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Vitro Cytotoxicity And Genotoxicity Testing Of Artemisinin, Digoxin And Silymarin

By: Saran Siddique | Dr. Aqeel Javeed.
Contributor(s): Prof. Dr. Muhammad Ashraf.
Material type: materialTypeLabelBookPublisher: 2013Subject(s): Department of Pharmaoclogy & ToxicologyDDC classification: 1568,T Dissertation note: The cytotoxicity and genotoxicity of three drugs artemisinin, digoxin and silymarin were evaluated against vero cell lines in this study. Thesolution of drugs was prepared in phosphate buffer saline(PBS) after dissolving in DMSO. For cytoticity dilutions of these drugs were applied in triplicate manner on Vero cells that were confluent in 96 well cell culture plates. MTT (3-[4.5-dimethylthiazol-2-yl]-2.5 diphenyltetrazolium bromide)assay was used for the cytotoxicity testing of these drugs and the cytotoxic doses of these drugs was 100µM for artemisinin, 100nM for digoxin and 380 µM for silymarin. After the cytotoxicity testing we also evaluated the genotoxic potential of these drugs against the same cell lines. For the genotoxicity testing we have used alkaline comet assay.For that base slides was prepared with normal melting agar and then a layer of pretreated cell suspension in low melting agar is used and after that another layer of low melting agar is coated on the last layer on the slides.Then lysis was carried out of the cells in lysing solution after that electrophoresis was done after that the slides was washed with neutralizing buffer and after that ethedium bromide stain is used and then slides were viewed under fluorescent microscope and we have observed that artemisinin showed genotoxic potential at 250µM, digoxin had shown genotoxic potential at 1000nM and silymarin have showngenotoxic potential at 500µM.
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The cytotoxicity and genotoxicity of three drugs artemisinin, digoxin and silymarin were evaluated against vero cell lines in this study. Thesolution of drugs was prepared in phosphate buffer saline(PBS) after dissolving in DMSO. For cytoticity dilutions of these drugs were applied in triplicate manner on Vero cells that were confluent in 96 well cell culture plates. MTT (3-[4.5-dimethylthiazol-2-yl]-2.5 diphenyltetrazolium bromide)assay was used for the cytotoxicity testing of these drugs and the cytotoxic doses of these drugs was 100µM for artemisinin, 100nM for digoxin and 380 µM for silymarin. After the cytotoxicity testing we also evaluated the genotoxic potential of these drugs against the same cell lines. For the genotoxicity testing we have used alkaline comet assay.For that base slides was prepared with normal melting agar and then a layer of pretreated cell suspension in low melting agar is used and after that another layer of low melting agar is coated on the last layer on the slides.Then lysis was carried out of the cells in lysing solution after that electrophoresis was done after that the slides was washed with neutralizing buffer and after that ethedium bromide stain is used and then slides were viewed under fluorescent microscope and we have observed that artemisinin showed genotoxic potential at 250µM, digoxin had shown genotoxic potential at 1000nM and silymarin have showngenotoxic potential at 500µM.

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