It is generally accepted that the quantities of available oxygen and glucose selleck chemicals llc can fluctuate considerably in con nective tissues such as articular cartilage, growth plates and the intervertebral disc. Articular chondro cytes consume less oxygen in comparison with most other cell types. Consequently, anaerobic glycolysis forms the principal source of cellular ATP in cartilage. The direct investigation of the function of exogenous NO production on articular chondrocytes has been ham pered by the lack of uniformity between the different types of NO donor compounds. Since the past decade, the diazeniumdiolates began to replace traditional donors, such as SIN 1, SNP, SNAP and S nitrosogluthathione, as sources of exogenous NO production, because have been shown to be reliable sources of NO under a variety of culture conditions.

The main advantages of these compounds are, known rates of NO generation, NO generation rates cov Inhibitors,Modulators,Libraries ering a wide range, spontaneity of NO generation and ten able generation of NO redox forms. For all these reasons, besides the classical donor SNP, Inhibitors,Modulators,Libraries we used a diazeniumdiolate, NOC 12, with a half life of 327 minutes, for exogenous NO pro duction, to further investigate the conditions in which NO is cytotoxic to chondrocytes, and compare Inhibitors,Modulators,Libraries the different effects induced by the two different types of NO donors. We wished to determine whether NO modulates the pathogenesis of OA, inducing apoptosis by means of the inhibition of mitochondrial function Materials and methods Cartilage acquisition and cell isolation Normal human cartilage from femoral heads and knees was obtained from 11 adult donors without history of joint disease and who had macroscopically normal cartilage, human OA cartilage was obtained from the femoral heads of 12 patients.

All patients and healthy donors have signed the informed consent and the project was approved by the Regional Ethical Committee from Galicia. Small cartilage fragments were digested as previously described. Primary culture of chondrocytes Chondrocytes were recovered and plated at high Inhibitors,Modulators,Libraries density in DMEM supple mented with 100 units ml penicillin, 100 ug ml strepto mycin, 1% glutamine, and 10% FCS. Chondrocytes were incubated at 37 C in a humidified gas mixture containing 5% CO2 balanced with air. Chondrocytes were used at weeks 2 to 3 at confluence in primary culture. Cell viability was assessed by trypan blue dye exclusion, and stained cells were discarded to carry out experiments. General procedure and NO donor compounds employed NO donor compounds were added from 60 mM stock solution dissolved in 0. 1 M NaOH and 10 mM stock solution dissolved in medium. This NO donor compound Inhibitors,Modulators,Libraries was freshly prepared before each experiment selleck compound but NOC 12 was stored as 60 mM stock solutions in 0. 1 mM NaOH at 20 C.

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