Human cultured epidermis accelerates wound healing regardless of its viability in a diabetic mouse model
Allogeneic cultured epidermis (allo-CE) is a cultured keratinocyte sheet manufactured from donor cells and promotes wound healing when used in deep dermal burns, donor sites, and chronic ulcers and serves as a wound dressing. Allo-CE is usually cryopreserved to be ready to use. However, the cryopres...
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| creator | Sakamoto, Michiharu Ogino, Shuichi Shimizu, Yoshihiro Inoie, Masukazu Lee, Sunghee Yamanaka, Hiroki Tsuge, Itaru Saito, Susumu Morimoto, Naoki |
| description | Allogeneic cultured epidermis (allo-CE) is a cultured keratinocyte sheet manufactured from donor cells and promotes wound healing when used in deep dermal burns, donor sites, and chronic ulcers and serves as a wound dressing. Allo-CE is usually cryopreserved to be ready to use. However, the cryopreservation procedure will damage the cell viability, and the influence of Allo-CE, according to its viability or wound healing process, has not been evaluated sufficiently. In this study, we aimed to prove the influence of keratinocyte viability contained in allo-CEs on wound healing. We prepared CEs with Green's method using keratinocytes obtained from a polydactyly patient and then prepared four kinds of CEs with different cell viabilities [fresh, cryopreserved, frozen, and FT (freeze and thaw)]. The cell viabilities of fresh, cryopreserved, frozen, and FT CEs were 95.7%, 59.9%, 16.7%, and 0.0%, respectively. The four CEs had homogeneous characteristics, except for small gaps found in the FT sheet by transmission electron microscopy observation. The four CEs were applied on the full-thickness skin defect of diabetic mice (BKS.Cg-Dock 7.sup.m +/+ Lepr.sup.db /Jcl), and the wound area and neoepithelium length were evaluated on days 4, 7, and 14. As a result, FT CEs without viable cells similarly promoted epithelialization on days 4 and 7 (p |
| doi_str_mv | 10.1371/journal.pone.0237985 |
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Allo-CE is usually cryopreserved to be ready to use. However, the cryopreservation procedure will damage the cell viability, and the influence of Allo-CE, according to its viability or wound healing process, has not been evaluated sufficiently. In this study, we aimed to prove the influence of keratinocyte viability contained in allo-CEs on wound healing. We prepared CEs with Green's method using keratinocytes obtained from a polydactyly patient and then prepared four kinds of CEs with different cell viabilities [fresh, cryopreserved, frozen, and FT (freeze and thaw)]. The cell viabilities of fresh, cryopreserved, frozen, and FT CEs were 95.7%, 59.9%, 16.7%, and 0.0%, respectively. The four CEs had homogeneous characteristics, except for small gaps found in the FT sheet by transmission electron microscopy observation. The four CEs were applied on the full-thickness skin defect of diabetic mice (BKS.Cg-Dock 7.sup.m +/+ Lepr.sup.db /Jcl), and the wound area and neoepithelium length were evaluated on days 4, 7, and 14. As a result, FT CEs without viable cells similarly promoted epithelialization on days 4 and 7 (p<0.05) and accelerated wound closure on day 7 (p<0.01) as fresh CEs compared with the control group. In conclusion, the promoting effect of allo-CE on wound healing does not depend on cell viability. Lyophilized CEs may be a suitable wound dressing with a long storage period at room temperature.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0237985</identifier><identifier>PMID: 32822395</identifier><language>eng</language><publisher>San Francisco: Public Library of Science</publisher><subject>Biology and Life Sciences ; Burns ; Care and treatment ; Cell viability ; Cryopreservation ; Diabetes ; Diabetes mellitus ; Electron microscopy ; Epidermis ; Freeze-thawing ; Health aspects ; Keratinocytes ; Medicine ; Medicine and Health Sciences ; Methods ; Microscopy ; Polydactyly ; Reconstructive surgery ; Research and Analysis Methods ; Room temperature ; Skin ; Skin grafts ; Tissue culture ; Tissue engineering ; Transmission electron microscopy ; Ulcers ; Wound care ; Wound healing</subject><ispartof>PloS one, 2020-08, Vol.15 (8), p.e0237985-e0237985</ispartof><rights>COPYRIGHT 2020 Public Library of Science</rights><rights>2020 Sakamoto et al. 