03. Науково-дослідний інститут експериментальної та клінічної медицини ХНМУ
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Item Changes in cell membranes of white blood cells treated with a common food additive E407a(2021) Tkachenko, Anton; Onishchenko, Anatolii; Posokhov, Yevgen; Roshal, Alexander; Myasoedov, Valeriy; Nakonechna, OksanaObjectives: To estimate the state of phospholipid bilayer of rats WBCs exposed to a common food additive E407a, which is used as a thickener and emulsifier, during 4 h using a fluorescent probe − ortho-hydroxy derivative of 2,5-diaryl-1,3-oxazole. Materials and methods: Steady-state fluorescence spectroscopy: a study by the environment-sensitive fluorescent probe − 2-(2′-hydroxy-phenyl)-5-phenyl-1,3-oxazole (probe O1O). Results: Changes are detected in the spectra of the fluorescent probe bound to rat WBCs treated with the solutions of E407a of various concentrations in comparison with the corresponding spectra of the probe incubated with the untreated leukocytes. The decrease in polarity and protondonor ability is observed in the lipid membranes of leukocytes in the region, where the probe locates. Conclusions: Our findings suggest a higher dehydration of leukocyte membranes of rats treated with a common food additive E407a at high concentrations and, thus, indicate that exposure to high doses of E407a leads to the increase in the lipid order (i.e. to decrease in fluidity) of the membranes of rat WBCs.Item Experimental Evaluation of Food-Grade Semi-Refined Carrageenan Toxicity(2021-10-16) Pogozhykh, Denys; Posokhov, Yevgen; Myasoedov, Valeriy; Gubina-Vakulyck, Galina; Chumachenko, Tetyana; Knigavko, Oleksandr; Polikarpova, Hanna; Kalashnyk-Vakulenko, Yuliia; Sharashydze, Ketino; Nakonechna, Oksana; Prokopyuk, Volodymyr; Onishchenko, Anatolii; Tkachenko, AntonAbstract: The safety of food additives E407 and E407a has raised concerns in the scientific community. Thus, this study aims to assess the local and systemic toxic effects of the common food additive E407a in rats orally exposed to it for two weeks. Complex evaluations of the effects of semi-refined carrageenan (E407a) on rats upon oral exposure were performed. Local effects of E407a on the intestine were analyzed using routine histological stains and CD68 immunostaining. Furthermore, circulating levels of inflammatory markers were assessed. A fluorescent probe O1O (2- (20-OH-phenyl)-5-phenyl- 1,3-oxazole) was used for evaluating the state of leukocyte cell membranes. Cell death modes of leukocytes were analyzed by flow cytometry using Annexin V and 7-aminoactinomycin D staining. Oral administration of the common food additive E407a was found to be associated with altered small and large intestinal morphology, infiltration of the lamina propria in the small intestine with macrophages (CD68+ cells), high systemic levels of inflammation markers, and changes in the lipid order of the phospholipid bilayer in the cell membranes of leukocytes, alongside the activation of their apoptosis. Our findings suggest that oral exposure to E407a through rats results in the development of intestinal inflammation.Item Effects of E407a on the viability, metabolic and functional activity of dermal fibroblasts(2021) Tkachenko, Anton; Prokopiuk, Volodymyr; Onishchenko, Anatolii; Shevchenko, MariaAim: To evaluate the effects of semi-refined carrageenan (E407a) on the viability, metabolic and functional activity of skin fibroblasts in vitro. Material and methods: Various concentrations of E407a (0 – 10 mg/ml) were incubated for 24 h with skin fibroblast cultures isolated from rat embryos. The viability of cells exposed to semi-refined carrageenan was assessed by the neutral red uptake assay. MTT test was selected to estimate the metabolic activity of cells. In addition, scratch assay was used to determine the functional activity of dermal fibroblasts. Results: Direct exposure to semi-refined carrageenan did not affect the viability of rat dermal fibroblasts, evidenced by the results of MTT and neutral red uptake assays. However, experimental data of MTT and scratch assays revealed that high concentrations of this food additive increased the metabolic activity of fibroblasts and diminished the motility of these cells, suggesting that E407a is cytotoxic at high concentrations. Conclusion: Food-grade semi-refined carrageenan is cytotoxic towards skin fibroblasts at high concentrations.