toxreviewТоксикологический вестникToxicological Review0869-79223034-4611Federal Scientific Center of Hygiene named after F.F. Erisman10.36946/0869-7922-2019-4-16-22toxreview-211Research ArticleСтатьиНОВЫЕ ДАННЫЕ К ВОПРОСУ ОБ ИНФОРМАТИВНОСТИ ЭКСПЕРИМЕНТОВ НА КЛЕТОЧНЫХ КУЛЬТУРАХ ДЛЯ ОЦЕНКИ СРАВНИТЕЛЬНОЙ И КОМБИНИРОВАННОЙ ТОКСИЧНОСТИ МЕТАЛЛООКСИДНЫХ НАНОЧАСТИЦNEW DATA ON THE QUESTION OF INFORMATIVENESS OF EXPERIMENTS ON CELL CULTURES FOR ASSESSMENT OF COMPARATIVE AND COMBINED TOXICITY OF METAL OXIDE NANOPARTICLESБушуеваТ. В.BushuevaT. V.

Бушуева Татьяна Викторовна - кандидат медицинских наук, заведующая Научно-производственным отделом Лабораторно-диагностических технологий.

Bushueva Tatiana Victorovna

620014, Ekaterinburg

bushueva@ymrc.ruМинигалиеваИ. А.MinigalievaI. A.

Минигалиева Ильзира Амировна - кандидат биологических наук, старший научный сотрудник, заведующий лабораторией промышленной токсикологии.

Minigalieva Ilzira Amirovna

620014, Ekaterinburg

ilzira-minigalieva@yandex.ruПановВ. Г.PanovV. G.

Панов Владимир Григорьевич - кандидат физико-математических наук, старший научный сотрудник лаборатории математического моделирования в экологии и медицине.

620990, Екатеринбург

 

Panov Vladimir Grigorievich

620990, Ekaterinburg

vpanov@ecko.uran.ruКузнецоваА. Н.KuznetsovaA. N.

Кузнецова Алиса Николаевна - лаборант-исследователь НПО Лабораторно-диагностических технологий.

620014, Екатеринбург

Kuznetsova Alisa Nikolaevna

620014, Ekaterinburg

alisa@ymrc.ruНаумоваА. С.NaumovaA. S.

Наумова Анна Сергеевна - лаборант-исследователь НПО Лабораторно-диагностических технологий.

620014, Екатеринбург

Naumova Anna Sergeevna

620014, Ekaterinburg

naumova@ymrc.ruCутунковаМ. П.SutunkovaM. P.

Сутункова Марина Петровна - кандидат медицинских наук, заведующий лабораторией токсикологии среды обитания.

620014, Екатеринбург

Sutunkova Marina Petrovna

620014, Ekaterinburg

sutunkova@ymrc.ruШурВ. Я.ShurV. Ya.

Шур Владимир Яковлевич - доктор физико-математических наук, профессор, директор Центра коллективного пользования «Современные нанотехнологии».

620000, Екатеринбург

Shur Vladimir Yakovlevich

620000, Ekaterinburg

vladimir.shur@urfu.ruШишкинаЕ. В.ShishkinaЕ. V.

Шишкина Екатерина Владимировна - кандидат физико-математических наук, старший научный сотрудник Центра коллективного пользования «Современные нанотехнологии».

Shishkina Ekaterina Vladimirovna

620000, Ekaterinburg

ekaterina.shishkina@labfer.usu.ruГурвичВ. Б.GurviсhV. B.

Гурвич Владимир Борисович - доктор медицинских наук, директор.

620014, Екатеринбург

Gurvich Vladimir Borisovich

620014, Ekaterinburg

gurvich@ymrc.ruКацнельсонБ. А.KatsnelsonB. A.

Кацнельсон Борис Александрович - доктор медицинских наук, профессор, заслуженный деятель науки РФ, заведующий отделом токсикологии и биопрофилактики.

