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CarbonBlack – Prediction of the human-toxicological effect of synthetic carbon black nanoparticles

CarbonBlack was a joint project aimed at establishing a test system with test models of increasing complexity to verify the toxicological effect of modified and well-characterised carbon black nanoparticles (CBNP) in the respiratory tract and in the lung. The multi-stage test system included simple cell culture models, tissue culture models, and verification methods based on inhalation studies within animal models.

It has yet to be determined whether the potential risk of carbon black nanoparticles is determined primarily by the particle size and geometry or by the surface chemistry. Therefore, the Carbon Black project consortium analysed and compared unmodified carbon black nanoparticles (Printex90) with CBNPs coated with polycyclic aromatic hydrocarbons (PAHs). The PAH-containing group of CBNPs include Printex®90 nanoparticles, which were coated with benzo[a]pyrene or 9-nitroanthracene and acetylene black particles, which are equipped with a mixture of various PAHs on the particle surface. Basis for all analyses of the corresponding biological effects was a thorough chemical and physical characterisation of the carbon black nanoparticles. The nanoparticles were suspended according to a standardised protocol and then send to all project partners for testing in the various model systems of increasing complexity.

The overall results of the CarbonBlack project clearly demonstrated that the effects of carbon black nanoparticles in biological systems are largely determined by the chemical nature of their surface. The strongest biological effects were seen in the in vitro and ex vivo systems when exposed to acetylene black particles. After treatment with CBNPs, the observed effects were largely associated with the induced mRNA expression of cytochrome-P450-oxidoreductases CYP1A1 and CYP1B1 together with an increased cytotoxicity particularly in cells with secretory function. Acetylene black proved to be the only carbonaceous nanoparticles, which stimulated the mRNA expression of proinflammatory cytokines / chemokines.

In the mouse model, the impact of the carbon black nanoparticles on the respiratory epithelium showed a decrease from the trachea down to the bronchioles. The induction of proinflammatory cytokines in the lung parenchyma was more pronounced in the tissues of rats and humans compared to the mouse, which was confirmed in comparative experiments on precision cut lung slices (PCLS). After oropharyngeal aspiration of suspensions with carbon black nanoparticles, only acetylene black particles caused an increase of alveolar epithelial cells type II, as well as an altered mRNA expression of key enzymes responsible for synthesis of alveolar surfactant.

The findings of the in vitro and ex vivo assay systems correlated well with the performed in vivo inhalation study. Exposure of rats to carbon black nanoparticles using the OECD test guideline 412 showed no significant toxic effects with only the acetylene black particles causing a very moderate inflammation.

The results of this project suggest that the biological / toxic effects of carbon black nanoparticles can be enhanced by functionalising the particle surface with polycyclic aromatic hydrocarbons (PAH) provided that the exposed cells are able to react sensitive to PAHs by expression of cytochrome-P450-oxidoreductases. In addition, these findings also suggest that the acute pulmonary toxicity of unmodified carbon black nanoparticles and PAH-coated carbon black nanoparticles is very low.


Grant Number: BMBF - FKZ 03X0093
Duration: 01.08.2010 - 31.07.2013 (extended to 31.12.2013)

Project Lead

FZ BOrstel Logo
Prof. Dr. Heinz G. Fehrenbach, Research Centre Borstel, Leibniz-Center for Medicine and Biosciences , Borstel (DE)

Project Partners

FZ BOrstel Logo
Research Centre Borstel, Leibniz-Center for Medicine and Biosciences, Borstel (DE)
Universität Marburg Logo
Zellbiologie der Lunge, Philipps University Marburg, Marburg (DE)
KIT Logo English
Engler-Bunte-Institute - Division of Combustion Technology, Karlsruhe Institute of Technology (KIT), Karlsruhe (DE)
Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM) Logo
Fraunhofer-Institute for Toxicologiy and Experimental Medicine (ITEM), Hannover (DE)
Universität Lübeck Anatomie Logo
Institute of Anatomy, AG Barrier Organs, University Lübeck, Luebeck (DE)



  • Lindner K., Strobele M., Schlick S., Webering S., Jenckel A., Kopf J., Danov O., Sewald K., Buj C., Creutzenberg O., Tillmann T., Pohlmann G., Ernst H., Ziemann C., Huttmann G., Heine H., Bockhorn H., Hansen T., Konig P., Fehrenbach H. (2017). Biological effects of carbon black nanoparticles are changed by surface coating with polycyclic aromatic hydrocarbons. Part Fibre Toxicol, 14(1): 8.


  • Schreiber N., Strobele M., Hochscheid R., Kotte E., Weber P., Bockhorn H., Muller B. (2016). Modifications of carbon black nanoparticle surfaces modulate type II pneumocyte homoeostasis.J Toxicol Environ Health A, 79(4): 153-164.


