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Carcinogenesis Advance Access originally published online on August 29, 2003
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Carcinogenesis, Vol. 24, No. 11, 1793-1800, November 2003
© 2003 Oxford University Press

CARCINOGENESIS

In vivo genotoxicity of hard metal dust: induction of micronuclei in rat type II epithelial lung cells

Marlies De Boeck1,*, Peter Hoet2,*, Noömi Lombaert1, Benoit Nemery2, Micheline Kirsch-Volders1 and Dominique Lison3,4

1 Vrije Universiteit Brussel, Laboratorium voor Cellulaire Genetica, Pleinlaan 2, 1050 Brussel, 2 Katholieke Universiteit Leuven, Laboratorium voor Pneumologie, Eenheid Longtoxicologie, Herestraat 49, B-3000 Leuven and 3 Université catholique de Louvain, Unité de Toxicologie industrielle et Médecine du Travail, Clos Chapelle aux Champs 30.54, B-1200 Bruxelles, Belgium

Inhalation of hard metal dust (WC-Co particles) has been associated with an increased risk for lung cancer in occupational settings. In vitro, WC-Co was genotoxic in human lymphocytes producing DNA strand breaks and micronuclei. The aim of the present study was to evaluate the in vivo genotoxic effects of WC-Co dust in rat type II pneumocytes. DNA breaks/alkali-labile sites (alkaline comet assay) and chromosome/genome mutations (micronucleus test) were assessed after a single intra-tracheal (i.t.) instillation of WC-Co, including dose-effect and time trend relationships. In addition, the alkaline comet assay was performed on cells obtained after broncho-alveolar lavage (BAL) and on peripheral blood mononucleated cells (PBMC). As pulmonary toxicity parameters, protein content, lactate dehydrogenase activity, total and differential cell count in BAL fluid were evaluated in parallel. In type II pneumocytes, WC-Co induced a statistically significant increase in tail DNA (12 h time point) and in micronuclei (72 h) after a single treatment with 16.6 mg WC-Co/kg body wt, a dose that produced mild pulmonary toxicity. This observation provides the first evidence of the in vivo mutagenic potential of hard metal dust. In PBMC, no increase in DNA damage or micronuclei was observed. This study indicates the potential to detect chromosome/genome mutations (micronuclei) in relevant target cells (type II pneumocytes) after i.t. instillation of a particle mixture.


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