Development of dual-fluorescence cell-based biosensors for detecting the influence of environmental factors on...

With the expanding use of engineered nanoparticles (NPs), development of a high-throughput, sensitive method for evaluating NP safety is important. In this study, we developed cell-based biosensors to efficiently and conveniently monitor NP toxicity. The biosensor cells were obtained by transiently transfecting human cells with biosensor plasmids containing a mCherry gene regulated by an inducible promoter [an activator protein 1 (AP-1) promoter, an interleukin 8 (IL8) promoter, or a B cell translocation gene 2 (BTG2) promoter], with an enhanced green-fluorescent protein gene driven by the cytomegalovirus promoter as the internal control. After optimizing flow cytometric analysis, these dual-fluorescence cell-based biosensors were capable of accurately and rapidly detecting NP toxicity. We found that the responses of AP-1, BTG2, and IL8 biosensors in assessing the toxicity of silver nanoparticles (Ag NPs) showed good dose-related increases after exposure to Ag NPs and were consistent with data acquired by conventional assays, such as western blot, real-time polymerase chain reaction, and immunofluorescence. Further investigation of the effects of environmental factors on Ag NP toxicity revealed that aging in water, co-exposure with fulvic acid, and irradiation with ultraviolet A light could affect Ag NP-induced biosensor responses. These results indicated that these novel dual-fluorescence biosensors can be applied to accurately and sensitively monitor NP toxicity.