the knee that leads to GAG loss from both lateral and medial femoral cartilage. In cartilage with a high GAG content (long T1(Gd)), more GAG is released into the synovial fluid, suggesting that cartilage quality is a factor to consider when interpreting cartilage biomarkers of metabolism.

Purpose: To establish the reproducibility of a standardized region of interest (ROI) drawing procedure in delayed gadolinium-enhanced magnetic resonance imaging (MRI) of cartilage (dGEMRIC)., Material and Methods: A large ROI in lateral and medial femoral weight-bearing cartilage was drawn in images of 12 healthy male volunteers by 6 investigators with different skills in MRI. The procedure was done twice, with a 1-week interval. Calculated T1-values were evaluated for intra- and interobserver variability., Results: The mean interobserver variability for both compartments ranged between 1.3% and 2.3% for the 6 different investigators without correlation to their experience in MRI. Post-contrast intra-observer variability was low in both the lateral and the medial femoral cartilage, 2.6% and 1.5%, respectively. The larger variability in lateral than in medial cartilage was related to slightly longer and thinner ROIs., Conclusion: Intra-observer variability and interobserver variability are both low when a large standardized ROI is used in dGEMRIC. The experience of the investigator does not affect the variability, which further supports a clinical applicability of the method.

It is well known that exercise, alone or combined with weight reduction, reduces pain and improves function in patients with osteoarthritis. The knowledge of the effects of exercise on cartilage is limited and needs to be improved however. It seems as cartilage adapts to loading as other biological tissues like bone and muscle, and moderate loading seems to be beneficial both for prevention and treatment of osteoarthritis. Too high loads, like elite soccer or repeated knee bendings several hours daily, are associated with increased risk of osteoarthritis. Too high or too low mechanical load decrease the proteoglycan content of the cartilage, indicating not only elite sports but also physical inactivity being a possible risk factor for osteoarthritis development. Muscle weakness may proceed osteoarthritis. Prevention and treatment of osteoarthritis should include regular loading of the cartilage, keeping muscles strong and maintaining normal body weight.

Delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) is a new imaging technique to estimate joint cartilage glycosaminoglycan content by T1-relaxation time measurements after penetration of the hydrophilic contrast agent Gd-DTPA(2-). This study compares dGEMRIC in age-matched healthy volunteers with different levels of physical activity: Group 1 (n = 12): nonexercising individuals; Group 2 (n = 16): individuals with physical exercise averaging twice weekly; Group 3 (n = 9): male elite runners. dGEMRIC was performed 2 hr after an intravenous injection of Gd-DTPA(2-) at 0.3 mmol/kg body weight. T1 differed significantly between the three different levels of physical exercise. T1 values (mean of medial and lateral femoral cartilage) for Groups 1, 2, and 3 were: 382 +/- 33, 424 +/- 22 and 476 +/- 36, respectively (ms, mean +/- SD) (P = 0.0004, 1 vs. 2 and 0.0002, 2 vs. 3). Irrespective of the exercise level, T1 was longer in lateral compared to medial femoral cartilage (P = 0.00005; n = 37). In conclusion, this cross-sectional study indicates that human knee cartilage adapts to exercise by increasing the glycosaminoglycan content. Furthermore, results suggest a compartmental difference within the knee with a higher glycosaminoglycan content in lateral compared to medial femoral cartilage. A higher proportion of extracellular water, i.e., larger distribution volume, may to some extent explain the high T1 in the elite runners., (Copyright 2004 Wiley-Liss, Inc.)

Delayed contrast-enhanced MRI of cartilage (dGEMRIC) is a noninvasive technique to study cartilage glycosaminoglycan (GAG) content in vivo. This study evaluates dGEMRIC in patients with preradiographic degenerative cartilage changes. Seventeen knees in 15 patients (age 35-70) with arthroscopically verified cartilage changes (softening and fibrillations) in the medial or lateral femoral compartment, knee pain, and normal weight-bearing radiography were included. MRI (1.5 T) was performed precontrast and at 1.5 and 3 hr after an intravenous injection of Gd-DTPA(2-) at 0.3 mmol/kg body weight. T(1) measurements were made in regions of interest in medial and lateral femoral cartilage using sets of five turbo inversion recovery images. Precontrast, R(1) (R(1) = 1/T(1), 1/s) was slightly lower in diseased compared to reference compartment, indicating increased hydration (P = 0.01). Postcontrast, R(1) was higher in diseased than in reference compartment at 1.5 hr, 3.45 +/- 0.90 and 2.64 +/- 0.58 (mean +/- SD), respectively (P < 0.01), as well as at 3 hr, 2.94 +/- 0.60 and 2.50 +/- 0.37, respectively (P = 0.01). The washout of the contrast medium was faster in diseased cartilage as shown by a higher R(1) at 1.5 than at 3 hr in the diseased but not in the reference compartment. In conclusion, dGEMRIC can identify GAG loss in early stage cartilage disease with a higher sensitivity at 1.5 than 3 hr., (Copyright 2003 Wiley-Liss, Inc.)

