Differential matrix degradation and turnover in early cartilage lesions of human knee and ankle joints

OBJECTIVE: To determine whether there are differences in matrix turnover within early cartilage lesions of the ankle (talocrural) joint compared with the knee (tibiofemoral) joint that may help explain differences in the prevalence of osteoarthritis in these 2 joints. METHODS: Cartilage removed from lesions of the tali and femoral condyles was analyzed for type IIB collagen messenger RNA, C-terminal type II procollagen propeptide (CPII), the collagenase cleavage neoepitope (Col2-3/4C(short)), and the denaturation epitope (Col2-3/4m). The content of collagen, glycosaminoglycan, and epitope 846 of aggrecan was quantitated. RESULTS: In ankle lesions, there was an up-regulation of markers of synthesis (CPII [P = 0.07]; epitope 846 [P < or = 0.0001]), but these were down-regulated in the knee (CPII [P = 0.1]; epitope 846 [P = 0.004]). In lesions of the knee, but not the ankle, there was an up-regulation of collagen degradation markers (P = 0.008). On a molar basis, there was 24 times more cleavage epitope than denaturation epitope in knee lesions compared with ankle lesions. CONCLUSION: The up-regulation of matrix turnover that is seen in early cartilage lesions of the ankle would appear to represent an attempt to repair the damaged matrix. The increase in collagen synthesis and aggrecan turnover seen in ankle lesions is absent from knee lesions. Instead, there is an increase in type II collagen cleavage. Together with the differences in collagen denaturation, these changes point to an emphasis on matrix assembly during early lesion development in the ankle and to degradation in the knee, resulting in fundamental differences in matrix turnover in these lesions.     

Matrix homeostasis in aging normal human ankle cartilage

OBJECTIVE: To study age-related (as opposed to arthritis-related) changes in collagen and proteoglycan turnover. METHODS: Macroscopically nondegenerate normal ankle cartilage obtained from 30 donors (ages 16-75 years) was processed for in situ hybridization to detect messenger RNA (mRNA) of type IIB collagen (CIIB); antibodies to the C-propeptide of type II collagen (CPII), to the type II collagen (CII) collagenase-generated cleavage neoepitope (Col2-3/4C(short)), and to the CII denaturation product (Col2-3/4m) were used for immunohistochemistry analysis and immunoassay. In addition, immunoblotting was used to detect the 4 collagenases. Assays were also performed to detect glycosaminoglycan (GAG) content and the 846 epitope of aggrecan. RESULTS: There were no significant changes in CII, GAG, and the content of the 846 epitope after the age of 30 years. Both mRNA for CIIB and the CPII were present in all zones, and CPII content did not change significantly with age. While the collagenase-cleaved CII showed a trend to increase with age, the denatured collagen did not. However, the molar ratio of cleaved versus denatured collagen was positively correlated with age. All 4 collagenases were detectable in the ankle cartilage but showed no identifiable changes in content with age. CONCLUSION: Synthesis and degradation of CII is associated with the pericellular matrix and is maintained at a steady state throughout life. The contents of CII and proteoglycan did not change. There was a significant reduction in the denaturation of CII with age, relative to collagenase-mediated cleavage. These observations reveal that, in aging of the intact ankle articular cartilage, there is no evidence of molecular degenerative changes of the kind observed in osteoarthritis, thereby distinguishing aging from the osteoarthritis process.     

Type II collagen fragment HELIX-II is a marker for early cartilage lesions but does not predict the progression of cartilage destruction in human knee joint synovial fluid

