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High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents HPHC
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Optimized High-Content Imaging Screening Quantifying Micronuclei Formation in Polymer-Treated HaCaT Keratinocytes
Fariba Saadati1,2, Walison Augusto da Silva Brito1,3, Steffen Emmert2
1ZIK plasmatis, Leibniz Institute for Plasma Science and Technology (INP), Felix-Hausdorff-Str. 2, 17489 Greifswald, Germany.
Nanomaterials (Basel, Switzerland)
|December 23, 2022
Summary
Most nano- and micro-plastic particles (NMPPs) are neither toxic nor genotoxic, failing to induce micronuclei (MN) formation. Only high concentrations of specific polystyrene particles showed effects, highlighting the need for careful risk assessment in environmental toxicity studies.
Area of Science:
- Environmental Toxicology
- Nanotechnology
- Cell Biology
Background:
- Nano- and micro-plastic particles (NMPPs) pose potential human health risks.
- Genotoxic effects, like micronuclei (MN) formation, have been inconsistently reported for NMPPs.
- Clarifying NMPP genotoxicity is crucial for accurate risk assessment.
Purpose of the Study:
- To investigate the genotoxic potential of various polystyrene (PS) and poly methyl methacrylate (PMMA) particles.
- To evaluate NMPP effects on micronuclei (MN) formation in HaCaT keratinocytes under acute and chronic exposure.
- To establish and apply an optimized high-content imaging workflow for quantitative NMPP toxicity analysis.
Main Methods:
- Exposure of HaCaT cells to PS and PMMA particles of different sizes and concentrations.
- Acute (1-day) and chronic (5-week) exposure protocols.
- OECD-compliant high-content imaging with algorithm-based MN analysis, using accutase for cell preparation.
- Positive controls included cytotoxic amino-modified NMPPs and genotoxic methyl methanesulfonate (MMS).
Main Results:
- Cytotoxic amino-modified particles were not genotoxic; MMS demonstrated both toxicity and genotoxicity.
- PS and PMMA particles generally showed no toxicity or increased MN formation.
- An exception was 1000 nm PS particles at 100 µg/mL, which did not increase MN but decreased ROS formation.
- Non-charged polymer particles were neither toxic nor genotoxic; aminated particles were toxic but not genotoxic.
Conclusions:
- Most non-charged NMPPs (PS, PMMA) are neither toxic nor genotoxic to HaCaT cells.
- Aminated NMPPs exhibit toxicity but not genotoxicity.
- The study presents a robust imaging workflow for NMPP toxicity assessment, indicating limited genotoxic risk from common plastic particles at tested conditions.