The negatively charged contrast agent Gd-DTPA2- distributes inversely to the cartilage fixed charged density. This enables structural cartilage examinations by contrast-enhanced MRI. In line with the development of a clinically applicable protocol for such examinations, this study describes the temporal pattern of Gd-DTPA2- distribution in femoral knee cartilage at three different doses in healthy volunteers. Nineteen volunteers (ages 21-28 years) were examined with a 1.5T MRI system. Quantitative relaxation rate measurements were made in weight-bearing central parts of femoral cartilage using sets of five turbo inversion recovery images with different inversion times. The cartilage was analyzed before and four times (1-4 h) after an intravenous injection of Gd-DTPA2- at single, double, and triple doses: 0.1, 0.2, and 0.3 mmol/kg body weight, respectively. The increase in R1 postcontrast was linearly dose-related at all times. The highest R1 values were registered at 2 and 3 h postcontrast, suggesting 2 h to be optimal in the clinical situation. The triple dose indicated a subtle compartmental difference in men, with higher contrast distribution medially than laterally. Results suggest that the triple dose is needed to detect minor cartilage matrix differences., (Copyright 2001 Wiley-Liss, Inc.)

Objective: To compare interleukin-1alpha (IL-1alpha)-induced degradation of nasal and articular cartilages in terms of proteoglycan loss and type II collagen cleavage, denaturation, and release; to examine the temporal relationship of these changes; and to investigate the effects of an inhibitor of collagenase 2 and collagenase 3 on these catabolic processes., Methods: Discs of mature bovine nasal and articular cartilages were cultured with or without human IL-1alpha (5 ng/ml) with or without RS102,481, a selective synthetic inhibitor of collagenase 2 and collagenase 3 (matrix metalloproteinase 8 [MMP-8] and MMP-13, respectively) but not of collagenase 1 (MMP-1). Immunoassays were used to measure collagenase-generated type II collagen cleavage neoepitope (antibody COL2-3/4C(short)) and denaturation (antibody COL2-3/4m), as well as total type II collagen content (antibody COL2-3/4m) in articular cartilage and culture media. A colorimetric assay was used to measure total proteoglycan concentration (principally of aggrecan) as sulfated glycosaminoglycans (sGAG)., Results: IL-1alpha initially induced a decrease in tissue proteoglycan content in nasal cartilage. A progressive loss of proteoglycan was noted during culture in articular cartilages, irrespective of the presence of IL-1alpha. In both cartilages, proteoglycan loss was followed by IL-1alpha-induced cleavage of type II collagen by collagenase, which was often reflected by increased denaturation. The inhibitor RS102,481 had no clear effect on the reduction in proteoglycan content (measured by sGAG) and collagen denaturation in either cartilage, but at 10 nM it inhibited the enhanced cleavage of type II collagen, partially in nasal cartilage and completely in articular cartilage., Conclusion: IL-1alpha-induced cleavage and denaturation of type II collagen is observed in both hyaline cartilages and is secondary to proteoglycan loss. It probably involves different collagenases, since there is no evidence of a rate-limiting role for collagenase 1 in articular cartilage, unlike the case for nasal cartilage. Inhibitors of this kind may be of value in the treatment of cartilage damage in arthritis. Also, the ability to detect the release of type II collagen collagenase-generated fragments from degraded cartilage offers the potential to monitor cartilage collagen damage and its control in vivo.

