Cited by 152Nanoparticles: synthesis and applicationsOccupational and environmental lung diseasesImmunotoxicology and immune responses

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An in vitro alveolar macrophage assay for predicting the short-term inhalation toxicity of nanomaterials

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Wiemann, Martin, Vennemann, Antje, Sauer, Ursula G., Wiench, Karin, Ma-Hock, Lan, Landsiedel, Robert·2016·Journal of Nanobiotechnology

DOI 10.1186/s12951-016-0164-2PMC PMC4779246

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Abstract

BACKGROUND: Most in vitro studies investigating nanomaterial pulmonary toxicity poorly correlate to in vivo inhalation studies. Alveolar macrophages (AMs) play an outstanding role during inhalation exposure since they effectively clear the alveoli from particles. This study addresses the applicability of an in vitro alveolar macrophage assay to distinguish biologically active from passive nanomaterials. METHODS: Rat NR8383 alveolar macrophages were exposed to 18 inorganic nanomaterials, covering AlOOH, BaSO4, CeO2, Fe2O3, TiO2, ZrO2, and ZnO NMs, amorphous SiO2 and graphite nanoplatelets, and two nanosized organic pigments. ZrO2 and amorphous SiO2 were tested without and with surface functionalization. Non-nanosized quartz DQ12 and corundum were used as positive and negative controls, respectively. The test materials were incubated with the cells in protein-free culture medium. Lactate dehydrogenase, glucuronidase, and tumour necrosis factor alpha were assessed after 16 h. In parallel, H2O2 was assessed after 1.5 h. Using the no-observed-adverse-effect concentrations (NOAECs) from available rat short-term inhalation studies (STIS), the test materials were categorized as active (NOAEC < 10 mg/m(3)) or passive. RESULTS: In vitro data reflected the STIS categorization if a particle surface area-based threshold of <6000 mm(2)/mL was used to determine the biological relevance of the lowest observed significant in vitro effects. Significant effects that were recorded above this threshold were assessed as resulting from test material-unspecific cellular 'overload'. Test materials were assessed as active if ≥2 of the 4 in vitro parameters undercut this threshold. They were assessed as passive if 0 or 1 parameter was altered. An overall assay accuracy of 95 % was achieved. CONCLUSIONS: The in vitro NR8383 alveolar macrophage assay allows distinguishing active from passive nanomaterials. Thereby, it allows determining whether in vivo short-term inhalation testing is necessary for hazard assessment. Results may also be used to group nanomaterials by biological activity. Further work should aim at validating the assay.

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Fig. 1 Test material sedimentation and uptake by NR8383 rat alveolar macrophages. A Corundum; B Quartz DQ12; C TiO2 NM-105; D CeO2 NM-212; E SiO2.naked; F BaSO4 NM-220; G DPP Orange N; H Graphite nanoplatelets. Phase contrast micrographs show settled particles at the bottom of cell-free wells in 96-well plates (a1–h1 on the left side of the individual images) and corresponding particle-laden NR8383 cells at the end of the 16-h incubation period with the same particle concentration a2–h2 on the right side of the individual images). Note that the uptake of particles (loaded with 90 μg/mL) appears complete as viewed by light microscopy, except for SiO2.naked (loaded with 22.5 μg/mL), where settled particles are hardly visibleCC BY · PubMed Central

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Authors: Wiemann, M., Vennemann, A., Sauer, U. G., Wiench, K., Ma-Hock, L., & Landsiedel, R.
Affiliations: BASF (Germany) (DE)
Corresponding author: Wiemann, Martin
ORCID: Wiemann, Martin
Vennemann, Antje
Sauer, Ursula G.
Wiench, Karin
Landsiedel, Robert
Venue: Journal of Nanobiotechnology
Publisher: Springer Science and Business Media LLC
Published: 2016-03-05
Volume: 14
Issue: 1
Pages: 16-16
ISSN: 1477-3155 (ISSN-L)
DOI: 10.1186/s12951-016-0164-2
PMC: PMC4779246
OpenAlex: W2290248210
Cited by: 152
References: 120
Citations by year: 2026: 6 · 2025: 13 · 2024: 12 · 2023: 12 · 2022: 18 · 2021: 14 · 2020: 18 · 2019: 18 · 2018: 17 · 2017: 18 · 2016: 6
FWCI: 8.76
Type: journal article
Language: en
Open access: gold
doi.org
Licence: cc-by
Locations: Journal of Nanobiotechnology (publishedVersion, cc-by)
PubMed Central (publishedVersion, cc-by)
PubMed (publishedVersion)
Funders: Bundesministerium für Bildung und Forschung (03X0105, 0315483A)
Topics: Physical Sciences › Materials Science › Materials Chemistry › Nanoparticles: synthesis and applications
Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Occupational and environmental lung diseases
Life Sciences › Immunology and Microbiology › Immunology › Immunotoxicology and immune responses
Keywords: alveolar macrophages, NR8383 cells, in vitro-in vivo correlation, zirconia nanoparticles, amorphous silica nanoparticles, surface functionalization, hazard assessment, cytotoxicity, oxidative stress
MeSH terms: Animals, Cell Line, Ferric Compounds, Graphite, Hydrogen Peroxide, Lung, Particle Size, Silicon Dioxide, Titanium, Macrophages, Alveolar, Inhalation Exposure, Nanostructures, Rats

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An in vitro alveolar macrophage assay for predicting the short-term inhalation toxicity of nanomaterials

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