Studies designed to investigate the environmental or biological interactions of nanoscale materials frequently rely on the use of ultrasound (sonication) to prepare test suspensions. However, the inconsistent application of ultrasonic treatment across laboratories, and the lack of process standardization can lead to significant variability in suspension characteristics. At present, there is widespread recognition that sonication must be applied judiciously and reported in a consistent manner that is quantifiable and reproducible; current reporting practices generally lack these attributes. The objectives of the present work were to: (i) Survey potential sonication effects that can alter the physicochemical or biological properties of dispersed nanomaterials (within the context of toxicity testing) and discuss methods to mitigate these effects, (ii) propose a method for standardizing the measurement of sonication power, and (iii) offer a set of reporting guidelines to facilitate the reproducibility of studies involving engineered nanoparticle suspensions obtained via sonication.
Stable aqueous suspensions of colloidal C60 fullerenes free of toxic organic solvents were prepared by two methods: ethanol to water solvent exchange (EthOH/nC60 suspensions) and extended mixing in water (aqu/nC60 suspensions). The extended mixing method resulted in the formation of larger (dp approximately 178 nm) and less negatively charged (zeta approximately -13.5 mV) nC60 colloids than nC60 prepared by ethanol to water solvent exchange (dp approximately 122 nm, zeta approximately -31.6 mV). Genotoxicity of these suspensions was evaluated with respect to human lymphocytes using single-cell gel electrophoresis assay (Comet assay). The assay demonstrated genotoxicity for both types of suspensions with a strong correlation between the genotoxic response and nC60 concentration, and with genotoxicity observed at concentrations as low as 2.2 microg/L for aqu/nC60 and 4.2 microg/L for EtOH/nC60. The Olive tail moments (OTM) for these two concentrations were 1.54 +/- 0.24 and 1.34 +/- 0.07, respectively, which in comparison to the negative control OTM of 0.98 +/- 0.17 is statistically different with a p value of at least 0.05. Aqu/nC60 suspensions elicited higher genotoxic response than EthOH/nC60 for the same nC60 concentration. The results represent the first genotoxicity data for colloidal fullerenes produced by simple mixing in water.
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