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

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

doi:10.1016/j.impact.2022.100439

Cited by 18 publications

(7 citation statements)

References 37 publications

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“…This suggests an effective aerosol to cell delivery of the Cloud α AX12 and the impact of physiological CS on the uptake of NPs. Moreover, an inter-laboratory study conducted in collaboration with 7 different laboratory groups recently confirmed the homogenous dispersion of nanoparticles and consistent dose deposition profile using the VITROCELL ® Cloud 12 system ( Bannuscher et al, 2022 ). This study is a significant advancement in the standardization of in vitro toxicity studies using ALI culture conditions and advanced cloud exposure devices for regulatory purposes.…”

Section: Discussionmentioning

confidence: 80%

A multiplex inhalation platform to model in situ like aerosol delivery in a breathing lung-on-chip

et al. 2023

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Prolonged exposure to environmental respirable toxicants can lead to the development and worsening of severe respiratory diseases such as asthma, chronic obstructive pulmonary disease (COPD) and fibrosis. The limited number of FDA-approved inhaled drugs for these serious lung conditions has led to a shift from in vivo towards the use of alternative in vitro human-relevant models to better predict the toxicity of inhaled particles in preclinical research. While there are several inhalation exposure models for the upper airways, the fragile and dynamic nature of the alveolar microenvironment has limited the development of reproducible exposure models for the distal lung. Here, we present a mechanistic approach using a new generation of exposure systems, the Cloud α AX12. This novel in vitro inhalation tool consists of a cloud-based exposure chamber (VITROCELL) that integrates the breathing AXLung-on-chip system (AlveoliX). The ultrathin and porous membrane of the AX12 plate was used to create a complex multicellular model that enables key physiological culture conditions: the air-liquid interface (ALI) and the three-dimensional cyclic stretch (CS). Human-relevant cellular models were established for a) the distal alveolar-capillary interface using primary cell-derived immortalized alveolar epithelial cells (AXiAECs), macrophages (THP-1) and endothelial (HLMVEC) cells, and b) the upper-airways using Calu3 cells. Primary human alveolar epithelial cells (AXhAEpCs) were used to validate the toxicity results obtained from the immortalized cell lines. To mimic in vivo relevant aerosol exposures with the Cloud α AX12, three different models were established using: a) titanium dioxide (TiO2) and zinc oxide nanoparticles b) polyhexamethylene guanidine a toxic chemical and c) an anti-inflammatory inhaled corticosteroid, fluticasone propionate (FL). Our results suggest an important synergistic effect on the air-blood barrier sensitivity, cytotoxicity and inflammation, when air-liquid interface and cyclic stretch culture conditions are combined. To the best of our knowledge, this is the first time that an in vitro inhalation exposure system for the distal lung has been described with a breathing lung-on-chip technology. The Cloud α AX12 model thus represents a state-of-the-art pre-clinical tool to study inhalation toxicity risks, drug safety and efficacy.

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Section: Discussionmentioning

confidence: 80%

A multiplex inhalation platform to model in situ like aerosol delivery in a breathing lung-on-chip

et al. 2023

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“…Using ALI exposures is however more time-consuming and less high-throughput. Additionally, robustness of deposited dose after nebulization in an ALI setup may be low, depending on the NM used [ 219 ].…”

Section: Evaluation Of In Vitro Methods For Sbd Hazard Testingmentioning

confidence: 99%

The State of the Art and Challenges of In Vitro Methods for Human Hazard Assessment of Nanomaterials in the Context of Safe-by-Design

