Nazish Matti

Evaluation Of Cytotoxic, Antiviral And Mutagenic Potential Of Micronutrient Combination (Iodin, Potassium Iodide AndAscorbic Acid) In Vitro Cell Culture - 2013

Iodine has been proved over an extended period of time as antimicrobial especially antiviral and is employed extensively as skin disinfectant. Ascorbic acid also possesses antioxidant and antiviral activities. A micronutrient combination consisting of iodine, ascorbic acid, potassium iodide and excipients have been developed to cope with increasing burden of viral infections in Pakistan and particularly against Hepatitis C Virus clinically. The objective of this project was to analyze antiviral activity, cytotoxic and mutagenic potential of micronutrient combinationthrough H9N1 influenza virus strain antiviral infectivity assay, MTT cell viability assay and Ames Salmonella test, respectively. Commercial preparation of micronutrient combination i.e. MTI Renessans, comparable self-prepared micronutrient UVAS Renessans, pharmaceutical grade individual ingredientsand standard antiviral Ribavirin, Amantadine and Oseltamivir were tested.Renessan's concentrations of 375, 187.5, 93.75, 46.87, 23.43, 11.71, 5.86, 2.93, 1.46, and 0.73ug/ml were employed in these tests while comparable concentrations of UVAS-Renessans and individual micronutrients were analyzed. Ribavirin, Amantadine and Oseltamivir had doubling dilution from 100ug/ml concentrations 14 in descending manner. In MTT assay, confluent monolayer of Chicken Embryo Fibroblasts grown in 96-well cell culture plates were treated with ten concentrations of each extract in triplicate manner and was used to determine the viability of the cells and cell survival percentage was calculated.Antiviral efficacy was determined against influenza virus H9N1 strain by virus infection and subsequent cell viability assay. Cytotoxicity was measured by cell survival analysis after tetrazolium reduction assay (MTT assay) while mutagenicity was measured by bacterial reverse mutation analysis by Amestestusing two strains of Salmonella typhimurium TA100 and TA 98 with and without S9 and after appropriate incubation, number of revertant colonies per plate were counted in triplicate manner and Mutagenic Index MI was calculated. Our combination showed cytotoxicity at doses higher than 93 ug/ml while showed significant antiviral efficacy at concentrations of 93ug/ml, 46ug/ml, 23ug/ml, 11.5 ug/ml, 5.86ug/ml and 2.93ug/mlwhich faded away at lower dilutions. Renessans was found to be non-mutagenic in our experiment. The results are well supported by previous studies and may show a paradigm shift in systemic use of high dose of iodine as antiviral compounds.



Department of Pharmaoclogy & Toxicology

1825,T


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