Katsnelson Boris Aleksandrovich

620014, Ekaterinburg

bkaznelson@etel.ruФБУН «Екатеринбургский медицинский научный центр профилактики и охраны здоровья рабочих промпредприятий» РоспотребнадзораРоссияMedical Research Center for Prophylaxis and Health Protection in Industrial WorkersRussian FederationИнститут промышленной экологии, УрО РАНРоссияInstitute of Industrial Ecology, Urals Branch of the Russian Academy of SciencesRussian FederationИнститут естествознания и математики, Уральский федеральный университетРоссияSchool of Natural Sciences and Mathematics, Ural Federal UniversityRussian Federation201928082019041622Copyright © Бушуева Т.В., Минигалиева И.А., Панов В.Г., Кузнецова А.Н., Наумова А.С., Cутункова М.П., Шур В.Я., Шишкина Е.В., Гурвич В.Б., Кацнельсон Б.А., 20192019Бушуева Т.В., Минигалиева И.А., Панов В.Г., Кузнецова А.Н., Наумова А.С., Cутункова М.П., Шур В.Я., Шишкина Е.В., Гурвич В.Б., Кацнельсон Б.А.Bushueva T.V., Minigalieva I.A., Panov V.G., Kuznetsova A.N., Naumova A.S., Sutunkova M.P., Shur V.Y., Shishkina Е.V., Gurviсh V.B., Katsnelson B.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.toxreview.ru/jour/article/view/211

Изолированное и комбинированное повреждающее действие наночастиц (НЧ) PbO и CuO оценены на стабильной линии человеческих фибробластов с помощью трёх различных показателей цитотоксичности, основанных на снижении (а) дегидрогеназной активности клеток (МТТ-тест); (б) содержания АТФ (тест на жизнеспособность культуры по интенсивности люминесценции); (в) интегральной оценки клеточной пролиферации, распластывания и прикрепления к поверхности по электрическому импедансу («нормализованный клеточный индекс»). По всем этим показателям продемонстрирована для обоих видов метало-оксидных наночастиц чёткая зависимость повреждения клетки от их концентрации, адекватно описываемая гиперболической функцией, в то время как при одном и том же уровне воздействия количественные характеристики цитотоксичности PbO-НЧ в сравнении с CuO-НЧ схожи. Последнее ранее наблюдалось и в субхроническом эксперименте на крысах. Математически описанная с помощью методологии построения поверхности отклика комбинированная цитотоксичность наночастиц in vitro найдена неоднозначной, что также согласуется с выводами из эксперимента на крысах с теми же наночастицами.

Isolated and combined damaging effects of PbO and CuO nanoparticles have been estimated on an established line of human fibroblasts using three different cytotoxicity indices based on reduction in: (a) the cellular dehydrogenase activity (MTT Assay), (b) the ATP content (Luminescent Cell Viability Assay), (c) the cellular proliferation, viability, spreading, and attachment to substrate evaluated integrally by continuous impedance-based measurement of the Normalized Cell Index. For all these indicators a clear dependence of cell damage on concentration of metal oxide nanoparticles has been demonstrated for both types of metal oxide nanoparticles, which is adequately described by the hyperbolic function, while at the same level of exposure the quantitative characteristics of cytotoxicity of PbO-NPs in comparison with CuO-NPs are similar. The latter was previously observed in the subchronic experiment on rats. The combined in vitro cytotoxicity of nanoparticles has been also described mathematically using the response surface construction methodology and found to be ambiguous, which is also consistent with the conclusions from the experiment on rats with the same nanoparticles.

