Advanced in vitro exposure systems.

9. Sep. 2016

21st Century Toxicology & In Vitro-Based Testing Methodologies

Julia Hoeng, Anita Iskandar, Filippo Zanetti

Philip Morris International


The studies used human cells grown in three-dimensional culture systems. The cultures were grown on top of an artificial membrane at the air-liquid interface, allowing them to develop ‘organotypic’ tissue complexity. Testcigarette is the Tobacco Heating System 2.2 (THS), a new technology that heats tobacco without burning it. The endpoints are shown in measurements of cytotoxicity, alterations in tissue morphology (histology), impact on processes involved in the metabolism of toxicants (xenobiotic metabolism response), secretion of pro-inflammatory mediators, and perturbation of genome-wide gene profiles. The nasal study also looked at ciliary function.



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8. Sep. 2016

A lab scale measurement technique for the air-liquid interface exposure of human lung cell cultures towards particulate emissions from combustion processes

11th International Particle Toxicology Conference, Sep 26 – 30, 2016 in Singapore


1S. Mülhopt, 3M. Berger, 1C. Schlager, 2M. Dilger, 2S. Diabaté, 3T. Krebs, 4,5Ralf Zimmermann, 4Jeroen Buters, 4Sebastian Oeder, 2C. Weiss, 1H.-R. Paur

1Institute for Technical Chemistry
2Institute of Toxicology and Genetics

3Vitrocell Systems GmbH, Germany
4Helmholtz CentreMunich, Germany
5University of Rostock, Germany
 

The Karlsruhe Exposure System is a lab scale measurement system for the air-liquid interface exposure of human lung cell cultures towards air borne nanoparticles under well defined conditions.

 

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1. Sep. 2016

Aerosol Flow in the Vitrocell 24/48 Exposure System: Flow Mixing and Aerosol Coalescence

Applied In Vitro Toxicology. September 2016, Vol. 2, No. 3: 165-174 

Arkadiusz K. Kuczaj,1,2 Markus Nordlund,1 Santhosh Jayaraju,3 Ed Komen,3 Tobias Krebs,4 Manuel C. Peitsch,1 and Julia Hoeng1
1Philip Morris International Research & Development, Philip Morris Products S.A., Neuchâtel, Switzerland.
2Multiscale Modeling & Simulation, Department of Applied Mathematics, University of Twente, Enschede, The Netherlands.
3NRG, Petten, The Netherlands.
4Vitrocell Systems GmbH, Waldkirch, Germany.
 

In this work, the flow-mixing efficiency of the dilution unit in the Vitrocell® 24/48 (VC24/48) exposure system and the role of thermal coalescence is presented . The dilution unit of the VC24/48 system guarantees efficient and localized mixing/dilution of the aerosol and the thermal coalescence effects do not influence the aerosol droplet size distribution.

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14. Aug. 2016

Copper oxide nanoparticles: Impact on alveolar epithelial-like cells following air-liquid interface exposure

EEMGS Annual Meeting: 14 – 18 August 2016, Copenhagen, Denmark

doi:10.1007/s00204-015-1621-7

Matthias Hufnagel, Sarah Schoch, Bettina Fischer, Andrea Hartwig
Karlsruhe Institute of Technology, Institute of Applied Biosciences, Department of Food Chemistry and Toxicology

This is work is part of ProCycle (No. 03XP0009), a project funded by the German Federal Ministry of Education and Research.and shows the exposition of copper oxide (CuO) nanoparticles on A549 cells at different doses in the VITROCELL® Cloud, equipped with a quartz crystal microbalance. The effects were determined using the colony formation assay (CFA) and high-throughput RT-qPCR gene expression analyses.

