Cytotoxicity and Potential Carcinogenicity of Two Anthraquinone Dye Mixtures

Abstract

To evaluate the potential health hazard of two anthraquinone dye mixtures to be used in smoke grenades, five short-term tests were conducted using Fischer 344 rat pulmonary cells in culture. The five tests were designed to measure dye effects on (1) rat tracheal epithelial cell colony forming efficieny (2) rat tracheal epithelial cell neoplas-tic transformation, (3) rat pulmonary alveolar macrophage trypan blue exclusion, (4) release of lactate dehydrogenase activity from pulmonary alveolar macrophages, and (5) pulmonary alveolar macrophage intercellular adenosine triphosphate activity In the rat tracheal epithelial assays, freshly isolated rat tracheal epithelial cells in primary culture were exposed for 24 h to both the red and the violet dye mixes. Then the cultures were scored for either colony-forming efficiency or for the appearance of morphologically altered transformed cell colonies after 30 days. Both dye mixes decreased the colony-forming efficiency of rat tracheal epithelial cells following exposure to concentrations of approximately 1–12 μg/ml. In the transformation assays both tests showed that the red and violet dyes were positive for neoplastic transformation of rat tracheal epithelial cells at concentrations of 0.03 and 0.7 μg/ml, respectively. freshly isolated pulmonary alveolar macrophages were exposed for 2 h to either dye, then assayed. A high percentage of pulmonary alveolar macrophages were unable to exclude trypan blue dye following exposure to 700–200 μg/ml of the dye mixtures. Rat pulmonary macrophages released abnormally high amounts of lactate deyhdrogenase enzyme following exposure to similar concentrations of both dyes. Measurement of intercellular adenosine triphosphate levels following dye exposure indicated that significant amounts of energy were lost due to internal cellular damage or membrane leakage. The results indicate that both the red and violet dye mixes have the potential for acute and chronic pulmonary cell damage.