оксид свинцаоксид мединаночастицытоксичность in vitrolead oxidecopper oxidenanoparticlesin vitro toxicityReferencesMinigalieva, I.A., Bushueva, T.V., Froehlich, E., Meindl C., Panov V.G, Varaksin A.N, Shur V. Ya, Shishkina E.V., Gurvivh V.B., Katsnelson B.A. Are in vivo and in vitro assessments of comparative and combined toxicity of the same metallic nanoparticles compatible, or contradictory, or both? A juxtaposition of data obtained in respective experiments with NiO and Mn3O4 nanoparticles. Food Chem. Toxicol. 2017; 109: 393-4Minigalieva, I.A., Bushueva, T.V., Froehlich, E., Meindl C., Panov V.G, Varaksin A.N, Shur V. Ya, Shishkina E.V., Gurvivh V.B., Katsnelson B.A. Are in vivo and in vitro assessments of comparative and combined toxicity of the same metallic nanoparticles compatible, or contradictory, or both? A juxtaposition of data obtained in respective experiments with NiO and Mn3O4 nanoparticles. Food Chem. Toxicol. 2017; 109: 393-4Минигалиева И.А., Бушуева Т. В., Панов В. Г., Вараксин А.Н., Шур В.Я., Шишкина Е.В., Гурвич В.Б., Кацнельсон Б.А. Некоторые аспекты оценки токсичности металло- оксидных наночастиц на клеточных культурах (на примере NiO и Mn3O4). Токсикологический вестник. 205: 35-43.Минигалиева И.А., Бушуева Т. В., Панов В. Г., Вараксин А.Н., Шур В.Я., Шишкина Е.В., Гурвич В.Б., Кацнельсон Б.А. Некоторые аспекты оценки токсичности металло- оксидных наночастиц на клеточных культурах (на примере NiO и Mn3O4). Токсикологический вестник. 205: 35-43.Minigalieva I.A., Bushueva, T.V., Panov V.G., Shur V. Ya, Shishkina E.V., Gurvivh V.B., Katsnelson B.A. Some aspects of metal oxide nanoparticles toxicity assessment on cell cultures as exemplified by NiO and Mn3OToxicological Review. 205: 35-43 (in Russian).Minigalieva I.A., Bushueva, T.V., Panov V.G., Shur V. Ya, Shishkina E.V., Gurvivh V.B., Katsnelson B.A. Some aspects of metal oxide nanoparticles toxicity assessment on cell cultures as exemplified by NiO and Mn3OToxicological Review. 205: 35-43 (in Russian).Katsnelson B.A., Privalova L.I., Sutunkova M.P., Minigalieva IA, Gurvich V.B., Shur V.Y., et al. Experimental research into metallic and metal oxide nanoparticle toxicity in vivo, In: B. Yan, H. Zhou, J. Gardea-Torresdey (Eds.). “Bioactivity of Engineered Nanoparticles", Springer; 2017; Chapter 11: 259-319.Katsnelson B.A., Privalova L.I., Sutunkova M.P., Minigalieva IA, Gurvich V.B., Shur V.Y., et al. Experimental research into metallic and metal oxide nanoparticle toxicity in vivo, In: B. Yan, H. Zhou, J. Gardea-Torresdey (Eds.). “Bioactivity of Engineered Nanoparticles", Springer; 2017; Chapter 11: 259-319.Karlsson, H.L., Cronholm, P., Gustafsson, J., Moller, L. Copper oxide nanoparticles are highly toxic: A comparison between metal oxide nanoparticles and carbon nanotubes. Chem. Res. Toxicol. 2008; 21: 1726-1732.Karlsson, H.L., Cronholm, P., Gustafsson, J., Moller, L. Copper oxide nanoparticles are highly toxic: A comparison between metal oxide nanoparticles and carbon nanotubes. Chem. Res. Toxicol. 2008; 21: 1726-1732.Studer, A.M., Limbach, L.K., van Duc, L., Krumeich, F., Athanassiou, E.K., Gerber, L.C., Moch, H., Stark, W.J. Nanoparticle cytotoxicity depends on intracellular solubility: Comparison of stabilized copper metal and degradable copper oxide nanoparticles. Toxicol. 