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 Federation10.15825/1995-1191-2026-2-117-127vtio-2143Research ArticleРЕГЕНЕРАТИВНАЯ МЕДИЦИНА И КЛЕТОЧНЫЕ ТЕХНОЛОГИИREGENERATIVE MEDICINE AND CELL TECHNOLOGIESНаноструктурный анализ фибробластов мыши 3Т3, культивированных на тканевых скаффолдах на основе натурального шелка в условиях in vitroNanostructural analysis of 3T3 mouse fibroblasts cultured on natural silk-based tissue scaffolds in vitrohttps://orcid.org/0000-0002-0769-301XЕфимовА. Е.EfimovA. E.

Ефимов Антон Евгеньевич.

Москва

Moscow

antefimov@gmail.comhttps://orcid.org/0009-0002-7064-2570ПодболотоваЕ. И.PodbolotovaE. I.

Подболотова Екатерина Игоревна.

Москва; Долгопрудный, Московская область

Moscow

podbolotova.ei@phystech.eduДосиО.DosiO.

Доси Онур.

Долгопрудный, Московская область

Moscow

dosi.o@phystech.eduhttps://orcid.org/0000-0002-4507-1852АгаповаО. И.AgapovaO. I.

Агапова Ольга Игоревна.

Москва

Moscow

olya.agape@gmail.comhttps://orcid.org/0000-0002-0273-4601АгаповИ. И.AgapovI. I.

Агапов Игорь Иванович.

123182, Москва, ул. Щукинская, д. 1

Тел. (499) 190-66-19

Igor I. Agapov.

1, Shchukinskaya str., Moscow, 123182

Phone: (499) 190-66-19

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 FederationФГБУ «Национальный медицинский исследовательский центр трансплантологии и искусственных органов имени академика В.И. Шумакова» Минздрава России; ФГАОУ ВО «Московский физико-технический институт (Национальный исследовательский университет)»РоссияShumakov National Medical Research Center of Transplantology and Artificial Organs; Moscow Institute of Physics and TechnologyRussian FederationФГАОУ ВО «Московский физико-технический институт (Национальный исследовательский университет)»РоссияMoscow Institute of Physics and TechnologyRussian Federation202628062026282117127Copyright © Ефимов А.Е., Подболотова Е.И., Доси О., Агапова О.И., Агапов И.И., 20262026Ефимов А.Е., Подболотова Е.И., Доси О., Агапова О.И., Агапов И.И.Efimov A.E., Podbolotova E.I., Dosi O., Agapova O.I., Agapov I.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.transpl.ru/vtio/article/view/2143

Цель: провести наноструктурный анализ особенностей взаимодействия фибробластов мыши 3Т3 с тканевыми скаффолдами на основе натурального шелка в условиях in vitro с использованием технологии сканирующей зондовой нанотомографии. Материалы и методы. Для исследования были получены образцы двух типов тканевых скаффолдов из натурального шелка: «Фиброплен-Атлас» и «Фиброплен-Газ», а также их модифицированные версии «Фиброплен-Атлас 80» и «Фиброплен-Газ 80». Фибробласты мыши 3Т3 культивировались на тканевых скаффолдах в течение 7 суток. Препараты тканевых скаффолдов с культивированными фибробластами были фиксированы и залиты в эпоксидную смолу. Исследование поверхности полученных образцов после среза ультрамикротомом производилось методом сканирующей зондовой нанотомографии. Анализ полученных данных позволяет количественно определить важные морфологические параметры клеток, тканевых скаффолдов и интерфейсов между ними. Результаты. Анализ полученных изображений указывает на утончение и разобщение микроволокон в нитях модифицированных скаффолдов «Фиброплен-Атлас 80» и «Фиброплен-Газ 80». В отличие от немодифицированных версий скаффолдов в данном случае фибробласты в равной мере взаимодействуют как с внутренними, так и с внешними поверхностями микроволокон нитей шелковых скаффолдов, что приводит к значительному (до 40%) росту параметра, отражающего долю площади поверхности клеток, взаимодействующую с поверхностью скаффолда. Заключение. Наноструктурный анализ фибробластов мыши 3Т3, культивированных на тканевых скаффолдах на основе натурального шелка, позволил получить данные о топологических и морфологических особенностях взаимодействия клеток с модифицированными и немодифицированными тканевыми скаффолдами. Взаимодействие клеток с микроволокнами внутри нитей модифицированных скаффолдов может служить ускорению биодеградации данных скаффолдов. Совокупность полученных наноструктурных данных и результатов клеточных экспериментов in vitro позволяет сделать вывод о том, что разработанные скаффолды поддерживают адгезию клеток 3T3 и формирование клеточно-матриксных контактов.

Objective: to perform a nanostructural analysis of the interaction between mouse 3T3 fibroblasts and natural silk-based tissue scaffolds in vitro using scanning probe nanotomography (SPN). Materials and methods. Two types of natural silk tissue scaffolds were investigated: Fibroplen-Atlas and Fibroplen-Gaz, along with their modified variants, Fibroplen-Atlas 80 and Fibroplen-Gaz 80. Mouse 3T3 fibroblasts were cultured on the scaffolds for 7 days. Following incubation, scaffold–cell constructs were fixed and embedded in epoxy resin. Ultrathin sections were prepared using an ultramicrotome, and the sample surfaces were analyzed by SPN. The acquired data enabled quantitative assessment of key morphological parameters of the cells, the scaffolds, and the interfaces between them. Results. Analysis of the acquired images revealed thinning and fragmentation of microfibers within the strands of the modified scaffolds (Fibroplen-Atlas 80 and Fibroplen-Gaz 80). In contrast to the unmodified scaffolds, fibroblasts in the modified matrices interacted with both the inner and outer surfaces of the microfibers. This enhanced interaction resulted in a marked increase (up to 40%) in the proportion of the cell surface area in contact with the scaffold surface. Conclusion. Nanostructural analysis of mouse 3T3 fibroblasts cultured on natural silk-based tissue scaffolds provided detailed insights into the topological and morphological features of cell–scaffold interactions in both modified and unmodified matrices. Enhanced interaction between cells and microfibers within the modified scaffold strands may promote accelerated scaffold biodegradation. The combined nanostructural findings and in vitro cell culture results indicate that the developed scaffolds effectively support 3T3 cell adhesion and the formation of stable cell–matrix contacts.

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