morphoМорфологические ведомостиMorphological newsletter1812-31712686-8741Private Medical University REAVIZ10.20340/mv-mn.2016.24(4):36-45morpho-215Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯRESEARCH ARTICLESСРАВНИТЕЛЬНЫЙ АНАЛИЗ ЭФФЕКТИВНОСТИ ПРОСТОГО И МНОГОКОМПОНЕНТНОГО МЕТОДОВ ЩЕЛОЧНОЙ ДЕЦЕЛЛЮЛЯРИЗАЦИИ НА ПРИМЕРЕ ОЧИСТКИ ВОЛОКНИСТОГО ВНЕКЛЕТОЧНОГО МАТРИКСА ДЕРМЫThe comparative efficiency analysis of simple and multicomponent alkaline decellularization on the example of purification of the fibrous extracellular matrix of the dermaКалмыковаНина ВладимировнаKalmykovaN. V.k.nina.v@mail.ruДемьяненкоИлья АлександровичDemyanenkoI. A.ilya871@rambler.ruШевлягинаНаталья ВладимировнаShevlyaginaN. V.microbanatomy@gmail.comАндреевскаяСветлана ГеоргиевнаAndreevskayaS. G.hacaranda@yandex.ruСусловАнатолий ПетровичSuslovA. P.Anatoly.Suslov@gmail.comФНИЦЭМ им. Н.Ф.ГамалеиRussian Federation2016301220162443645Copyright © Калмыкова Н.В., Демьяненко И.А., Шевлягина Н.В., Андреевская С.Г., Суслов А.П., 20162016Калмыкова Н.В., Демьяненко И.А., Шевлягина Н.В., Андреевская С.Г., Суслов А.П.Kalmykova N.V., Demyanenko I.A., Shevlyagina N.V., Andreevskaya S.G., Suslov A.P.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.morpholetter.com/jour/article/view/215

Щелочная децеллюляризация является одним из наиболее распространенных методов получения очищенного внеклеточного матрикса (BKM). Тем не менее, в литературе отсутствуют работы, посвященные сравнительному анализу эффективности различных методов щелочной очистки BKM. B настоящей работе было проведено сравнительное исследование влияния простого метода децеллюляризации раствором гидроксида натрия и многкомпонентного метода несколькими различными растворами на основе гидроксида натрия и сульфата натрия на биохимические, морфологические и термомеханические характеристики волокнистого внеклеточного матрикса дермы крупного рогатого скота (KPC), а именно быка домашнего Bos taurus taurus. Показано, что оба метода вызывают эффективное удаление клеточных компонентов из BKM. Однако в образцах, очищенных мнокомпонентньм методом выявлено меньшее количество остаточной дцДНК. Результаты морфологического и термомеханического исследований свидетельствуют о том, что применение многокомпонентной децеллюляризации по сравнению с простой обработкой гидроксидом натрия приводит к уменьшению дезорганизации волокнистой сети BKM с сохранением структуры коллагеновых волокон, а также к увеличению эластичности материала. Полученные данные указывают на большую сохранность нативной структуры децеллюляризированного BKM дермы KPC при применении щєлочной очистки с использованием растворов, содержащих соли сульфата. По резуль татам работы можно заключить, что использование многокомпонентного метода является более предпочтительным для осуществления щелочной децеллюляризации дермы KPC.

Alkaline decellularization represents one of the most common methods for obtaining acellular matrix (ECM). However, there is no published data concerning the effectiveness of different alkaline ECM decellularization protocols. In the present study comparative efficiency analysis of the simple decellularization protocol by sodium hydroxide solution and complex protocol by several different solutions based on sodium hydroxide and sodium sulfate was carried out. The biochemical, morphological, and thermo-mechanical characteristics of the acellular fibrous extracellular matrix of the bovine (Bos taurus taurus) dermis were evaluated. It was shown that both protocols effectively remove cellular components from the ECM. However, applying complex protocol led to the reduction in the amount of residual dsDNA in the ECM. The results of morphological and thermomechanical studies suggest that the use of a complex protocol in comparison with a simple treatment with sodium hydroxide leads to decrease in the ECM fiber network disorganization, retaining structure of collagen fibers, and also to an increase in the elasticity of the material. Thus, the use of alkaline treatment with solutions containing sulfate better preserve the native structure of the decellularized ECM from bovine dermis. According to the obtained data it can be concluded that the use of the complex protocol for the alkaline decellularization of bovine dermis is more preferable.

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