2010; Lett. 1: 169-174.Studer, A.M., Limbach, L.K., van Duc, L., Krumeich, F., Athanassiou, E.K., Gerber, L.C., Moch, H., Stark, W.J. Nanoparticle cytotoxicity depends on intracellular solubility: Comparison of stabilized copper metal and degradable copper oxide nanoparticles. Toxicol. 2010; Lett. 1: 169-174.Cronholm, P., Karlsson, H.L., Hedberg, J., Lowe, T.A., Winnberg, L., Elihn, K., Wallinder, I.O., Moller, L. Intracellular uptake and toxicity of Ag and CuO nanoparticles: A comparison between nanoparticles and their corresponding metal ions. Small. 2013; 8: 970-982.Cronholm, P., Karlsson, H.L., Hedberg, J., Lowe, T.A., Winnberg, L., Elihn, K., Wallinder, I.O., Moller, L. Intracellular uptake and toxicity of Ag and CuO nanoparticles: A comparison between nanoparticles and their corresponding metal ions. Small. 2013; 8: 970-982.Cuillel, M., Chevallet, M., Charbonnier, P., Fauquant, C., Pignot-Paintrand, I., Arnaud, J., Cassio, D., Michaud-Soret, I., Mintz, E. Interference of CuO nanoparticles with metal homeostasis in hepatocytes under sub-toxic conditions. Nanoscale. 2014; 16: 1707-1715.Cuillel, M., Chevallet, M., Charbonnier, P., Fauquant, C., Pignot-Paintrand, I., Arnaud, J., Cassio, D., Michaud-Soret, I., Mintz, E. Interference of CuO nanoparticles with metal homeostasis in hepatocytes under sub-toxic conditions. Nanoscale. 2014; 16: 1707-1715.Privalova L.I., Katsnelson B.A., Loginova N.V., Gurvich V.B., Shur V.Y., Valamina I.E., et al. Subchronic Toxicity of Copper Oxide Nanoparticles and Its Attenuation with the Help of a Combination of Bioprotectors. Int J Mol Sci. 2014; 15: 12379-12406.Privalova L.I., Katsnelson B.A., Loginova N.V., Gurvich V.B., Shur V.Y., Valamina I.E., et al. Subchronic Toxicity of Copper Oxide Nanoparticles and Its Attenuation with the Help of a Combination of Bioprotectors. Int J Mol Sci. 2014; 15: 12379-12406.Bondarenko, O., Ivask, A., Kakinen, A., Kahru A. Sub-toxic effects of CuO nanoparticles on bacteria: Kinetics, role of Cu ions and possible mechanisms of action. Environ. Pollut. 2012; 169: 81-89.Bondarenko, O., Ivask, A., Kakinen, A., Kahru A. Sub-toxic effects of CuO nanoparticles on bacteria: Kinetics, role of Cu ions and possible mechanisms of action. Environ. Pollut. 2012; 169: 81-89.Pang, C., Selck, H., Misra, S.K., Berhanu, D., Dybowska, A., Valsami-Jones, E., Forbes, V.E. Effects of sediment-associated copper to the deposit-feeding snail, Potamopyrgus antipodarum: A comparison of Cu added in aqueous form or as nano- and micro-CuO particles. Aquat. Toxicol. 2012; 15: 114-122.Pang, C., Selck, H., Misra, S.K., Berhanu, D., Dybowska, A., Valsami-Jones, E., Forbes, V.E. Effects of sediment-associated copper to the deposit-feeding snail, Potamopyrgus antipodarum: A comparison of Cu added in aqueous form or as nano- and micro-CuO particles. Aquat. Toxicol. 2012; 15: 114-122.Magaye, R., Zhao, J., Bowman, L., Ding, M. Genotoxicity and carcinogenicity of cobalt-, nickel- and copper-based nanoparticles. Exp. Ther. Med. 2012; 4: 551-561.Magaye, R., Zhao, J., Bowman, L., Ding, M. Genotoxicity and carcinogenicity of cobalt-, nickel- and copper-based nanoparticles. Exp. Ther. Med. 2012; 4: 551-561.Liao, M., Liu, H. Gene expression profiling of nephrotoxicity from copper nanoparticles in rats after repeated oral administration. Environ. Toxicol. Pharmacol. 