Role of in vitro Factors in Ozone Toxicity for Cultured Rat Lung Fibroblasts

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Abstract
Ozone toxicity for cultured rat lung fibroblasts was concentration dependent and was affected by the manner in which ozone was delivered to the cells, i.e. cultures were either rotated with a thin moving overlay of medium or were stationary with a fixed layer of medium between the cells and the gas phase. The influence of culture medium components and culture dish composition on the toxicity of ozone were also investigated. Cell viability, used to measure ozone toxicity, was quantified by the chromium-51 release assay, and by a viability index calculated from the percentage of cells stained with a vital dye combined with the decrease in cell number as determined by DNA measurements. During stationary ozone exposure, toxicity appeared to be mediated primarily by hydrogen peroxide and could be inhibited by catalase or fetal bovine serum when measured by the viability index. During rotated exposure, catalase and fetal bovine serum provided no protection when measured by the viability index, however, when measured by the chromium-51 release assay, fetal bovine serum was partially protective. The effect of ozone on the fibroblasts was not influenced by whether culture dishes were glass or plastic, or whether the culture medium was balanced salt solution or complete chemically-defined medium.

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