Advanced in vitro exposure systems.

9. Dec. 2021

Iota-carrageenan extracted from red algae is a potent inhibitor of SARS-CoV-2 infection in reconstituted human airway epithelia

https://doi.org/10.1016/j.bbrep.2021.101187

David Bovard b, Marco van der Toorn b, Walter K. Schlage a, Samuel Constant c, Kasper Renggli b, Manuel C. Peitsch b, Julia Hoeng b,
a Biology Consultant, Max-Baermann-Str. 21, 51429, Bergisch Gladbach, Germany
b PMI R&D, Philip Morris Products S.A, Quai Jeanrenaud 5, 2000, Neuchaˆtel, Switzerland
c Epithelix Sarl, 18 Chemin des Aulx, Plan-les-Ouates, 1228, Geneva, Switzerland

 

This study successfully demonstrates the exposure of human bronchial epithelial cultures to defined doses of nebulized Iota-carrageenan which were reproducibly generated and administered using the VITROCELL Cloud 12 system. 
The aim of this study was to further investigate the efficacy and safety of IC treatment on SARS-CoV-2 infection by using advanced in vitro models of human respiratory epithelium, the primary target and entry port of SARS-CoV-2. The experimental models were 3D cultures of reconstituted bronchial and nasal epithelia, representing the surface of the human upper respiratory tract. This apical exposure of reconstructed epithelia more closely mimics real-life exposure conditions and the absence of toxicity or any functional or structural impairment of the bronchial mucociliary epithelium demonstrates that topical treatment with nebulized IC is well tolerated at the effective concentrations. 

 

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15. Jun. 2022

Characterization of Aerosol Deliveries from Combustible Cigarettes, Heated Tobacco Products, and Electronic Nicotine Delivery Systems Using the Vitrocell Ames 48

DOI: 10.1089/aivt.2022.0001

Brian M. Keyser,1 Robert Leverette,1 Michael Hollings,2 Adam Seymour,2 Randy J. Weidman,3 Carlton J. Bequette,3 and Kristen Jordan1
1 Scientific & Regulatory Affairs, RAI Services Company, Winston-Salem, North Carolina, USA.
2 Labcorp Early Development Laboratories Ltd., Harrogate, North Yorkshire, United Kingdom.
3 RJ Reynolds Tobacco Company, Winston-Salem, North Carolina, USA

 

This study utilized a Vitrocell Ames 48 exposure module climatic chamber. Cigarette smoke and eHTP/ENDS aerosol was generated using a Vitrocell VC10. Different concentrations of whole smoke or aerosol were achieved by altering the diluting airflow using mass flow controllers  The dosimetry measure used for assessing both combustible and NGTP aerosols in this characterization exercise was the laser photometer purchased from Vitrocell Systems and harmonized for each tobacco product type used in this study. To evaluate the multiple dosimetry techniques, pairwise comparisons were made between the two Vitrocell Ames 48 exposure modules for each of the three test articles. 

 

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15. Apr. 2022

Aerosol–Cell Exposure System Applied to Semi-Adherent Cells for Aerosolization of Lung Surfactant and Nanoparticles Followed by High Quality RNA Extraction

https://doi.org/10.3390/nano12081362


Mélanie M. Leroux 1 , Romain Hocquel 1, Kevin Bourge 1 , Boštjan Kokot 2 , Hana Kokot 2 , Tilen Kokliˇc 2, Janez Štrancar 2, Yaobo Ding 3 , Pramod Kumar 3, Otmar Schmid 3 , Bertrand H. Rihn 1, Luc Ferrari 1 and Olivier Joubert 1
1 Institut Jean Lamour, UMR CNRS 7198, Université de Lorraine, CNRS, IJL, F-54000 Nancy, France
2 Jožef Stefan Institute, Department of Condensed Matter Physics, 1000 Ljubljana, Slovenia
3 Institute of Lung Health and Immunity, Helmholtz Zentrum München, German Research Center for Environmental Health, 85764 Neuherberg, Germany

The goal of this work was to refine an ALI pre-established protocol for the VITROCELL® Cloud 6 system. Therefore, after addressing each crucial step, (i.e., the use of very low concentrations of NP, the deposition of a homogeneous layer of surfactant, the specific use of semi-adherent cells, and finally the enhancement of the extraction yield of RNA to carry out transcriptomic studies), we present here for the first time a robust almost ready-to-use protocol. This one will be helpful for nanotoxicologists interested in developing such methodologies.

 

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27. Mar. 2022

Nrf2 Responses of Commercial Cigarette Whole Smoke and Aerosol from Six Electronic Nicotine Delivery Systems (ENDS) in a 3D Human Airway Model

Poster SOT 2022, 61st Annual Meeting and ToxExpo, March 27-31, San Diego, USA

Brian M. Keyser1, John Wertman1, Michael Hollings2, Robert Bedford2, and Kristen Jordan1
1 Scientific & Regulatory Affairs, RAI Services Company, Winston-Salem, NC; 
2 Labcorp Early Development Laboratories Ltd., Harrogate, UK

In this study whole smoke from a marketed combustible cigarette and whole aerosol from four different ENDS were evaluated on cell viability and Nrf2 response in a 3D human airway model transfected with a luciferase Nrf2 promoter.
Whole aerosol was generated using a Vitrocell ® VC10® Smoke Exposure System. The tissues were exposed to whole smoke from a market combustible generated under Health Canada Intense regime.These data show that the 3D Nrf2 EpiAirway™ in vitro model can be used to assess and discriminate responses from a biomarker (oxidative stress) for disease pathways associated with tobacco product usage.

