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PubMed1990

Ultrasonic propagation properties of articular cartilage at 100 MHz.

A pilot study on articular cartilage assessed the contributions of individual matrix components to ultrasound propagation. The influence of collagen fibril orientation and collagen cross linking was also assessed. Sections of adult bovine articular cartilage cut both parallel and perpendicular to the articular surface were examined using the scanning laser acoustic microscope (SLAM) operating at an ultrasonic frequency of 100 MHz. A set of samples was evaluated that had been sequentially treated by enzymes to (1) remove 85% of the chondroitin sulfate; (2) remove remaining glycosaminoglycans, glycoproteins, and other noncollagen proteins, leaving only the collagen fibril network; and (3) disrupt the collagen intermolecular cross links. Two striking observations were made: a profound effect of the "preferred" collagen fibril orientation on ultrasonic speed and a marked increase in attenuation coefficient when intermolecular cross links were broken in the collagen.

Animals↗
PubMed2001

Use of an antineoepitope antibody for identification of type-II collagen degradation in equine articular cartilage.

OBJECTIVE: To develop an antibody that specifically recognizes collagenase-cleaved type-II collagen in equine articular cartilage. SAMPLE POPULATION: Cartilage specimens from horses euthanatized for problems unrelated to the musculoskeletal system. PROCEDURE: A peptide was synthesized representing the carboxy- (C-) terminus (neoepitope) of the equine type-II collagen fragment created by mammalian collagenases. This peptide was used to produce a polyclonal antibody, characterized by western analysis for reactivity to native and collagenase-cleaved equine collagens. The antibody was evaluated as an antineoepitope antibody by ELISA, using peptides +/- an amino acid at the C-terminus of the immunizing peptide. Collagen cleavage was assayed from equine articular cartilage cultured with interleukin-1 (IL-1), +/- a synthetic MMP inhibitor, BAY 12-9566. Cartilage specimens from osteoarthritic and nonarthritic joints were compared for antibody staining. RESULTS: An antibody, 234CEQ, recognized only collagenase-generated 3/4-length fragments of equine type-II collagen. This was a true antineoepitope antibody, as altering the C-terminus of the immunizing peptide significantly decreased competition for binding in an inhibition ELISA. The IL-1-induced release of type-II collagen fragments from articular cartilage was prevented with the MMP inhibitor. Cartilage from an osteoarthritic joint of a horse had increased staining with the 234CEQ antibody, compared with normal articular cartilage. CONCLUSIONS AND CLINICAL RELEVANCE: We generated an antineoepitope antibody recognizing collagenase-cleaved type-II collagen of horses. This antibody detects increases in type-II collagen cleavage in diseased equine articular cartilage. The 234CEQ antibody has the potential to aid in the early diagnosis of arthritis and to monitor treatment responses.

Animals↗
PubMed1991

The effects of hydrostatic pressure on matrix synthesis in articular cartilage.

The direct effects of hydrostatic pressure on matrix synthesis in articular cartilage can be studied independently of the other factors that change during loading. We have found that the influence of hydrostatic pressure on incorporation rates of 35SO4 and [3H]proline into adult bovine articular cartilage slices in vitro depends on the pressure level and on the time at pressure. Pressures in the "physiological" range (5-15 MPa) applied for 20 s or for 5 min could stimulate tracer incorporation (30-130%) during the following 2 h, but higher pressures (20-50 MPa) had no effect on incorporation rates. The degree of stimulation in cartilage obtained from different animals was found to vary; in some animals none was seen. Stimulation also varied with position along the joint. Physiological pressures (5-10 MPa) applied continuously for the 2-h incubation period also stimulated incorporation rates, but pressures greater than 20 MPa always produced a decrease that was related to the applied pressure and that was reversible. These results suggests that the hydrostatic pressure that occurs during loading is a signal that can stimulate matrix synthesis rates in articular cartilage.

Animals↗
PubMed2001

In situ compressive stiffness, biochemical composition, and structural integrity of articular cartilage of the human knee joint.

