title: Development of dual-fluorescence cell-based biosensors for detecting the influence of environmental factors on nanoparticle toxicity.
abstract: 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.
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, China Academy of Science, Hefei, Anhui, PR China; School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, PR China; Key Laboratory of Environmental Toxicology and Pollution Control Technology of Anhui Province, Hefei, Anhui, PR China.
authors.forename: Junting
authors.initials: J
authors.lastname: Liang
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, China Academy of Science, Hefei, Anhui, PR China; Key Laboratory of Environmental Toxicology and Pollution Control Technology of Anhui Province, Hefei, Anhui, PR China.
authors.forename: Xuanyu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, China Academy of Science, Hefei, Anhui, PR China; Key Laboratory of Environmental Toxicology and Pollution Control Technology of Anhui Province, Hefei, Anhui, PR China.
authors.forename: Luzhi
authors.initials: L
authors.lastname: Li
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, China Academy of Science, Hefei, Anhui, PR China; Key Laboratory of Environmental Toxicology and Pollution Control Technology of Anhui Province, Hefei, Anhui, PR China.
authors.forename: Shengmin
authors.initials: S
authors.lastname: Xu
authors.affiliation: Department of Surgery-Transplant and Mary & Dick Holland Regenerative Medicine Program, University of Nebraska Medical Center, Omaha, NE 68198, United States.
authors.forename: Jiang
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, China Academy of Science, Hefei, Anhui, PR China; Key Laboratory of Environmental Toxicology and Pollution Control Technology of Anhui Province, Hefei, Anhui, PR China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Wu
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, China Academy of Science, Hefei, Anhui, PR China; Key Laboratory of Environmental Toxicology and Pollution Control Technology of Anhui Province, Hefei, Anhui, PR China. Electronic address: gpz@ipp.ac.cn.
authors.forename: Guoping
authors.initials: G
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Hefei Institutes of Physical Science, China Academy of Science, Hefei, Anhui, PR China; Key Laboratory of Environmental Toxicology and Pollution Control Technology of Anhui Province, Hefei, Anhui, PR China. Electronic address: spchen035@ipp.ac.cn.
authors.forename: Shaopeng
authors.initials: S
authors.lastname: Chen
mesh_terms: D015374:Biosensing Techniques; D004777:Environment; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006801:Humans; D053768:Metal Nanoparticles / Q000633:toxicity*; D012834:Silver / Q000737:chemistry / Q000633:toxicity*
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D012834:Silver
keywords: Biosensor; Environmental factor; Flow cytometry; Fluorescent protein; Nanoparticles