mesЭкстремальная биомедицинаExtreme Medicine3033-89643033-8972Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2023.057mes-81Research ArticleОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕORIGINAL RESEARCHИсследование биораспределения биомедицинского клеточного продукта на основе хондроцитов человека при имплантации мышам линии Balb/c NudeAssessing biodistribution of biomedical cellular product based on human chondrocytes following implantation to Balb/c Nude miceПикинаА. С.PikinaA. S.

Арина Сергеевна Пикина

ул. Малая Пироговская, д. 1а, г. Москва, 119435

Moscow

ГолубинскаяП. А.GolubinskayaP. A.

Москва

Moscow

РучкоЕ. C.RuchkoE. S.

Москва

Moscow

КоженевскаяЕ. В.KozhenevskayaE. V.

Нижний Новгород

Nizhny Novgorod

ПоспеловА. Д.PospelovA. D.

Нижний Новгород

Nizhny Novgorod

БабаевА. А.BabayevA. A.

Нижний Новгород

Nizhny Novgorod

ЕремеевА. В.EremeevA. V.

Москва

Moscow

Федеральный научно-клинический центр физико-химической медицины имени Ю. М. ЛопухинаРоссияLopukhin Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological AgencyRussian FederationИнститут биологии развития имени Н. К. Кольцова Российской Академии НаукРоссияKoltzov Institute of Developmental Biology of Russian Academy of SciencesRussian FederationНижегородский государственный университет имени Н. И. ЛобачевскогоРоссияLobachevsky State University of Nizhny NovgorodRussian FederationНижегородский государственный университет имени Н. И. ЛобачевскогоРоссияLobachevsky State University of Nizhny Novgorod,Russian FederationФедеральный научно-клинический центр физико-химической медицины имени Ю. М. Лопухина; Институт биологии развития имени Н. К. Кольцова Российской Академии НаукРоссияLopukhin Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency; Koltzov Institute of Developmental Biology of Russian Academy of SciencesRussian Federation202324102024254123130Copyright © Пикина А.С., Голубинская П.А., Ручко Е.C., Коженевская Е.В., Поспелов А.Д., Бабаев А.А., Еремеев А.В., 20242024Пикина А.С., Голубинская П.А., Ручко Е.C., Коженевская Е.В., Поспелов А.Д., Бабаев А.А., Еремеев А.В.Pikina A.S., Golubinskaya P.A., Ruchko E.S., Kozhenevskaya E.V., Pospelov A.D., Babayev A.A., 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://www.extrememedicine.ru/jour/article/view/81

Несмотря на перспективность подхода клеточной терапии повреждений хряща человека с помощью аутологичных хондроцитов, подобные технологии только начинают внедрять в медицинскую практику в Российской Федерации. В связи с этим разработка биомедицинских клеточных продуктов (БМКП) для восстановления хрящевой ткани достаточно актуальна, а использование органоидных технологий наиболее приближено к условиям нативной ткани. Согласно требованиям законодательства РФ, в рамках доклинических исследований необходимо изучение биораспределения, характеризующего миграционный потенциал клеток, их тропность к тканям организма при имплантации. Целью работы было исследовать биораспределение нового БМКП на основе хондроцитов человека в виде хондросфер после подкожной имплантации мышам линии Balb/c nude. На первом этапе осуществляли имплантацию 12 мышам, а также введение физиологического раствора 12 контрольным животным. В течение 90 дней проводили взвешивание и наблюдение, а затем выводили мышей из эксперимента для получения образцов органов и тканей для гистологического анализа импланта, оценки его состоятельности, интеграции. На втором этапе изучали биораспределение методом ПЦР для выявления ДНК человека в образцах тканей и органов. Хондросферы успешно интегрировались в окружающие ткани зоны инокуляции, формировали хрящевую ткань. Статистически значимых (p < 0,05) изменений в весе не зафиксировали. В образцах из зоны имплантации хондросфер была выявлена ДНК человека, которую не обнаруживали в других органах и тканях. БМКП через 90 дней после имплантации демонстрировал отсутствие биораспределения в другие ткани и органы мышей, что свидетельствует о безопасности разрабатываемого продукта.

Despite the prospects of the approach to cell therapy of cartilage damage in humans involving autologous chondrocytes, similar technologies are just beginning to be introduced into medical practice in the Russian Federation. In this regard, the development of biomedical cell products (BCPs) for cartilage tissue repair is quite topical, while the use of organoid technology is the most close to the native tissue conditions. According to requirements of legislation of the Russian Federation, it is necessary to assess biodistribution characterizing migration potential of the cells, their tropism for body tissues following implantation within the framework of preclinical trials. The study was aimed to assess biodistribution of novel BCP based on human chondrocytes in the form of chondrospheres after subcutaneous implantation in Balb/c nude mice. Implantation to 12 mice was performed during the first phase, along with administration of saline to 12 control animals. Weighting and follow-up were conducted for 90 days. Then mice were withdrawn from the experiment to collect samples of organs and tissues for histological analysis of the implant, estimation of its viability, integration. During the second phase biodistribution was assessed by PCR in order to detect human DNA in the organ and tissue samples. Chondrospheres successfully integrated in the tissues surrounding the inoculation zones and formed cartilage tissue. No significant (p < 0.05) changes in weight were reported. No human DNA found in chondrosphere implantation zones was detected in the samples collected from other organs and tissues. BCP demonstrated no biodistribution across other tissues and organs of mice 90 days after implantation, which suggested that the product developed was safe. Keywords: biomedical cellular product, chondrocytes, biodistribution, preclinical trials, chondrospheres

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