OBJECTIVE: Reduction of compressive stiffness of articular cartilage has been reported as one of the first signs of cartilage degeneration. For the measurement of in situ compressive stiffness, a hand-held indentation probe has recently been developed and baseline data for macroscopically normal knee joint cartilage were provided. However, the histological stage of degeneration of the measured cartilage was not known. The purpose of this study was to investigate whether there is a relationship between the in situ measured compressive stiffness, the histological stage of degeneration, and the biochemical composition of articular cartilage. DESIGN: Instantaneous compressive stiffness was measured for the articular cartilage of 24 human cadaver knees. Additionally, biochemical composition (total proteoglycan and collagen content) and histological appearance (according to the Mankin score) were assessed for each measurement location. RESULTS: Despite visually normal surfaces, various histological signs of degeneration were present. A high correlation between Mankin score and cartilage stiffness was observed for the lateral patellar groove (R(2)=0.81), the medial (R(2)=0.83) and the lateral femoral condyle (R(2)=0.71), whereas a moderate correlation was found for the medial patellar groove (R(2)=0.44). No correlation was observed between biochemical composition and cartilage compressive stiffness. CONCLUSIONS: Our results are in agreement with others and show that the instantaneous compressive stiffness is primarily dependent on the integrity of the extracellular matrix, and not on the content of the major cartilage constituents. The high correlation between stiffness and Mankin score in mild osteoarthrosis suggests that the stage of cartilage degeneration can be assessed quantitatively with the hand-held indentation probe. Moderate and severe case of osteoarthrosis remains to be investigated.

Adult↗
PubMed1994

Localization of freezing injury in articular cartilage.

In order to improve techniques for cryopreservation of articular cartilage, a study has been carried out to assess localization of cryoinjury in intact articular cartilage. Osteochondral dowels taken from the femoral condyles of sheep were subjected to graded freezing in the presence and absence of a cryoprotectant (10% DMSO). The graded freezing technique involves slow cooling (1 degree C/min) to various subzero temperatures before either rapid warming or rapid cooling by plunging in liquid nitrogen. This protocol allows assessment of the separate effects of rapid and slow freezing which damage cells in different ways, and the effects of cryoprotectants on the different types of damage. To assay damage, thin slices of cartilage were cut with a vibratome, which allows viable cells within the matrix to be observed microscopically. Injury was assessed by staining with fluorescent dyes to indicate damage to the plasma membrane. In general, tissue response was similar to that of cell suspensions, showing at least two mechanisms of injury acting on the cells: one at slow cooling rates and another at rapid cooling rates. The primary effect of DMSO was to reduce injury due to slow cooling. When the location of injury within the tissue was examined, it was found that chondrocytes of the intermediate layer were injured more extensively than those of either the deep or superficial layers.

Animals↗
PubMed2002

The biochemical content of articular cartilage: an original MRI approach.

The MR aspect of articular cartilage, that reflects the interactions between protons and macromolecular constituents, is affected by the intrinsic tissue structure (water content, the content of matrix constituents, collagen network organization), imager characteristics, and acquisition parameters. On the T1-weighted sequences, the bovine articular cartilage appears as an homogeneous tissue in high signal intensity, whatever the age of animals considered, whereas on the T2-weighted sequences, the articular bovine cartilage presents variations of its imaging pattern (laminar appearance) well correlated to the variations of its histological and biochemical structure. The T2 relaxation time measurement (T2 mapping), which reflects quantitatively the signal intensity variations observed on T2 weighted sequences, is a way to evaluate more precisely the modifications of cartilage structure during the aging and maturation processes (rat's study). This technique so far confined to experimental micro-imagers is now developed on clinical imagers. Consequently, it may permit to depict the early stages of osteoarthritic disease (OA) or to evaluate the chondroprotective effect of drugs.

Aging↗
PubMed2000

Comparison of the degradation of type II collagen and proteoglycan in nasal and articular cartilages induced by interleukin-1 and the selective inhibition of type II collagen cleavage by collagenase.

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.

Animals↗
PubMed1995

Magnetic resonance imaging of articular cartilage: the state of the art.

Recent advances in the treatment of articular cartilage abnormalities have created a need for an accurate noninvasive imaging method for the evaluation of articular cartilage. Although its role remains undefined in this regard, magnetic resonance imaging (MRI) appears to be the most promising method. We review the current status and possible future directions of MRI of articular cartilage.

Cartilage, Articular↗
PubMed1990

The effect of experimental trypsin on the regeneration of hyaline articular cartilage.

