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An inter-laboratory effort to harmonize the cell-delivered in vitro dose of aerosolized materials

Anne Bannuscher, Otmar Schmid, Barbara Drasler, Alain Rohrbasser, Hedwig M. Braakhuis, Kirsty Meldrum, Edwin P. Zwart, Eric R. Gremmer, Barbara Birk, Manuel Rissel, Robert Landsiedel, Elisa Moschini, Stephen Evans Orcid Logo, Pramod Kumar, Sezer Orak, Ali Doryab, Johanna Samulin Erdem, Tommaso Serchi, Rob J. Vandebriel, Flemming R. Cassee, Shareen Doak Orcid Logo, Alke Petri-Fink, Shanbeh Zienolddiny, Martin Clift Orcid Logo, Barbara Rothen-Rutishauser

NanoImpact, Volume: 28, Start page: 100439

Swansea University Authors: Kirsty Meldrum, Stephen Evans Orcid Logo, Shareen Doak Orcid Logo, Martin Clift Orcid Logo

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DOI (Published version): 10.1016/j.impact.2022.100439

Abstract

Air-liquid interface (ALI) lung cell models cultured on permeable transwell inserts are increasingly used for respiratory hazard assessment requiring controlled aerosolization and deposition of any material on ALI cells. The approach presented herein aimed to assess the transwell insert-delivered do...

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Published in: NanoImpact
ISSN: 2452-0748
Published: Elsevier BV 2022
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URI: https://cronfa.swan.ac.uk/Record/cronfa62077
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Abstract: Air-liquid interface (ALI) lung cell models cultured on permeable transwell inserts are increasingly used for respiratory hazard assessment requiring controlled aerosolization and deposition of any material on ALI cells. The approach presented herein aimed to assess the transwell insert-delivered dose of aerosolized materials using the VITROCELL® Cloud12 system, a commercially available aerosol-cell exposure system. An inter-laboratory comparison study was conducted with seven European partners having different levels of experience with the VITROCELL® Cloud12. A standard operating procedure (SOP) was developed and applied by all partners for aerosolized delivery of materials, i.e., a water-soluble molecular substance (fluorescence-spiked salt) and two poorly soluble particles, crystalline silica quartz (DQ12) and titanium dioxide nanoparticles (TiO2 NM-105). The material dose delivered to transwell inserts was quantified with spectrofluorometry (fluorescein) and with the quartz crystal microbalance (QCM) integrated in the VITROCELL® Cloud12 system. The shape and agglomeration state of the deposited particles were confirmed with transmission electron microscopy (TEM). Inter-laboratory comparison of the device-specific performance was conducted in two steps, first for molecular substances (fluorescein-spiked salt), and then for particles. Device- and/or handling-specific differences in aerosol deposition of VITROCELL® Cloud12 systems were characterized in terms of the so-called deposition factor (DF), which allows for prediction of the transwell insert-deposited particle dose from the particle concentration in the aerosolized suspension. Albeit DF varied between the different labs from 0.39 to 0.87 (mean (coefficient of variation (CV)): 0.64 (28%)), the QCM of each VITROCELL® Cloud 12 system accurately measured the respective transwell insert-deposited dose. Aerosolized delivery of DQ12 and TiO2 NM-105 particles showed good linearity (R2 > 0.95) between particle concentration of the aerosolized suspension and QCM-determined insert-delivered particle dose. The VITROCELL® Cloud 12 performance for DQ12 particles was identical to that for fluorescein-spiked salt, i.e., the ratio of measured and salt-predicted dose was 1.0 (29%). On the other hand, a ca. 2-fold reduced dose was observed for TiO2 NM-105 (0.54 (41%)), which was likely due to partial retention of TiO2 NM-105 agglomerates in the vibrating mesh nebulizer of the VITROCELL® Cloud12. This inter-laboratory comparison demonstrates that the QCM integrated in the VITROCELL® Cloud 12 is a reliable tool for dosimetry, which accounts for potential variations of the transwell insert-delivered dose due to device-, handling- and/or material-specific effects. With the detailed protocol presented herein, all seven partner laboratories were able to demonstrate dose-controlled aerosolization of material suspensions using the VITROCELL® Cloud12 exposure system at dose levels relevant for observing in vitro hazard responses. This is an important step towards regulatory approved implementation of ALI lung cell cultures for in vitro hazard assessment of aerosolized materials.
Keywords: Aerosol-cell exposure; Nanoparticles; Nanomaterials; Inter-laboratory comparison; Standard operating procedure (SOP); VITROCELL® Cloud12 system; DQ12; TiO2 NM-105
College: Faculty of Medicine, Health and Life Sciences
Funders: Partners AMI, RIVM, SU, and BASF SE acknowledge the funding by the PATROLS project, European Union' Horizon 2020 Research and Innovation Programme under grant agreement No: 760813. HMGU has also received funding from the European Union Horizon 2020 research and innovation program under grant agreement No. 953183 (HARMLESS project). Anne Bannuscher, Barbara Drasler, Alain Rohrbasser, Alke Petri-Fink and Barbara Rothen-Rutishauser also acknowledge the support by the Adolphe Merkle Foundation. Johanna Samulin Erdem and Shanbeh Zienolddiny acknowledge the support by the National Institute of Occupational Health, Norway. STAMI would like to acknowledge the work of Oda Haarr Foss for excellent technical assistance.
Start Page: 100439

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