Abstract

Aim. To study the viability of the tissue-engineered graft (TEG) based on the devitalized tracheal scaffold (DTS), seeded by mesenchymal stromal and epithelial cells in the experiment on rabbits with the assessment of the cytocompatibility and biocompatibility in vivo. Materials and methods. Syngeneic bone marrow-derived mesenchymal stromal cells (BM-MSCs) and lung epithelial cells of rabbit were obtained. Morphology and phenotype of the BMSCs culture were confirmed by immunofluorescent staining on CD90 and CD271 markers. Pulmonary epithelium cells obtained by enzymatic treatment of shredded lung tissue of rabbit were stained with CKPan, CK8/18 and CK14 markers of epithelial cells. The donor trachea was devitalized in three freeze-thawing cycles. Double-layer cell seeding of DTS was performed under static and dynamic culturing. Orthotopic implantation of TEG was performed at the site of the anterolateral wall defect in the rabbit trachea formed as a result of tracheal resection over four rings. The results were evaluated by computed tomography, histological and immunohisto- chemical analyses. Results. TEG implant based on DTS with two-layer cell cultures of rabbit BM-MSCs and epithelial cells was obtained. Three months after the implantation, TEG engraftment was observed, no tracheal wall stenosis was observed, but a slight narrowing of the lumen in the implantation area was observed. Viability of the tissue-engineered graft was confirmed by histological method 6 months after implantation. Epithelialization and vascularization of the tracheal wall, absence of signs of purulent inflammation and aseptic necrosis were shown. Small narrowing of the tracheal lumen was caused by chronic inflammation produced by irritation of the mucous suture material. Conclusion. The new model for evaluating the viability of a tissue engineering implant during critical airway defect closure was obtained. Two-layer vitalization of DTS with lung epithelial cells and BM-MSCs allows to create a tissue engineered structure for replacement of non-long tracheal defects in the experiment in vivo. The use of the tracheal tissue-engineered graft for orthotopic implantation showed a biocompatibility with minimal tissue reaction.

Highlights

  • Experimental orthotopic implantation of the tissue‐engineered graft of trachea based on devitalized scaffold seeded with mesenchymal and epithelial cells

  • tissue-engineered graft (TEG) implant based on devitalized tracheal scaffold (DTS) with two-layer cell cultures of rabbit bone marrow stromal cells (BMSCs) and epithelial cells was obtained

  • The article was submitted to the journal on 14.10.2019

Read more

Summary

РЕГЕНЕРАТИВНАЯ МЕДИЦИНА И КЛЕТОЧНЫЕ ТЕХНОЛОГИИ

Aim. To study the viability of the tissue-engineered graft (TEG) based on the devitalized tracheal scaffold (DTS), seeded by mesenchymal stromal and epithelial cells in the experiment on rabbits with the assessment of the cytocompatibility and biocompatibility in vivo. TEG implant based on DTS with two-layer cell cultures of rabbit BMSCs and epithelial cells was obtained. Two-layer vitalization of DTS with lung epithelial cells and BMSCs allows to create a tissue engineered structure for replacement of non-long tracheal defects in the experiment in vivo. Однако ранее не проводилась оценка в эксперименте in vivo жизнеспособности ТИК на основе девитализированной трахеальной стенки с большим остаточным содержанием экзогенных маркеров при заселении такого матрикса комбинацией из двух типов клеток. Изучить жизнеспособность тканеинженерной конструкции (ТИК) на основе девитализированного трахеального матрикса (ДТМ), заселенного мезенхимальными стромальными и эпителиальными клетками, на модели оценки жизнеспособности тканеинженерного имплантата при закрытии критического дефекта дыхательных путей у кроликов. Оценить потенциал ТИК к поддержанию стабильного просвета трахеи в области имплантации

Лабораторные животные
Получение культуры МСК КМ
Получение культуры эпителиальных клеток
Оценка фенотипа МСК КМ и эпителия трахеи
Девитализация трахеи кролика
Оценка качества девитализации матрикса
Оценка цитотоксичности ДТМ
Создание ТИК методом заселения клеточных культур
Проведение ортотопической имплантации ТИК
Мультиспиральная компьютерная томография
Морфологический анализ
Статистическая обработка результатов
Фенотипирование первичной культуры клеток
Девитализация трахеальной ткани
Цитотоксичность ДТМ и жизнеспособность заселенных клеток
Приживление ТИК после имплантации
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call