There is evidence from other studies that some degree of cartilage healing may take place after the initiation of an inflammatory response. It is postulated that the induction of the platelet-cartilage interaction may eventuate in cartilage repair. The treatment of fresh articular cartilage with proteolytic enzymes rendered the tissue active as a platelet aggregant. During platelet aggregation a host of active substances are released which are known to play a role in the inflammatory response (Thompson 1975). This study was undertaken to evaluate the effects of trypsin on the surface injury of rabbit hyaline cartilage. The results were as follows: 1) Hyaline cell regeneration was observed only in the group treated with trypsin and blood; 2) Hyaline cartilage regeneration did not occur in the group treated with a single injection of trypsin or blood; 3) There was no significant damage to the healthy articular cartilage by the single injection of trypsin or blood, or both; and 4) Platelets do not adhere to cartilage and superficial damaged cartilage does not induce platelet aggregation.

Animals↗
PubMed1999

Influence of different exercise levels and age on the biochemical characteristics of immature equine articular cartilage.

This study aimed to examine whether biochemical characteristics of juvenile articular cartilage are changing during the first year post partum and whether they can be influenced by exercise at young age. Water, glycosaminoglycan (GAG), DNA, total collagen, hydroxylysine and hydroxylysylpyridinoline (HP) content were measured in articular cartilage of 43 foals that were subdivided into 3 groups (n = 15, 14 and 14) which were subjected to different exercise regimens from one week after birth to age 5 months. At the age of 5 months all foals were weaned and 8 foals were selected randomly from each exercise group and subjected to euthanasia. The remaining foals (n = 19) were grouped and subjected to a similar exercise regimen for an additional 6 months. Differences were tested by student's t test (P<0.01). No effect of exercise on the water or DNA content was found. GAG content increased with increasing exercise in the 5 months group. These differences had disappeared after 6 months of similar exercise. No influence of exercise could be demonstrated on any of the collagen parameters. When comparing 5 months with 11 months group, all parameters except hydroxylysine changed significantly during these 6 months. Water, DNA and GAG content decreased during maturation. Collagen and HP content increased. It is hypothesised that juvenile equine articular cartilage may be seen as a dynamic, continuously remodelling tissue that is gradually taking on the biochemical characteristics it will have during the rest of the life of the animal. Moderate exercise does not influence the collagen component of the extracellular matrix. It has a beneficial, but reversible, effect on the glycosaminoglycan component.

Aging↗
PubMed1990

Articular cartilage defects of the knee: correlation between magnetic resonance imaging and gross pathology.

Magnetic resonance imaging (MRI) of the knee articular cartilage is possible owing to the contrast provided by different signal intensities of adjacent menisci and subchondral bone. The objective of this study was to determine the accuracy of MRI in quantitatively detecting thinning and focal defects of articular cartilage in vivo. High resolution MRI was performed followed by dissection of the knee within one hour of amputations above the knee of eight patients (62-89 years) with peripheral vascular disease. Articular cartilage was examined for erosions, surface irregularities, and appearance. Mean thicknesses of femoral and tibial articular cartilage sagittal sections from MRI were statistically indistinguishable from matched gross thicknesses. In those joints in which cartilage erosions, thinning, or irregularities were detected by MRI the same defects were apparent by gross examination. Cartilage that appeared normal by MRI had a normal gross appearance by gross examination. Thus high resolution MRI can accurately predict gross articular cartilage appearance and thickness, allowing an objective, quantitative, noninvasive assessment of eroded cartilage.

Aged↗
PubMed2006

Effects of rapid cooling on articular cartilage.

In order to improve the technique and protocols of cryopreservation of articular cartilage, a study was carried out to assess the effects of rapid cooling on the intact articular cartilage. Cartilage slices with a thickness ranging from 0.2 to 0.5 mm taken from bovine metacarpal-phalangeal joints were subjected to rapid cooling by immersing them in liquid nitrogen with and without treatment of the VS55 cryoprotective agent (CPA). The ultrastructure, chondrocyte viability, swelling property, and glycosaminoglycan (GAG) content were then examined before and after cryopreservation to give qualitative and quantitative evaluation on the functional state of both chondrocytes and extracellular matrix. The transmission electron microscopy study demonstrated that damage to chondrocytes without CPA was far more pronounced than those with VS55 protection while the structure of the extracellular matrix altered little in either group. The cell viability assay showed that although the exposure to VS55 led to about 36% chondrocytes losing membrane integrity, the VS55 could provide protection to chondrocytes during rapid cooling and thawing, with approximately 51% of the cells having survived rapid cooling compared to fewer than 5% in the absence of CPA. There were no significant differences in degrees of swelling or the GAG contents of cartilage slices after cryopreservation indicating rapid freezing caused little damage to the matrix. Future research activities include searching improved CPA formulation, optimising the treatment protocol and investigating the long-term effects of rapid cooling on articular cartilage.

