ATLA::Alternatives to Laboratory Animals

Volume 29, Number 2

The red blood cell phototoxicity test (photohaemolysis and haemoglobin oxidation): EU/COLIPA validation programme on phototoxicity (phase II).

ATLA 29, 163-177, March/April 2001

Emil Rudolf, Jan Peychl and Miroslav Cervinka

Department of Medical Biology and Genetics, Charles University in Prague, Faculty of Medicine in Hradec Králové, Simkova 870, Hradec Králové I 500 OA, Czech Republic

SUMMARY

Many human activities, particularly industrial ones, result in an ever-growing production of toxic waste materials. The dynamics of the toxic effects of chromium acetate hydroxide, which is found in high concentrations in a waste sediment produced in the Czech Republic, were assessed by using a battery of in vitro tests carried out on two cell lines: L-929 (mouse fibroblasts) and Hep 2 (human laryngeal cells). Various markers of cell damage were assessed by phase-contrast, video and fluorescence microscopy, fluorometry, and DNA analysis. Chromium acetate hydroxide, over a concentration range of 1-0.02 mol/l induced immediate cell death by fixation, whereas, at 0.00 2mol/l, the treated cells died in a much slower, more discrete manner. All the detected markers of cell damage, whether immediate or slow, clearly demonstrated that the cells died by necrosis. On the other hand, test concentration of 0.001 mol/l appeared to constitute a threshold at which no pathological changes of Hep 2 cells were observed over 96 hours. We conclude that chromium acetate hydroxide has a high toxic potential in vitro, which should be considered when studying the toxicity of waste materials containing it.

Keywords: chromium acetate hydroxide, in vitro, toxicity, necrosis, fixation, microscopy