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In vitro models as physiologically relevant tools to investigate pulmonary and intestinal toxicity
Abstracts / Toxicology Letters 229S (2014) S40–S252
most marked in the juvenile earthworms. The CV in the juvenile earthworm A. caliginosa were measured at 0, 1, 3, 6, 8, 24, and 48 h following exposure to AgNP and AgNO3 concentrations of LC0.001 , LC0.01 , LC0.1 , LC1 , LC2 , and L. variegatus to concentrations of LC1 , LC5 , LC10 , and LC20 . MGF CV progressively decreased in L. variegatus and A. caliginosa exposed to both AgNP and AgNO3 up to 8 h and then gradually increased. To eliminate possible degradation of the Ag compounds, the AgNP and AgNO3 solutions were renewed every 3 h and CV measurements taken at hourly intervals for 7 h. Based on our experience with P-glycoprotein (P-gp) transporters in oligochaetes, it is proposed that the oligochaetes adapt to excessive exposure of AgNP and AgNO3 via the elimination of chemicals via P-gp transporters. In conclusion, MGF CV measurements in the earthworms (monitor soil pollution) and aquatic worms (monitor aquatic pollution) can be used as a novel, relevant, non-invasive, sensitive, and early warning biomarker of exposure to NP. However, interpretation of chronic exposure requires further evaluation in view of the role of P-gp transporters in the excretion of excess chemicals from the body. http://dx.doi.org/10.1016/j.toxlet.2014.06.647 P-3.135 Effect of the presence of aquatic humic substances on the toxicity of chitosan/tripolyphosphate nanoparticles containing paraquat Renato Grillo 1,2,∗ , Zaira Clemente 3 , Victor Chalupe 3 , Claudio Jonsson 3 , Renata Lima 4 , Gabriela Sanches 4 , Caroline Nishisaka 4 , Kathleen Oehlke 5 , Ralf Greiner 5 , Leonardo Fraceto 1,2 1
State University of Campinas, Campinas, SP, Brazil, 2 São Paulo State University, Sorocaba, SP, Brazil, 3 Embrapa Environmental, Jaguariúna, SP, Brazil, 4 University of Sorocaba, Sorocaba, SP, Brazil, 5 Max Rubner-Institut, Karlsruhe, Germany
There is increasing production and use of nanoparticles, which can then be released into the environment. It is therefore important to understand the risks and behavior of these nanomaterials. However, the knowledge on stability of nanoparticles in environmental media containing humic substances is limited. Humic substances are known to interact among others with metals and organic compounds resulting in an alteration of their behavior in the environment. The objective of this work was the investigation of the effect of the presence of aquatic humic substances (AHS) on the toxicity of polymeric chitosan/tripolyphosphate nanoparticles (CS:TPP) containing the herbicide paraquat. Nanoparticles were prepared using a chitosan polymer (CS) cross-linked with the tripolyphosphate (TPP) anion and the amount of the herbicides associated with the nanoparticles were measured using the ultrafiltration/centrifugation method by HPLC. Allium cepa genotoxicity tests and ecotoxicity assays with the alga Pseudokirchneriella subcapitata were used to determine the toxicity of the systems containing nanoparticles and AHS. The encapsulation efficiency of CS:TPP nanoparticles containing paraquat was 62.66 ± 0.77%. Association of the nanoparticles with the AHS resulted in decreased genotoxicity, due to competition and/or binding of paraquat with free binding sites in the humic substance molecules. The ecotoxicity assays also showed a decreased toxicity of the herbicide when associated with the AHS. The findings presented here provide a basis for the establishment of protocols for the commercial use of these systems.
