Genetic Toxicology

© Fraunhofer ITEM

Elucidating the genotoxic potential is of pivotal importance in the toxicological assessment of environmentally and occupationally relevant noxious substances, and also in the notification and registration procedures stipulated for chemicals and drugs.

The Working Group on Genetic Toxicology performs in-vitro and in-vivo genotoxicity studies to evaluate the mutagenic potential of chemicals, pharmaceuticals and particles for industry and public authorities. On behalf of our clients, we conduct the standard tests prescribed by the regulatory authorities, in compliance with international regulations (OECD, EU, EPA, FDA) and GLP guidelines. Special emphasis is placed at the Institute on investigating the toxic effects of airborne substances. Through interdisciplinary cooperation within the Institute, in-vivo and in-vitro studies on the mutagenic potential of gases, aerosols and complex mixtures can thus also be performed. Such investigations are important, for example, for aspects of occupational safety and for environmental analyses.

The Working Group on Genetic Toxicology adapts the standard methods of genetic toxicology to each client's specific problem, develops them further and, if need be, uses complementary methods to elucidate the mechanism of action. The in-house expertise in the area of cell biology/cell isolation provides the basis for investigating the genotoxic potential in relevant target cells.

Methods

Exposure modalities:

Cell exposure via the culture medium

Cell exposure at the air/liquid interface (gases, aerosols)

Treatment of rodents: by inhalation (nose-only, whole-body), intratracheal, oral, i.p.

Methods:

Micronucleus assay in vitro and in vivo

Chromosomal aberration test (e.g. V79, human lymphocytes)

Mammalian cell gene mutation tests (mouse lymphoma tk assay, HPRT test)

Bacterial reverse mutation assay (Ames test)

Comet assay (alkaline, neutral, acelluar, modified protocol for detection of crosslinks)

32P postlabeling for detection of DNA adducts

8-OHdG and dG measurements by HPLC for demonstration of oxidative DNA damage

Demonstration of apoptotic DNA fragmentation

Live cell count and analysis of adhesion efficiency

When dealing with complex problems, the above methods can be complemented by other techniques from the fields of analytical chemistry, in-vitro toxicology, clinical chemistry, immunotoxicology, molecular toxicology, and pathology. The Institute's Working Group on Analytical Chemistry, for example, offers different options for chemical analysis of test substance solutions right up to structure elucidation.

Test systems:

Bacteria

Adherent cell lines (e.g. V-79, RPMI-2650, A-549)

Suspension cell lines (e.g. L5178Y/TK+/-, HL-60)

Primary cells (e.g. human lymphocytes, hepatocytes,alveolar macrophages, alveolar type-II cells)

Rodents (mouse, rat)

Projects

In-vitro investigations into the toxic and genotoxic potentials of quartz-containing ceramic dusts  

Comet assay-based evaluation of the genotoxicity of quartz-containing ceramic fibers

Mechanistic investigations into the genotoxicity and toxicity of nitrostyrene derivatives, taking into account oxidative DNA damage

Comparative investigation of the DNA-damaging potential of formaldehyde in V79 cells, primary nasal epithelial cells, and the nasal epithelial tumor cell line RPMI 2650

Mechanistic investigations into the genotoxic potential of an initiator from among dental filling materials, giving particular consideration to reactive oxygen species

In-vitro and in-vivo genotoxicity testing of a novel antimalarial drug

Further development and validation of an in-vitro model for genotoxicity testing of inhalable gases as required for registration

Publications

2013

Rittinghausen, S.; Bellmann, B.; Creutzenberg, O.; Ernst, H.; Kolling, A.; Mangelsdorf, I.; Kellner, R.; Beneke, S.; Ziemann, C.: Evaluation of immunohistochemical markers to detect the genotoxic mode of action of fine and ultrafine dusts in rat lungs
In: Toxicology 303 (2013), p. 177-186

2012

Ziemann, C.; Reamon-Büttner, S.-M.; Schlichting, A.; Brockmeyer, H.; Rahmer, H.; Bellmann, B.: Human pleural mesothelial MeT-5A cells are a limited in vitro model system in determining potential asbestos-like effects of multiwall carbon nanotubes.
in: Naunyn-Schmiedebergs archives of pharmacology 385 (2012), Supplement 1, p. 107, Abstract 473. ISSN: 0028-1298