Animals↗
PubMed2003

Measurement of the mechanical condition of articular cartilage with an ultrasonic probe: quantitative evaluation using wavelet transformation.

OBJECTIVE: To develop a new diagnostic technology to evaluate articular cartilage quantitatively by introducing an ultrasonic probe into the knee joint under arthroscopy and analyzing the A-mode echogram by means of wavelet transformation. DESIGN: Quantitative evaluation using comparison of two indices on the wavelet map and macroscopic evaluation using the Outerbridge classification. As the quantitative indices on the wavelet map, the maximum magnitude and the echo duration which was defined as the length of time that included 95% of echo signal were selected. BACKGROUND: Quantitative evaluation of articular cartilage in situ is required for new tissue-engineered cartilage but an evaluation system that fully meets this requirement has yet to be established for clinical use. METHODS: Human articular cartilage specimens were analyzed using an ultrasonic probe after macroscopic evaluation and the cartilage characteristics on the echo duration-maximum magnitude graph were examined. RESULTS: There were significant differences between grade 1 and 3, and grade 2 and 3 in the echo duration and maximum magnitude. The cartilage specimens had a L-shaped distribution in echo duration-maximum magnitude graph. CONCLUSIONS: The present study is the new quantitative evaluation system of the articular cartilage in situ and a clinical trial under arthroscopy is presently underway. The ultrasonic measurement is highly reproducible, so we can expect this system to be suitable for in situ reliable examination under arthroscopy. RELEVANCE: Precise evaluation of articular cartilage is of particular importance for longitudinal clinical trials for determination of best surgical option or effect of new chondroprotective drugs in arthritic disease and rheumatoid arthritis.

Adult↗
PubMed1990

Indentation stiffness of young canine knee articular cartilage--influence of strenuous joint loading.

The indentation stiffness of knee articular cartilage subjected to strenuous physical training (SPT: treadmill running 20 km day-1 for 15 weeks, n = 6) of young Beagles was tested and compared to that obtained from age-matched (55 weeks, n = 9) controls. The mathematical solution for the shear modulus, as determined from indentation of an elastic layer bonded to a rigid half space, was extended to small Poisson's ratios and applied to the analysis of cartilage response after a step stress (0.39 MPa) application. In these measurements with an impervious, plane-ended indenter, the equilibrium deformation was systematically greater than values predicted from the instant response by the linear biphasic theory. Therefore, the accurate determination of Poisson's ratio from the creep curves was not possible. The mean shear modulus (calculated by using the deformation at 900 s after load application and assuming a constant Poisson's ratio of 0.40 for the matrix) of canine knee articular cartilage was 0.37 MPa. While the cartilage thickness was not affected by SPT, the cartilage of the lateral tibial plateau was stiffer (13.3%, p less than 0.05) than that in controls. However, in the femoral condyles, the stiffness was at the control level or even below. Our results on cartilage structure and properties suggest that SPT, in contrast to our previous findings with moderate training, does not necessarily improve the biological properties of articular cartilage in young animals.

Animals↗
PubMed1989

A mechanical apparatus with microprocessor controlled stress profile for cyclic compression of cultured articular cartilage explants.

An apparatus was designed for mechanical compression of cultured articular cartilage explants with acylindrical plain-ended loading head (diameter 2-5 mm) driven by a stepping motor. A load cell under the culture dish was applied for feedback regulation utilizing a microprocessor-based control unit. The operating programs allowed either continuous or cyclic loading, the latter with adjustable loading/resting ratio. The improvements in the present design compared with previously described apparatuses for similar purposes include: (1) the accurately controlled compression by a load cell and a rapid feedback circuit; (2) the wide range of selectable stresses (25 kPa-12.5 MPa) with both continuous and cyclic loading modes; (3) the ability to handle cycles as short as 1 s with 15 ms peak loading phase. Using a 4 s cycle and 0.5 MPa load for 1.5 h resulted in a significantly enhanced incorporation of radiosulphate in cultured bovine articular cartilage explants, suggesting a stimulation of proteoglycan synthesis. Light and scanning electron microscopic examinations revealed a slight depression and superficial alterations in cartilage structure at the impact site following high pressures. We expect that this apparatus will help in revealing how articular cartilage tissue and chondrocytes respond to external mechanical stimuli.

