scbmtБИОМЕДИЦИНАJournal Biomed2074-59822713-0428Scientific center of biomedical technologies of Federal Medical and Biological Agency10.33647/2713-0428-20-3E-160-175scbmt-1643Research ArticleДОКЛИНИЧЕСКИЕ ИССЛЕДОВАНИЯ В БИОМЕДИЦИНЕNON-CLINICAL RESEARCH IN BIOMEDICINEИсследование биораспределения тканеинженерного продукта на основе хондроцитов человека различного происхождения при имплантации мышам линии Balb/c NudeStudy of the Biodistribution of a Tissue-Engineered Product Based on Human Chondrocytes of Various Sources After Implantation into Balb/c Nude MiceГолубинскаяП. А.GolubinskayaP. A.

Голубинская Полина Александровна, к.м.н. 

119435, Москва, ул. Малая Пироговская, 1а

Polina A. Golubinskaya, Cand. Sci. (Med.) 

119435, Moscow, Malaya Pirogovskaya Str., 1a

polinapigeon@gmail.comПикинаА. С.PikinaA. S.

Пикина Арина Сергеевна 

119435, Москва, ул. Малая Пироговская, 1а

Arina S. Pikina 

119435, Moscow, Malaya Pirogovskaya Str., 1a

arina.pikina@yandex.ruРучкоЕ. C.RuchkoE. S.

Ручко Евгений Сергеевич 

119435, Москва, ул. Малая Пироговская, 1а

Eugene S. Ruchko 

119435, Moscow, Malaya Pirogovskaya Str., 1a

ruchkoevgeny@yandex.ruКоженевскаяЕ. В.KozhenevskayaE. V.

Коженевская Евгения Владимировна 

603022, Нижний Новгород, просп. Гагарина, 23

Evgeniya V. Kozhenevskaya 

603022, Nizhny Novgorod, Gagarina Ave., 23

kozhenevskaya@neuro.nnov.ruПоспеловА. Д.PospelovA. Dz.

Поспелов Антон Джонович 

603022, Нижний Новгород, просп. Гагарина, 23

Anton Dz. Pospelov 

603022, Nizhny Novgorod, Gagarina Ave., 23

eso103163@gmail.comБабаевА. А.BabaevA. A.

Бабаев Алексей Александрович, к.б.н. 

603022, Нижний Новгород, просп. Гагарина, 23

Aleksej A. Babaev, Cand. Sci. (Biol.) 

603022, Nizhny Novgorod, Gagarina Ave., 23

babaev@unn.ruИвановВ. А.IvanovV. A.

Иванов Виктор Андреевич 

119435, Москва, ул. Малая Пироговская, 1а

Viktor A. Ivanov 

119435, Moscow, Malaya Pirogovskaya Str., 1a

vanov.va@inbox.ruБеспятыхЮ. А.BespyatykhJu. A.

Беспятых Юлия Андреевна, к.б.н. 

119435, Москва, ул. Малая Пироговская, 1а

Julia A. Bespyatykh, Cand. Sci. (Biol.) 

119435, Moscow, Malaya Pirogovskaya Str., 1a

JuliaBes@rcpcm.orgШнайдерЛ. С.ShnayderL. S.

Шнайдер Лев Сергеевич, к.м.н. 

111123, Москва, ул. Новогиреевская, 1, корп. 1

Lev S. Shnayder, Cand. Sci. (Med.) 

111123, Moscow, Novogireyevskaya Str., 1, Building 1

shnayderlev@gmail.comЕремеевА. В.EremeevA. V.

Еремеев Артём Валерьевич, к.б.н. 

119435, Москва, ул. Малая Пироговская, 1а

Artem V. Eremeev, Cand. Sci. (Biol.) 

119435, Moscow, Malaya Pirogovskaya Str., 1a

art-eremeev@yandex.ruФГБУ «Федеральный научно-клинический центр физико-химической медицины им. Ю.М. Лопухина ФМБА России»РоссияFederal Scientific and Clinical Center of Physical and Chemical Medicine named after Yu.M. Lopukhin of the Federal Medical and Biological Agency of RussiaRussian FederationФГАОУ ВО «Нижегородский государственный университет им. Н.И. Лобачевского»РоссияLobachevsky State University of Nizhny NovgorodRussian FederationГБУЗ «Московский клинический научный центр им. А.С. Логинова» ДЗМРоссияMoscow Clinical Scientific Center named after A.S. Loginov of the Moscow Health Care DepartmentRussian Federation202419112024203E160175Copyright © Голубинская П.А., Пикина А.С., Ручко Е.C., Коженевская Е.В., Поспелов А.Д., Бабаев А.А., Иванов В.А., Беспятых Ю.А., Шнайдер Л.С., Еремеев А.В., 20242024Голубинская П.А., Пикина А.С., Ручко Е.C., Коженевская Е.В., Поспелов А.Д., Бабаев А.А., Иванов В.А., Беспятых Ю.А., Шнайдер Л.С., Еремеев А.В.Golubinskaya P.A., Pikina A.S., Ruchko E.S., Kozhenevskaya E.V., Pospelov A.D., Babaev A.A., Ivanov V.A., Bespyatykh J.A., Shnayder L.S., Eremeev A.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.scbmt.ru/jour/article/view/1643

Целью данной работы было исследовать биораспределение разработанного нами продукта тканевой инженерии (ПТИ) на основе хондроцитов различного генеза в виде 3D-структур (хондросфер) после подкожной имплантации иммунодефицитным мышам линии Balb/c Nude. В начале имплантировали хондросферы на основе хондроцитов человека, хондроцитов из дифференцированных индуцированных плюрипотентных стволовых клеток (ИПСК), в т. ч. линии с нокаутом гена β2m. В течение 9 мес. проводили наблюдение за животными. Далее от мышей после эвтаназии получали образцы органов и тканей для гистологического анализа, оценки состоятельности имплантата, его интеграции и исследования биораспределения методом ПЦР. Хондросферы из дифференцированных производных ИПСК успешно интегрировались в окружающие ткани в зоны инокуляции, формировали хрящевую ткань. В образцах около имплантационной зоны опытных групп животных не было выявлено человеческой ДНК, которую обнаруживали в образцах органов контрольных групп (введение MDA231 и мезенхимальных стволовых клеток). Таким образом, исследуемые образцы ПТИ через 3 и 9 мес. после имплантации демонстрировали отсутствие биораспределения в другие ткани и органы мышей, что свидетельствует в пользу безопасности разрабатываемого препарата.

In this research, we develop a tissue-engineered product (TEP) based on chondrocytes of various genesis in the form of 3D structures (chondrospheres) after subcutaneous implantation in immunodeficient Balb/c Nude mice and investigate its biodistribution profile. Initially, chondrospheres based on chondrocytes and chondrocytes from differentiated induced pluripotent stem cells (iPSCs), including lines with a knockout of the β2m gene, were implanted. The animals were monitored for nine months. Further, after euthanasia, organ and tissue samples were obtained for histological analysis, evaluation of the viability of the implant, its integration and biodistribution research by PCR. Chondrospheres from differentiated iPSCs derivatives of both types successfully integrated into the surrounding tissues in the inoculation zones and formed cartilage tissue. In the samples near the implantation zone of the experimental groups of animals, no human DNA was detected. Human DNA was found in the samples of organs of the control groups (introduction of MDA231 and mesenchymal stem cells). Thus, three and nine months after implantation, the studied TEP samples demonstrated the absence of biodistribution to other tissues and organs of mice, which indicates the safety of the drug being developed.

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