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ToxSci Advance Access published online on August 24, 2005
Toxicological Sciences, doi:10.1093/toxsci/kfi297
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© The Author 2005. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oupjournals.org
Received May 2, 2005
Accepted May 18, 2005

Genetic Toxicology

Genotoxicity of nicotine in mini-organ cultures of human upper aerodigestive tract epithelia

Andrea W. Sassen 1, Elmar Richter 2, Marzell P. Semmler 1, Ulrich A. Harréus 3, Fernando Gamarra 4, and Norbert H. Kleinsasser 1*

1 Otolaryngology-Head and Neck Surgery, University of Regensburg, Germany
2 Walther Straub Institute of Pharmacology and Toxicology, Ludwig-Maximilians University Munich, Germany
3 Otolaryngology-Head and Neck Surgery, Ludwig-Maximilians University Munich, Germany
4 Internal Medicine-Pneumology, Ludwig-Maximilians University Munich, Germany

* To whom correspondence should be addressed.
Norbert H. Kleinsasser, E-mail: norbert.kleinsasser{at}klinik.uni-regensburg.de


  Abstract

The direct role of nicotine in tobacco carcinogenesis is still controversial. Recently, DNA damage by nicotine has been demonstrated in isolated human tonsillar tissue cells. Presently, these effects were investigated using mini-organ cultures (MOC) of human nasal epithelia. Intact MOC were repeatedly exposed to 2 and 4 mM nicotine for 1 hour on culture days 7, 9, and 11. MNNG served as a positive control. DNA damage was examined by Comet assay either directly after exposure or following a 24 h recovery period. Cell viability was not reduced by any treatment. On day 7, one hour exposure to 2 and 4 mM nicotine caused a significant dose-dependent 3.3- and 5.6-fold increase in DNA damage compared to solvent controls. Although there was no evidence of significant repair within 24 h recovery, DNA damage wasnot further increased by nicotine on days 9 and 11. After double and triple exposure to 4 mM nicotine a significant reduction in DNA damage following 24 h recovery was observed. In contrast, treatment with MNNG resulted in a highly significant and cumulative increase in DNA migration up to 110- bfold compared to controls. During recovery periods, MNNG-induced DNA damage was significantly repaired leading to a 1.5- to 1.8-fold reduction in DNA migration within 24 hours. These results conferm genotoxic effects of nicotine on human nasal epithelia. Further studies are needed to explain the lack of cumulative DNA-damaging effects of nicotine and the absence of significant DNA repair. These studies should include a battery of assays with multiple end points.

Keywords: genotoxicity; nicotine; Comet assay; mini-organ cultures; human epithelia.
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