et al. 2023

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The Safe-by-Design (SbD) concept aims to facilitate the development of safer materials/products, safer production, and safer use and end-of-life by performing timely SbD interventions to reduce hazard, exposure, or both. Early hazard screening is a crucial first step in this process. In this review, for the first time, commonly used in vitro assays are evaluated for their suitability for SbD hazard testing of nanomaterials (NMs). The goal of SbD hazard testing is identifying hazard warnings in the early stages of innovation. For this purpose, assays should be simple, cost-effective, predictive, robust, and compatible. For several toxicological endpoints, there are indications that commonly used in vitro assays are able to predict hazard warnings. In addition to the evaluation of assays, this review provides insights into the effects of the choice of cell type, exposure and dispersion protocol, and the (in)accurate determination of dose delivered to cells on predictivity. Furthermore, compatibility of assays with challenging advanced materials and NMs released from nano-enabled products (NEPs) during the lifecycle is assessed, as these aspects are crucial for SbD hazard testing. To conclude, hazard screening of NMs is complex and joint efforts between innovators, scientists, and regulators are needed to further improve SbD hazard testing.

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“…All SOPs can be found in the PATROLS SOP Handbook (PATROLS, 2022). Details of the aerosolisation using the VITROCELL® Cloud12 system are specified in the PATROLS SOPs "Guidance Document for ENMs Aerosolization using VITROCELL® Cloud System" (#3601); and "Vitrocell dry powder system" (#3206; see also (Bannuscher et al, 2022)). Details of the materials and methods for testing transferability and reproducibility of the Calu-3 model with and without macrophages are given in the PATROLS SOPs "Guidance Document for co-culture of a lung epithelial cell-line (Calu-3), and macrophages derived from peripheral blood monocytes" (#3603) and "Guidance Document for coculture of an lung epithelial cell-line (Calu-3), and macrophages derived from a monocytic cell line (dTHP-1)" (#3604).…”

Section: Methodsmentioning

confidence: 99%

“…For aerosolized NM exposure at the ALI, the VITROCELL® Cloud12 system was used. The dosimetrically accurate NM deposition using the Vitrocell® Cloud12 system has been established among the different partners in a first phase of this interlaboratory study (Bannuscher et al, 2022). This first phase comprised the same laboratories as the ones performing the interlaboratory study described in the present manuscript (second phase).…”

Section: Nm Exposurementioning

confidence: 99%

97 Transferability and Reproducibility of Exposed Air-Liquid Interface co-Culture Lung Models

Braakhuis

Gremmer

Bannuscher

et al. 2023

Annals of Work Exposures and Health

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The establishment of reliable and robust in vitro models for hazard assessment, moving away from animal testing, requires evaluation of model transferability and reproducibility. Lung models that can be exposed via the air, by means of an air-liquid interface are promising in vitro models for evaluating the safety of nanomaterials (NMs) after inhalation exposure. We performed an inter-laboratory comparison study to evaluate the transferability and reproducibility of a lung model consisting of the human bronchial cell line Calu-3 as monoculture and, to increase the relevance of the model, also as co-culture with macrophages (derived from the THP-1 monocyte cell line or from human blood monocytes). Exposure employed the VITROCELL® Cloud12 system at physiologically relevant doses. Overall, the results of the seven participating laboratories are quite similar. No effects of lipopolysaccharide (LPS), quartz (DQ12) or titanium dioxide (TiO2) NM-105 exposure on cell viability and barrier integrity were seen in the monoculture and co-culture models. Most laboratories showed that LPS induced cytokine release in the co-culture models, but not the monoculture model. Exposure to DQ12 or TiO2 did not increase cytokine release, probably due to too low deposited doses. Interlaboratory variability indicated acceptable variation for cell viability measurements and relatively high variation for cytokine measurements. Based on the transferability and reproducibility of the model and its air exposure we are confident that, based on detailed SOPs, performing a new round of interlaboratory comparison, and increasing the deposited doses of NMs, these models may be taken further towards a possible OECD guideline.

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

Cited by 18 publications

References 37 publications

A multiplex inhalation platform to model in situ like aerosol delivery in a breathing lung-on-chip

A multiplex inhalation platform to model in situ like aerosol delivery in a breathing lung-on-chip

The State of the Art and Challenges of In Vitro Methods for Human Hazard Assessment of Nanomaterials in the Context of Safe-by-Design

97 Transferability and Reproducibility of Exposed Air-Liquid Interface co-Culture Lung Models

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