S191
Acknowledgements: FAPESP (processes 2013/12322-2 and 2011/01872-6), CNPq (Bilateral Brazil-Germany Cooperation Program – CNPq/BMBF/IB-DLR), and FUNDUNESP. http://dx.doi.org/10.1016/j.toxlet.2014.06.648 P-3.136 Dermal absorption of Zn from ZnO particles in sunscreens applied to humans Brian Gulson 1,∗ , Maxine McCall 2 , Fiona Larner 3 , Yalchin Oytam 2 , Laura Gomez 1,2 , Brent Baxter 4 1
Macquarie University, North Ryde NSW, Australia, 2 CSIRO Advanced Materials TCP (Nanosafety)/Animal, Food & Health Sciences, North Ryde NSW, Australia, 3 Earth Science & Engineering, Imperial College, London, UK, 4 Baxter Laboratories, Victoria, Australia Zinc oxide (ZnO) nanoparticles (NP) in sunscreens provide protection against UV exposure. Most dermal penetration studies of NP concluded that the NP reside near the surface of the skin. However, most of these studies have been short, used animal models or in vitro assays, and/or not taken into account the effects of UV exposure or skin flexing. Use of ZnO with a highly enriched stable 68Zn isotope (>99%), coupled with measurements of Zn isotopic ratios by multiple-collector inductively coupled plasma mass spectrometry, is highly sensitive in tracing 68Zn from 68ZnO particles in a complex matrix against a high Zn background. Using this approach, a study in Australia detected small increases of 68Zn in the blood of human females following dermal and UV exposure over 5 days to a sunscreen containing 68ZnO NP (∼30 nm) compared with a sunscreen with larger 68ZnO particles (∼100 nm). There was no difference in 68Zn concentrations in the blood of males exposed to either sunscreen. In an earlier pilot study over 5 days of 3 adults with minimal UV exposure, employing 68ZnO enriched to only 51% in a different formulation, small increases in 68ZnO in blood were also detected. These investigations require the resolution of subtle differences in Zn isotopic ratios. To independently confirm the above outcomes, blood samples were transferred from Australia to Imperial College, London. Data from the two studies performed 3 years apart, using similar techniques and instrumentation, but in different laboratories and analysts, show good agreement (r = 0.98, n = 13). http://dx.doi.org/10.1016/j.toxlet.2014.06.649 P-3.137 In vitro models as physiologically relevant tools to investigate pulmonary and intestinal toxicity Sebastian G. Klein 1,2 , Anastasia Georgantzopoulou 1 , Tommaso Serchi 1 , Sebastien Cambier 1 , Celine C. Leclercq 1 , Jenny Renaut 1 , Marcin Kruszewski 3 , Anna Lankoff 3 , Esther Lentzen 1 , Patrick Grysan 1 , Jean-Nicolas Audinot 1 , Cedric Guignard 1 , Andreas Krein 1 , Jürgen Junk 1 , Sylvain Legay 1 , Lucien Hoffmann 1 , Brunhilde Blömeke 1 , Arno C. Gutleb 1,∗ 1
CRP – Gabriel Lippmann, Belvaux, Luxembourg, 2 University of Trier, Trier, Germany, 3 Institute of Nuclear Chemistry and Technology, Warsawa, Poland The aim of our studies is to establish physiologically relevant in vitro models to investigate the response of two organs playing a key role in exposure to chemicals and nanomaterials. These two in vitro models reproduce the alveoli (3D-tetraculture system) and small
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Abstracts / Toxicology Letters 229S (2014) S40–S252
intestine barriers (co-culture) and there is surfactant secretion in the tetraculture system at the air-liquid interface (ALI) and mucus production in the intestinal coculture. The tetraculture model was exposed to different realistic amounts of diesel exhaust particulate matter (80 ng/cm2 , or 240 ng/cm2 ). A clear dose-dependent translocation of the transcription factor Nrf2, which regulates gene expression in response to oxidative stress, was observed after 4 h of incubation in the endothelial cells without reduction of cell viability. The alveolar model is able to detect secondary induced toxicity upon realistic exposure to environmental relevant concentration of particulate matter. The