Direct nanomaterial-DNA contact effects on DNA and mutation induction.

Toxicology letters (2014-02-08)
P Thongkumkoon, K Sangwijit, C Chaiwong, S Thongtem, P Singjai, L D Yu
ABSTRACT

The toxicity of nanomaterials has been well known, but mechanisms involved have been little known. This study was aimed at looking at direct interaction between nanomaterials and naked DNA for some fundamental understanding. Two different types of nanomaterials, carbon nanotubes (CNTs) and tungsten trioxide (WO₃) nanoplates, were simply mixed with naked DNA plasmid, respectively, in two different contact modes, dry or wet (in solution), for varied time periods. DNA topological forms were analyzed for changes using gel electrophoresis and fluoro-spectrometry. The nanomaterial-contacted DNA was transferred into bacteria Escherichia coli (E. coli) cells for mutation observation. Certain types and degrees of DNA damage were observed, such as single strand break and double strand break, and bacterial mutation was confirmed. The DNA damage increased with the contacting time in an exponential manner and increased more rapidly in the initial stage for the wet contact. The nanomaterials-contacted DNA transferred bacteria had about less than 10% survival but almost 100% mutation for the surviving cells. The CNTs were more offensive than the metal oxide nanomaterials. The mutation spectrum from the DNA sequencing analysis showed that DNA point mutation was dominated by transversion, which was dominated by guanine changes in the wet contact condition while by cytosine changes in the dry contact condition. The point mutation occurrence in the wet contact was more than in the dry contact, confirming the wet contact more active and thus dangerous than dry contact. This experiment, although as a model study, revealed that direct simple contacts between nanomaterials and DNA could cause DNA changes and thus induce mutations which might potentially lead to cancers, diseases and genetic changes. This could be a mechanism for nanomaterial genotoxicity to the cells and also provided a caution to applications in using nanomaterials for DNA delivery.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Tungsten, powder, 12 μm, 99.9% trace metals basis
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Tungsten, wire reel, 5m, diameter 0.005mm, gold plated, 99.95%
Tungsten, wire reel, 25m, diameter 0.025mm, clean, 99.95%
Tungsten, wire reel, 10m, diameter 0.3mm, clean, 99.95%
Tungsten, wire, straight, 300mm, diameter 0.10mm, 99.9+%
Tungsten, wire, straight, 300mm, diameter 0.075mm, 99.9+%
Tungsten, wire reel, 100m, diameter 0.025mm, clean, 99.95%
Tungsten, insulated wire, 2m, conductor diameter 0.025mm, insulation thickness 0.005mm, polyimide insulation, 99.9%
Tungsten, wire reel, 1m, diameter 0.125mm, gold plated, 99.95%
Tungsten, wire reel, 10m, diameter 0.035mm, clean, 99.95%
Tungsten, wire reel, 10m, diameter 0.025mm, clean, 99.95%
Tungsten, foil, 100x100mm, thickness 0.125mm, as rolled, 99.95%
Tungsten, foil, 100x100mm, thickness 0.15mm, as rolled, 99.95%
Tungsten, foil, 100x100mm, thickness 0.1mm, as rolled, 99.95%
Tungsten, foil, 100x100mm, thickness 0.2mm, as rolled, 99.95%
Tungsten, foil, 100x100mm, thickness 0.3mm, as rolled, 99.95%
Tungsten, foil, 100x100mm, thickness 0.5mm, as rolled, 99.95%
Tungsten, foil, 100x100mm, thickness 1.0mm, as rolled, 99.95%
Tungsten, foil, 100x100mm, thickness 2.0mm, as rolled, 99.95%
Tungsten, foil, 150x150mm, thickness 0.125mm, as rolled, 99.95%

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