Creutzenberg, O. H.; Ziemann, C.; Hansen, T.; Ernst, H.; Schaudien, D.; Schuchardt, S.; Wiench, K.; Bellmann, B.: Subacute and subchronic inhalation toxicity and dermal absorption of the nanoscaled zinc oxide Z-COTE HP1 in the rat.
In: The Toxicologist 126 (2012), Nr. 1, p. 142-143, Abstract 658. ISSN: 0731-9193

Heinrich, U.; Schlichting, A.; Ziemann, C.; Seyock, S.; Haase, D.; Leonhardt, A.; Bellmann, B.: Suspension of MWCNT for toxicological in vivo and in vitro studies.
In: The Toxicologist 126 (2012), Nr. 1, p. 143, Abstract 661. ISSN: 0731-9193

2011

Ziemann, C.; Apitius, F.; Elend, M.; Baumgartner, S.; Volk, J.; Ritter, D.; Steinfelder, H. J.; Schuchardt, S.: Discrepancy between high clastogenic activity of trans-beta-nitrostyrene in vitro and lack in carcinogenicity might be related to inactivating metabolism and its high reactivity towards thiol groups.
In: Berlin: Springer, 2011 (Naunyn-Schmiedeberg's Archives of Pharmacology 383.2011, Supplement 1), p. 12-13, Abstract V48. ISSN: 0028-1298

Parker, C. M.; Schreiner, C. A.; Hallmark, N.; Kriech, A. J.; Osborn, L. V.; Fuhst, R.; Buschmann, J.; Ernst, H.; Hansen, T.; Pohlmann, G.; Preiß, A.; Ziemann, C.: Evaluation of reproductive/developmental and repeated dose (subchronic) toxicity and cytogenetic effects in rats of a roofing asphalt fume condensate by nose-only inhalation.
In: Regulatory toxicology and pharmacology 59 (2011), Nr. 3, p. 445-453. ISSN: 0273-230

2010

Fitz-Maier, H.; Volk, J.; Leyhausen, G.; Ziemann, C.; Geurtsen, W.: Combinatorial effects of dental composite components on glutathione and ROS levels in oral epithelial cells.
In: Deutsche Gesellschaft für Experimentelle und Klinische Pharmakologie und Toxikologie -DGPT: 51. Jahrestagung 2010. Deutsche Gesellschaft für Experimentelle und Klinische Pharmakologie und Toxikologie. Abstracts: 23. - 25. März 2010, Mainz.
Berlin: Springer, 2010 (Naunyn-Schmiedeberg's Archives of Pharmacology 381.2010, Supplement 1), p. 79. ISSN: 0028-1298

Heinemann, D.; Ritter, D.; Kasper, C.; Eger, K.; Steinfelder, H. J.; Ziemann, C.: Comparative mechanistical investigations on the genotoxic potential of nitrovinyluracil derivatives.
In: Deutsche Gesellschaft für Experimentelle und Klinische Pharmakologie und Toxikologie -DGPT: 51. Jahrestagung 2010. Deutsche Gesellschaft für Experimentelle und Klinische Pharmakologie und Toxikologie. Abstracts: 23. - 25. März 2010, Mainz.
Berlin: Springer, 2010 (Naunyn-Schmiedeberg's Archives of Pharmacology 381.2010, Supplement 1), p. 74. ISSN: 0028-1298

Volk, J.; Leyhausen, G.; Beckedorf, A.; Weitz, A.; Fitz-Maier, H.; Ziemann, C.; Geurtsen, W.: Genotoxic potential of TEGDMA in oral epithelial cells.
In: Journal of dental research 89 (2010), Special Issue B, Abstr. 719. ISSN: 1544-0591; ISSN: 0022-0345

Licht, O.; Gerdes, H.; Ziemann, C.; Bitsch, A.; Mangelsdorf, I.: Genotoxicity of 1-propanol. Is there a problem?
In: Deutsche Gesellschaft für Experimentelle und Klinische Pharmakologie und Toxikologie -DGPT-: 51. Jahrestagung 2010. Deutsche Gesellschaft für Experimentelle und Klinische Pharmakologie und Toxikologie. Abstracts: 23. - 25. MÄrz 2010, Mainz.
Berlin: Springer, 2010 (Naunyn-Schmiedeberg's Archives of Pharmacology 381.2010, Supplement 1), p. 85. ISSN: 0028-1298