intestinal coculture model was used to evaluate effects of Ag 20 and 200 nm particles on the metabolic activity, oxidative stress and pro-inflammatory cytokine release. AgNO3 induced a reduction in metabolic activity in a dose dependent manner whereas no reduction was observed for both Ag particles. Ag was found to be homogenously distributed in the cell with aggregates observed for Ag 20 with a 5-fold increase in IL8 release. The proteomic data revealed that both Ag particles induced oxidative stress pathways and affected cytoskeleton, but regulated different sets of proteins compared to AgNO3 . Overall these two systems may become valuable tools for toxicological studies. http://dx.doi.org/10.1016/j.toxlet.2014.06.650 P-3.138 Protein carbonylation as a marker of oxidative stress induced by nanoparticles: Analysis of 16 inorganic nanoparticles Marc D. Driessen 1 , Rainer Ossig 2 , Jürgen Schnekenburger 2 , Antje Vennemann 3 , Martin Wiemann 3 , Andreas Luch 1 , Andrea Haase 1,∗ 1
German Federal Institute for Risk Assessment (BfR), Berlin, Germany, Biomedical Technology Center, Westfälische Wilhelms-Universität, Muenster, Germany, 3 IBE R&D gGmbH, Muenster, Germany 2
The BMBF-funded project “nanoGEM” follows a systematic approach to understand hazards associated with different types of nanoparticles (NP). Oxidative stress is considered to be a major paradigm to explain NP toxicity. Here we focused on protein carbonylation as a consequence of oxidative stress for a set of 16 different nanoparticles, used as either plane materials or with different surface coatings. In parallel several in vitro and in vivo toxicity endpoints have been analyzed. We used NP of 10 nm (ZrO2 ), 15 nm (SiO2 ) and 50 nm or 200 nm (Ag), furnished either with acidic, basic or polymeric functionalities and TiO2 , ZnO, BaSO4 and AlOOH as references. In a screening approach we studied time- and dose-dependent carbonylation of all 16 NP in NRK-52E cells via 1D immunoblots. Data were correlated with cytotoxicity (WST-8, LDH assay) and ROS formation (DCFDA assay). Furthermore we applied a 2D proteomics approach combined with MALDI-MS/MS to identify the proteins modified. Finally, for several NP we analyzed lung tissues after in vivo instillation in rats. Eight out of 16 NP induced protein carbonylation in NRK-52E cells. Observed protein carbonylation correlated well with overall toxicity. The 2D approach revealed a complex and distinct pattern of carbonyls. Modified proteins were identified as cytoskeleton proteins, heat shock proteins or proteins of major cellular pathways (i.e. glycolysis). We also observed carbonyl modifications in lung tissue homogenates of rats intratracheally instilled with the same NP.
Taken together, analysis of protein carbonylation is a useful tool for the analysis and mechanistic understanding of ROS dependent NM toxicity. http://dx.doi.org/10.1016/j.toxlet.2014.06.651 P-3.139 Nanoparticle-induced oxidative stress alters phospho-tyrosine patterns in mammalian cells: Results of an SH2 profiling approach Marc D. Driessen 1 , Rainer Ossig 2 , Jürgen Schnekenburger 2 , Antje Vennemann 3 , Martin Wiemann 3 , Andreas Luch 1 , Peter Nollau 4 , Andrea Haase 1,∗ 1
German Federal Institute for Risk Assessment (BfR), Berlin, Germany, Biomedical Technology Center, Westfälische Wilhelms-Universität, Muenster, Germany, 3 IBE R&D gGmbH, Muenster, Germany, 4 Forschungsinstitut Kinderkrebs-Zentrum Hamburg, University Medical Center Hamburg-Eppendorf, Hamburg, Germany 2
