A Novel Exposure System Termed NAVETTA for In Vitro Laminar Flow Electrodeposition of Nanoaerosol and Evaluation of Immune Effects in Human Lung Reporter Cells

Abstract

A new prototype air-liquid interface (ALI) exposure system, a flatbed aerosol exposure chamber termed NAVETTA, was developed to investigate deposition of engineered nanoparticles (NPs) on cultured human lung A549 cells directly from the gas phase. This device mimics human lung cell exposure to NPs due to a low horizontal gas flow combined with cells exposed at the ALI. Electrostatic field assistance is applied to improve NP deposition efficiency. As proof-of-principle, cell viability and immune responses after short-term exposure to nanocopper oxide (CuO)-aerosol were determined. We found that, due to the laminar aerosol flow and a specific orientation of inverted transwells, much higher deposition rates were obtained compared to the normal ALI setup. Cellular responses were monitored with postexposure incubation in submerged conditions, revealing CuO dissolution in a concentration-dependent manner. Cytotoxicity was the result of ionic and nonionic Cu fractions. Using the optimized inverted ALI/postincubation procedure, pro-inflammatory immune responses, in terms of interleukin (IL)-8 promoter and nuclear factor kappa B (NFκB) activity, were observed within short time, i.e. One hour exposure to ALI-deposited CuO-NPs and 2.5 h postincubation. NAVETTA is a novel option for mimicking human lung cell exposure to NPs, complementing existing ALI systems.

Time-resolved characterization of the mechanisms of toxicity induced by silica and amino-modified polystyrene on alveolar-like macrophages

“…Time-resolved approaches such as we show here can be used to determine the series of events triggered in cells following exposure to nanoparticles, and in this way determine the mechanisms involved and differences in response to different materials. Additionally, the inclusion of more representative routes for delivering the nanoparticles to the model cells, e.g., by air-liquid interface exposure (Frijns et al 2017), could be beneficial to evaluate more realistically the interaction of nanoparticles and cells.…”

“…In the context of nanoparticle-induced toxicity, the lung and its resident phagocytes, the alveolar macrophages, are undoubtedly a relevant model, as the lung constitutes one of the major potential routes of exposure (Schlinkert et al 2015;Wohlleben et al 2016;Frijns et al 2017). Recently, murine Max Planck Institute (MPI) cells have been introduced as novel GM-CSF-dependent, continuously growing, non-transformed macrophages, derived from fetal liver.…”

When Would Immunologists Consider a Nanomaterial to be Safe? Recommendations for Planning Studies on Nanosafety

“…Complex models have been developed; for example several systems for alveolar exposure to NMs have become available during the past years. [69,[145][146][147] Other advanced cellular models include macrophages and dendritic cells to monitor responses in a realistic interplay of these cells. [148] Including polarized macrophages (M1 vs M2) in such models enables a deeper look into immune modulation.…”

Open-Source Digitally Replicable Lab-Grade Scales

“…Digital scales are used in a wide range of scientific applications including chemical research, genomics, drug discovery, and proteomics. Scales are classified by precision; where a precision balance has a measurement resolution of 0.001 g, an analytical balance has one of 0.0001 g and a micro balance has one of 0.0000001 g. On the high-end of scales, an open-source quartz crystal microbalance (OpenQCM) [55] uses an open-source Arduino Nano and is already well established in the scientific literature [56][57][58][59][60].…”