2012; 34: 67-80.Liao, M., Liu, H. Gene expression profiling of nephrotoxicity from copper nanoparticles in rats after repeated oral administration. Environ. Toxicol. Pharmacol. 2012; 34: 67-80.Xu, J., Li, Z., Xu, P., Xiao, L., Yang, Z. Nanosized copper oxide induces apoptosis through oxidative stress in podocytes. Arch. Toxicol. 2013; 87: 1067-1073.Xu, J., Li, Z., Xu, P., Xiao, L., Yang, Z. Nanosized copper oxide induces apoptosis through oxidative stress in podocytes. Arch. Toxicol. 2013; 87: 1067-1073.Sizova, E., Miroshnikov, S., Polyakova, V., Gluschenko, N., Skalny, A. Copper nanoparticles as modulators of apoptosis and structural changes in tissues. J. Biomater. Nanobiotechnol. 2012; 3: 97-104.Sizova, E., Miroshnikov, S., Polyakova, V., Gluschenko, N., Skalny, A. Copper nanoparticles as modulators of apoptosis and structural changes in tissues. J. Biomater. Nanobiotechnol. 2012; 3: 97-104.Gomes, T., Araujo, O., Pereira, R., Almeida, A.C., Cravo, A., Bebianno, M.J. Genotoxicity of copper oxide and silver nanoparticles in the mussel Mytilus galloprovincialis. Mar. Environ. Res. 2013; 84: 51-59.Gomes, T., Araujo, O., Pereira, R., Almeida, A.C., Cravo, A., Bebianno, M.J. Genotoxicity of copper oxide and silver nanoparticles in the mussel Mytilus galloprovincialis. Mar. Environ. Res. 2013; 84: 51-59.Alarifi, S., Ali, D., Verma, A., Alakhtani, S., Ali, B.A. Cytotoxicity and genotoxicity of copper oxide nanoparticles in human skin keratinocytes cells. Int. J. Toxicol. 2013; 32: 296-307.Alarifi, S., Ali, D., Verma, A., Alakhtani, S., Ali, B.A. Cytotoxicity and genotoxicity of copper oxide nanoparticles in human skin keratinocytes cells. Int. J. Toxicol. 2013; 32: 296-307.Minigalieva, I.A., Katsnelson, B.A., Panov, V.G., Privalova, L.I., Varaksin, A.N., Gurvich, V.B., Sutunkova, M.P., Shur, V.Ya., Shishkina, E.V., Valamina, I.E., Zubarev, I.V., Makeyev, O.H., Meshtcheryakova, E.Y., Klinova, S.V. In vivo toxicity of copper oxide, lead oxide and zinc oxide nanoparticles acting in different combinations and its attenuation with a complex of innocuous bio-protectors. Toxicology. 2017; 380: 72-93.Minigalieva, I.A., Katsnelson, B.A., Panov, V.G., Privalova, L.I., Varaksin, A.N., Gurvich, V.B., Sutunkova, M.P., Shur, V.Ya., Shishkina, E.V., Valamina, I.E., Zubarev, I.V., Makeyev, O.H., Meshtcheryakova, E.Y., Klinova, S.V. In vivo toxicity of copper oxide, lead oxide and zinc oxide nanoparticles acting in different combinations and its attenuation with a complex of innocuous bio-protectors. Toxicology. 2017; 380: 72-93.Amiri, A., Mohammadi, M., Shabani, M. Synthesis and Toxicity Evaluation of Lead Oxide (PbO) Nanoparticles in Rats. Electronic J Biol. 2016; 12(2): 110-114.Amiri, A., Mohammadi, M., Shabani, M. Synthesis and Toxicity Evaluation of Lead Oxide (PbO) Nanoparticles in Rats. Electronic J Biol. 2016; 12(2): 110-114.Dumkova, J., Smutna, T., Vrlikova, L., Le Coustumer, P., Vecera, Z., Docekal, B., Mikuska, P., Capka, L., Fictum, P., Hampl, A., Buchtova M. Sub-chronic inhalation of lead oxide nanoparticles revealed their broad distribution and tissue-specific subcellular localization in target organs. Part Fibre Toxicol. 2017; 14(1):Dumkova, J., Smutna, T., Vrlikova, L., Le Coustumer, P., Vecera, Z., Docekal, B., Mikuska, P., Capka, L., Fictum, P., Hampl, A., Buchtova M. Sub-chronic inhalation of lead oxide nanoparticles revealed their broad distribution and tissue-specific subcellular localization in target organs. Part Fibre Toxicol. 