The BMBF-funded project “nanoGEM” follows a systematic approach to understand hazards associated with different types of nanoparticles (NP). Oxidative stress is considered to be a major paradigm to explain NP toxicity and can result in altered levels of tyrosine phosphorylation, probably by protein tyrosine phosphatase inhibition which are major components of various cellular signaling pathways. Here we focused on global changes of tyrosine phosphorylation resulting from treatment with 16 NP bearing different surface coatings. In parallel several well established in vitro and in vivo toxicity endpoints were analyzed. We used ZrO2 , SiO2 Ag NP, either plane or acidic, basic or polymeric functionalisation and in addition TiO2 , ZnO, BaSO4 and AlOOH as references. We analyzed NP treated NRK-52E cells by far western blot analysis with a set of 70 src homolgy 2 (SH2) domains known to differentially bind phospho-tyrosine sites. From these 70 SH2 domains, a set of 9 domains was selected and cellular reactions after SiO2 and Ag NP treatment were studied in detail. Furthermore we used these 9 SH2 domains to study changes in signaling pathways in lung tissue lysates after in vivo instillation in rats. Most NP caused alterations in phospho-tyrosine dependent cell signaling. Strongest responses were observed for cytotoxic NP (SiO2 naked, ZnO), fitting very well to overall observed toxicity, additionally enabling us to detect subtle changes in phosphotyrosine dependent signaling also for non-toxic NP. SH2 profiling appears to be a powerful tool to study cellular responses after NP treatment and opens insights into signaling mechanisms underlying toxicty. http://dx.doi.org/10.1016/j.toxlet.2014.06.652 P-3.140 Critical role of interleukin 1 for the initiation and resolution of pulmonary inflammation induced by carbon nanomaterials Nunja C. Habel 1,∗ , David Kutschke 1 , Oliver Eickelberg 1,2 , Silke Meiners 1 , Tobias Stoeger 1 1 Comprehensive Pneumology Center, Institute of Lung Biology and Disease, Helmholtz Zentrum München, Munich, Germany, 2 Institute of Experimental Pneumology, Munich, Germany
Inhalation of carbon nanoparticles has been shown to cause acute lung inflammation, and particle shape has been related to inflammasome and downstream Il1-R1 receptor signaling
most marked in the juvenile earthworms. The CV in the juvenile earthworm A. caliginosa were measured at 0, 1, 3, 6, 8, 24, and 48 h following exposure to AgNP and AgNO3 concentrations of LC0.001 , LC0.01 , LC0.1 , LC1 , LC2 , and L. variegatus to concentrations of LC1 , LC5 , LC10 , and LC20 . MGF CV progressively decreased in L. variegatus and A. caliginosa exposed to both AgNP and AgNO3 up to 8 h and then gradually increased. To eliminate possible degradation of the Ag compounds, the AgNP and AgNO3 solutions were renewed every 3 h and CV measurements taken at hourly intervals for 7 h. Based on our experience with P-glycoprotein (P-gp) transporters in oligochaetes, it is proposed that the oligochaetes adapt to excessive exposure of AgNP and AgNO3 via the elimination of chemicals via P-gp transporters. In conclusion, MGF CV measurements in the earthworms (monitor soil pollution) and aquatic worms (monitor aquatic pollution) can be used as a novel, relevant, non-invasive, sensitive, and early warning biomarker of exposure to NP. However, interpretation of chronic exposure requires further evaluation in view of the role of P-gp transporters in the excretion of excess chemicals from the body. http://dx.doi.org/10.1016/j.toxlet.2014.06.647 P-3.135 Effect of the presence of aquatic humic substances on the toxicity of chitosan/tripolyphosphate nanoparticles containing paraquat Renato Grillo 1,2,∗ , Zaira Clemente 3 , Victor Chalupe 3 , Claudio Jonsson 3 , Renata Lima 4 , Gabriela Sanches 4 , Caroline Nishisaka 4 , Kathleen Oehlke 5 , Ralf Greiner 5 , Leonardo Fraceto 1,2 1
State University of Campinas, Campinas, SP, Brazil, 2 São Paulo State University, Sorocaba, SP, Brazil, 3 Embrapa Environmental, Jaguariúna, SP, Brazil, 4 University of Sorocaba, Sorocaba, SP, Brazil, 5 Max Rubner-Institut, Karlsruhe, Germany
