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ToxSci Advance Access originally published online on June 19, 2007
Toxicological Sciences 2007 99(1):277-288; doi:10.1093/toxsci/kfm166
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© The Author 2007. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Internalization of Libby Amphibole Asbestos and Induction of Oxidative Stress in Murine Macrophages

David J. Blake*, Celeste M. Bolin{dagger}, David P. Cox{dagger}, Fernando Cardozo-Pelaez{dagger} and Jean C. Pfau{dagger},1

* Division of Biological Sciences and Department of Biomedical & Pharmaceutical Sciences {dagger} Center for Environmental Health Sciences, University of Montana, Missoula, Montana 59812

1 To whom correspondence should be addressed at Center for Environmental Health Sciences, University of Montana, Missoula, MT 59812. Fax: (406) 243-2807. E-mail: jean.pfau{at}umontana.edu.

Received April 4, 2007; accepted June 14, 2007


  Abstract

The community members of Libby, MT, have experienced significant asbestos exposure and developed numerous asbestos-related diseases including fibrosis and lung cancer due to an asbestos-contaminated vermiculite mine near the community. The form of asbestos in the contaminated vermiculite has been characterized in the amphibole family of fibers. However, the pathogenic effects of these fibers have not been previously characterized. The purpose of this study is to determine the cellular consequences of Libby amphibole exposure in macrophages compared to another well-characterized amphibole fiber; crocidolite asbestos. Our results indicate that Libby asbestos fibers are internalized by macrophages and localize to the cytoplasm and cytoplasmic vacuoles similar to crocidolite fibers. Libby asbestos fiber internalization generates a significant increase in intracellular reactive oxygen species (ROS) as determined by dichlorofluorescein diacetate and dihydroethidine fluorescence indicating that the superoxide anion is the major contributing ROS generated by Libby asbestos. Elevated superoxide levels in macrophages exposed to Libby asbestos coincide with a significant suppression of total superoxide dismutase activity. Both Libby and crocidolite asbestos generate oxidative stress in exposed macrophages by decreasing intracellular glutathione levels. Interestingly crocidolite asbestos, but not Libby asbestos, induces significant DNA damage in macrophages. This study provides evidence that the difference in the level of DNA damage observed between Libby and crocidolite asbestos may be a combined consequence of the distinct chemical compositions of each fiber as well as the activation of separate cellular pathways during asbestos exposure.

Key Words: asbestos; Libby amphibole; murine macrophage; oxidative stress; DNA damage.


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