Nanoparticles: Synthesis, characterisation, and cellular effects
Pharmacology
Physical Chemistry of Molecules, Liquids and Interfaces, Biophysical Chemistry
Final Report Abstract
Quantification of the influence of the properties of nanoparticles on the induction of cellular responses requires substantial knowledge in the fields of nanoparticle synthesis, physical and chemical characterisation of nanoparticles, defined application of nanoparticles to biological systems, and determination of biological responses to nanoparticles in cell systems. The cluster NANO-SYNCC of the KIT institutes Engler-Bunte-Institute (EBI-VBT), Institute of Meteorology and Climate Research (IMK-AAF), Institute for Technical Chemistry (ITC), and Institute of Toxicology and Genetics (ITG) has achieved the following major progress: Metaloxide NPs (SiO2, TiO2, Fe2O3) with tailored properties were prepared by low pressure flame and by microwave plasma (MWP) synthesis. For the first time a complete model of the particle formation of metal oxides integrating gas- and particle phase processes was developed and verified by extensive measurements with a particle mass spectrometer. Based on low pressure flame synthesis, modelling studies were performed. Through this model particle formation and particle size distributions from Fe(CO)5 could be simulated and compared to measured values. All particles were extensively characterized with respect to biologically relevant properties such as particle size, degree of agglomerationand surface characteristics. Agglomerates of metal oxide NPs collected in flame and MWP-synthesis showed high stabilities circumventing a deagglomeration with reasonable measures to beyond 60-80 nm agglomerate size in mobility equivalent diameter. Development of a novel collection system for NP directly into a stabilising aqueous suspension yielded in first tests stable suspensions of uncoated, non-agglomerated 15 nm SiO2-Particles. Suspended metal oxide particles are reproducibly transferred to the aerosol phase by electrospray. The novel Air Liquid Interface Deposition-System (ALIDA) was developed, integrating an electrospray generator, humidification and an air liquid interface exposure system. By this system nanoparticles are directly deposited onto human lung cells (e. g. A549), thereby avoiding artefacts due to interaction of nanoparticles with biological media and allowing for exact dose determination at the same time. To monitor the amount of NPs deposited online a quartz crystal microbalance (QCM) was developed and transferred to industry. The exposure system was equipped with an electrostatic deposition enhancement increasing deposition efficiencies by an order of magnitude. Systematic exposure studies with electrosprayed metal oxide NPs (SiO2, TiO2, Fe2O3) were performed at the air-liquid interface. A screening of defined metal-oxide nanoparticles (Fe2O3, SiO2, TiO2) has been performed by in vitro experiments. The particles were synthesised and characterized by the cluster members EBI-VBT (Fe2O3, SiO2, and TiO2 NPs by flame synthesis) and ITC-TAB (SiO2 NPs in microwave plasma) and compared with commercial NPs, which are similar with respect to chemical composition and synthesis routes. SiO2-NPs were the most effective with regard to cytotoxicity, induction of anti-oxidative and inflammatory genes, although ROS production was not observed. RAW264.7 macrophages were more responsive than A549 cells and presence of serum significantly reduced the adverse effects. Preliminary results on the effects of TiO2 provided as an aerosol at the air-liquid interface in the new ALIDA system show no adverse effects at the used deposition dose. SiO2 NPs however, only induced cytotoxicity at very high doses. When compared to conventional submerged exposure conditions cells exposed at the ALI might be less sensitive to SiO2 NP induced toxicity.
Publications
- (2008). The response of a co-culture lung model to fine and ultrafine particles of incinerator fly ash at the air-liquid interface. Altern. Lab Anim 36, 285- 298
Diabaté, S., Mülhopt, S., Paur, H.R., and Krug, H.F.
- In vitro exposure systems and bioassays for the assessment of toxicity of nanoparticles to the human lung. Journal für Verbraucherschutz und Lebensmittelsicherheit, 3 (2008) S. 319-29
Paur, H.R.; Mülhopt, S.; Weiss, C.; Diabaté, S.
- (2009). Combustion Generated Fine Carbonaceous Particles. KIT Scientific Publishing, Karlsruhe
Bockhorn, H., A. D’Andrea, A. Sarofim, H. Wang
- (2009). Vorrichtung zur Messung von Feinstpartikelmassen. Deutschland patent application DE-OS 10 2007 013 938
Mülhopt, S., Paur, H.R., & Wäscher, T.
- (2011). Toxizität und pro-inflammatorisches Potenzial verschiedener Metalloxid Nanopartikel in Lungenzellen. Dissertation, Karlsruher Institut für Technologie
Panas, A.
- 2011. In-vitro cell exposure studies for the assessment of nanoparticle toxicity in the lung - A dialogue between aerosol science and biology. J. Aerosol Sci. 42, 668-692
Paur, H.R., Fissan, H., Rothen-Rutishauser, B., Teeguarden, J.G., Diabaté, S., Aufderheide, M., Kreyling, W., Cassee, F.R., Hänninen, O., Kasper, G., Riediker, M., Schmid, O.
- 2011. Uptake and intracellular localization of submicron and nanosized SiO(2) particles in HeLa cells. Arch. Toxicol. 85, 813-826
Al-Rawi, M., Diabaté, S., Weiss, C.
- (2012). Identification of serum proteins bound to industrial nanomaterials. Toxicol. Lett. 208, 41-50
Ruh, H., Kühl, B., Brenner-Weiss, G., Hopf, C., Diabaté, S., Weiss, C.