There is increasing production and use of nanoparticles, which can then be released into the environment. It is therefore important to understand the risks and behavior of these nanomaterials. However, the knowledge on stability of nanoparticles in environmental media containing humic substances is limited. Humic substances are known to interact among others with metals and organic compounds resulting in an alteration of their behavior in the environment. The objective of this work was the investigation of the effect of the presence of aquatic humic substances (AHS) on the toxicity of polymeric chitosan/tripolyphosphate nanoparticles (CS:TPP) containing the herbicide paraquat. Nanoparticles were prepared using a chitosan polymer (CS) cross-linked with the tripolyphosphate (TPP) anion and the amount of the herbicides associated with the nanoparticles were measured using the ultrafiltration/centrifugation method by HPLC. Allium cepa genotoxicity tests and ecotoxicity assays with the alga Pseudokirchneriella subcapitata were used to determine the toxicity of the systems containing nanoparticles and AHS. The encapsulation efficiency of CS:TPP nanoparticles containing paraquat was 62.66 ± 0.77%. Association of the nanoparticles with the AHS resulted in decreased genotoxicity, due to competition and/or binding of paraquat with free binding sites in the humic substance molecules. The ecotoxicity assays also showed a decreased toxicity of the herbicide when associated with the AHS. The findings presented here provide a basis for the establishment of protocols for the commercial use of these systems.
S191
Acknowledgements: FAPESP (processes 2013/12322-2 and 2011/01872-6), CNPq (Bilateral Brazil-Germany Cooperation Program – CNPq/BMBF/IB-DLR), and FUNDUNESP. http://dx.doi.org/10.1016/j.toxlet.2014.06.648 P-3.136 Dermal absorption of Zn from ZnO particles in sunscreens applied to humans Brian Gulson 1,∗ , Maxine McCall 2 , Fiona Larner 3 , Yalchin Oytam 2 , Laura Gomez 1,2 , Brent Baxter 4 1
Macquarie University, North Ryde NSW, Australia, 2 CSIRO Advanced Materials TCP (Nanosafety)/Animal, Food & Health Sciences, North Ryde NSW, Australia, 3 Earth Science & Engineering, Imperial College, London, UK, 4 Baxter Laboratories, Victoria, Australia Zinc oxide (ZnO) nanoparticles (NP) in sunscreens provide protection against UV exposure. Most dermal penetration studies of NP concluded that the NP reside near the surface of the skin. However, most of these studies have been short, used animal models or in vitro assays, and/or not taken into account the effects of UV exposure or skin flexing. Use of ZnO with a highly enriched stable 68Zn isotope (>99%), coupled with measurements of Zn isotopic ratios by multiple-collector inductively coupled plasma mass spectrometry, is highly sensitive in tracing 68Zn from 68ZnO particles in a complex matrix against a high Zn background. Using this approach, a study in Australia detected small increases of 68Zn in the blood of human females following dermal and UV exposure over 5 days to a sunscreen containing 68ZnO NP (∼30 nm) compared with a sunscreen with larger 68ZnO particles (∼100 nm). There was no difference in 68Zn concentrations in the blood of males exposed to either sunscreen. In an earlier pilot study over 5 days of 3 adults with minimal UV exposure, employing 68ZnO enriched to only 51% in a different formulation, small increases in 68ZnO in blood were also detected. These investigations require the resolution of subtle differences in Zn isotopic ratios. To independently confirm the above outcomes, blood samples were transferred from Australia to Imperial College, London. Data from the two studies performed 3 years apart, using similar techniques and instrumentation, but in different laboratories and analysts, show good agreement (r = 0.98, n = 13). http://dx.doi.org/10.1016/j.toxlet.2014.06.649 P-3.137 In vitro models as physiologically relevant tools to investigate pulmonary and intestinal toxicity Sebastian G. Klein 1,2 , Anastasia Georgantzopoulou 1 , Tommaso Serchi 1 , Sebastien Cambier 1 , Celine C. Leclercq 1 , Jenny Renaut 1 , Marcin Kruszewski 3 , Anna Lankoff 3 , Esther Lentzen 1 , Patrick Grysan 1 , Jean-Nicolas Audinot 1 , Cedric Guignard 1 , Andreas Krein 1 , Jürgen Junk 1 , Sylvain Legay 1 , Lucien Hoffmann 1 , Brunhilde Blömeke 1 , Arno C. Gutleb 1,∗ 1
