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Comparison of in vitro methods for the study of cytotoxicity
Eliášová, Pavla ; Maixnerová, Jana (advisor) ; Bárta, Pavel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Pavla Eliášová Supervisor: RNDr. Jana Maixnerová, Ph.D. Title of diploma thesis: Comparison of in vitro methods for the study of cytotoxicity People are exposed to a growing number of toxic substances from the environment. Endocrine disrupting chemicals (EDCs) are a broad category of molecules that are thought to cause adverse effects on the endocrine system by interfering with the synthesis, transport, degradation or action of endogenous ligands. One of the aims of this thesis was to determine the in vitro toxicity of 17 selected endocrine disruptors on the human hepatocellular carcinoma HepG2 cell line. Cell viability was determined using the CellTiter96® AQueous One Solution Cell Proliferation Assay colorimetric method, the principle of which is the reduction of MTS to the colored product formazan by mitochondria in viable cells. The cytotoxic potential of the compounds was expressed by using the toxicological parameter IC50, which was measured in three time intervals (6, 12 and 24 hours). For 14 substances: Atrazine, DHEP, Bisphenol A, Carbofuran, 3-hydroxycarbofuran, Cypermethrin, DDE, DES, MEHP, PCB 118, PCB 153, PFOA, PFOS, Propiconazole, IC50 > 100 µM (respectively > 250 µM) was...
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Determination of toxicity using a cell model.
Fojkesová, Karolína ; Maixnerová, Jana (advisor) ; Bárta, Pavel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Karolína Fojkesová Supervisor: RNDr. Jana Maixnerová, Ph.D. Title of diploma thesis: Determination of toxicity using a cell model The subject of the diploma thesis was measure and evaluate the cytotoxicity in vitro of 22 newly synthesized substances with potential use for their antifungal, antibacterial and antimycobacterial activity on the standard HepG2 cell line. The tested substances were synthetized at the Department of Organic and Bioorganic Chemistry at the Faculty of Pharmacy in Hradec Králové, Charles University and their chemical structure is mafenide derivative. The compounds are MAF, MAF-4, MAF-8, MAF-9, MAF-10, MAF-11, MAF-12, MAF-13, MAF- 14, MAF-16, A-5-F, A-3,5-F2, A-3-Br-5-Cl, A-3,5-Cl2, A-3,5-Br2, A-5-Br, A-THIOF, A-SA, A-3,5-I2, A-5-Cl, A-3-I-5-Cl, A-5-I. Cytotoxicity has been measured using the CellTiter 96® AQueous One Solution Cell Proliferation Assay, which is a colorimetric method based on the reduction of the tetrazolium salt of MTS to the colored soluble formazan, and substances have been compared and evaluated according to the IC50 parameter. In case of 9 substances, specifically MAF, MAF-8, MAF-11, MAF-12, MAF-16, A-5- F, A-3,5-F2, A-SA, A-5-Cl, the IC50 was greater than...
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Determination of potential cytotoxic effect of experimental substances in vitro.
Osinková, Barbora ; Maixnerová, Jana (advisor) ; Smutná, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Barbora Osinková Supervisor: RNDr. Jana Maixnerová Ph.D. Title of diploma thesis: Determination of potential cytotoxic effect of experimental substances in vitro The subject of this master's thesis was to evaluate the cytotoxic potential of newly synthesized substances in vitro in the cell model HepG2, which is cell line obtained from a human liver carcinoma. These substances were synthesized within the Department of Organic and Bioorganic Chemistry at the Faculty of Pharmacy in Hradec Králové, Charles University (GUAM, GUAM-NH2, THIOSEMIK, GUAM-2, GUAM-6, GUAM- 11, GUAM-12, GUAM-18, GUAM-19, GUAM-2-1, GUAM-6-3). These substances are being investigated for assumed antimicrobial activity against G+ bacteria and fungi or for their ability to inhibit cholinesterases. The commercial colorimetric method CellTiter 96® AQueous One Solution Cell Proliferation Assay was used for in vitro determination of cytotoxicity, the principle of which is to measure the metabolic activity of cells based on the reduction of the tetrazolium salt of MTS to coloured formazan. The half-maximal inhibitory concentration IC50 was used to determine the number of viable cells and for evaluation of cytotoxic potential of...
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Study on cytotoxicity of compounds in vitro.
Vašková, Lucie ; Maixnerová, Jana (advisor) ; Kuchařová, Monika (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Studentka: Lucie Vašková Školitel: RNDr. Jana Maixnerová, Ph.D. Název diplomové práce: Study on cytotoxicity of compounds in vitro The subject of this diploma thesis was to assess the effect of newly synthesized antimycobacterial substances on the viability of human hepatocellular carcinoma (HepG2) cells. The tested substances were esters (HE-nMe, HE-4PHOPH, HE-KARVA, HE-2NAFT, HE-METRO, HE-CH2PY, HE-8CHIN) and thioesters (HES-4H, HES- nETH) of antituberculotic isoniazid. Experiments performed with these substances have shown, that like isoniazid, the substances inhibit InhA enzyme in mycobacteria and therefore interfere with cell wall biosynthesis. Isoniazid is a drug standardly used in the first line of TB treatment. Together with other first-line antituberculotics, some hepatotoxic potential has been reported during treatment. To assess the possible cytotoxic effect of the tested isoniazid derivatives, the standard human hepatocyte cell line HepG2 was chosen as the cell model. Cell viability was assessed by a colorimetric method that measures the metabolic activity of cells based on the reduction of the tetrazolium compound MTS. Obtained values were quantitatively compared using the toxicological...
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Evaluation of liver toxicity in vitro
Kafuňková, Kateřina ; Maixnerová, Jana (advisor) ; Bárta, Pavel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Kateřina Kafuňková Supervisor: RNDr. Jana Maixnerová, Ph.D. Title of diploma thesis: Evaluation of liver toxicity in vitro The subject of the diploma thesis is toxicity evaluation of newly synthesised substances on a cellular model representing a hepatocyte. The tested substances have been provided by the Department of Organic and Bioorganic Chemistry at the Faculty of Pharmacy in Hradec Králové, Charles University, as potential antifungal pharmaceuticals and medicine effective against Methicillin-resistant Staphylococcus aureus (MK-NO2-1, MK-NO2-2, DAB-5-K, PABA-Me-5, PABA-Et-5, MK-F-1, PABAN- 3, PABAN-5, MK-F-2, MK-CF3-1, MK-CF3- 2). In order to determine the toxicity we have implemented two methods. The first method is based upon measuring metabolic activity of cells by means of reducing tetrazolium to a colored product. The second method detects the amount of LDH released as a marker of toxicity. The human hepatoma cell line HepG2was used as a model. . The IC50 and EC50 parameters were used to assess the degree of viability and cytotoxicity. The final values obtained from the first method indicated all tested substances showed a certain level of toxicity to the hepatic tissue. MK-CF3-2 is the most...
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