vtioВестник трансплантологии и искусственных органовRussian Journal of Transplantology and Artificial Organs1995-1191Academician V.I.Shumakov National Medical Research Center of Transplantology and Artificial Organs", Ministry of Health of the Russian Federation10.15825/1995-1191-2020-4-115-122vtio-1272Research ArticleРегенеративная медицина и клеточные технологииRegenerative Medicine and Cell TechnologiesИсследование микро- и наноструктуры клеток печени, культивированных на биодеградируемых скаффолдах на основе фиброина шелка, методом сканирующей зондовой оптической нанотомографииInvestigation of the micro- and nano-structure of liver cells cultured on biodegradable silk fibroin-based scaffolds using scanning probe optical nanotomographyАгаповаО. И.AgapovaO. I.ЕфимовА. Е.EfimovA. E.СафоноваЛ. А.SafonovaL. A.БоброваМ. М.BobrovaM. M.АгаповИ. И.AgapovI. I.

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igor_agapov@mail.ruФГБУ «Национальный медицинский исследовательский центр трансплантологии и искусственных органов имени академика В.И. Шумакова» Минздрава РоссииРоссияShumakov National Medical Research Center of Transplantology and Artificial OrgansRussian Federation202026012021224115122Copyright © Агапова О.И., Ефимов А.Е., Сафонова Л.А., Боброва М.М., Агапов И.И., 20212021Агапова О.И., Ефимов А.Е., Сафонова Л.А., Боброва М.М., Агапов И.И.Agapova O.I., Efimov A.E., Safonova L.A., Bobrova M.M., Agapov I.I.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.transpl.ru/vtio/article/view/1272Цель

Цель. Провести анализ трехмерной микро- и наноструктуры и количественных морфологических параметров клеток печени, культивированных на биодеградируемых  скаффолдах в виде пленок на основе фиброина шелка.

Материалы и методы

Материалы и методы. Для исследования, были получены образцы биодеградируемых скаффолдов на основе фиброина шелка с культивированными  клетками печени крысы породы Wistar. Изучение трехмерной структуры клеток печени, культивированных на скаффолдах, было выполнено методом сканирующей зондовой  оптической нанотомографии при помощи экспериментальной установки, объединяющей ультрамикротом и сканирующий зондовый микроскоп в корреляции с флуоресцентной микроскопией.

Результаты

Результаты. Получены наномасштабные изображения и трехмерные  нанотомографические реконструкции участков клеток печени крысы, культивированных на скаффолде. Определены морфологические параметры клеток печени: средняя  шероховатость, удельная эффективная площадь. Установлено, что средняя шероховатость поверхности клеток печени Ra составляет 124,8 ± 8,2 нм, в то время как  эффективная площадь поверхности σ составляет 1,13 ± 0,02. Анализ объемного распределения липидных капель показал, что они занимают 28% объема клетки.

Выводы

Выводы. Полученные в результате исследования данные демонстрируют, что технология сканирующей зондовой оптической нанотомографии позволяет успешно анализировать наноструктуру и определять количественные характеристики  наноморфологии клеток печени, культивированных на биодеградируемых скаффолдах.

Objective

Objective: to analyze the 3D micro- and nano-structure and quantitative morphological parameters of liver cellscultured on biodegradable silk fibroin-based film scaffolds.

Materials and methods

Materials and methods. Samples of biodegradable silk fibroin-based scaffolds with cultured Wistar rat liver cells were obtained for the study. The 3D structure of liver cells cultivated on the scaffolds was studied by scanning probe optical nanotomography using an experimental setup combining an ultramicrotome and a scanning probe  microscope in correlation with fluorescence  microscopy.

Results

Results. Nanoscale images and 3D nanotomographic reconstructions of rat liver cells cultured on scaffold were obtained. The morphological parameters of liver cells (average roughness, specific effective area) were determined. The average surface roughness of the liver cells Ra was found to be 124.8 ± 8.2 nm, while the effective surface area σ was 1.13 ± 0.02. Analysis of the volume distribution of lipid droplets showed that they occupy 28% of the cell volume.

Conclusion

Conclusion. Scanning probe optical nanotomography can successfully analyze the nanostructure and quantify the nanomorphology of liver cells cultured on biodegradable scaffolds.

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