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An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
Published on: May 13, 2019
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Exploring Flow Cytometry-Based Micronucleus Scoring for Reliable Nanomaterial Genotoxicity Assessment.
Pauline Franz1, Alexander Bürkle2, Peter Wick1
1Laboratory for Particles-Biology Interactions, Empa - Swiss Federal Laboratories for Materials Science and Technology, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland.
Chemical Research in Toxicology
|September 18, 2020
Summary
Engineered nanomaterials like SiO2 and TiO2 showed no genotoxicity in human intestinal cells. Flow cytometry-based micronuclei scoring offers a faster, scalable, and reliable method for nanogenotoxicity testing.
Area of Science:
- Nanotechnology
- Toxicology
- Genetics
Background:
- Engineered nanomaterials (ENM) like silicon dioxide (SiO2) and titanium dioxide (TiO2) are increasingly used in food products, raising health concerns.
- Investigating ENM-induced genotoxicity is critical due to its link to carcinogenesis, but current data are contradictory.
- There is a need for robust, scalable in vitro methods for nanogenotoxicity assessment.
Purpose of the Study:
- To evaluate the suitability of flow cytometry-based micronuclei (MN) scoring for reliable nanogenotoxicity assessment in human intestinal cells.
- To assess the genotoxicity of various SiO2 and TiO2 nanomaterials using established assays.
- To identify and mitigate ENM-induced interferences in flow cytometric scoring.
Main Methods:
- Genotoxicity testing of different SiO2 and TiO2 nanomaterials using the alkaline single cell gel electrophoresis (Comet assay) and the micronucleus (MN) assay.
- Flow cytometric readout for the MN assay was employed for high-throughput analysis.
- Development and application of interference controls to ensure data reliability in flow cytometry.
Main Results:
- Physiologically relevant doses of tested SiO2 and TiO2 nanomaterials did not induce genotoxicity as assessed by both Comet and MN flow cytometry assays.
- Specific ENM-induced interferences with flow cytometric scoring were identified and addressed using control experiments.
- The study demonstrated the applicability of these controls for various nanomaterials and cell lines.
Conclusions:
- Flow cytometry-based MN scoring is a promising, rapid, scalable, and less operator-dependent method for nanogenotoxicity testing.
- This method enhances data reliability by effectively detecting and managing ENM-induced false-positive or false-negative results.
- The findings support the safety of tested ENM at relevant doses and highlight an improved methodology for future assessments.