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12. Jul. 2016

Systems Toxicology Assessment of the Biological Impact of a Candidate Modified Risk Tobacco Product on Human Organotypic Oral Epithelial Cultures

DOI:10.1021/acs.chemrestox.6b00174

Chem. Res. Toxicol.2016, 29, 1252−1269

Filippo  Zanetti, Alain  Sewer, Carole  Mathis, Anita  R.  Iskandar, Radina  Kostadinova, Walter K. Schlage, Patrice Leroy, Shoaib Majeed, Emmanuel Guedj, Keyur Trivedi, Florian Martin, Ashraf  Elamin, Celine  Merg, Nikolai  V.  Ivanov, Stefan  Frentzel, Manuel  C.  Peitsch, and  Julia  Hoeng*,

Philip Morris International Research and Development, Quai Jeanrenaud 5, 2000 Neuchatel, Switzerland
Biology Consultant, Max-Baermann-Str. 21, 51429 Bergisch Gladbach, Germany

 
The tobacco heating system (THS) 2.2 and the 3R4F reference cigarette smoke were exposed to human organotypic oral epithelial tissue cultures (EpiOral, MatTek)at the Vitrocell 24/48 exposure system. The results are shown by the evaluation of cytotoxicity and cytochrome P450 activity assays, mRNA and miRNA purification, measurements of secreted proin flammatory markers and histopathological analysis.

 

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7. Jul. 2016

3-D Nasal Cultures: Systems Toxicological Assessment of a Candidate Modified-Risk Tobacco Product

doi: 10.14573/altex.1605041

Anita R. Iskandar, Carole Mathis, Florian Martin, Patrice Leroy, Alain Sewer, Shoaib Majeed, Diana Kuehn, Keyur Trivedi, Davide Grandolfo, Maciej Cabanski, Emmanuel Guedj, Celine Merg, Stefan Frentzel, Nikolai V. Ivanov, Manuel C. Peitsch and Julia Hoeng

Philip Morris International R&D, Philip Morris Products S.A., Neuchâtel, Switzerland

Human nasal epithelium is the first barrier against inhaled toxicants. The air-liquid interface human nasal culture MucilAir™ offers a more physiologically relevant and robust systems for studying the effects of exposure. This cells were exposed to 3R4F cigarettes and THS2.2 tobacco sticks
in the the Vitrocell® 24/48 exposure system. Results have been evaluated by measuring the cytotoxicity, histology processing, immunostaining, cilia beating frequency (CBF), cytochrome P450 (CYP) activity, luminex-based measurement of secreted analytes, RNA/miRNA purification and mRNA microarray.

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27. Jun. 2016

Metabolic Profiling as Well as Stable Isotope Assisted Metabolic and Proteomic Analysis of RAW 264.7 Macrophages Exposed to Ship Engine Aerosol Emissions

Different Effects of Heavy Fuel Oil and Refined Diesel Fuel

DOI:10.1371/journal.pone.0157964

Sean C. Sapcariu1,12, Tamara Kanashova2,12, Marco Dilger3,4,12, Silvia Diabaté3,12, Sebastian Oeder5,6,12,13, Johannes Passig7,12, Christian Radischat7,12, Jeroen Buters5,6,12,13, Olli Sippula8,12, Thorsten Streibel7,9,12, Hanns-Rudolf Paur4,12, Christoph Schlager4,12, Sonja Mülhopt4,12, Benjamin Stengel10,12, Rom Rabe10,12, Horst Harndorf10,12, Tobias Krebs11,12, Erwin Karg9, Thomas Gröger9, Carsten Weiss3,12, Gunnar Dittmar2,12, Karsten Hiller1,12, Ralf Zimmermann7,9,12

