Study On The Cytotoxic Effects Of Silver Nanoparticles On C6 Glioma Cells

ObjectiveThe killing effect and mechanism of silver nanoparticles on C6 glioma cells were investigated by different concentration of silver nanoparticles,including toxicity,cell morphology,apoptosis and cell cycle.Method1.The toxic effects of silver nanoparticles on C6 glioma cells were evaluated by CCK-8 assay.According to the concentration of Nano silver in C6 glioma cell culture board,the group of 0.1 mg/ml,0.2 mg/ml,0.3 mg/ml,0.4 mg/ml,0.5 mg/ml,0.6 mg/ml,were detected,respectively.2.After the action of silver nanoparticles,the intercellular fusion of C6 gliomas is sparse,there is no aggregation growth,the cell density decreases obviously,and the cell density is distributed and growing,while the cell density decreases with the increase of silver nanoparticles concentration;The cell morphology was reduced from a naturally occurring fusiform to a smaller circle,with no obvious cell morphology and black particles in the cell,and few proliferating mitotic cells were observed.And C6 glioma cell is very easy to fall off from the bottom of the culture plate,the number of suspension cells increased,its admixture growth ability decreased obviously.3.Observation of cell membrane damage in C6 cells by means of Anaxin V and dimethyliodide(PI)detection: different concentrations of nano-silver in C6 glioma cell cultures were divided into 0.025 mg/ml group,0.05 mg/ml group,0.1 mg/ml group and 0.2 mg/ml group,control group.After cultivation of 24 h,add FITC Annex V and Propidium Iodide.Double variable flow cytometer was used to detect the fluorescence intensity and analyze the data in 1h.4.Flow cytometry to detect the effect of silver nanoparticles on the cell cycle:According to the different concentrations of silver nanoparticles in the cell cultures of C6 gliomas: 0.2 mg/ml group,0.4 mg/ml group,0.6 mg/ml group,0.8 mg/ml group,1.0 mg/ml group,control group.After culture of 24 h,the red fluorescence was detected by flow cytometry with Ranseye iodinated propanil,and Jianceguang scattering was also detected by using flow cytometry.Result1.The proliferation rate of C6 glioma cells decreased significantly after treatment of C6 glioma cells with different concentrations of silver,and the viability of C6 glioma cells decreased gradually with the increase of the concentration of silver nanoparticles and the increase of the time of action.2.After the action of silver nanoparticles,the intercellular fusion of C6 gliomas is sparse,there is no aggregation growth,the cell density decreases obviously,and the cell density is distributed and growing,while the cell density decreases with the increase of silver nanoparticles concentration;The cell morphology was reduced from a naturally occurring fusiform to a smaller circle,with no obvious cell morphology and black particles in the cell,and few proliferating mitotic cells were observed.And C6 glioma cell is very easy to fall off from the bottom of the culture plate,the number of suspension cells increased,its admixture growth ability decreased obviously.3.Early and late apoptosis rates in the control group were 6.95 %,0.025 mg mg L were 19.08 %,0.05 mg/ml was 33 %,0.1 mg/ml was 59.18 %,and 0.2 mg/ml was 27.22 %.The effect of different concentrations of silver nanoparticles on apoptosis was dependent on the concentration,the higher the concentration,the higher the induced death rate.4.Flow cytometry to detect the effect of silver nanoparticles on the cell cycle:The results showed that the number of cells in the G0/ G1 phase decreased when the S phase increased,but at what concentration the G2 /M phase showed little change in cell count compared with the S phase,with a large number of damaged cells blocked.ConclusionIn this study,the morphology of C6 glioma cells was observed by differentconcentrations of silver nanoparticles.The toxicity of C6 glioma cells was observed.With the increase of silver concentration and time,the viability of C6 glioma cells decreased,and the inhibition of C6 glioma cells by silver nanoparticles showed concentration and time dependence.The effect of different concentrations of silver nanoparticles on apoptosis was concentration dependent.The higher the concentration,the higher the death rate.