 

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27. Mar. 2022

Cytotoxicity Assessment of Electronic Nicotine Delivery Systems (ENDS) and Combustible Cigarette Aerosols Utilizing Standard and Whole Aerosol Exposure Approaches in the Neutral Red Uptake Assay

Poster SOT 2022, 61st Annual Meeting and ToxExpo, March 27-31, San Diego, USA

Robert Leverette1, Thomas Shutsky1, John Wertman1, Katarina Aleksa2, Dhatri Lakshmanan2, Rebecca Payne3, Kristen Jordan1
1Scientific & Regulatory Affairs, RAI Services Company, Winston-Salem, NC, USA; 
2Labstat International Inc. Kitchener, ON, Canada;
3Labcorp Early Development Laboratories Ltd., Harrogate, UK

This poster shows whole aerosol exposures of combustible and next generation tobacco products, including Electronic Nicotine Delivery Systems. A Vitrocell® VC10® robot generated and delivered aerosols to the Mammalian 6/48 aerosol dilution and exposure system, with up to 7 concurrent doses plus a clean air control. Dosimetry module allowed the capture and quantification of deposited aerosol constituents (nicotine, glycerol and carbonyls). The study utilized the Neutral Red Uptake (NRU) assay in which mammalian cells were exposed to either combined TPM + GVP (submerged culture) or Whole Aerosol (ALI).

 

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27. Mar. 2022

In Vitro Cytotoxicity Assessment of 3D Human Airway Tissue Following Exposure to Whole Aerosol/Smoke Generated from Electronic and Combustible Cigarettes

Poster SOT 2022, 61st Annual Meeting and ToxExpo, March 27-31, San Diego, USA

Thomas Shutsky1, Brian M. Keyser1, Kristen Jordan1, Michael Hollings2, and Emma Rothwell2
1Scientific & Regulatory Affairs, RAI Services Company, Winston-Salem, NC; 
2Labcorp Early Development Laboratories Ltd., Harrogate, UK

In this study, EpiAirway™ tissues were exposed to whole aerosol/smoke generated from six different Vuse Alto electronic nicotine delivery system test articles with various nicotine concentrations and a combustible cigarette. A Vitrocell® VC10® robot was used to generate whole aerosol/smoke with either a modified ISO or Health Canada Intense smoking regimen. Whole smoke/aerosol was puffed with clean air with a series of different air flows to achieve the delivered dose range. 

 

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1. Mar. 2022

The priming effect of diesel exhaust on native pollen exposure at the air-liquid interface

https://doi.org/10.1016/j.envres.2022.112968


Joana Candeiasa, Elias J. Zimmermannbc, Christoph Bisigb, Nadine Gawlittabc, Sebastian Oederb, Thomas Grögerb, Ralf Zimmermannbc, Carsten B. Schmidt-Webera, Jeroen Butersa
a Center Allergy & Environment (ZAUM), Member of the German Center for Lung Research (DZL), Technical University Munich / Helmholtz Center Munich, Germany
b Joint Mass Spectrometry Center (JMSC) at Comprehensive Molecular Analytics (CMA), Helmholtz Center Munich, Ingolstädter Landstraße 1, D-85764, Neuherberg, Germany
c Joint Mass Spectrometry Center (JMSC) at Analytical Chemistry, Institute of Chemistry, University of Rostock, Dr. Lorenzweg 2, D-18051, Rostock, Germany

 

Human bronchial epithelial BEAS-2B cells were exposed to native birch pollen (real life intact pollen, not pollen extracts) at the air-liquid interface (pollen-ALI). BEAS-2B cells were also pre-exposed in a diesel-ALI to diesel CAST for 2 h (a model for diesel exhaust) and then to pollen in the pollen-ALI 24 h later. Effects were analysed by genome wide transcriptome analysis after 2 h 25 min, 6 h 50 min and 24 h. Selected genes were confirmed by qRT-PCR.

