Measurement of reactive species production by nanoparticles prepared in biologically relevant media

Exposure to nanoparticles may pose a risk to health and this hypothesis is currently being investigated by toxicologists. Although the mechanism of nanoparticle toxicity has been shown to be mediated, in part, by oxidative stress, the precise mechanism and molecules involved are still unknown. In light of this, the evaluation of the oxidative potential of nanoparticles is an important consideration in measuring their toxicity. The aim of this study was to examine the use of a fluorogenic probe, 2′,7′-dichlorofluorescin (DCFH), in a cell-free assay system and to assess the relationship between the results obtained with this method and with the reactive species formation observed in cells. In order to obtain a well-dispersed nanoparticle suspension, bovine serum albumin (BSA) and dipalmitoyl phosphatidyl choline (DPPC) addition in suspension medium was investigated. Both 1% BSA and 0.025% DPPC added to the medium significantly improved the stability of the nanoparticle suspension, decreasing the extent of particle agglomeration and settling over time. In a cell-free system, reactive oxygen species (ROS) production by 14 nm carbon black particles (CB) suspended in DPPC was higher than that measured with the other suspensions (saline or 1% BSA). A greater ROS production was observed in MonoMac 6 cells (MM6) following treatment with 14 nm CB suspended in medium containing BSA and/or DPPC compared to medium alone. In conclusion, 1% BSA and 0.025% DPPC solution was the most efficient for the preparation of a nanoparticle suspension and to measure their oxidative potential.