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Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure

Decellularized tissues are promising materials that mainly consist of extracellular matrices (ECMs) obtained by removing all cells from organs and tissues. High hydrostatic pressure (HHP) has been used for decellularization to remove cells physically from organs or tissues rather than by chemical me...

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Detalles Bibliográficos
Autores principales: Zemmyo, Daiki, Yamamoto, Masashi, Miyata, Shogo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7696941/
https://www.ncbi.nlm.nih.gov/pubmed/33203164
http://dx.doi.org/10.3390/mi11111008
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author Zemmyo, Daiki
Yamamoto, Masashi
Miyata, Shogo
author_facet Zemmyo, Daiki
Yamamoto, Masashi
Miyata, Shogo
author_sort Zemmyo, Daiki
collection PubMed
description Decellularized tissues are promising materials that mainly consist of extracellular matrices (ECMs) obtained by removing all cells from organs and tissues. High hydrostatic pressure (HHP) has been used for decellularization to remove cells physically from organs or tissues rather than by chemical methods. However, ultrahigh pressure induces denaturation of the ECM structure. In this study, we examined the effects of cyclic HHP at low and high pressures on the cell membrane structure to establish a novel decellularization method that enables decellularization without the denaturation of the ECM. A decellularization device using cyclic HHP (maximum pressure: 250 MPa, cycle number: 5) was developed. NB1RGB cell suspension was injected into a plastic bag to be subjected to cyclic HHP. After applying cyclic HHP, the amount of DNA inside the cells and the morphological changes of the cells were evaluated. As a result, the amount of DNA inside the cells decreased after the cyclic HHP compared to the static HHP. In addition, cyclic HHP was suggested to promote the destruction of the cell and nuclear membrane. In conclusion, it was revealed that the cell structure could be denatured and destroyed by cyclic HHP at a lower level than that of previous approaches.
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spelling pubmed-76969412020-11-29 Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure Zemmyo, Daiki Yamamoto, Masashi Miyata, Shogo Micromachines (Basel) Article Decellularized tissues are promising materials that mainly consist of extracellular matrices (ECMs) obtained by removing all cells from organs and tissues. High hydrostatic pressure (HHP) has been used for decellularization to remove cells physically from organs or tissues rather than by chemical methods. However, ultrahigh pressure induces denaturation of the ECM structure. In this study, we examined the effects of cyclic HHP at low and high pressures on the cell membrane structure to establish a novel decellularization method that enables decellularization without the denaturation of the ECM. A decellularization device using cyclic HHP (maximum pressure: 250 MPa, cycle number: 5) was developed. NB1RGB cell suspension was injected into a plastic bag to be subjected to cyclic HHP. After applying cyclic HHP, the amount of DNA inside the cells and the morphological changes of the cells were evaluated. As a result, the amount of DNA inside the cells decreased after the cyclic HHP compared to the static HHP. In addition, cyclic HHP was suggested to promote the destruction of the cell and nuclear membrane. In conclusion, it was revealed that the cell structure could be denatured and destroyed by cyclic HHP at a lower level than that of previous approaches. MDPI 2020-11-15 /pmc/articles/PMC7696941/ /pubmed/33203164 http://dx.doi.org/10.3390/mi11111008 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zemmyo, Daiki
Yamamoto, Masashi
Miyata, Shogo
Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure
title Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure
title_full Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure
title_fullStr Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure
title_full_unstemmed Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure
title_short Fundamental Study of Decellularization Method Using Cyclic Application of High Hydrostatic Pressure
title_sort fundamental study of decellularization method using cyclic application of high hydrostatic pressure
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7696941/
https://www.ncbi.nlm.nih.gov/pubmed/33203164
http://dx.doi.org/10.3390/mi11111008
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