Ziemann, C.; Hansen, T.; Pohlmann, G.; Farrar, D.; Pohlenz-Michel, C.; Tillmann, T.; Mangelsdorf, I.: Genotoxicity testing of sulfur dioxide (SO2) in a mouse bone marrow micronucleus test complemented with hematological endpoints.
In: Mutation research. Genetic toxicology and environmental mutagenesis 697 (2010), Nr. 1-2, p. 38-46. ISSN: 1383-5718; ISSN: 1388-2120

Ziemann, C.; Bellmann, B.; Serwatzki, K.; Creutzenberg, O.; Ernst, H.; Kolling, A.; Beneke, S.; Rittinghausen, S.: Quantification of local genotoxicity in rat lungs after 3 months of particle exposure by immunohistochemical detection of 8-OH-dG, gamma-H2AX, and poly(ADP-ribose).
In: Deutsche Gesellschaft für Experimentelle und Klinische Pharmakologie und Toxikologie -DGPT: 51. Jahrestagung 2010. Deutsche Gesellschaft für Experimentelle und Klinische Pharmakologie und Toxikologie. Abstracts: 23. - 25. März 2010, Mainz.
Berlin: Springer, 2010 (Naunyn-Schmiedeberg's Archives of Pharmacology 381.2010, Supplement 1), p. 65, Abstract 300. ISSN: 0028-1298

Switalla, S.; Knebel, J.; Ritter, D.; Krug, N.; Braun, A.; Sewald, K.: Effects of acute in vitro exposure of murine precision-cut lung slices to gaseous nitrogen dioxide and ozone in an air-liquid interface (ALI) culture.
In: Toxicology letters 196 (2010), Nr. 2, p. 117-124. ISSN: 0378-4274

Ritter, D.; Switalla, S.; Sewald, K.; Knebel, J.: PCLS (Precision Cut Lung Slices) as a putative model for genotoxicity testing by application of the comet assay.
In: Mutagenesis 25 (2010), Nr. 1, p. 2. ISSN: 0267-8357; ISSN: 1464-3804

Pirow, R.; Smirnova, L.; Liebsch, M.; Tharmann, J.; Luch, A.; Graebsch, C.; Bauer, M.; Linsel, G.; Siemers, R.; Otto, C.; Berger-Preiß, E.; Kock, H.; Oertel, A.; Ritter, D.; Knebel, J.: Testing of the toxicity of volatile compounds on human lung cells using the Air/Liquid Interface (ALI) culturing and exposure technique: A prevalidation study.
In: Alternatives to animal experimentation: ALTEX 27 (2010), Suppl. 2, p. 101. ISSN: 1868-596X; ISSN: 0946-7785

2009

Creutzenberg, O.; Ziemann, C.; Hansen, T.; Ernst, H.; Jackson, P.; Cartlidge, D.; Brown, R.: In vivo study with quartz-containing ceramic dusts: Inflammatory effects of two factory samples in lungs after intratracheal instillation in a 28-day study with rats
In: Journal of Physics: Conference Series [Elektronische Ressource] 151 (2009), 012031, 5 p. 

Ziemann, C.; Brockmeyer, H.; Reddy, S. B.; Prihoda, T. J.; Kuster, N.; Tillmann, T.; Dasenbrock, C.: Absence of genotoxic potential of 902 MHz (GSM) and 1747 MHz (DCS) wireless communication signals: In vivo two-year bioassay in B6C3F1 mice
In: International Journal of Radiation Biology (2009), 11 p. [First article]

Ziemann, C.; Class, P.; Bellmann, B.; Creutzenberg, O.; Brown, R.: Effects of heating on free crystalline silica content, appearance, and biological activity of devitrified alkaline earth insulation wools
In: Naunyn-Schmiedebergs Archives of Pharmacology 379 (2009), Suppl. 1, p. 74, No. 366

Ziemann, C.; Jackson, P.; Brown, R.; Attik, G.; Rihn, B. H.; Creutzenberg, O.: Quartz-containing ceramic custs: in vitro screening of the cytotoxic, genotoxic and pro-inflammatory potential of 5 factory samples
In: Journal of Physics: Conference Series [Elektronische Ressource] 151 (2009), 012067, 6 p.