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Fig. 2 | Particle and Fibre Toxicology

Fig. 2

From: SiO2 and TiO2 nanoparticles synergistically trigger macrophage inflammatory responses

Fig. 2

SiO2 and TiO2 NPs induce different cellular stress pathways in BMDMs. a LPS-primed BMDMs were stimulated with rhodamine-labeled SiO2 NPs and/or FITC-labeled TiO2 NPs (10 μg/cm3 each) for 30 min at 37 °C. Cells were analyzed by flow cytometry. b LPS-primed BMDMs were untreated or treated rhodamine-labeled SiO2 NPs and FITC-labeled TiO2 NPs (10 μg/cm3 each) for 30 min at 37 °C, and were stained with Hoechst 33482. After fixation with 4% paraformaldehyde, cells were analyzed by confocal microscopy. White thick bars indicate 10 μm. A higher magnification of the white square in the middle image is shown in the rightmost image. c, d LPS-primed BMDMs were stained with LysoTracker-Red and were untreated or stimulated with FITC-labeled SiO2 NPs or FITC-labeled TiO2 NPs (10 μg/cm3 each) for 30 min at 37 °C. After fixation with 4% paraformaldehyde, cells were analyzed by confocal microscopy. White thick bars indicate 10 μm in (c). FITC and LysoTracker signal intensities in area indicated by white thin arrows were measured and shown in (d). e LPS-primed BMDMs were untreated (white bars) or pretreated with YVAD-CHO (20 μM: black bars), Bafilomycin A1 (250 nM: gray bars), or BHA (100 μM: red bars) for 1 h at 37 °C, and then BMDMs were stimulated with SiO2 NPs and TiO2 NPs (10 μg/cm3 each) or ATP (1 mM) for 4 h at 37 °C. The amount of IL-1β in culture supernatants was measured by ELISA. Data are indicated as the mean + S.D. f LPS-primed BMDMs were untreated (white bars) or pretreated with YVAD-CHO (20 μM: black bars) or BHA (100 μM: red bars) for 1 h at 37 °C, and then were treated with DHR123 (1 μM) and NPs (10 μg/cm3 each) for 4 h at 37 °C. Percent DHR123-positive cells in propidium iodide-negative cells (Percent ROS-producing cells in live cells) was calculated by flow cytometry. Data are indicated as the mean + S.D. Similar results were obtained in at least three independent experiments

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