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Aspirin aggravates the degeneration of canine joint cartilage caused by immobilization.

The effect of aspirin on the degeneration of knee cartilage caused by immobilization was examined. If dogs were fed aspirin daily (serum salicylate = 20-25 mg/dl) for 6 weeks while one hind limb was immobilized in a cast, the decreases in uronic acid content and net proteoglycan synthesis in cartilage from the immobilized knee were significantly greater than the decreases in cartilage from immobilized knees of dogs that had not received aspirin (P less than 0.01). Furthermore, neither aspirin administration nor immobilization alone affected the extractability of proteoglycans from the cartilage. However, in organ cultures of cartilage from the immobilized knee of dogs fed aspirin, the proportion of the total 35S-proteoglycans present in the culture medium was nearly twice that from cultures of cartilage of the contralateral knee. Also, more than twice as many of the total tissue proteoglycans (uronic acid) were extractable with 0.4 M guanidinium chloride, a nondissociating solvent (P less than 0.01). Regardless of whether the dogs received aspirin, the in vitro interaction of proteoglycans with hyaluronic acid from cartilage of the immobilized knee was diminished, apparently due to an abnormality in the hyaluronate-binding region of the core protein. Although these results indicate that aspirin had an adverse effect in vivo on articular cartilage of immobilized joint, aspirin administration did not preclude reversal of all of the above changes if the dog was allowed to walk about in a pen for 3 weeks after cast removal. If, however, the dog was run daily on a treadmill for 3 weeks after cast removal, the decrease in uronic acid content in cartilage from the immobilized knee persisted and was more profound if the animal had received aspirin than if it had not (P less than 0.01).

Animals↗

Heterogeneity of proteoglycans extracted before and after collagenase treatment of human articular cartilage. I. Physical properties related to age.

Proteoglycans were isolated from young and mature human articular cartilage 4 different ways: by direct extraction with 4M guanidine hydrochloride (GuHCl); after digestion of the residue from this first extraction with collagenase, by extraction with 4M GuHCl; associatively with 0.5M GuHCl after digestion of the cartilage with collagenase; and dissociatively with 4M GuHCl after digestion of the cartilage with collagenase. The structural properties of these proteoglycans were compared. Proteoglycan aggregates and monomers isolated from second extractions and from young cartilage were of larger hydrodynamic size than proteoglycans isolated from first extractions and mature cartilage, respectively. The same applied to the chondroitin sulfate chain lengths of these proteoglycans. The proteoglycan fraction from second extractions of cartilage contained a larger proportion of monomers than the fraction from first extractions. Associative extraction of mature collagenase-digested cartilage yielded mainly proteoglycan monomers, whereas an appreciable amount of proteoglycan aggregate was also liberated from young collagenase-digested cartilage. Our results indicate that, because of their larger size, proteoglycans from second extractions of cartilage are more entrapped in the collagen network. These large proteoglycans can only be liberated from the matrix after extraction of the smaller proteoglycans, followed by digestion of the residue with collagenase. This indicates that proteoglycans overlap and entangle with the collagen and protect it from degradation by collagenase.

Cartilage, Articular↗

Heterogeneity of proteoglycans extracted before and after collagenase treatment of human articular cartilage. II. Variations in composition with age and tissue source.

Proteoglycans (A1 fractions) were extracted with 4M guanidine hydrochloride (GuHCl) from human articular cartilage samples of a wide age range. Distinctions were made between hip and knee, and upper and lower layers. The residues of these extractions were digested with purified collagenase, and a second extraction with 4M GuHCl was performed, which yielded appreciable amounts of proteoglycans. When proteoglycans from second extractions were compared with those from first extractions, the following changes were observed: an increase in chondroitin sulfate; a relative decrease in keratan sulfate; a decrease in protein content; and a decrease in the ratio of chondroitin 6-sulfate to chondroitin 4-sulfate. The same changes were found when nonaggregating proteoglycans were compared with proteoglycan aggregates, when proteoglycans from young cartilage were compared with those from mature cartilage, when proteoglycans from knee cartilage were compared with those from hip cartilage, and when proteoglycans from upper layers of cartilage were compared with those from deeper layers. It is suggested that the differences found between first and second extractions of cartilage, between upper and lower layers of cartilage, and between knee and hip cartilage are caused by variations in the relative amount of nonaggregating proteoglycans and/or variations in proteoglycan size.

