Cellular effects of an anionic surfactant detected in V79 fibroblasts by different cytotoxicity tests

Several cytotoxicity tests were employed to detect the cellular effects of low concentrations of the anionic surfactant linear alkylbenzene sulphonate (LAS). When added to growth medium containing 5% foetal calf serum, LAS did not affect V79 cell growth, nor did it alter the permeability of cell membranes. The inactivity depended on the serum component of the medium. When treatments were carried out in serum-free saline, LAS inhibited cell proliferation, made the plasma membrane permeable to otherwise-undiffusible compounds, and reduced the uptake of tritiated thymidine. The alterations in membrane permeability were evaluated from the release of cytoplasmic molecules of different size (lactate dehydrogenase, adenine nucleotides, RNA) into the medium. The sensitivity of the spectrophotometric lactate dehydrogenase assay was inadequate for the conditions of treatment required to detect the cytotoxicity of LAS. In cultures pre-incubated with tritiated adenine instead, the release of labelled ATP pool components was time and dose dependent and allowed discrimination between levels of membrane damage causing the same degree of trypan blue staining. Also, macromolecular nucleic acids were detected outside the treated cells at doses of 4-6 mg LAS/litre, which indicated severe membrane damage.