biopreparatБИОпрепараты. Профилактика, диагностика, лечениеBiological Products. Prevention, Diagnosis, Treatment2221-996X2619-1156Scientific Centre for Expert Evaluation of Medicinal Products10.30895/2221-996X-2024-24-2-172-187biopreparat-574Research ArticleТЕМА НОМЕРА: ВЫСОКОТЕХНОЛОГИЧНЫЕ ЛЕКАРСТВЕННЫЕ ПРЕПАРАТЫISSUE TOPIC: ADVANCED THERAPY MEDICINAL PRODUCTSОпыт производства и клинического применения биомедицинского клеточного продукта Изитенс® для восстановления повреждений хрящевой ткани коленного сустава человекаExperience in the production and clinical application of the cell-based medicinal product Easytense® for the repair of cartilage defects of the human kneehttps://orcid.org/0009-0000-2851-217XЗоричеваА. С.ZorichevaA. S.

Зоричева Анастасия Сергеевна

ул. Заводская, стр. 273, пос. Вольгинский, Петушинский район, Владимирская область, 601125

Anastasia S. Zoricheva

273 Zavodskaya St., Volginsky, Petushinsky District, Vladimir Region 601125

https://orcid.org/0000-0002-7152-6881ЗвоноваЕ. А.ZvonovaE. A.

Звонова Елизавета Александровна, канд. биол. наук

ул. Тестовская, д. 10, Москва, 123112

Elizaveta A. Zvonova, Cand. Sci (Biol.)

10 Testovskaya St., Moscow 123112

https://orcid.org/0009-0004-3727-7406АгаповаЛ. С.AgapovaL. S.

Агапова Лариса Степановна, канд. биол. наук

ул. Заводская, стр. 273, пос. Вольгинский, Петушинский район, Владимирская область, 601125

Larissa S. Agapova, Cand. Sci (Biol.)

273 Zavodskaya St., Volginsky, Petushinsky District, Vladimir Region 601125

https://orcid.org/0009-0006-2061-321XЛыковаМ. С.LykovaM. S.

Лыкова Мария Сергеевна, канд. биол. наук

ул. Заводская, стр. 273, пос. Вольгинский, Петушинский район, Владимирская область, 601125

Mariya S. Lykova, Cand. Sci (Biol.)

273 Zavodskaya St., Volginsky, Petushinsky District, Vladimir Region 601125

https://orcid.org/0000-0002-1179-3881МарковаО. А.MarkovaO. A.

Маркова Оксана Анатольевна

ул. Тестовская, д. 10, Москва, 123112

Oksana A. Markova

10 Testovskaya St., Moscow 123112

https://orcid.org/0009-0003-5863-0267ЛеоновВ. С.LeonovV. S.

Леонов Вячеслав Сергеевич, канд. биол. наук

ул. Заводская, стр. 273, пос. Вольгинский, Петушинский район, Владимирская область, 601125

Vyacheslav S. Leonov, Cand. Sci (Biol.)

273 Zavodskaya St., Volginsky, Petushinsky District, Vladimir Region 601125

leonov@generium.ruАкционерное общество «ГЕНЕРИУМ»РоссияGENERIUM JSCRussian Federation202430052024242Высокотехнологичные лекарственные препараты172187Copyright © Зоричева А.С., Звонова Е.А., Агапова Л.С., Лыкова М.С., Маркова О.А., Леонов В.С., 20242024Зоричева А.С., Звонова Е.А., Агапова Л.С., Лыкова М.С., Маркова О.А., Леонов В.С.Zoricheva A.S., Zvonova E.A., Agapova L.S., Lykova M.S., Markova O.A., Leonov V.S.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.biopreparations.ru/jour/article/view/574