To determine whether there is a direct correlation between the concentration of type II collagen fragment HELIX-II in synovial fluid and the severity of cartilage damage at the knee joint, 83 patients who had undergone knee arthroscopy or total knee replacement were enrolled in this study (49% women, mean ± SD age 49.5 ± 19). The content of HELIX-II in the synovial fluid samples was measured by enzyme-linked immunosorbent assay (ELISA). Cartilage damage at the knee joint was classified during arthroscopy or direct surgical observation, using the Outerbridge cartilage damage scoring system. The maximum damage score was defined as the highest score among the six areas of the knee joint, and the cumulative score was defined as the sum of the scores of the six areas of the knee joint. The intra-assay and inter-assay variations of the HELIX-II ELISA were lower than 13 and 15%, respectively. The level of HELIX-II in the severely damaged cartilage groups (cumulative scores = 11–24 or maximum score = 2–4) was much higher than in the slightly damaged cartilage groups (cumulative scores = 0–10 or maximum score = 0–1). The level of HELIX-II in cartilage from severely damaged cartilage groups was significantly higher than in the slightly damaged groups, but no significant difference was detected in the level of HELIX-II among the severely damaged cartilage sub-groups. There was a significant correlation between the HELIX-II concentration in the synovial fluid and the cumulative (r = 0.807) and maximum scores (r = 0.794). Thus, elevated HELIX-II level is correlated with early cartilage lesions, but does not have the sensitivity to predict the progression of severity of cartilage damage in the knee joint.     

Amyloid deposits in human knee and hip joints

Summary A systematic search for articular amyloidosis was carried out on both knees of 53 autopsy cases and on 26 femoral heads resected during surgery for hip prosthesis. Typical amyloid deposits exhibiting green apple birefringence following Congo red staining and thioflavin T fluorescence were found in 58.5% and 29% of the cases, in the knee and hip joint respectively. They occurred more frequently in articular cartilage than in the synovium, and in elderly subjects more than young ones. In the knee joint, the osteoarthritic changes and to a minor extent synovial inflammation appear to be positively correlated for the presence of intraarticular amyloid deposits. Such a correlation was not observed for the presence of calcium pyrophosphate dihydrate crystal deposits.     

Validity of histopathological grading of articular cartilage from osteoarthritic knee joints

OBJECTIVES: To determine the validity of the histological-histochemical grading system (HHGS) for osteoarthritic (OA) articular cartilage. METHODS: Human articular cartilage was obtained from macroscopically normal (n = 13) and OA (n = 21) knee joints. Sections of central and peripheral regions of normal samples were produced. Sections of regions containing severe, moderate, and mild OA changes were produced from each OA sample. A total of 89 sections were graded by means of the HHGS (0-14) twice by three observers. RESULTS: Average scores for regions designated severe (8.64) and moderate (5.83) OA were less than the expected (10-14 and 6-9, respectively) according to the HHGS, whereas average scores for the region designated mild (5.29) OA and central and peripheral regions (2.19) of normal cartilage were higher than expected (2-5 and 0-1, respectively). The HHGS was capable of differentiating between articular cartilage from macroscopically normal and OA joints and between the region designated severe OA and other regions. However, the HHGS did not adequately differentiate between regions designated mild and moderate OA. Values for sensitivity, specificity, and efficiency for all regions varied considerably. CONCLUSION: The HHGS is valid for normal and severe OA cartilage, but does not permit distinction between mild and moderate OA changes in articular cartilage.     

Effect of dehydroepiandrosterone on cartilage and synovium of knee joints with osteoarthritis in rabbits

The aim of this study was to investigate the effects of intra-articular injection of dehydroepiandrosterone (DHEA) on cartilage and synovium of knee joints with osteoarthritis (OA) in rabbits and the underlying mechanism. Forty rabbits underwent unilateral anterior cruciate ligament transaction and were divided into two groups. Rabbits were injected with 100 μmol/l DHEA dissolved in the dimethylsulphoxide (DMSO) in the knee joints 5 weeks after transaction, once a week for 5 weeks. Rabbits injected with DMSO under the same condition were served as a control. All rabbits were killed 1 week after the last injection. The knee joints were evaluated by gross morphology, histology, and gene expression analysis. Gross morphologic inspection and histological evaluation showed that the DHEA group appeared less damage in cartilage and synovium as compared with the control. Gene expression analysis revealed that the mRNA expression of matrix metalloproteinase-3 (MMP-3) in cartilage and synovium decreased significantly in the DHEA group and that of tissue inhibitor of metalloproteinase-1 (TIMP-1) increased. No significant difference of interleukin-1 beta (IL-1β) mRNA expression was found in the cartilage between two groups while the mRNA expression of IL-1β in the synovium was largely suppressed in the DHEA group. The study suggests that DHEA plays a protective role against cartilage degradation and synovium inflammation in rabbits with OA. This role may be achieved through the regulation of the MMP-3, TIMP-1, and IL-1β gene expression in the cartilage and synovium.     