Animals↗
PubMed1997

The effect of intraarticular hydrocortisone injection on the articular cartilage of rabbits.

We investigated the effect of hydrocortisone on the articular cartilage of the knee in rabbits. 27 New Zealand white rabbits were injected intraarticularly with 25, 50 or 100 mg betamethasone acetate in 2 or 4 weekly intervals. Control animals were injected with normal saline and demonstrated no histological changes in the articular cartilage. Hydrocortisone administration was associated with increased cell size, as well as an increased stain density in the cytoplasm surrounding vacuoles. In addition, loss of cell organelles was also observed. High dose of hydrocortisone was associated with an obvious loss of cell shape and distortion of the cell membrane and nucleus. The magnitude of histological changes, found under light and electron microscopy, were proportional to the amount of hydrocortisone injected. Our findings strongly indicate that intraarticular injection of hydrocortisone alters the shape of articular cartilage chondrocytes, producing abnormal changes in the cytoplasm and nucleus and leading to cell degeneration.

Animals↗
PubMed2003

Articular cartilage functional histomorphology and mechanobiology: a research perspective.

The histomorphogenesis of articular cartilage is regulated during skeletal development by the intermittent forces and motions imposed at diarthrodial joints. A key feature in this development is the formation of the superficial, transitional, radial, and calcified cartilage zones through the cartilage thickness. The histomorphological, biological, and mechanical characteristics of these zones can be correlated with the distributions of pressures, deformations, and pressure-induced fluid flow that are created in vivo. In a mature joint, cyclic loads produce cyclic hydrostatic fluid pressure through the entire cartilage thickness that is comparable in magnitude to the applied joint pressure. Prolonged physical activity can cause the total cartilage thickness to decrease about 5%, although the consolidation strains vary tremendously in the different zones. The superficial zone can experience significant fluid exudation and consolidation (compressive strains) in the range of 60% while the radial zone experiences relatively little fluid flow and consolidation. The topological variation in the histomorphologic appearance of articular cartilage is influenced by the local mechanical loading of chondrocytes in the different zones. Patterns of stress, strain, and fluid flow created in the joint result in spatial and temporal changes in the rates of synthesis and degradation of matrix proteins. When viewed over the course of a lifetime, even subtle difference in these cellular processes can affect the micro- and macro-morphology of articular cartilage. This hypothesis is supported by in vivo and ex vivo experiments where load-induced changes in matrix synthesis and catabolism, gene expression, and signal transduction pathways have been observed.

Animals↗
PubMedDate not supplied

Articular cartilage changes seen with magnetic resonance imaging-detected bone bruises associated with acute anterior cruciate ligament rupture.

Occult osteochondral lesions (bone bruises) have been documented on magnetic resonance images in more than 80% of patients sustaining acute anterior cruciate ligament ruptures. Despite the high prevalence of these lesions, little is known about the histologic changes in the adjacent articular cartilage. Ten patients with acute anterior cruciate ligament ruptures who had a preoperatively documented (by magnetic resonance imaging) geographic bone bruise at the sulcus terminalis on the lateral femoral condyle underwent a 3-mm diameter trephine biopsy of the articular cartilage and subchondral bone overlying the bone bruise at the time of anterior cruciate ligament reconstruction. Biopsy samples of the articular cartilage and subchondral bone were stained with hematoxylin and eosin and toluidine blue. All patients had significant arthroscopic and histologic articular cartilage irregularity in the area overlying the bone bruise. Arthroscopic findings of the articular cartilage included softening (dimpling), fissuring, or overt chondral fracture. Histologic examination revealed degeneration of the chondrocytes and loss of toluidine blue staining in the articular cartilage (loss of proteoglycan). There was necrosis of osteocytes in the subchondral bone, and empty lacuna were visible. This study defines the exact histologic changes of the articular cartilage overlying a geographic bone bruise secondary to an acute anterior cruciate ligament tear. Our findings suggest that a geographic bone bruise found on magnetic resonance imaging indicates substantial damage to normal articular cartilage homeostasis.

Adolescent↗