1 Luxembourg Centre for Systems Biomedicine 6, avenue du Swing, L-4362 Esch-sur-Alzette, Luxembourg,
2 Mass Spectrometry Core Unit, Max Delbrück Center for Molecular Medicine Berlin-Buch, Berlin, Germany,
3 Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany,
4 Institute for Technical Chemistry (ITC), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany,
5 Center of Allergy and Environment (ZAUM), Helmholtz Zentrum München and Technische Universität München, Munich, Germany,
6 CK-CARE, Christine Kühne Center for Allergy Research and Education, Davos, Switzerland,
7 Joint Mass Spectrometry Centre, Division of Analytical and Technical Chemistry, Institute of Chemistry, University Rostock, Rostock, Germany,
8 University of Eastern Finland, Department of Environmental Science, P.O. Box 1627, FI-70211 Kuopio, Finland,
9 Joint Mass Spectrometry Centre, CMA – Comprehensive Molecular Analytics, Helmholtz Zentrum München, Neuherberg, Germany,
10 Chair of Piston Machines and Internal Combustion Engines, University Rostock, Rostock, Germany,
11 Vitrocell GmbH, Waldkirch, Germany,
12 HICE – Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health – Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg,
13 German Center for Lung Research (DZL), Munich, Germany

An automated ALI exposure system station with 18 exposure positions was used as the interface for mouse macrophage RAW 264.7 cell line exposures of the diesel engine exhaust. The evaluation is devided in LDH release assay, Metabolite extraction and GC-MS processing, Stable isotope labeling by amino acids in cell culture (SILAC), Proteome extraction and LC-MS/MS analysis of peptides and Proteomics Data An

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24. Jun. 2016

An improved aerosol deposition device to assess safety and efficacy of dry powder formulations at the air/liquid interface

M Hittinger1, S Barthold2, A Gress3, B Wiegand1, L Siebenbürger1, C Börger1, M Berger4, T Krebs4, CM Lehr1, 2, 5, H Groß1

1PharmBioTec GmbH, Saarbrücken, Germany
2Saarland University, Department of Pharmacy, Saarbrücken, Germany
3Max Planck Institute for Informatics, Saarbrücken, Germany
4Vitrocell Systems GmbH, Waldkirch, Germany
5Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), HZI, Saarbrücken, Germany

This poster shows a newly developed instrument which can be used with commercial dry powder inhaler devices and offers flexible adjustment of the flow rate and other process parameters. Salbutamol sulfate dry powder formulation and sodium fluorescein were investigated in the device. The Results were estimated by electron microscop and the deposition of salbutamol sulfate was quantified by HPLC-MS analysis.

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16. Jun. 2016

Nanoparticles on human lung cells

The Vitrocell Exposure System for cell cultures at the air-liquid interface

Sonja Mülhopt1, Tobias Krebs2, Dr Silvia Diabaté3, Christoph Schlager1, Dr Hanns-Rudolf Paur1

1 Institute for Technical Chemistry, Karlsruhe Institute of Technology, 
2 Vitrocell Systems GmbH, 
3 Institute of Toxicology and Genetics, Karlsruhe Institute of Technology

 

This article shows the development of an automated exposure system which allows both reproducible sampling and conditioning of aerosols and exposure of the cell cultures under conditions imitating those of the human lung. Bioassays were developed and used for toxicological analysis of particulate emissions from the industry as well as of nanoparticles.

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30. Apr. 2016

Assessing the mutagenic activities of smoke from different cigarettes in direct exposure experiments using the modified Ames Salmonella assay

Shinkichi Ishikawa, Yuki Kanemaru, Hidenori Nara, Kazuo Erami, Yasufumi Nagata

Scientific Product Assessment Center, R&D Group, Japan Tobacco Inc., 6-2 Umegaoka, Aoba-ku, Yokohama, Kanagawa 227-8512, Japan

 

This article shows the successful exposition exposure of cigarette smoke on two different Ames bacteria strains by using the Vitrocell exposure system including the VC10 smoking robot. Conventional 1 mg tar commercial cigarettes (CC1), 1R5F reference cigarettes (University of Kentucky,Lexington, KY, USA), and 3R4F reference cigarettes (University of Kentucky) were compared with a prototype heated cigarette (HC).

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