ВВЕДЕНИЕ. Существующие клеточные методы восстановления хрящевой ткани, такие как аутологичная трансплантация хондроцитов, недостаточно эффективны, а хирургическое вмешательство болезненно и травматично. Поэтому требуется разработка более эффективного продукта для клеточной терапии с малоинвазивной хирургической процедурой его имплантации пациенту.ЦЕЛЬ. Разработка технологии производства аутологичного биомедицинского клеточного продукта (БМКП), содержащего трехмерные структуры (3D-сфероиды) на основе хондроцитов, выделенных из хрящевой ткани пациента, и оценка клинической эффективности его применения.МАТЕРИАЛЫ И МЕТОДЫ. Аутологичные хондроциты, выделенные из биоптата хрящевой ткани пациента, культивировали в монослойной культуре для получения необходимого количества клеток. Дальнейшее культивирование хондроцитов проводили с использованием неадгезивного покрытия для формирования 3D-сфероидов. Все этапы производства БМКП проводили в асептических условиях в изоляторах клеточных культур. Для характеристики фенотипа хондроцитов на разных этапах культивирования использовали фазово-контрастную микроскопию и иммуногистохимическое окрашивание с применением специфических флуоресцентно меченных антител. Генетическую стабильность контролировали кариологическим методом. В рамках клинического исследования (КИ) проводили оценку эффективности БМКП Изитенс® при помощи специализированных функциональных тестов и по Шкале оценки восстановления хрящевой ткани по результатам магнитно-резонансной томографии (MOCART). Основным критерием эффективности было изменение общего балла по Шкале оценки исхода травмы и остеоартроза коленного сустава (KOOS).РЕЗУЛЬТАТЫ. Разработана технология производства аутологичного БМКП Изитенс® из собственных клеток пациента без применения сыворотки животных, ростовых факторов, цитокинов и других добавок. С использованием кариологического метода подтверждено сохранение генетической стабильности хондроцитов на протяжении 3 пассажей культивирования в монослойной культуре. При 3D-культивировании в виде сфероидов хондроциты продуцируют характерные для матрикса хрящевой ткани белки (коллаген типа II, аггрекан), приобретая таким образом способность к восстановлению поврежденного хряща. В ходе КИ продемонстрировано статистически значимое улучшение состояния хрящевой ткани коленных суставов через 12 мес. после трансплантации 3D-сфероидов из аутологичных хондроцитов. Среднее изменение общего балла KOOS составило 23,8±15,9.ВЫВОДЫ. Продемонстрирована высокая эффективность БМКП Изитенс® для восстановления хрящевой ткани. На основании результатов клинических исследований данный БМКП зарегистрирован на территории Российской Федерации, введен в клиническую практику и может стать альтернативой эндопротезированию и дорогостоящим операциям за рубежом.

INTRODUCTION. The current cell-based cartilage repair methods, such as autologous chondrocyte transplantation, are not sufficiently effective, and the surgery is painful and traumatic. Therefore, there is a need for a more effective cell therapy product with a minimally invasive surgical procedure for its implantation into the patient.AIM. This study aimed to develop a manufacturing technology for the production of an autologous cell-based medicinal product (CBMP) comprising three-dimensional structures (3D-spheroids) based on chondrocytes isolated from the patient’s cartilage tissue, as well as to evaluate its clinical efficacy.MATERIALS AND METHODS. Autologous chondrocytes isolated from the patient’s cartilage biopsy were propagated in monolayer culture to obtain the required number of cells. Subsequently, the chondrocytes were cultivated on plates with a non-adhesive coating to form 3D spheroids. All CBMP production steps were performed under aseptic conditions in cell culture isolators. The authors used phase-contrast microscopy and immunohistochemical staining with specific fluorescence-labelled antibodies to characterise chondrocyte phenotypes at different stages of cultivation. Genetic stability was controlled by karyotyping. The efficacy of Easytense® was evaluated in a clinical trial using specialised functional tests and the Magnetic Resonance Observation of Cartilage Repair Tissue (MOCART) score. The primary efficacy endpoint was a change in the overall score on the Knee Injury and Osteoarthritis Outcome Score (KOOS).RESULTS. A manufacturing technology without using animal sera, growth factors, cytokines, or other additives was developed for the production of the autologous CBMP Easytense®. Karyological data confirmed that the chondrocytes retained genetic stability for 3 passages in monolayer culture. When cultured as 3D spheroids, the chondrocytes produced cartilage extracellular matrix proteins (type II collagen, aggrecan), thus acquiring the ability to repair damaged cartilage. The clinical trial demonstrated a statistically significant improvement in knee cartilage 12 months after the transplantation of 3D spheroids derived from autologous chondrocytes. The mean change in the overall KOOS score was 23.8±15.9.CONCLUSIONS. The clinical trial results indicate that Easytense® is highly effective for cartilage repair. Based on these results, the CBMP has been granted marketing authorisation and introduced into clinical practice in the Russian Federation. Easytense® has the potential to replace endoprosthetics and expensive surgeries abroad.

Изитенс®биомедицинский клеточный продуктБМКПхрящевая ткань коленного суставахондроцитыаутологичные хондроцитысфероидытрансплантациягенетическая стабильностьконтроль качества БМКПклинические исследования БМКПEasytense®cell-based medicinal productCBMPknee cartilagechondrocytesautologous chondrocytesspheroidstransplantationgenetic stabilityquality control of CBMPsclinical trials of CBMPsРабота выполнена в рамках клинических исследований биомедицинского клеточного продукта Изитенс®, спонсируемых АО «ГЕНЕРИУМ».This study was conducted as part of clinical trials of the cell-based medicinal product Easytense® funded by GENERIUM JSC.ReferencesBrittberg M, Recker D, Ilgenfritz J, Saris DBF, SUMMIT Extension Study Group. Matrix-applied characterized autologous cultured chondrocytes versus microfracture: five-year follow-up of a prospective randomized trial. 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The authors declare that there are no conflicts of interest present.