Objective: To examine whether type II collagen cleavage by collagenase and loss of proteoglycan are excessive in human osteoarthritic (OA) articular cartilage compared with nonarthritic articular cartilage, and whether this can be inhibited by a selective synthetic inhibitor that spares collagenase 1 (matrix metalloproteinase 1 [MMP-1])., Methods: Articular cartilage samples were obtained during surgery from 11 patients with OA and at autopsy from 5 adults without arthritis. The articular cartilage samples were cultured in serum-free medium. A collagenase-generated neoepitope, which reflects cleavage of type II collagen, and proteoglycan glycosaminoglycan (GAG), which predominantly reflects aggrecan release, were assayed in culture media. In addition, cultures were performed using either of 2 synthetic MMP inhibitors, both of which inhibited collagenase 2 (MMP-8) and collagenase 3 (MMP-13), but one of which spared collagenase 1. Cultures were also biolabeled with 3H-proline in the presence and absence of these inhibitors to measure collagen synthesis (as tritiated hydroxyproline) and incorporation in articular cartilage., Results: As a group, cleavage of type II collagen by collagenase was significantly increased in OA cartilage samples. In contrast, proteoglycan (GAG) release was not increased. This release of a collagenase-generated epitope was inhibited by both MMP inhibitors in 2 of 5 nonarthritic samples and in 9 of 11 OA cartilage samples. The inhibitor that spared collagenase 1 was generally more effective and inhibited release from 4 of 5 nonarthritic cartilage samples and the same OA cartilage samples. Group analyses revealed that the inhibition of collagenase neoepitope release by both inhibitors was significant in the OA patient cartilage, but not in the nonarthritic cartilage. Proteoglycan loss was unaffected by either inhibitor. Newly synthesized collagen (predominantly, type II) exhibited increased incorporation in OA cartilage, but only in the presence of the inhibitor that arrested collagenase 1 activity., Conclusion: These results further indicate that the digestion of type II collagen by collagenase is selectively increased in OA cartilage, and that this can be inhibited in the majority of cases by a synthetic inhibitor that can inhibit collagenases 2 and 3, but not collagenase 1. The results also suggest that in OA, newly synthesized collagen is digested, but in a different manner than that of resident molecules. Proteoglycan release was not increased in OA cartilage and was unaffected by these inhibitors. Inhibitors of this kind may be of value in preventing damage to type II collagen in human arthritic articular cartilage.

Objective: To determine the within- and between-patient variability in the concentrations of synovial fluid, serum and urine markers of joint tissue metabolism in a cohort of patients with knee pain and cartilage changes consistent with early-stage knee osteoarthritis., Design: Samples of synovial fluid, serum, and urine were obtained from 52 patients on eight different occasions during 1 year, as part of a clinical trial in patients with cartilage abnormalities and knee pain. In joint fluid, aggrecan fragments were quantified by dye precipitation and enzyme-linked immunosorbent assay (ELISA), and matrix metalloproteinases-1 and -3, and tissue inhibitor of metalloproteinases-1 by sandwich ELISAs. In serum, keratan sulfate was quantified by ELISA. Type I collagen N-telopeptide cross-links in urine were determined by ELISA., Results: The degree of cross-sectional variability in marker concentrations did not vary between the different sampling occasions, and did not differ between the periods of weeks 0 (baseline), 1-4 (treatment) and 13-26 (follow-up). Both between-patient and within-patient coefficients of variation varied for markers in different body fluid compartments, with the lowest variability for serum keratan sulfate, followed by urine type I collagen N-telopeptide crosslinks, and the highest for synovial fluid markers. For synovial fluid, aggrecan fragments showed the least variability, and matrix metalloproteinases the highest. One patient with septic arthritis showed a fivefold peak increase in joint fluid aggrecan fragment concentrations, while the concentration of matrix metalloproteinase-3 increased 100-fold., Conclusions: Molecular markers of joint tissue metabolism have been suggested as, for example, outcome measures for clinical trials of disease-modifying drugs in osteoarthritis. This report is the first to present data on between- and within-patient variability for such molecular markers in three different body fluid compartments in stable cohort of patients. The availability of such data enables calculations to determine the number of patients needed in prospective studies using these markers as outcome measures.

We demonstrate the direct involvement of increased collagenase activity in the cleavage of type II collagen in osteoarthritic human femoral condylar cartilage by developing and using antibodies reactive to carboxy-terminal (COL2-3/4C(short)) and amino-terminal (COL2-1/4N1) neoepitopes generated by cleavage of native human type II collagen by collagenase matrix metalloproteinase (MMP)-1 (collagenase-1), MMP-8 (collagenase-2), and MMP-13 (collagenase-3). A secondary cleavage followed the initial cleavage produced by these recombinant collagenases. This generated neoepitope COL2-1/4N2. There was significantly more COL2-3/4C(short) neoepitope in osteoarthritis (OA) compared to adult nonarthritic cartilages as determined by immunoassay of cartilage extracts. A synthetic preferential inhibitor of MMP-13 significantly reduced the unstimulated release in culture of neoepitope COL2-3/4C(short) from human osteoarthritic cartilage explants. These data suggest that collagenase(s) produced by chondrocytes is (are) involved in the cleavage and denaturation of type II collagen in articular cartilage, that this is increased in OA, and that MMP-13 may play a significant role in this process.