 

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1. Feb. 2022

Automation and Standardization—A Coupled Approach Towards Reproducible Sample Preparation Protocols for Nanomaterial Analysis

https://doi.org/10.3390/molecules27030985

Jörg Radnik 1, Vasile-Dan Hodoroaba 1, Harald Jungnickel 2, Jutta Tentschert 2, Andreas Luch 2, Vanessa Sogne 3, Florian Meier 3, Loïc Burr 4, David Schmid 4, Christoph Schlager 5, Tae Hyun Yoon 6,7, Ruud Peters 8, Sophie M. Briffa 9 and Eugenia Valsami-Jones 9


1 Division 6.1, Federal Institute for Material Testing and Research (BAM), Unter den Eichen 44–46, 12203 Berlin, Germany;
2 Department of Chemical & Product Safety, German Federal Institute for Risk Assessment, Max-Dohrn-Strasse 8–10, 10589 Berlin, Germany; 
3 Postnova Analytics GmbH, Rankine-Strasse 1, 86899 Landsberg, Germany;
4 Centre Suisse d’Electronique et de Microtechnique (CSEM), Bahnhofstrasse 1, 7302 Landquart, Switzerland;
5 Vitrocell Systems GmbH, Fabrik Sonntag 3, 78193 Waldkirch, Germany; c.schlager@vitrocell.com
6 Department of Chemistry, College of Natural Sciences, Hanyang University, Seoul 04763, Korea;
7 Institute of Next Generation Material Design, Hanyang University, Seoul 04673, Korea
8 Wageningen Food Safety Research, Wageningen University & Research, Akkermaalsbos 2, 6708 Wageningen, The Netherlands;
9 School of Geography, Earth and Environmental Sciences, University of Birmingham, Birmingham B15 2TT, UK;

 

The aim of this publication is to discuss how and when automated preparation can enhance the quality of the measurement results. For modern apparatuses, the measurement conditions are recorded and saved in the metadata automatically. As a result, the main reason for varying results is the different sample preparation. An actual interlaboratory comparison is ongoing to investigate the effect of different preparation methods systematically for ToF-SIMS. An air–liquid interface was developed to show that automation is possible for rather complex samples. Biological samples can be prepared in a reproducible manner, under exactly the same conditions on a TEM grid for the analysis of size and shape. Furthermore, chemical analysis can be performed by means of mass spectrometry.

 

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27. Nov. 2021

Agglomeration State of Titanium-Dioxide (TiO2) Nanomaterials Influences the Dose Deposition and Cytotoxic Responses in Human Bronchial Epithelial Cells at the Air-Liquid Interface

https://doi.org/10.3390/nano11123226

Sivakumar Murugadoss 1, Sonja Mülhopt 2, Silvia Diabaté 3, Manosij Ghosh 1, Hanns-Rudolf Paur 2,
Dieter Stapf 2, Carsten Weiss 3, and Peter H. Hoet 1,

1 Laboratory of Toxicology, Unit of Environment and Health, Department of Public Health and Primary Care, KU Leuven, 3000 Leuven, Belgium
2 Institute for Technical Chemistry, Karlsruhe Institute of Technology, 76021 Karlsruhe, Germany
3 Institute of Biological and Chemical Systems—Biological Information Processing, Karlsruhe Institute of Technology, 76021 Karlsruhe, Germany

 

In this study, they investigated the influence of agglomeration on the deposition and cytotoxic potency of TiO2 NMs at the ALI. Our results indicate that dose deposition and the cytotoxic potential are influenced by agglomeration, particularly for nano-sized TiO2 particles.

 

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27. Jun. 2021

Impact of Nanocomposite Combustion Aerosols on A549 Cells and a 3D Airway Model

https://doi.org/10.3390/nano11071685

Matthias Hufnagel 1, Nadine May 2, Johanna Wall 1, Nadja Wingert 3, Manuel Garcia-Käufer 3, Ali Arif 3, Christof Hübner 4, Markus Berger 5, Sonja Mülhopt 2, Werner Baumann 2, Frederik Weis 6, Tobias Krebs 5, Wolfgang Becker 4, Richard Gminski 3, Dieter Stapf 2, and Andrea Hartwig 1,


1 Department of Food Chemistry and Toxicology, Institute of Applied Biosciences, Karlsruhe Institute of Technology, 76131 Karlsruhe, Germany;
2 Institute for Technical Chemistry, Karlsruhe Institute of Technology, 76344 Eggenstein-Leopoldshafen, Germany; 
3 Institute for Infection Prevention and Hospital Epidemiology, Medical Center—University of Freiburg, Faculty of Medicine, University of Freiburg, 79110 Freiburg, Germany;
4 Fraunhofer Institute of Chemical Technology, 76327 Pfinztal, Germany; 
5 Vitrocell® Systems GmbH, 79183 Waldkirch, Germany; 
6 Palas GmbH, 76229 Karlsruhe, Germany; 

 

This study was the first to investigate the toxicological effects of well characterized aerosols released during combustion of thermoplastic nanocomposites using an air–liquid interface exposure system. Even though studies on the toxicological potential of combustion-generated particulate matter as well as VOCs have been published, none of them was designed to investigate the effect of the native aerosol using appropriate realistic lung cell culture models. In the current study we investigated the combustion behavior of PE-based nanocomposites on a lab-scale burner. As nanoscaled fillers TiO2 NP, CuO NP, as well as CNT were chosen for this study, with TiO2 NP representing a commonly used insoluble and inert nanomaterial, CuO NP as a known in vitro cyto- as well as genotoxic nanomaterial, and CNT as a fiber-shaped nanomaterial.

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