Adolescent↗

Relationship between arthroscopic evidence of cartilage damage and radiographic evidence of joint space narrowing in early osteoarthritis of the knee.

We examined the relationship between articular cartilage degeneration, as visualized arthroscopically, and joint space narrowing (JSN) in standing anteroposterior knee radiographs of 161 patients with chronic knee pain. The majority of these patients had radiographic findings of mild osteoarthritis. Twenty-five (33%) of the 76 patients in the series whose radiographs showed tibiofemoral JSN had grossly normal articular cartilage in both tibiofemoral compartments at arthroscopy (false-positive). The specificity of medial JSN for the presence of medial compartment articular cartilage degeneration was 0.61, i.e., only 61% of patients with normal (grade 0) medial compartment cartilage had a normal medial joint space. Of 22 patients with greater than 50% medial JSN, 9 (41%) had normal articular cartilage in the medial compartment at arthroscopy. Of 6 patients with greater than 50% lateral JSN, 3 (50%) had normal lateral compartment articular cartilage at arthroscopy. Among 36 patients with greater than 25% JSN who had neither medial nor lateral compartment articular cartilage degeneration, JSN was associated with articular cartilage degeneration in the patellofemoral compartment in 8 (22%), with meniscus degeneration in 18 (50%), and with both in 8 (22%). Thus, in these patients with chronic knee pain, radiographic evidence of JSN in the tibiofemoral compartment did not permit confident prediction of the status of the articular cartilage.

Adolescent↗

Platelet-activating factor inhibits proteoglycan synthesis and enhances neutrophil-mediated proteoglycan degradation in cartilage explants.

OBJECTIVE: Platelet-activating factor (PAF), which stimulates the release of tissue-destructive enzymes and reactive oxygen metabolites from neutrophils, was investigated for its role in neutrophil-mediated cartilage breakdown. METHODS: Bovine cartilage explants were incubated with or without human neutrophils, PAF, and other reagents. Cartilage damage was measured as either proteoglycan degradation (percent release of 35S-labeled proteoglycan from 35S-labeled cartilage) or inhibition of proteoglycan synthesis (rate of incorporation of 35S into proteoglycan). RESULTS: PAF increased neutrophil-mediated proteoglycan degradation in the 2-20 microM range. Three specific PAF-receptor antagonists, WEB2086, CV3988, and CV6209, reversed this effect of PAF. These antagonists also reduced the enhancement of neutrophil-mediated cartilage damage caused by granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor alpha (TNF alpha). The results suggest that there may be a positive feedback mechanism whereby cytokine-primed neutrophils produce PAF, which amplifies the release of other tissue-damaging substances from neutrophils. In the absence of neutrophils, PAF (2-20 microM) inhibited the synthesis of proteoglycan by bovine cartilage. Neutrophils also inhibit proteoglycan synthesis, but PAF probably is not involved in this effect of neutrophils because the PAF receptor antagonists had no consistent effect. CONCLUSION: PAF increases neutrophil-mediated cartilage proteoglycan degradation in vitro. GM-CSF and TNF alpha enhancement of neutrophil damage to cartilage is partly due to PAF. PAF alone inhibits cartilage proteoglycan synthesis.

Animals↗

Tenascin distribution in articular cartilage from normal subjects and from patients with osteoarthritis and rheumatoid arthritis.