CRP – Gabriel Lippmann, Belvaux, Luxembourg, 2 University of Trier, Trier, Germany, 3 Institute of Nuclear Chemistry and Technology, Warsawa, Poland The aim of our studies is to establish physiologically relevant in vitro models to investigate the response of two organs playing a key role in exposure to chemicals and nanomaterials. These two in vitro models reproduce the alveoli (3D-tetraculture system) and small
S192
Abstracts / Toxicology Letters 229S (2014) S40–S252
intestine barriers (co-culture) and there is surfactant secretion in the tetraculture system at the air-liquid interface (ALI) and mucus production in the intestinal coculture. The tetraculture model was exposed to different realistic amounts of diesel exhaust particulate matter (80 ng/cm2 , or 240 ng/cm2 ). A clear dose-dependent translocation of the transcription factor Nrf2, which regulates gene expression in response to oxidative stress, was observed after 4 h of incubation in the endothelial cells without reduction of cell viability. The alveolar model is able to detect secondary induced toxicity upon realistic exposure to environmental relevant concentration of particulate matter. The intestinal coculture model was used to evaluate effects of Ag 20 and 200 nm particles on the metabolic activity, oxidative stress and pro-inflammatory cytokine release. AgNO3 induced a reduction in metabolic activity in a dose dependent manner whereas no reduction was observed for both Ag particles. Ag was found to be homogenously distributed in the cell with aggregates observed for Ag 20 with a 5-fold increase in IL8 release. The proteomic data revealed that both Ag particles induced oxidative stress pathways and affected cytoskeleton, but regulated different sets of proteins compared to AgNO3 . Overall these two systems may become valuable tools for toxicological studies. http://dx.doi.org/10.1016/j.toxlet.2014.06.650 P-3.138 Protein carbonylation as a marker of oxidative stress induced by nanoparticles: Analysis of 16 inorganic nanoparticles Marc D. Driessen 1 , Rainer Ossig 2 , Jürgen Schnekenburger 2 , Antje Vennemann 3 , Martin Wiemann 3 , Andreas Luch 1 , Andrea Haase 1,∗ 1
German Federal Institute for Risk Assessment (BfR), Berlin, Germany, Biomedical Technology Center, Westfälische Wilhelms-Universität, Muenster, Germany, 3 IBE R&D gGmbH, Muenster, Germany 2
The BMBF-funded project “nanoGEM” follows a systematic approach to understand hazards associated with different types of nanoparticles (NP). Oxidative stress is considered to be a major paradigm to explain NP toxicity. Here we focused on protein carbonylation as a consequence of oxidative stress for a set of 16 different nanoparticles, used as either plane materials or with different surface coatings. In parallel several in vitro and in vivo toxicity endpoints have been analyzed. We used NP of 10 nm (ZrO2 ), 15 nm (SiO2 ) and 50 nm or 200 nm (Ag), furnished either with acidic, basic or polymeric functionalities and TiO2 , ZnO, BaSO4 and AlOOH as references. In a screening approach we studied time- and dose-dependent carbonylation of all 16 NP in NRK-52E cells via 1D immunoblots. Data were correlated with cytotoxicity (WST-8, LDH assay) and ROS formation (DCFDA assay). Furthermore we applied a 2D proteomics approach combined with MALDI-MS/MS to identify the proteins modified. Finally, for several NP we analyzed lung tissues after in vivo instillation in rats. Eight out of 16 NP induced protein carbonylation in NRK-52E cells. Observed protein carbonylation correlated well with overall toxicity. The 2D approach revealed a complex and distinct pattern of carbonyls. Modified proteins were identified as cytoskeleton proteins, heat shock proteins or proteins of major cellular pathways (i.e. glycolysis). We also observed carbonyl modifications in lung tissue homogenates of rats intratracheally instilled with the same NP.
