In vitro rabbit articular cartilage organ model I. Morphology and glycosaminoglycan metabolism

Self Cite

Articular cartilage slice explants were stored under various conditions, including freezing-thawing at various rates by using dimethyl sulfoxide (DMSO) as a cryoprotective agent, incubating in standard tissue culture medium (MEM Eagle:NCTC 13515% fetal calf serum) in 5% CO, and air at 4", 21", and 37"C, and incubating in standard tissue culture medium containing 200 pg/ml a-tocopherol (vitamin E) at 37°C after first ascertaining a dose-response curve of vitamin E. Results indicated that articular cartilage slice explants did not survive freezing or storage at 4" and 21OC as measured by "S uptake. When stored at 37°C in standard tissue culture in 5% CO, and air, the slice explants remained viable for up to 60 days. The addition of a-tocopherol to the medium resulted in significantly less release of previously incorporated in stored cartilage slices and significantly less reduction of the amount of hexosamine present in the stored explants. a-Tocopherol in the m e dium also preserved safranin 0 staining. Thus, the a p plication of tissue culture techniques to the storage of articular cartilage made it possible to preserve cartilage slice explants in a viable, biochemically "normal" state.

Section: Discussionsupporting

confidence: 74%

Articular cartilage preservation and storage

Brighton

Shadle

Jimenez

et al. 1979

Self Cite

“…Following culture, explants pulsed with labeled sulfate were dialyzed against repeated changes of distilled water for 48-72 hours, lyophilized, and alkaline hydrolyzed (23). Aliquots were: 1) directly counted as previously described (8), 2) the macro precipitability of label determined by cetyl trimethyl ammonium bromide precipitation using cold chondroitin sulfate as a carrier (8), and 3) protein content quantitated by Lowry analysis (24).…”

Section: Methodsmentioning

confidence: 99%

Lymphokine‐mediated suppression of chondrocyte glycosaminoglycan and protein synthesis

Herman

Nutman

Nozoe

et al. 1981

Spontaneously released and T cell mitogen augmented lymphokine produced by human mononuclear cells has been shown to induce a concentration dependent reversible suppression of chondrocyte glycosaminoglycan and protein synthesis without significantly enhancing chondrocyte catabolic activity. The modulatory factor(s) is of T cell origin and is trypsin, pronase, and heat sensitive. Prostaglandin inhibitors failed to influence factor formation or activity. Although eluting from Sephadex G-100 over a wide range, peak activity had an approximate molecular weight of 53,000 and appeared distinct from recognized forms of lymphotoxin.Mounting evidence, albeit largely circumstantial, has implicated cell mediated immunologic mechanisms in the pathogenesis of human and experimental animal models of articular disease. Thus the predominant mononuclear cell infiltrating synovial membranes and that present in synovial fluids secured from distinct

“…The articular cartilage model used in this study was described in a previous article (7). "Slice" and "plug" explants were removed aseptically from the distal femoral condyles of adult male New Zealand white rabbits and were cultured in MEM Eagle/ NCTC 109/10~o fetal calf serum at 37°C and saturation humidity in 5% carbon dioxide and in 1% (7.6 mm Hg), 5% (38 mm Hg), 21% (160 mm Hg), 35% (266 mm Hg), 60% (456 mm Hg) and 90% (684 mm Hg) oxygen with nitrogen making up the balance of the gas.…”

Section: Methodsmentioning

confidence: 99%

“…A 0.2-ml aliquot was realkalinized and assayed for DNA with indole (7,9). A 0.1-ml aliquot was assayed for protein (7,lO).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

In vitro rabbit articular cartilage organ model II. ³⁵S incorporation in various oxygen tensions

Brighton

Lane

Koh

1974

Self Cite

Adult rabbit articular cartilage slice and plug explants were cultured in MEM/NCTC 109/fetal calf serum at 37°C in various oxygen tensions. In both the slice and plug explants, 85S04 incorporation exhibited a broad tolerance to oxygen tensions varying from 5 (38 mm. Hg) to 60 (456 mm Hg) per cent. Significantly (P 0.01) less 35S04 was incorporated in 1 (7.6 mm Hg) and 90% (684 mm Hg) oxygen than in 21 % (160 mm Hg) oxygen. No histologic or autoradiographic differences, and only slight histochemical differences, were discernible after seven days culture in the various oxygen tensions. It is suggested that low synovial fluid oxygen tension is detrimental to articular cartilage glycosaminoglycan metabolism. suggest that articular cartilage follows an anaerobic metabolic pathway (1-3). Glycolytic enzymes and high concentrations of lactic acid (1) are found in articular cartilage, and articular cartilage is little affected by short periods of oxygen deprivation (2), is relatively tolerant to high concentrations of potassium cyanide(3), and is very sensitive to monoiodoacetate (3), an agent that prevents glycolysis. Recent investigations (4-6) indicate that synovial fluid oxygen tensions decrease to very low levels in inflammatory joint diseases. These last studies present the possibility that synovial fluid hypoxia might cause chondrocyte damage and death.This report describes a series of experiments in which an in vitro articular cartilage model was cultured in various oxygen tensions. The results indicate that while articular cartilage sulfate incorporation is quite tolerant to rather wide variations in the prevailing oxygen tension, sulfate incorporation is significantly diminished at 1% and 90% oxygen.