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 Federation10.15825/25/1995-1191-2021-4-119-131vtio-1420Research ArticleРегенеративная медицина и клеточные технологииRegenerative Medicine and Cell TechnologiesБиологически активное покрытие для тканеинженерной конструкции кровеносных сосудов малого диаметраBioactive coating for tissue-engineered smalldiameter vascular graftsСургученкоВ. А.SurguchenkoV. A.

Сургученко Валентина Александровна

123182, Москва, ул. Щукинская, д. 1

Тел. (499) 196-26-61

Valentina Surguchenko

1, Shchukinskaya str., Moscow, 123182, Russian Federation

Phone: (499) 196-26-61

valent.egorova@gmail.comНемецЕ. А.NemetsE. A.

Москва

Moscow

БеловВ. Ю.BelovV. Yu.

Москва

Moscow

СевастьяновВ. И.SevastianovV. I.

Москва

Moscow

ФГБУ «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; АНО «Институт медико-биологических исследований и технологий»РоссияShumakov National Medical Research Center of Transplantology and Artificial Organs; Institute of Biomedical Research and TechnologyRussian Federation202112112021234119131Copyright © Сургученко В.А., Немец Е.А., Белов В.Ю., Севастьянов В.И., 20222022Сургученко В.А., Немец Е.А., Белов В.Ю., Севастьянов В.И.Surguchenko V.A., Nemets E.A., Belov V.Y., Sevastianov V.I.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.transpl.ru/vtio/article/view/1420Цель работы

Цель работы: разработка способа модифицирования композитных пористых трубчатых биополимерных каркасов малого диаметра на основе бактериального сополимера поли(3-гидроксибутирата-со-3-гидроксивалерата) и желатина двухслойным биологически-активным покрытием на основе гепарина и лизата тромбоцитов, способствующим адгезии и пролиферации клеточных культур.

Материалы и методы

Материалы и методы. Композитные пористые трубчатые биополимерные каркасы с внутренним диаметром 4 мм изготавливали методом электроспиннинга из смеси 1 : 2 (по объему) 10% раствора сополимера поли(3-гидроксибутирата-со-3-гидроксивалерата) и 10% раствора желатина соответственно в гексафтор-2-пропаноле. Структуру каркасов стабилизировали парами глутарового альдегида. Каркасы модифицировали биологически-активным покрытием на основе гепарина и лизата тромбоцитов человека. Морфологию поверхности образцов анализировали с применением сканирующей электронной микроскопии. Биологическую безопасность модифицированных каркасов in vitro (гемолиз, цитотоксичность) оценивали согласно ГОСТ ISO 10993. Взаимодействие с культурами эндотелиальных клеток человека линии EA.hy926 и мезенхимальных стромальных клеток из жировой ткани человека исследовали с применением витальных красителей.

Результаты

Результаты. Разработан способ модифицирования композитных пористых трубчатых биополимерных каркасов малого диаметра, полученных методом электроспиннинга из смеси поли(3-гидроксибутирата-со- 3-гидроксивалерата) и желатина, двухслойным биологически-активным покрытием на основе ковалентно иммобилизованного гепарина и лизата тромбоцитов человека. In vitro доказано отсутствие цитотоксичности и гемолитической активности образцов модифицированных каркасов. Показано, что разработанное покрытие способствует адгезии и пролиферации мезенхимальных стромальных клеток жировой ткани человека на внешней поверхности и эндотелиальных клеток пупочной вены человека линии EA.hy926 на внутренней поверхности композитных пористых трубчатых биополимерных каркасов в условиях in vitro.

Заключение

Заключение. Полученные результаты позволяют прийти к заключению о возможности использования разработанного покрытия для формирования in vivo тканеинженерной конструкции кровеносных сосудов малого диаметра.

Objective

Objective: to develop a method for modifying composite small-diameter porous tubular biopolymer scaffolds based on bacterial copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and gelatin modified with a double-layered bioactive coating based on heparin (Hp) and platelet lysate (PL) that promote adhesion and proliferation of cell cultures.

Materials and methods

Materials and methods. Composite porous tubular biopolymer scaffolds with 4 mm internal diameter were made by electrospinning from a 1 : 2 (by volume) mixture of a 10% solution of poly(3-hydroxybutyrateco- 3-hydroxyvalerate) copolymer, commonly known as PHBV, and a 10% solution of gelatin, respectively, in hexafluoro-2-propanol. The structure of the scaffolds was stabilized with glutaraldehyde vapor. The scaffolds were modified with a bioactive Hp + PL-based coating. The surface morphology of the samples was analyzed using scanning electron microscopy. Biological safety of the modified scaffolds in vitro (hemolysis, cytotoxicity) was evaluated based on the GOST ISO 10993 standard. Interaction with cultures of human endothelial cell line (EA. hy926) and human adipose-derived mesenchymal stem cells (hADMSCs) was studied using vital dyes.

Results

Results. We developed a method for modifying small-diameter composite porous tubular biopolymer scaffolds obtained by electrospinning from a mixture of PHBV and gelatin modified with double-layered bioactive coating based on covalently immobilized Hp and human PL. The modified scaffold was shown to have no cytotoxicity and hemolytic activity in vitro. It was also demonstrated that the developed coating promotes hADMSC adhesion and proliferation on the external surface and EA.hy926 on the internal surface of the composite porous tubular biopolymer scaffolds in vitro.

Conclusion

Conclusion. The developed coating can be used for the formation of in vivo tissueengineered small-diameter vascular grafts.

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