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Oznaczanie aktywności przeciwutleniającej metodami in-vitro

Malinowski Jacek, Drzeżdżon Joanna, Chmurzyński Lech, Jacewicz Dagmara

Wiadomości Chemiczne

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2019

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[Z] 73, 9-10

523--542

EN

In - vitro methods of determination of the antioxidant activity of complex compounds are very interesting and not fully investigated areas of knowledge from the borderline of chemistry and biology. Methods used for determination of the activity of antioxidant complex compounds are modified due to the conditions of the experiments in which they should be carried out, e.g. reactions at physiological pH. Civilization diseases, stress related to the fast pace of life and increasing requirements of our lives cause the formation of free radicals in our body, i.e. particles characterized by a high reactivity. The methods of determination of the antioxidant activity of complexes discussed in this work apply tests carried out in laboratory conditions - in - vitro.

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Cytotoxicity of Resorcinol Under Short- and Long-Term Exposure in Vitro

Skowroń J., Zapór L.

International Journal of Occupational Safety and Ergonomics

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2004

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Vol. 10, No. 2

147--156

EN

Cytotoxicity of resorcinol to 3T3 fibroblast in short- (3 hrs) and long-term (72 hrs or 6 weeks) exposure was investigated. The effects of resorcinol on cell viability (neutral red uptake, NRU assay), mitochondrial function (MTT assay) and total cell protein (Kenacid Blue assay) were estimated. As a model for long-term exposure an INTEGRA CL 6-WELL bioreactor was used. The concentrations of resorcinol producing 20, 50 and 80% inhibition of cell growth in the NRU test were lower than in the MTT test after 3 hrs of exposure. The use of an INTEGRA CL 6-WELL bioreactor allows continuous culturing and exposure to test chemical of cells for several weeks, but the strong adhesiveness of fibroblast and forming aggregates make it difficult to remove them from chambers. Resorcinol in concentration of 1 μ/cm³ did not decrease the viability of cells to 50% of control in long-term exposure in the bioreactor.

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The Cytotoxicity of Some Organic Solvents on Isolated Hepatocytes in Monolayer Culture

Zapór L., Skowroń J., Gołofit-Szymczak M.

International Journal of Occupational Safety and Ergonomics

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2002

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Vol. 8, No. 1

121--129

EN

The cytotoxic effects of volatile and water-insoluble organic solvents (ethylbenzene, tetrachloroethylene, n-hexane) were tested on isolated hepatocytes in monolayer culture by using the 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) reduction assay. All of the tested compounds inhibited metabolic activity of hepatocytes and this effect depended on the concentration of solvents in the incubatory medium. The presence of fetal calf serum in the medium did not change the cytotoxicity of xenobiotics. IC50 values calculated on the basis of the MTT assay indicated that ethylbenzene was more cytotoxic than tetrachloroethylene and n-hexane. Using hepatocyte monolayer culture and the MTT assay to assess cytotoxicity of organic solvents causes many technical problems. It seems that it cannot be used as a rapid, cheap, and credible method.

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Połączenie dwóch metod testowania biomateriałów in vitro

Kudelska-Mazur D., Lewandowska-Szumieł M., Komender J., Benke G.

Inżynieria Biomateriałów

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2000

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R. 3, nr 9

26--29

PL

W pracy przedstawiono połączenie dwóch metod testowania biomateriałów w układzie in vitro. W doświadczeniu wykorzystano osteoblasty i fibroblasty ludzkie otrzymane z hodowli pierwotnych. Komórki wysiano na powierzchnię próbek materiałów: hydroksyapatytu, tlenku glinu i stali chirurgicznej. Zbadano żywotność, liczbę i rozpłaszczenie komórek. Żywotność oznaczono w teście XTT (EL/SA). Komórki poddano znakowaniu barwnikiem fluorescencyjnym Hoechst oraz barwieniu z użyciem przeciwciał anty-a-tubulinie metodą pośrednią. Następnie próbki obserwowano w mikroskopie fluorescencyjnym. Liczbę i rozpłaszczenie komórek zmierzono za pomocą programu do analizy obrazu Image- ProPlus. Wyniki testów żywotności porównano z wynikami pomiarów rozpłaszczenia i liczby komórek. Zauważono korelację pomiędzy rozpłaszczeniem i żywotnością osteoblastów i fibroblastów hodowanych na hydroksyapatycie, tlenku glinu i stali chirurgicznej.

EN

We present a combination of two methods of biomaterial testing in vitro. Primary culture human osteoblasts and fibroblasts seeded on the samples of hydroxyapatite, alumina and surgical steel were used in the experiments. Celt viability, number and spreading were investigated. Viability was estimated in XTT test (ELISA). Anti-a-tubulin antibodies were used for indirect immunofluorescent labelling of cells and Hoechst fluorescent dye for staining of nuclei. Subsequently the samples were observed in a fluorescence microscope. Cell number and spreading were measured by means of Image ProPtus software. The results of viability test were compared with calculations of cell number and spreading, It was observed that correlation existed between spreading and viability of osteoblasts and fibroblasts cultured on hydroxyapatite, alumina and surgical steel.