Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment
Decellularized tissues are considered superior scaffolds for cell cultures, preserving the microstructure of native tissues and delivering many kinds of cytokines. High hydrostatic pressure (HHP) treatment could remove cells physically from biological tissues rather than chemical methods. However, t...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-11-01
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| Series: | Micromachines |
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| Online Access: | https://www.mdpi.com/2072-666X/12/12/1486 |
| _version_ | 1797502423008280576 |
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| author | Daiki Zemmyo Masashi Yamamoto Shogo Miyata |
| author_facet | Daiki Zemmyo Masashi Yamamoto Shogo Miyata |
| author_sort | Daiki Zemmyo |
| collection | DOAJ |
| description | Decellularized tissues are considered superior scaffolds for cell cultures, preserving the microstructure of native tissues and delivering many kinds of cytokines. High hydrostatic pressure (HHP) treatment could remove cells physically from biological tissues rather than chemical methods. However, there are some risks of inducing destruction or denaturation of extracellular matrices (ECMs) at an ultrahigh level of HHP. Therefore, efficient decellularization using moderate HHP is required to remove almost all cells simultaneously to suppress tissue damage. In this study, we proposed a novel decellularization method using a moderate HHP with supercooling pretreatment. To validate the decellularization method, a supercooling device was developed to incubate human dermal fibroblasts or collagen gels in a supercooled state. The cell suspension and collagen gels were subjected to 100, 150, and 200 MPa of HHP after supercooling pretreatment, respectively. After applying HHP, the viability and morphology of the cells and the collagen network structure of the gels were evaluated. The viability of cells decreased dramatically after HHP application with supercooling pretreatment, whereas the microstructures of collagen gels were preserved and cell adhesivity was retained after HHP application. In conclusion, it was revealed that supercooling pretreatment promoted the denaturation of the cell membrane to improve the efficacy of decellularization using static application of moderate HHP. Furthermore, it was demonstrated that the HHP with supercooling pretreatment did not degenerate and damage the microstructure in collagen gels. |
| first_indexed | 2024-03-10T03:32:52Z |
| format | Article |
| id | doaj.art-01401f0792104aa1beeece3e7c58ff5a |
| institution | Directory Open Access Journal |
| issn | 2072-666X |
| language | English |
| last_indexed | 2024-03-10T03:32:52Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Micromachines |
| spelling | doaj.art-01401f0792104aa1beeece3e7c58ff5a2023-11-23T09:36:04ZengMDPI AGMicromachines2072-666X2021-11-011212148610.3390/mi12121486Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling PretreatmentDaiki Zemmyo0Masashi Yamamoto1Shogo Miyata2Graduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Yokohama 223-8522, JapanGraduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Yokohama 223-8522, JapanDepartment of Mechanical Engineering, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Yokohama 223-8522, JapanDecellularized tissues are considered superior scaffolds for cell cultures, preserving the microstructure of native tissues and delivering many kinds of cytokines. High hydrostatic pressure (HHP) treatment could remove cells physically from biological tissues rather than chemical methods. However, there are some risks of inducing destruction or denaturation of extracellular matrices (ECMs) at an ultrahigh level of HHP. Therefore, efficient decellularization using moderate HHP is required to remove almost all cells simultaneously to suppress tissue damage. In this study, we proposed a novel decellularization method using a moderate HHP with supercooling pretreatment. To validate the decellularization method, a supercooling device was developed to incubate human dermal fibroblasts or collagen gels in a supercooled state. The cell suspension and collagen gels were subjected to 100, 150, and 200 MPa of HHP after supercooling pretreatment, respectively. After applying HHP, the viability and morphology of the cells and the collagen network structure of the gels were evaluated. The viability of cells decreased dramatically after HHP application with supercooling pretreatment, whereas the microstructures of collagen gels were preserved and cell adhesivity was retained after HHP application. In conclusion, it was revealed that supercooling pretreatment promoted the denaturation of the cell membrane to improve the efficacy of decellularization using static application of moderate HHP. Furthermore, it was demonstrated that the HHP with supercooling pretreatment did not degenerate and damage the microstructure in collagen gels.https://www.mdpi.com/2072-666X/12/12/1486decellularizationhigh hydrostatic pressurecyclic hydrostatic pressuresupercoolingcollagen structurepreservation of structure |
| spellingShingle | Daiki Zemmyo Masashi Yamamoto Shogo Miyata Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment Micromachines decellularization high hydrostatic pressure cyclic hydrostatic pressure supercooling collagen structure preservation of structure |
| title | Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment |
| title_full | Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment |
| title_fullStr | Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment |
| title_full_unstemmed | Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment |
| title_short | Efficient Decellularization by Application of Moderate High Hydrostatic Pressure with Supercooling Pretreatment |
| title_sort | efficient decellularization by application of moderate high hydrostatic pressure with supercooling pretreatment |
| topic | decellularization high hydrostatic pressure cyclic hydrostatic pressure supercooling collagen structure preservation of structure |
| url | https://www.mdpi.com/2072-666X/12/12/1486 |
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