Objective: To examine if unilateral knee injury affects the synovial fluid concentrations of aggrecan fragments, cartilage oligomeric matrix protein (COMP) fragments, stromelysin-1, and tissue inhibitor of metalloproteinases-1 (TIMP-1) in the contralateral uninjured knee., Methods: Synovial fluids from the injured and uninjured knees were obtained at different times in a group of patients after unilateral knee trauma. Serum samples were obtained on the same occasion. Concentrations of aggrecan fragments were determined by precipitation with Alcian Blue; those of COMP fragments, stromelysin-1, and TIMP-1 were measured by immunoassay. Concentrations were compared with those in a reference group of 10 healthy volunteers., Results: Immediately after knee injury, concentrations of aggrecan fragments, COMP fragments, stromelysin-1 and TIMP-1 were increased in the synovial fluid of the injured knee. However, concentrations of aggrecan and COMP fragments, and stromelysin-1 increased also in the contralateral uninjured knee immediately after injury, but less than in the injured knee. Subsequently, the concentrations of all markers decreased in the synovial fluid of the injured knee, but remained unchanged in the uninjured knee. The concentration of aggrecan fragments in the injured knee decreased to less than that in the uninjured knee in the chronic phase. Serum concentrations of COMP were much smaller than those in synovial fluid., Conclusions: The increased concentrations of aggrecan and COMP fragments and stromelysin-1 in the joint fluid of the contralateral, uninjured knee following unilateral knee injury, compared with concentrations in healthy reference knees, suggest changes in joint cartilage metabolism in both knees following unilateral knee injury. The mechanisms for these changes are unclear. The low serum concentration of COMP makes it less likely that there is any significant 'exchange' of molecular makers between the knees. A further consequence of these findings is that the contralateral knee cannot be recommended as the only control joint in studies of matrix metabolism in patients with unilateral knee injury.

This is the first study which quantifies aggrecan fragments, stromelysin-1 and tissue inhibitor of metalloproteinases-1 (TIMP-1) in SF samples prospectively obtained from the same patient at different time intervals after a cruciate ligament injury of the knee. Aggrecan fragment concentrations were determined by dye precipitation with Alcian Blue. Stromelysin-1 and TIMP-1 were analysed by immunoassay. Ten healthy volunteers formed the reference group. Immediately after knee injury, all marker concentrations were higher as compared to the reference group. The high marker concentrations decreased gradually with time, and in samples obtained between 6 months and 6 years after the injury, median concentrations of some of the markers were not different compared to reference levels. This was in contrast to results from previous cross-sectional studies, where chronic phase median concentrations of all markers were consistently higher than reference levels. In previous cross-sectional studies, however, the samples were obtained at arthroscopy done because of knee complaints at different times after a knee injury. In the present study, the knee injured patients visited the orthopaedic outpatient ward only for SF sampling, and they had no or only minor knee symptoms. We conclude that the temporal changes of marker concentrations in joint fluid after knee injury, suggested from cross-sectional studies, have now been confirmed in a longitudinal, prospective cohort study. We further find that in patients with mild knee symptoms in the chronic phase after cruciate ligament injury, median SF levels of aggrecan fragments, stromelysin-1, and TIMP-1 are lower than in patients with significant knee complaints after the same type of injury.(ABSTRACT TRUNCATED AT 250 WORDS)

Osteoarthritis is a major cause of disability and early retirement. Yet we lack the means to diagnose the disease in its early stages or to monitor the effects of treatment on the target tissue, the joint cartilage. Neither can we identify the disease mechanisms at the tissue or cell level. Current research focuses on the use of markers of cartilage matrix metabolism in body fluids as a means to diagnose and monitor osteoarthritis. Cartilage proteoglycan, collagen and glycoprotein fragments, as well as proteinases and their inhibitors, are being suggested for this purpose. Structural information on matrix molecule fragments released into body fluids may also help to identify the enzymes active in the destruction of the cartilage, a central issue in osteoarthritis.