Microscopy-based high-throughput assays enable multi-parametric analysis to assess adverse effects of nanomaterials in various cell lines

Abstract

Manufactured nanomaterials (MNMs) selected from a library of over 120 different MNMs with varied compositions, sizes, and surface coatings were tested by four different laboratories for toxicity by high-throughput/-content (HT/C) techniques. The selected particles comprise 14 MNMs composed of CeO, Ag, TiO, ZnO and SiO with different coatings and surface characteristics at varying concentrations. The MNMs were tested in different mammalian cell lines at concentrations between 0.5 and 250 µg/mL to link physical-chemical properties to multiple adverse effects. The cell lines are derived from relevant organs such as liver, lung, colon and the immune system. Endpoints such as viable cell count, cell membrane permeability, apoptotic cell death, mitochondrial membrane potential, lysosomal acidification and steatosis have been studied. Soluble MNMs, Ag and ZnO, were toxic in all cell types. TiO and SiO MNMs also triggered toxicity in some, but not all, cell types and the cell type-specific effects were influenced by the specific coating and surface modification. CeO MNMs were nearly ineffective in our test systems. Differentiated liver cells appear to be most sensitive to MNMs, Whereas most of the investigated MNMs showed no acute toxicity, it became clear that some show adverse effects dependent on the assay and cell line. Hence, it is advised that future nanosafety studies utilise a multi-parametric approach such as HT/C screening to avoid missing signs of toxicity. Furthermore, some of the cell type-specific effects should be followed up in more detail and might also provide an incentive to address potential adverse effects in vivo in the relevant organ.

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

“…Though SYTOX staining does not allow to discriminate primary and secondary necrosis, the absence of clear Annexin V staining at 24 h (also visible in the double scatter plots shown in Supplementary Figure S1) suggested that, as opposed to what is usually observed with these nanoparticles in other cell models (Bexiga et al 2011;Wang et al 2013a, b), in MPI macrophages exposure to NH 2 -PS nanoparticles led to cell death, but not via apoptosis. This is in agreement with previous observations of necrotic cell death induced by the same nanoparticles in RAW 364.7 macrophages (Hansjosten et al 2018). On the contrary, SiO 2 nanoparticles displayed a clear apoptotic signature, with both an elevation of the percentage of apoptotic cells and increases in caspase 3/7 activity.…”

Surface Chemistry-Dependent Evolution of the Nanomaterial Corona on TiO2 Nanomaterials Following Uptake and Sub-Cellular Localization

“…Characterization of NM dispersions: Dynamic Light Scattering (DLS), zeta potential, and Inductively Coupled Plasma Mass Spectrometry (ICP-MS): NM stock suspensions were diluted as per the NanoMILE protocol for the dilution and dispersion of liquid suspension NMs in the relevant medium (ultrapure H 2 O, complete-serum containing medium (SCM), and artificial lysosomal fluid) to a concentration of 125 µg/mL [46]. Each NM suspension was then incubated at 37 • C in 5% CO 2 .…”

Ultraschnelle Synthese multivalenter radikalischer Nanopartikel durch ringöffnende Metathesepolymerisations‐induzierte Selbstorganisation

“…Eine erhöhte Freisetzung von LDH, einem im Zytoplasma vorhandenen Enzym, würde auf eine Schädigung der Plasmamembran der Zellen hinweisen. Sogar bis zu einer Konzentration von 100 μg mL −1 , was gut oberhalb klinisch relevanter Dosen liegt, verstärken ROMPISA‐NPs nicht die Freisetzung von LDH (Abbildung A, rote Balken), insbesondere im Unterschied zu positiven Kontrollproben wie aminofunktionalisierten Polystyrol‐NPs . In ähnlicher Weise zeigt der WST‐1‐Test eine hohe Biokompatibilität ohne Abnahme des zellulären Metabolismus an, wodurch die extrem niedrige Toxizität unserer ROMPISA‐Nanopartikel bestätigt wird (Abbildung A, grüne Balken).…”