OBJECTIVE: To determine whether tenascin is present in normal and diseased human cartilage. METHODS: Immunohistochemical and biochemical assays with a monoclonal antibody against all tenascin isoforms (BC-4) were used. RESULTS: Cartilage samples from osteoarthritis and rheumatoid arthritis patients contained increased amounts of tenascin compared with the levels in normal cartilage. Human fetal cartilage was also found to contain tenascin. In normal cartilage explants treated with interleukin-1 beta, tenascin was present in pericellular areas of all layers. Immunolocalization studies revealed that tenascin was most abundant in the superficial layers of osteoarthritic cartilage. Western blot analysis performed from dissociative extracts of diseased cartilage confirmed the presence of subunits of the native molecule. CONCLUSION: Tenascin is increased in arthritic cartilage and is weakly expressed in normal cartilage.

Adult↗

Inhibition of human fibroblast adhesion by cartilage surface proteoglycans.

OBJECTIVE: Recent studies from our laboratory have identified the nonaggregating, collagen-binding proteoglycans, fibromodulin (FM) and decorin, and fibronectin (Fn) and albumin, noncovalently bound at the articular surface of cartilage. The present studies were designed to investigate the interactions between these cartilage macromolecules and the underlying collagen matrix and their role as a barrier to cell adhesion in intact articular cartilage. METHODS: Cell adhesion studies were carried out with human skin fibroblasts incubated on the articular surface of bovine cartilage explants and on collagen-coated and/or Fn-coated plastic surfaces. Interactions of collagen and Fn with either FM or decorin were studied by radioimmunoassay of the same surfaces, using specific antibodies. RESULTS: The present studies show that 1) Fn is immunologically detectable at the intact articular surface of cartilage; 2) fibroblast adhesion to Fn is inhibited by cartilage surface extract proteins and by purified FM, but not by purified decorin; 3) FM has binding affinity for Fn; 4) FM interferes with the binding of a monoclonal antibody specific for the cell-binding domain of Fn; and 5) FM and decorin inhibit collagen-dependent fibroblast adhesion. CONCLUSION: These results indicate that the small proteoglycans at the normal articular surface may act as a barrier to cell adhesion. Since protective cartilage surface proteins break down readily after the induction of acute arthritis in experimental animals, and in rheumatoid cartilage specimens, it is postulated that proteolytic degradation of the surface proteoglycans may be responsible for increasing cell adhesion to, and subsequent pannus invasion of, articular cartilage in inflammatory arthritis.

Adolescent↗

Fibronectin synthesis in superficial and deep layers of normal articular cartilage.

OBJECTIVE: To study the distribution and synthesis of fibronectin (FN) in superficial and deep layers of normal articular cartilage. METHODS: Superficial and deep bovine and human articular cartilage slices were used to extract and quantitate FN by radioimmunoassay. Chondrocytes were also isolated by collagenase digestion for FN extraction and culture. Superficial and deep cartilage explants were cultured with and without stimulation by cytokines. Quantitation of newly synthesized FN was carried out by incubation with 35S-methionine. FN was purified on gelatin-agarose columns and further characterized by polyacrylamide gel electrophoresis. FN messenger RNA (mRNA) was quantitated by Northern blot analysis. RESULTS: Freshly isolated bovine chondrocytes from deep cartilage contained 2.3 +/- 0.2 times more FN than was found in superficial cells (P < 0.025). Deep cartilage explants contained 1.2 times more FN than was found in superficial tissue. Explants obtained from deep cartilage synthesized 2.4 times more FN per cell than did superficial tissues (P < 0.01). FN synthesis as a fraction of total protein synthesis was significantly greater in deep explants (P < 0.01) compared with superficial tissues. Isolated deep chondrocytes in culture synthesized 1.89 +/- 0.33-fold more FN than did superficial cells (P < 0.05). Cytokine-stimulated superficial cartilage explants failed to respond in terms of FN synthesis. FN mRNA quantitation showed no significant differences between superficial and deep populations. CONCLUSION: Since FN plays a major role in cell adhesion to damaged cartilage surfaces, our results suggest that modulation of FN synthesis near the articular surface of cartilage may be one of the factors that impede pannus invasion following an inflammatory insult to the joint.

