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Figure 3 | Particle and Fibre Toxicology

Figure 3

From: Amorphous nanosilica induce endocytosis-dependent ROS generation and DNA damage in human keratinocytes

Figure 3

Detection of oxidative stress induced by silica particle treatment in HaCaT cells. Detection of total ROS and hydroxyl radical induced by silica particle treatment in HaCaT cells. HaCaT cells were incubated with various concentrations of nSP70 (circles), nSP300 (squares), and mSP1000 (diamonds) for 3 h. (A) Total ROS induced by treatment with silica particles were expressed as relative fluorescence units in the DCFH assay.*P < 0.01 vs same dose of nSP300 and mSP1000. (B) Hydroxyl radical was measured by hydroxyphenyl fluorescein (HPF) assay. Data shown are means ± SD (n = 3).*P < 0.01 vs same dose of nSP300 and mSP1000. (C) Detection of 8-OH-dG induced by silica particle treatment in HaCaT cells. HaCaT cells were incubated with 10, 30 or 90 mg/ml nSP70, nSP300, or mSP1000, and As2O3 (positive control) for 3 h. Data shown are means ± SD (n = 3). *P < 0.01, **P < 0.05. (D and E) Effects of ROS inhibitor on DNA strand breaks induced by silica particle treatment in HaCaT cells. HaCaT cells were pretreated with 2 mM N-acetylcystein (NAC) for 30 min (NAC + nSP70) or nSP70 alone, prior to incubation with 90 mg/ml nSP70 for 3 h. As a positive control, HaCaT cells were treated with 0.2 mM H2O2 for 3 h. (D) Column height shows the tail length. (E) Column height shows the tail moment. Data shown are means ± SD of at least 16 cells per sample. Results shown are representative of more than three independent experiments. *P < 0.01.

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