  • Schreiber, N., M. Stroebele, J. Kopf, R. Hochscheid, E. Kotte, P. Weber, T. Hansen, H. Bockhorn, B. Mueller (2013). "Lung alterations following single or multiple low-dose carbon black nanoparticle aspirations in mice." J Toxicol Environ Health A 76(24): 1317-1332.

Alle verfügbaren Abschlussberichte des CarbonBlack Konsortiums finden Sie auf den Seiten der TIB Hannover


  • Danov O, Kopf J, Ströbele M, Bockhorn H, Fehrenbach H, Braun A, Sewald K, Hansen T (2014). Biological impact of modified inhalable carbon black nanoparticles assessed in cell and tissue culture models. Vortrag II-4a-455, p. 71; Proceedings of the 9th World Congress on Alternatives and Animal Use in the Life Sciences, 24-28. August 2014, Prag, Tschechien. [Vortrag ]


  • Fehrenbach, H., B. Mueller, P. Koenig, T. Hansen, H. Bockhorn (2013). "CarbonBlack: a joint research project to establish a test system for predicting human-toxicological effects of synthetic carbon black nanoparticles*." Pneumologie 67(12): A15. [Vortrag]
  • Hansen T, Kopf J, Danov O, Stroebele M, Braun A, Sewald K, Steinberg P and Fehrenbach H. (2013). Effects of carbon black nanoparticles on human pulmonary cell lines and precision cut lung slices. The Toxicologist 132 (1): 95 (abstract 444). [Poster ]
  • Hansen, T., J. Kopf, O. Danov, M. Stroebele, A. Braun, K. Sewald, H. Bockhorn, eH. Fehrnbach (2013). "In vitro and ex vivo toxicity screening for predicting toxicological effects of synthetic carbon black nanoparticles in humans." Pneumologie 67(12): A17. [Vortrag]
  • Schlick, S., M. Stroebele, J. Kopf, T. Hansen, H. Bockhorn, H. Fehrenbach (2013). "Airway region-specific effects of carbon black nanoparticles (CBNP)." Pneumologie 67(S 01): V361. [Vortrag]
  • Schlick, S., J. Kopf, M. Stroebele, H. Fehrenbach (2013). "Airway region-specific effects of carbon black nanoparticles (CBNP)." Pneumologie 67(12): A19. [Vortrag]
  • Schreiber, N., R. Hochscheid, E. Kotte, P. Weber, B. Mueller (2013). "Toxicological effects of functionalized Carbon Black Nanoparticles on Type II pneumocytes." Pneumologie 67(12): A20. [Vortrag]
  • Schreiber, N., M. Stroebele, J. Kopf, R. Hochscheid, E. Kotte, P. Weber, B. Mueller (2013). "Structural And Functional Changes Of The Lung Following Low Dose Single And Multiple Carbon Black Nanoparticle Applications." Am J Respir Crit Care Med 187: A5252. [Vortrag]
  • Stroebele, M. R., H. Bockhorn (2013). "Desorption behavior of polycyclic aromatic hydrocarbons (PAHs) on various carbon black nanoparticles (CBNPs)." Pneumologie 67(12): A14. [Vortrag]
  • Stroebele, M. R., H. Bockhorn (2013). "Specifically modified carbon black nanoparticles (CBNP) by gas-phase synthesis." Pneumologie 67(12): A16. [Vortrag]
  • Stroebele M., Henning Bockhorn, (2013). Effect of Synthesis Conditions on the Content of Polycyclic Aromatic Hydrocarbons on Carbon Black Nanoparticles. P5-52 in Proceedings of the European Combustion Meeting – 2013, June 25-28, 2013, Lund, Schweden. ISBN 978-91-637-2151-9. [Poster]
  • Weinhold, K., P. Koenig (2013). "Effects of surface-modified CBNP on the epithelium of the explanted mouse trachea (CarbonBlack sub-project 3)." Pneumologie 67(12): A18. [Vortrag]


  • Schlick, S., H. Fehrenbach (2012). "Airway region-specific effects of Carbon Black nanoparticles." Pneumologie 66 (7): A4. [Vortrag]
  • Schreiber, N., M. Stroebele, R. Hochscheid, E. Kotte, P. Weber, H. Bockhorn, B. Mueller (2012). "Low Dose Carbon Nanoparticle Application Induces Lung Inflammation." Am J Respir Crit Care Med 185 (2012): A1187. [Vortrag]
  • Weinhold, K., P. Koenig (2012). "Nanoparticles increase ciliary beat frequency in the murine trachea." Pneumologie 66 (6): A805. [Vortrag]

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