Animals↗

Induction of arthritis in HLA-DR4-humanized and HLA-DQ8-humanized mice by human cartilage proteoglycan aggrecan but only in the presence of an appropriate (non-MHC) genetic background.

OBJECTIVE: To determine whether the rheumatoid arthritis (RA)-predisposing class II molecules of the major histocompatibility complex (MHC) can present cartilage proteoglycan (PG) aggrecan, and if so, to determine the epitope repertoire of the human cartilage PG in HLA-transgenic mice and determine whether HLA-transgenic mice develop arthritis in response to immunization with human cartilage PG. METHODS: Mice transgenic for HLA-DR2.Ab(0), DR3.Ab(0), DR4.Ab(0), and DQ8.Ab(0), lacking their own (mouse) class II antigens (Ab(0)), on the original (arthritis-resistant) and the arthritis-susceptible BALB/c backgrounds, were immunized with human cartilage PG. The T cell epitope repertoire presented by these class II MHC alleles was determined using a synthetic peptide library (143 peptides of the core protein of human cartilage PG), and arthritis development was monitored and compared in wild-type and HLA-transgenic/congenic BALB/c mice. RESULTS: Mice of the 4 HLA-transgenic lines, either on the original mixed, arthritis-resistant background or DR4.Ab(0)- and DQ8.Ab(0)-transgenic/congenic mice on the arthritis-susceptible BALB/c genetic background, responded well to PG immunization (as assessed by T cell responses and antibody and cytokine production), and a number of T cell epitopes along the core protein of human cartilage PG were identified. DR4.Ab(0)- and DQ8.Ab(0)-transgenic mice immunized with human cartilage PG developed arthritis, but only when these class II MHC molecules were present on the arthritis-susceptible (BALB/c) genetic background. CONCLUSION: A number of human cartilage PG epitopes can be presented by HLA alleles that predispose to the development of RA, but the epitopes of the cartilage PG presented by HLA-DR4 or HLA-DQ8 can induce arthritis only in the presence of an appropriate genetic (non-MHC) background.

Aggrecans↗

Human beta-defensin 3 mediates tissue remodeling processes in articular cartilage by increasing levels of metalloproteinases and reducing levels of their endogenous inhibitors.

OBJECTIVE: Beta-defensins are broad-spectrum antimicrobial peptides (APs) that are components of innate immunity. Recent investigations showed the induction of beta-defensins in synovial membranes of osteoarthritic (OA) joints and suggested that they have functions other than the ability to kill microbes. As a result of these findings, we undertook this study to investigate the production of human beta-defensin 3 (HBD-3) in OA cartilage and to determine its influence on chondrocyte function. METHODS: Healthy and OA cartilage were assessed for HBD-3 expression by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. HBD-3 expression in C28/I2 chondrocytes after administration of tumor necrosis factor alpha (TNFalpha) and interleukin-1 (IL-1) was determined by real-time RT-PCR and immunodot blot. Enzyme-linked immunosorbent assay experiments were used to study the effects of HBD-3 in cultured articular chondrocytes and in healthy and OA cartilage discs. Immunohistochemical analyses were performed to study the expression of mouse beta-defensins (MBDs) in OA cartilage of STR/Ort mice. RESULTS: HBD-3 was induced in OA cartilage without bacterial challenge. Cytokines involved in the pathogenesis of OA, namely, TNFalpha and IL-1, were strong inducers of HBD-3 in cultured chondrocytes. Application of the recombinant HBD-3 protein to cultured chondrocytes and cartilage discs resulted in increased production of cartilage-degrading matrix metalloproteinases and in down-regulation of their endogenous regulators, tissue inhibitors of metalloproteinases 1 and 2. Furthermore, STR/Ort mice, which are genetically predisposed to develop OA-like lesions in the knee joint, demonstrated an increased expression of MBDs 3 and 4 in cartilage compared with that in healthy animals. CONCLUSION: These findings widen our knowledge of the functional spectrum of APs and demonstrate that HBD-3 is a multifunctional AP with the ability to link host defense mechanisms and inflammation with tissue-remodeling processes in articular cartilage. Moreover, our data suggest that HBD-3 is an additional factor in the pathogenesis of OA.