Relationship between the tensile properties of articular cartilage from the human knee and age.

The relationship between the tensile properties of articular cartilage and age has been investigated in vitro in the human knee joint. Specimens orientated parallel to the articular surface were excised from the superficial and deep zones of the femoral condyles of knee joints of persons in the age range from 8 to 91 years. The results showed that the tensile strength of the superficial zone increased with age to reach a maximum value in the third decade. Thereafter the strength decreased markedly with increasing age. The tensile strength of cartilage from the deep zone did not show an increase in the early years but decreased continuously with age. The tensile stiffness of the superficial layer at stresses of 5 MN/m2 and 10 MN/m2 increased to maximum values in the third decade and thereafter decreased with increasing age. The stiffness of the deep zone decreased continuously with age. It is suggested that these results reflect changes in the organisation of the collagen fibre mesh with age and possibly also changes in the collagen cross-links.     

Newly synthesized and endogenous proteoglycans in human osteoarthritic knee cartilage

Newly synthesized and endogenous proteoglycans of human osteoarthritic knee cartilages were studied in short term explant culture. There were more newly synthesized proteoglycans than endogenous uronic acid containing proteoglycans found in the A1 fraction generated by associative CsCl density gradient centrifugation. Thirty-five percent of the proteoglycans in the A1 fraction were shown to elute in the void volume on Sepharose CL-2B, under associative conditions. Reduction and alkylation of the A1 fraction indicated that the endogenous proteoglycan monomer was hydrodynamically smaller than the newly synthesized macromolecule. This was confirmed by a Sepharose CL-2B chromatography of the A1D1 fraction. In comparison to endogenous proteoglycans, the newly synthesized proteoglycan monomers were less capable of forming stable aggregates with exogenous hyaluronic acid. We also found a statistically significant decrease (p less than 0.05) in the amount of already existing proteoglycan subclass when compared to the amount of newly synthesized proteoglycans in the most dense A1D1 fraction. By contrast, a statistically significant increase (p less than 0.05) in the amount of endogenous vs newly synthesized proteoglycans was seen in the A1D2 fraction but not in fractions of a lower buoyant density.     

Prevalence of cartilage lesions in foot joints: a test of the concept of limited and progressive lesions.

Lesions in the articular cartilage of the joints of the first ray of feet amputated from 54 patients in the age range 10-79 years have been categorised by site, size, and stage of development. The large number of small lesions in early stages of development, appearing from an early age, and the few lesions with extensive bone exposure, occurring only in later years, lend corroboration to the concept of limited and progressive lesions, as established for the hip joint.     

Thickness of human articular cartilage in joints of the lower limb

OBJECTIVES: (a) To determine the topographical variations in cartilage thickness over the entire surfaces of cadaveric lower limb joints, and (b) to examine the correlations between: cartilage thickness and its site specific modulus; cartilage thickness and donor age, weight, height, and body mass index. METHODS: The cartilage thickness of 11 sets of cadaveric human joints each comprising an ankle, knee, and hip was measured using a needle probe technique. Statistical analysis was used to compare the cartilage thickness of the different lower limb joints and the differences in cartilage thickness over the surface of individual joints. It was further examined whether cartilage had a correlation with its stiffness, and any of the details of the specimen donors such as age, weight, height, and body mass index. RESULTS: The mean cartilage thickness of the knee was significantly greater than that of the ankle and hip (p < 0.001) in all 11 sets of joints, while the cartilage thickness of the hip was significantly greater than that of the ankle in 10 sets of joints (p < 0.001). The mass of specimen donors was found to correlate with the mean cartilage thickness of all three lower limb joints. A correlation was also found between the height of donors and the mean cartilage thickness of the knee and hip joints, while only in the ankle joint was a correlation found between the mean cartilage thickness and the body mass index of the specimen donors. A further correlation was found between cartilage thickness and its modulus; the thinner the cartilage, the higher the modulus. CONCLUSIONS: The thickness of articular cartilage seems to be related to the congruance of a joint; thin cartilage is found in congruent joints such as the ankle, whereas thick cartilage is found in incongruent joints such as the knee. The correlations in this study imply that the larger and heavier was a donor the thicker was the cartilage in the lower limb joints. The data further suggest the presence of an inverse relation between the mean cartilage thickness and mean compressive modulus in each of the joints examined.     

