First High Throughput Genotox Assay

Protocol for the high throughput Comet Assay. (A) Assembly of macrowell comet array. Agarose gel with microwells is sandwiched between a glass substrate and a bottomless 96-well plate and sealed with mechanical force. Approximately 300 arrayed microwells comprise the bottom of each macrowell. (B) Preparation of the nanoparticle suspension according to the protocol by Cohen at al. (C) Protocol for exposing the cells to the nanoparticles. (D) Loading of the exposed cell samples in the macrowells and running the microwell assay.

Protocol for the high throughput Comet Assay. (A) Assembly of macrowell comet array. Agarose gel with microwells is sandwiched between a glass substrate and a bottomless 96-well plate and sealed with mechanical force. Approximately 300 arrayed microwells comprise the bottom of each macrowell. (B) Preparation of the nanoparticle suspension according to the protocol by Cohen at al. (C) Protocol for exposing the cells to the nanoparticles. (D) Loading of the exposed cell samples in the macrowells and running the microwell assay.

Nanomaterials are part of daily life. Although there is a wide range of methods to evaluate their potential toxic effects, there is no way to evaluate gene damage.

Scientists at Harvard University in the School of Public Health in collaboration with the research Group of Bevin P. Engelward at MIT, have developed a screening assay to detect the genotoxic potential of nanomaterials. Metal oxide nanoparticles in biological systems can generate reactive oxygen species, which can overwhelm innate antioxidant defenses and cause oxidative stress. Oxidative stress, among other factors, has been associated with DNA damage and mutations, precursors to cancer. As more and more commercial products contain nanomaterials consisting of metal oxides such as titanium dioxide and zinc oxide, screening assays such as these are crucial to reducing potential health hazards. Christa Watson, postdoctoral research fellow at HSPH, suggests that accurate toxicity assessments of nanomaterials before they are incorporated into consumer products can help us prevent similar consequences that we are currently facing from asbestos exposures such as mesothelioma. Current efforts are ongoing to understand the novel toxicities nanomaterials may pose on public safety. This research was recently published in ACS Nano February 14, 2014 DOI: 10.1021/nn404871p