Adult↗

Immunohistochemical analysis of hip arthritis in ankylosing spondylitis: evaluation of the bone-cartilage interface and subchondral bone marrow.

OBJECTIVE: Previous histopathologic and magnetic resonance imaging studies suggest that the subchondral bone marrow might be the primary site of inflammation in patients with ankylosing spondylitis (AS) and that this might be reflected by inflammation found in hip joints. The aim of this study was to conduct an immunohistologic assessment of the bone-cartilage interface and subchondral bone marrow in AS patients with hip arthritis. METHODS: We collected femoral heads from patients with AS, osteoarthritis (OA), and rheumatoid arthritis (RA) who were undergoing hip replacement. The subchondral bone marrow and bone-cartilage interface were assessed immunohistochemically by evaluating infiltrating T cells, microvessel density, and osteoclasts. Areas of the femoral head surface with and without cartilage were assessed separately. RESULTS: At sites with surface cartilage, we found subchondral infiltration of CD3+ T cell aggregates at significantly higher numbers in AS patients as compared with OA patients, but not RA patients. At sites of complete cartilage destruction, the frequency of CD3+ T cell aggregates was significantly reduced as compared with sites with cartilage on the surface in AS patients, but not in RA patients. Similar differences were found for CD4+ and CD8+ T cells. Only at sites with surface cartilage, but not those without, angiogenesis and osteoclastic foci in the subchondral bone marrow in AS patients were significantly increased as compared with RA patients and with OA patients. CONCLUSION: These findings suggest that the subchondral bone marrow and bone-cartilage interface are primary sites of inflammation in AS and that cartilage might be necessary for the induction of inflammation.

Adult↗

Steroid hormones strongly support bovine articular cartilage integration in the absence of interleukin-1beta.

OBJECTIVE: Posttraumatic integration of articular cartilage at fracture sites is essential for mechanical stability of cartilage, and ruptured cartilage is a prerequisite for early osteoarthritis. This study was undertaken to investigate effects on articular cartilage integration mediated by steroid hormones, interleukin-1beta (IL-1beta), and combinations thereof. METHODS: Articular cartilage blocks were cultured in partial apposition for 2 weeks with ascorbic acid, testosterone, 17beta-estradiol, and dehydroepiandrosterone (DHEA), with or without IL-1beta. Mechanical integration was measured as adhesive strength, i.e., the maximum force at rupture of integrated cartilage blocks divided by the overlap area. Glycosaminoglycan content was used to study synthesized extracellular matrix. RESULTS: Culture in medium without supplements did not lead to integration (adhesive strength 0 kPa). With administration of ascorbic acid (100 microg/ml), the median adhesive strength was 49 kPa. In comparison with ascorbic acid alone, all steroid hormones induced a strong, concentration-dependent stimulation of integration (with maximum values observed with DHEA at 3 x 10(-5)M, testosterone at 10(-8)M, and 17beta-estradiol at 10(-11)M). For testosterone and 17beta-estradiol, this was also reflected by an increase of glycosaminoglycan content. Adhesive strength was increased with IL-1beta at 10 pg/ml, but not at 1 pg/ml or 100 pg/ml. In the presence of both IL-1beta and sex hormones, integration of articular cartilage was reduced. CONCLUSION: This is the first study to demonstrate that steroid hormones such as 17beta-estradiol, DHEA, and testosterone stimulate articular cartilage integration. This effect is abrogated by low concentrations of IL-1beta. In the absence of IL-1beta or after neutralization of IL-1beta, steroid hormones might be favorable adjuvant compounds to optimize cartilage integration.

