Figure 2

Protective effect of low-concentration sodium nitroprusside (SNP) on human articular chondrocytes. Cell death was induced by treating chondrocytes with 1 mM SNP for 24 hours. To protect them from cell death, chondrocytes were treated with 0.1 mM SNP or 1 mM dibutylyl guanosine-3',5'-cyclic monophosphate (DBcGMP) 14 hours prior to 1 mM SNP treatment. To inhibit cyclic guanylase, 1 μM Ly83583 was added with 0.1 mM of SNP. (a) Cell death was quantitated using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazdium bromide assay. Cell survival in control cultures was set at 100%. Data shown are the means and standard deviations of triplicate experiments from at least three different donors. * P < 0.05 versus control. (b) Cell death was verified by propidium iodide staining and fluorescence-activated cell sorting analysis. Chondrocytes were trypsinized after treatment and were sedimented. Cell pellets obtained were washed and stained in 100 μg/ml propidium iodide solution for 15 min. For each sample, 10⁴ cells were analyzed. Data are representative of samples from four different donors. Percentage values denote propidium iodide positive (dead) chondrocytes.