SupplementaryMaterial
The Acute Cytotoxicity of Bismuth Ferrite Nanoparticleson PC12 Cells
Qin Song1, Yongping Liu1,2, Ziyun Jiang1, Mingliang Tang1, Ning Li1, Fenfen Wei1,2 Guosheng Cheng1,2
1Suzhou Key Laboratory of Nanobiomedicine, Division of Nanobiomedicine, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou Industrial Park, Jiangsu 215123, P. R. China,
2University of Chinese Academy of Sciences, 19A Yuquan Road, Beijing 100049, P. R. China
Correspondence: Guosheng Cheng, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou Industrial Park, Jiangsu 215123, P. R. China, Tel: +86-512-62872557, Fax: +86-512-62872546,E-mail:
Running title: In Vitro Toxicity of Bismuth Ferrite Nanoparticles
Fig. S1Histograms illustrating cell cycle analysis of PC12 cells. (a, d, g) Control cells incubated for 3 h, 24 h and 48 h, (b, e, h) 50 μg/mL BFO-NP-treated cells incubated for 3 h, 24 h and 48 h, and (c, f, i) 200 μg/mL BFO-NP-treated cells incubated for 3 h, 24 h and 48 h.Cells treated with 50 μg/mL and 200 μg/mLBFO-NPs for 3 h show a decrease in S and G2/M population compared with control, indicating cell proliferation inhabition. However, cells treated with50 μg/mL and 200 μg/mL BFO-NPs for 48 h present an increase in S and G2/M population compared with control, suggesting cell proliferation promotion. The absence of sub-G1 population suggests the absence of apoptosis. Markers were drawn on regions of interest (sub-G1, G1, S and G2/M) to generate statistics of cells under each region. Corresponding statistics were generated in CellQuestPro software. The figures are representatives for three different experiments.
Fig. S2Dot plotspresenting apoptosis and necrosis analysis of PC12 cells. (a, d, g) Control cells incubated for 3 h, 24 h and 48 h, (b, e, h) 50 μg/mL BFO-NP-treated cells incubated for 3 h, 24 h and 48 h, and (c, f, i) 200 μg/mL BFO-NP-treated cells incubated for 3 h, 24 h and 48 h.Cells in Q1-Q4regionsrepresentnecrotic cells, late apoptosis cells, viable cells and apoptotic cells, respectively.Cells treated with 50 μg/mL and 200 μg/mL BFO-NPs for 3, 24 or 48 h show no increased apoptosis and necrosiscompared with control. The figures are representatives for three different experiments.
Fig. S3 Fluorescence microscopy images of PC12 cells stained with Hoechest 33258. PC12 cells were treated with 0, 50 and 200 μg/mL BFO-NPs for 3h (upper panel) and 48 h (lower panel). Both BFO-NP treated cells and control cells exhibit normal blue nuclei with organized cellular structures. The figures are representatives for three different experiments.
Fig. S4 The fluorescent images of comet assay. a) Untreated control and b) cells treated with 200 μg/ml BFO-NPs for 3h c) cells treated with 200 μg/ml BFO-NPs for 48h. The comet assay results were consistent with the apoptosis and necrosis assay. A small percentage (below 10%) of cells was undergoing mild DNA damage in all of the BFO-NP treated samples, which had no significant difference from untreated controls. The figures are representatives for three different experiments.