Animals↗

Evaluation of shark cartilage in patients with advanced cancer: a North Central Cancer Treatment Group trial.

BACKGROUND: Shark cartilage has been a popular complementary or alternative medicine intervention. The basis for this popularity is the claim that sharks rarely get cancer because of the high proportion of cartilage in the shark's body. However, early studies were equivocal. Therefore, a clinical trial was conducted to look at the impact of shark cartilage in patients with advanced cancer. The primary goal of this trial was to determine whether a shark cartilage product improved overall survival for patients with advanced cancer who were getting standard care. Secondary research goals were to evaluate toxicities, tolerability, and quality of life associated with this shark cartilage product. METHODS: The study was a two-arm, randomized, placebo-controlled, double-blind, clinical trial. Patients with incurable breast or colorectal carcinoma had to have good performance status and organ function. Patients could be receiving chemotherapy. Patients were all to receive standard care and then to be randomly selected to receive either a shark cartilage product or an identical-appearing and smelling placebo 3 to 4 times each day. RESULTS: Data on a total of 83 evaluable patients were analyzed. There was no difference in overall survival between patients receiving standard care plus a shark cartilage product versus standard care plus placebo. Likewise, there was no suggestion of improvement in quality of life for patients receiving the shark cartilage, compared with those receiving placebo. CONCLUSION: This trial was unable to demonstrate any suggestion of efficacy for this shark cartilage product in patients with advanced cancer.

Aged↗

Cartilage grafts--present status.

Cartilage grafts have been in use for almost a century and have proved their usefulness. Many questions about immunology, survival, growth, and role of perichondrium are still debated. We are presenting a review of the literature and of our experimental work on cartilage grafts. Ear cartilage was transplanted from 28 young New Zealand rabbits subcutaneously in the chest walls. The grafts were divided into 1) bare cartilage, 2) perichondrium, 3) cartilage covered with perichondrium on one side, and 4) cartilage covered with perichondrium on both sides. The grafts were measured in length and weight before transplantation, and at 2 and 4 months after transplantation; they were compared to a control piece of cartilage tagged in situ. Histologic examination was performed on all retrieved grafts. Transplanted cartilage survival was good in over 75% of cases; however, cartilage production from perichondrium was minimal and no growth was noticed in any of the grafts.

Animals↗

Tetrapyrroles as inhibitors of normal cartilage metabolism: relative potency of different compounds.

The present study was designed to explore the role of different tetrapyrroles as inhibitors of cartilage metabolism. We studied the effects of tetrapyrroles on the incorporation of [35S]sulfate into proteoglycans, [14C]-leucine into protein, and [3H]uridine into the RNA of normal cartilage from two different vertebrate classes using the embryonic chicken pelvic rudiment bioassay and the hypophysectomized rat costal cartilage bioassay, both very sensitive to cartilage growth factors and growth inhibitors. We compared the relative potencies of the following compounds: both metalloporphyrins (heme and chlorophyllin), linear tetrapyrroles (bilirubin), and heme proteins (hemoglobin, myoglobin, and cytochrome c). Hemoglobin and heme were the most potent inhibitors of rat cartilage metabolism, and bilirubin was a far more potent inhibitor of embryonic chick cartilage metabolism. Chlorophyllin had moderate inhibitory activity, especially on chick cartilage, whereas cytochrome c was inactive in these bioassays. Surprisingly, myoglobin was relatively ineffective despite its close similarity to heme and hemoglobin. The bilirubin-induced inhibition of sulfate incorporation into chick cartilage was partially prevented when glutathione was included in the incubation medium, suggesting that a free-radical mechanism may be involved. There were significant differences in the sensitivity of the two cartilages studied, indicating there may be species-dependent sensitivity to different tetrapyrroles.