Body composition and knee cartilage properties in healthy, community-based adults

BACKGROUND: Although obesity is widely accepted as a risk factor for knee osteoarthritis, whether weight per se or the specific components of body composition are the major determinants of properties of articular knee cartilage is unclear. OBJECTIVE: To examine associations between anthropometric and body composition measures and knee cartilage properties in healthy adults. METHODS: 297 healthy adults with no clinical knee osteoarthritis were recruited from an existing community-based cohort. Anthropometric measures and body composition, including fat-free mass and fat mass assessed using bioelectrical impedance analysis, were measured at baseline (1990-4) and current follow-up (2003-4). Tibial cartilage volume and tibiofemoral cartilage defects were assessed using MRI at follow-up. RESULTS: After adjustment for potential confounders, baseline and current fat-free mass, independent of fat mass, were positively associated with tibial cartilage volume (all p<0.001). Increased fat-free mass over the time period was positively associated with tibial cartilage volume (p<0.001). Current fat mass was negatively associated with tibial cartilage volume (p = 0.004). Baseline and current fat mass were weakly associated with increased tibiofemoral cartilage defects (p = 0.06 and p = 0.07, respectively), independent of fat-free mass. CONCLUSION: The findings suggest a beneficial effect of fat-free mass, but a deleterious effect of fat mass, on knee cartilage properties in healthy adults. This suggests that weight-loss programmes aimed at reducing fat mass but maintaining muscle mass may be important in preventing the onset and/or progression of knee osteoarthritis.     

Apoptosis and p53 expression in chondrocytes relate to degeneration in articular cartilage of immobilized knee joints

OBJECTIVE: We have reported that articular cartilage showed early stage degeneration at 7 and 14 days after immobilization, moderate degeneration at 28 days, and severe degeneration at 42 days in rabbits. To test whether apoptosis occurs in association with p53 expression in chondrocytes during the process of articular cartilage degeneration, we investigated the degree of cartilage degeneration, the frequency of apoptotic cells, and the levels of p53 mRNA in rabbits and mice after knee immobilization. METHODS: Right knees of male Japanese white rabbits were immobilized in full extension with fiberglass casts for up to 42 days. Similarly, right knees of male p53 wild-type [p53 (+/+)] and p53 null [p53 (-/-)] mice were immobilized in full extension with bandage tape for up to 84 days. Apoptotic cells were confirmed by TUNEL staining on the sections of knee joints. Total RNA of articular chondrocytes obtained from Day 0 or immobilized knees was analyzed semiquantitatively by RT-PCR using specific primers for p53. RESULTS: Articular cartilage degenerated after immobilization of p53 (+/+) mouse knees, but not after immobilization of p53 (-/-) knees. Apoptotic cells were observed in articular cartilage in the femur and tibia of rabbits and p53 (+/+) mice after immobilization. However, only a few apoptotic cells were observed at the same sites in p53 (-/-) mice. In RT-PCR analysis, the levels of p53 mRNA obtained from immobilized groups were significantly higher than those of Day 0 groups in rabbit and p53 (+/+) mouse knees. CONCLUSION: Apoptosis and p53 expression in chondrocytes relate to degeneration in articular cartilage of immobilized knee joints.     

