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The effects of nano-Silicon Dioxide on A549 cells was assessed in a series of experiments where cells were first seeded at 5×104 cells/ml into a series of 6 well plates (2 ml per well) that were incubated for 3 days until the cells were 70–80% confluence. The medium was then removed, the adherent cells washed with phosphate-buffer saline (PBS), before addition of culture medium containing between 0–10% fetal bovine serum and 0–1000 μg/ml nano-SiO2. The cells were then incubated for up to 24 h. The effect of nano-SiO2 on the cells was assessed at various time points by examining their morphology by light microscopy, flow cytometry, and the so-called MTT assay. Light microscopy was performed using an inverted microscope (Olympus CKX41) and images were captured using a QICam (QImaging, Surrey, BC, Canada). Flow cytometry was performed on cells that were recovered by trypsinisation as described above. The supernatant was removed and then the cells were suspended in 188 μl PBS containing 80 μM 3-hexyl-2-[3-(3-hexyl-2(3H)benzoxazolylidene)-1-propenyl]benzoxazolium iodide (DiOC6) and 240 μM propidium iodide, incubated at 37°C for 45 min in the dark and then analysed by flow cytometry (using a FACSCalibur, BD, Oxford, UK) recording DiOC6 and propidium iodide (PI) fluorescence. 

The MTT assay was performed in 96-well plates that were seeded with 0.1 ml 5×104 cells/ml and cultured as described above before addition of fresh culture medium containing either 0% or 1.25% fetal bovine serum and between 0–100 μg/ml nano-Silicon Dioxide. The cells were then incubated for up to 24 h after which 25 μl MTT reagent (5 mg/mL 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide diluted in PBS) was added to the wells and the plate incubated for a further 2 h at 37°C. Then, 100 μl of a solubilisation solution (10% SDS in 50% DMF) was added to the wells and the plate and incubated overnight at 37°C. The absorbance of the purple formazan product was measured at 595 nm using a microtitre plate reader.

Use Stober sol - gel method to synthesis monodisperse nano-Silicon Dioxide microspheres, and investigate the effects of temperature, ammonia concentration and TEOS concentration on synthetic silica.Use XRD, IR, SEM, TEM to test the characteristic of the samples. The results show that: the temperature increases to accelerate the hydrolysis was aged silica particles in the solution to cause agglomeration; increasing ammonia concentration increases the nucleation rate so that the hydrolysis rate increased, the particle size of the silica increases, the degree of agglomeration increases. Increasing the amount of TEOS also resulted in a slight increase in the silica particle size, agglomeration increased.



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