Direct Combination of Nanoparticle Fabrication and Exposure to Lung Cell Cultures in a Closed Setup as a Method To Simulate Accidental Nanoparticle Exposure of Humans

Abstract

The tremendous application potential of nanosized materials stays in sharp contrast to a growing number of critical reports of their potential toxicity. Applications of in vitro methods to assess nanoparticles are severely limited through difficulties in exposing cells of the respiratory tract directly to airborne engineered nanoparticles. We present a completely new approach to expose lung cells to particles generated in situ by flame spray synthesis. Cerium oxide nanoparticles from a single run were produced and simultaneously exposed to the surface of cultured lung cells inside a glovebox. Separately collected samples were used to measure hydrodynamic particle size distribution, shape, and agglomerate morphology. Cell viability was not impaired by the conditions of the glovebox exposure. The tightness of the lung cell monolayer, the mean total lamellar body volume, and the generation of oxidative DNA damage revealed a dose-dependent cellular response to the airborne engineered nanoparticles. The direct combination of production and exposure allows studying particle toxicity in a simple and reproducible way under environmental conditions.

No Evidence for Cerium Dioxide Nanoparticle Translocation in Maize Plants

“…The aerosol deposition and characterization was performed at identical conditions as reported in earlier work where ceria nanoparticles have been exposed to human cells (22). The nanoparticles and agglomerates were deposited homogeneously within the glove box as characterized previously by Rothen-Rutishauser et al (22).…”

Quantification of Gold Nanoparticle Cell Uptake Under Controlled Biological Conditions and Adequate Resolution

“…In each experiment, 1 ml of the supernatant was sampled for the lactate dehydrogenase (LDH) assay to determine cytotoxicity [43]. Briefly, Triton X (2% in RPMI, 30 min preincubation) was used for cell lysis and the positive control.…”

Investigations of the Biological Effects of Airborne and Inhalable Substances by Cell-Based In Vitro Methods: Fundamental Improvements to the ALI Concept

“…When setups in the "incubator-type" alignment [26,27] are used, but also in stagnation flow setups [25,31], the test atmosphere can readily diffuse into the culture medium and will therefore readily react with culture medium components, which include many reactive substances such as glutathione and others. Hence, the cell exposure is not only by way of air-liquid cell exposure but also by a route via the culture medium, which is normally not intended in these studies.…”

“…On the other hand, these setups always include noncommercial housings for the cultures in order to realise the stagnation flow arrangement. The other type of exposure setup is constructed as a small "incubator-type" arrangement [11,26,27] where cells in commercial multiwell plates are positioned in larger housings such as cubic exposure chambers or cell culture incubators. This is a very convenient and reasonable strategy from the point of view of applicability and use as well as mild cellular conditions.…”