Taken together, analysis of protein carbonylation is a useful tool for the analysis and mechanistic understanding of ROS dependent NM toxicity. http://dx.doi.org/10.1016/j.toxlet.2014.06.651 P-3.139 Nanoparticle-induced oxidative stress alters phospho-tyrosine patterns in mammalian cells: Results of an SH2 profiling approach Marc D. Driessen 1 , Rainer Ossig 2 , Jürgen Schnekenburger 2 , Antje Vennemann 3 , Martin Wiemann 3 , Andreas Luch 1 , Peter Nollau 4 , Andrea Haase 1,∗ 1
German Federal Institute for Risk Assessment (BfR), Berlin, Germany, Biomedical Technology Center, Westfälische Wilhelms-Universität, Muenster, Germany, 3 IBE R&D gGmbH, Muenster, Germany, 4 Forschungsinstitut Kinderkrebs-Zentrum Hamburg, University Medical Center Hamburg-Eppendorf, Hamburg, Germany 2
The BMBF-funded project “nanoGEM” follows a systematic approach to understand hazards associated with different types of nanoparticles (NP). Oxidative stress is considered to be a major paradigm to explain NP toxicity and can result in altered levels of tyrosine phosphorylation, probably by protein tyrosine phosphatase inhibition which are major components of various cellular signaling pathways. Here we focused on global changes of tyrosine phosphorylation resulting from treatment with 16 NP bearing different surface coatings. In parallel several well established in vitro and in vivo toxicity endpoints were analyzed. We used ZrO2 , SiO2 Ag NP, either plane or acidic, basic or polymeric functionalisation and in addition TiO2 , ZnO, BaSO4 and AlOOH as references. We analyzed NP treated NRK-52E cells by far western blot analysis with a set of 70 src homolgy 2 (SH2) domains known to differentially bind phospho-tyrosine sites. From these 70 SH2 domains, a set of 9 domains was selected and cellular reactions after SiO2 and Ag NP treatment were studied in detail. Furthermore we used these 9 SH2 domains to study changes in signaling pathways in lung tissue lysates after in vivo instillation in rats. Most NP caused alterations in phospho-tyrosine dependent cell signaling. Strongest responses were observed for cytotoxic NP (SiO2 naked, ZnO), fitting very well to overall observed toxicity, additionally enabling us to detect subtle changes in phosphotyrosine dependent signaling also for non-toxic NP. SH2 profiling appears to be a powerful tool to study cellular responses after NP treatment and opens insights into signaling mechanisms underlying toxicty. http://dx.doi.org/10.1016/j.toxlet.2014.06.652 P-3.140 Critical role of interleukin 1 for the initiation and resolution of pulmonary inflammation induced by carbon nanomaterials Nunja C. Habel 1,∗ , David Kutschke 1 , Oliver Eickelberg 1,2 , Silke Meiners 1 , Tobias Stoeger 1 1 Comprehensive Pneumology Center, Institute of Lung Biology and Disease, Helmholtz Zentrum München, Munich, Germany, 2 Institute of Experimental Pneumology, Munich, Germany
Inhalation of carbon nanoparticles has been shown to cause acute lung inflammation, and particle shape has been related to inflammasome and downstream Il1-R1 receptor signaling