2017; 14(1):Miri, A., Sarani, M., Hashemzadeh, A., Mardani Z., Darroudi, M., 20Biosynthesis and cytotoxic activity of lead oxide nanoparticles. Green Chemistry Letters and Reports. 2018; 11(4): 567-572.Miri, A., Sarani, M., Hashemzadeh, A., Mardani Z., Darroudi, M., 20Biosynthesis and cytotoxic activity of lead oxide nanoparticles. Green Chemistry Letters and Reports. 2018; 11(4): 567-572.Ng, D., Chu, Y., Tan, S., Wang, S., Lin, Y., Chu, C., Soo, Y., Song, Y., Chen, P. In vivo evidence of intestinal lead dissolution from lead dioxide (PbO2) nanoparticles and resulting bioaccumulation and toxicity in medaka fish. Environ. Sci.: Nano. 2019; 6: 580-5Ng, D., Chu, Y., Tan, S., Wang, S., Lin, Y., Chu, C., Soo, Y., Song, Y., Chen, P. In vivo evidence of intestinal lead dissolution from lead dioxide (PbO2) nanoparticles and resulting bioaccumulation and toxicity in medaka fish. Environ. Sci.: Nano. 2019; 6: 580-5Cimpan, M.R, Mordal, T., Scholermann J., Allouni Z.E., Pliquett U., Cimpan, E. An impedance-based high-throughput method for evaluating the cytotoxicity of nanoparticles. Journal of Physics: Conf. Ser. 2013; 429, 012026.Cimpan, M.R, Mordal, T., Scholermann J., Allouni Z.E., Pliquett U., Cimpan, E. An impedance-based high-throughput method for evaluating the cytotoxicity of nanoparticles. Journal of Physics: Conf. Ser. 2013; 429, 012026.Varaksin A.N., Katsnelson B.A., Panov V.G., Privalova L.I., Kireyeva E.P., Valamina I.E., et al. Some considerations concerning the theory of combined toxicity: a case study of subchronic experimental intoxication with cadmium and lead. Food Chem Toxicol. 2014; 64: 144-156.Varaksin A.N., Katsnelson B.A., Panov V.G., Privalova L.I., Kireyeva E.P., Valamina I.E., et al. Some considerations concerning the theory of combined toxicity: a case study of subchronic experimental intoxication with cadmium and lead. Food Chem Toxicol. 2014; 64: 144-156.Minigalieva I.A., Katsnelson B.A., Panov V.G., Varaksin A.N., Gurvich V.B., Privalova L.I., et al. Experimental study and mathematical modeling of toxic metals combined action as a scientific foundation for occupational and environmental health risk assessment. A summary of results obtained by the Ekaterinburg research team (Russia). Toxicol Rep. 2017; 4C: 194-201.Minigalieva I.A., Katsnelson B.A., Panov V.G., Varaksin A.N., Gurvich V.B., Privalova L.I., et al. Experimental study and mathematical modeling of toxic metals combined action as a scientific foundation for occupational and environmental health risk assessment. A summary of results obtained by the Ekaterinburg research team (Russia). Toxicol Rep. 2017; 4C: 194-201.Privalova, L.I., Katsnelson, B.A., Osipenko, A.B., Yushkov, B.H., Babushkina, L.G. Response of a phagocyte cell system to products of macrophage breakdown as a probable mechanism of alveolar phagocytosis adaptation to deposition of particles of different cytotoxicity. Environ. Health Perspect. 1980; 35: 205-218.Privalova, L.I., Katsnelson, B.A., Osipenko, A.B., Yushkov, B.H., Babushkina, L.G. Response of a phagocyte cell system to products of macrophage breakdown as a probable mechanism of alveolar phagocytosis adaptation to deposition of particles of different cytotoxicity. Environ. Health Perspect. 1980; 35: 205-218.

The authors declare that there are no conflicts of interest present.