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Cellular uptake and cytotoxic potential of respirable bentonite particles with different quartz contents and chemical modifications in human lung fibroblasts

  • Organ Toxicity and Mechanisms
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Abstract

Considering the biological reactivity of pure quartz in lung cells, there is a strong interest to clarify the cellular effects of respirable siliceous dusts, like bentonites. In the present study, we investigated the cellular uptake and the cytotoxic potential of bentonite particles (Ø< 10 μm) with an α-quartz content of up to 6% and different chemical modifications (activation: alkaline, acidic, organic) in human lung fibroblasts (IMR90). Additionally, the ability of the particles to induce apoptosis in IMR90-cells and the hemolytic activity was tested. All bentonite samples were tested for endotoxins with the in vitro-Pyrogen test and were found to be negative. Cellular uptake of particles by IMR90-cells was studied by transmission electron microscopy (TEM). Cytotoxicity was analyzed in IMR90-cells by determination of viable cells using flow cytometry and by measuring of the cell respiratory activity. Induced apoptotic cells were detected by AnnexinV/Propidiumiodide-staining and gel electrophoresis. Our results demonstrate that activated bentonite particles are better taken up by IMR90-cells than untreated (native) bentonite particles. Also, activated bentonite particles with a quartz content of 5–6% were more cytotoxic than untreated bentonites or bentonites with a quartz content lower than 4%. The bentonite samples induced necrotic as well as apoptotic cell death. In general, bentonites showed a high membrane-damaging potential shown as hemolytic activity in human erythrocytes. We conclude that cellular effects of bentonite particles in human lung cells are enhanced after chemical treatment of the particles. The cytotoxic potential of the different bentonites is primarily characterized by a strong lysis of the cell membrane.

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Acknowledgements

We are thankful to the Süd Chemie AG for providing us the bentonite dust, and for support. We thank Gabriele Zimmer and Ute Zimmermann for the excellent technical assistance and Prof. Dr. N. Seemayer for helpful discussions. We also thank Dr. K.H. Worm for support the DNA-analysis Prof. W.U. Müller for providing the Coulter Counter, Dr. D. Hoehr for performing the TEM analysis and Dr. C. A. Preston for carefully reading the manuscript.. This work was kindly supported by DFG (DO 332/1). Declaration: The experiments comply with the current laws of the country in which the experiments were performed.

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Authors and Affiliations

  1. Institute of Hygiene and Occupational Medicine, University Hospital, Hufelandstrasse 55, 45122, Essen, Germany

    Stefan Geh, Albert W. Rettenmeier & Elke Dopp

  2. Institute of Cell Biology (Cancer Research), University Hospital, Essen, Germany

    Raif Yücel

  3. Institute of Environmental Health Research (IUF), Düsseldorf, Germany

    Rodger Duffin, Catrin Albrecht & Paul J. A. Borm

  4. ELEGI COLT Lab, University of Edinburgh, Scotland, UK

    Rodger Duffin

  5. Verein für Technische Sicherheit und Umweltschutz e.V., Gotha, Germany

    Lorenz Armbruster

  6. Research Institute for Occupational Medicine of the Institutions for Statutory Accident Insurance and Prevention (BGFA), Bochum, Germany

    Monika Raulf-Heimsoth & Thomas Brüning

  7. Institute of Biology, Department of Cell Biology and Biosystems Technology, University of Rostock, Rostock, Germany

    Eik Hoffmann

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  1. Stefan Geh
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Correspondence to Elke Dopp.

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Geh, S., Yücel, R., Duffin, R. et al. Cellular uptake and cytotoxic potential of respirable bentonite particles with different quartz contents and chemical modifications in human lung fibroblasts. Arch Toxicol 80, 98–106 (2006). https://doi.org/10.1007/s00204-005-0013-9

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  • DOI: https://doi.org/10.1007/s00204-005-0013-9

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