Weight bearing controls glycosaminoglycan concentration and articualr cartilage thickness in the knee joints of young beagle dogs

Casting of the right knee (stifle) joints of young beagle dogs for 11 weeks caused up to 48% reduction in the glycosaminoglycan (GAG) concentration of the uncalcified articular cartilage, as assessed by a new microspectrophotometric method. The GAGs were depleted mainly in the superficial zone of the cartilage. Although the thickness of the uncalcified cartilage was not decreased, the calcified cartilage under the tidemark was thinned by 6–25% at the femoral condyles. The increased weight-bearing in the limb opposite the one in the splint caused uncalcified cartilage thickness to be augmented by 19% and GAG concentration by 25–35% in the intermediate, deep, and calcified zones of the summits of the femoral condyles; the changes were smaller in other, less loaded parts of the joint. It is concluded that in young dogs, increased weight-bearing augments local proteoglycan content of the articular cartilage matrix, while unloading reduces it.     

Correlation and sex differences between ankle and knee cartilage morphology determined by quantitative magnetic resonance imaging

OBJECTIVE: To study the correlation between ankle and knee cartilage morphology to test the hypothesis that knee joint cartilage loss in gonarthritis can be estimated retrospectively using quantitative MRI analysis of the knee and ankle and established regression equations; and to test the hypothesis that sex differences in joint surface area are larger in the knee than the ankle, which may explain the greater incidence of knee osteoarthritis in elderly women than in elderly men. METHODS: Sagittal MR images (3D FLASH WE) of the knee and hind foot were acquired in 29 healthy subjects (14 women, 15 men; mean (SD) age, 25 (3) years), with no signs joint disease. Cartilage volume, thickness, and joint surface area were determined in the knee, ankle, and subtalar joint. RESULTS: Knee cartilage volumes and joint surface areas showed only moderate correlations with those of the ankle and subtalar joint (r = 0.33 to 0.81). The correlations of cartilage thickness between the two joints were weaker still (r = -0.05 to 0.53). Sex differences in cartilage morphology at the knee and the ankle were similar, with surface areas being -17.5% to -23.5% lower in women than in men. CONCLUSIONS: Only moderate correlations in cartilage morphology of healthy subjects were found between knee and ankle. It is therefore impractical to estimate knee joint cartilage loss a posteriori in cross sectional studies by measuring the hind foot and then applying a scaling factor. Sex differences in cartilage morphology do not explain differences in osteoarthritis incidence between men and women in the knee and ankle.     

Long-distance running causes site-dependent decrease of cartilage glycosaminoglycan content in the knee joints of beagle dogs

Objective. To study the effects of a long-term (1-year) program of running exercise (up to 40 km/day) on the thickness and glycosaminoglycan (GAG) content of articular cartilage in the knee and humeral head cartilage of young dogs. Methods. Samples for histologic analysis were obtained from 12 different locations of the joints. We conducted a detailed, area-specific analysis, measuring the thickness of articular cartilage and analyzing the distribution of Safranin O stain that binds stoichiometrically to GAG as determined by quantitative microspectrophotometry. Results. Running exercise decreased the GAG content of the uncalcified articular cartilage in the weight-bearing summits of the femoral condyles by 5–13% (P < 0.05), while at margins of these areas the GAG content was equivalent to control levels. In the lateral condyle of the femur, the reduction was most prominent in the superficial zone (up to 28% decrease; P < 0.05), and extended into the intermediate zone (11% decrease; P < 0.05). GAG content was also significantly reduced in the superficial zone at the lateral condyle of the tibia and the head of the humerus, by 35% (P < 0.01) and 15% (P < 0.05), respectively. Running did not alter GAG concentration in the patellofemoral region. Conclusion. The GAG depletion caused by 40-km/day running exercise is restricted to prominent weight-bearing areas of the joint and begins from the superficial cartilage without signs of degeneration. The different degree and type of joint loading can explain the site-dependent cartilage response to long-distance running. The loss of GAGs was possibly due to breakdown of proteoglycans, which could not be compensated for by neosynthesis of molecules. With time, this may affect the condition of articular cartilage, especially if the joint is exposed to loading for lengthy periods.