Animals↗

Effect of gender on in vivo cartilage magnetic resonance imaging T2 mapping.

PURPOSE: To determine if gender is a significant variable for in vivo magnetic resonance imaging (MRI) T2-mapping of knee articular cartilage in young asymptomatic volunteers. MATERIALS AND METHODS: Cartilage MRI T2 mapping was performed in a young healthy population consisting of seven male and 10 female volunteers, 22 to 29 years of age. High-resolution in vivo T2 maps were obtained of patellar, tibial, and weight-bearing femoral articular cartilage. Spatial dependency of cartilage T2 between groups was evaluated through a comparison of cartilage T2 as a function of normalized distance from bone. RESULTS: Bulk cartilage T2 values were similar at all three anatomic sites, and between male and female volunteers. All volunteers demonstrated similar spatial variation in cartilage MRI T2 values, with a minimum located in the radial zone and increasing T2 values toward the articular surface. There was no difference in spatial dependency of cartilage T2 between males and females. CONCLUSION: In young, healthy volunteers, the magnitude and spatial dependency of cartilage T2 does not differ with gender.

Adult↗

The potential of adult human perichondrium to form hyalin cartilage in vitro.

The usefulness of adult human perichondrium for the restoration of articular cartilage defects depends on the potential to form hyalin cartilage. In order to evaluate the capacity of adult human perichondrium to form hyalin cartilage in vitro, perichondrium of the rib of eight adult human beings was cultured in vitro. After removal of residual cartilage, perichondrial explants were cultured for 7 or 10 days. The explants were histologically examined using specific stains to prove the presence of glycosaminoglycans (GAGs) normal for hyalin cartilage. Clear differentiation of perichondrial cells towards chondrocytes was noted. The chondrocytes synthesized new matrix substances normally present in hyalin cartilage. This investigation supports the usefulness of adult human rib perichondrium for the restoration of cartilage defects. Due to the enormous potential of the rib perichondrium to form hyalin cartilage in vitro, even defects in joints with a rather thick cartilage layer might be restored using this biological material.

Adult↗

Determination of fixed charge density in cartilage using nuclear magnetic resonance.

Many biomechanical and chemical properties of cartilage are dependent on the fixed charge density (FCD) of the extracellular matrix. In this study, nuclear magnetic resonance (NMR) spectroscopy was investigated as a nondestructive technique for determining FCD in cartilage. Sodium content was measured by NMR in cartilage explants and was compared with sodium content measured by inductively coupled plasma emission spectroscopy (ICP) in order to verify the total NMR visibility of sodium in cartilage. The ratio of NMR to ICP results was 1.02 +/- 0.04 (calf, mean +/- SD, n = 7) and 1.04 +/- 0.11 (adult bovine, n = 8). Sodium concentration as measured by NMR was then used with ideal Donnan theory to compute estimates of FCD. For calf articular cartilage (AC) near physiological conditions, calculated FCD was -0.28 +/- 0.03 M (n = 10). NMR measurements were then made for individual cartilage specimens sequentially equilibrated in baths of differing salt composition, pH, or ionic strength. For calf and adult AC, calculated FCD decreased dramatically between pH 3 and 2, with adult specimens becoming positively charged but calf tissue retaining a net negative charge. For calf AC equilibrated in 0.3-0.015 M NaCl, calculated FCD was observed to decrease slightly with decreasing bath ionic strength. For epiphyseal cartilage, FCD varied with the position of origin of the explant within the joint, ranging from -0.19 to -0.35 M in a manner that correlated with tissue glycosaminoglycan content. Preliminary NMR imaging experiments demonstrated similar variations of sodium concentration in intact ulnar epiphyseal cartilage. Collectively, these results demonstrate the ability of NMR to nondestructively follow FCD in cartilage. The technique is applicable to dynamic studies as well as to both in vitro and in vivo studies on living tissue.

Animals↗