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Allo-CE is usually cryopreserved to be ready to use. However, the cryopreservation procedure will damage the cell viability, and the influence of Allo-CE, according to its viability or wound healing process, has not been evaluated sufficiently. In this study, we aimed to prove the influence of keratinocyte viability contained in allo-CEs on wound healing. We prepared CEs with Green's method using keratinocytes obtained from a polydactyly patient and then prepared four kinds of CEs with different cell viabilities [fresh, cryopreserved, frozen, and FT (freeze and thaw)]. The cell viabilities of fresh, cryopreserved, frozen, and FT CEs were 95.7%, 59.9%, 16.7%, and 0.0%, respectively. The four CEs had homogeneous characteristics, except for small gaps found in the FT sheet by transmission electron microscopy observation. The four CEs were applied on the full-thickness skin defect of diabetic mice (BKS.Cg-Dock 7.sup.m +/+ Lepr.sup.db /Jcl), and the wound area and neoepithelium length were evaluated on days 4, 7, and 14. As a result, FT CEs without viable cells similarly promoted epithelialization on days 4 and 7 (p<0.05) and accelerated wound closure on day 7 (p<0.01) as fresh CEs compared with the control group. In conclusion, the promoting effect of allo-CE on wound healing does not depend on cell viability. Lyophilized CEs may be a suitable wound dressing with a long storage period at room temperature.</description><subject>Biology and Life Sciences</subject><subject>Burns</subject><subject>Care and treatment</subject><subject>Cell viability</subject><subject>Cryopreservation</subject><subject>Diabetes</subject><subject>Diabetes mellitus</subject><subject>Electron microscopy</subject><subject>Epidermis</subject><subject>Freeze-thawing</subject><subject>Health aspects</subject><subject>Keratinocytes</subject><subject>Medicine</subject><subject>Medicine and Health Sciences</subject><subject>Methods</subject><subject>Microscopy</subject><subject>Polydactyly</subject><subject>Reconstructive surgery</subject><subject>Research and Analysis Methods</subject><subject>Room temperature</subject><subject>Skin</subject><subject>Skin grafts</subject><subject>Tissue culture</subject><subject>Tissue engineering</subject><subject>Transmission electron 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one</jtitle><date>2020-08-21</date><risdate>2020</risdate><volume>15</volume><issue>8</issue><spage>e0237985</spage><epage>e0237985</epage><pages>e0237985-e0237985</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Allogeneic cultured epidermis (allo-CE) is a cultured keratinocyte sheet manufactured from donor cells and promotes wound healing when used in deep dermal burns, donor sites, and chronic ulcers and serves as a wound dressing. Allo-CE is usually cryopreserved to be ready to use. However, the cryopreservation procedure will damage the cell viability, and the influence of Allo-CE, according to its viability or wound healing process, has not been evaluated sufficiently. In this study, we aimed to prove the influence of keratinocyte viability contained in allo-CEs on wound healing. We prepared CEs with Green's method using keratinocytes obtained from a polydactyly patient and then prepared four kinds of CEs with different cell viabilities [fresh, cryopreserved, frozen, and FT (freeze and thaw)]. The cell viabilities of fresh, cryopreserved, frozen, and FT CEs were 95.7%, 59.9%, 16.7%, and 0.0%, respectively. The four CEs had homogeneous characteristics, except for small gaps found in the FT sheet by transmission electron microscopy observation. The four CEs were applied on the full-thickness skin defect of diabetic mice (BKS.Cg-Dock 7.sup.m +/+ Lepr.sup.db /Jcl), and the wound area and neoepithelium length were evaluated on days 4, 7, and 14. As a result, FT CEs without viable cells similarly promoted epithelialization on days 4 and 7 (p<0.05) and accelerated wound closure on day 7 (p<0.01) as fresh CEs compared with the control group. In conclusion, the promoting effect of allo-CE on wound healing does not depend on cell viability. Lyophilized CEs may be a suitable wound dressing with a long storage period at room temperature.</abstract><cop>San Francisco</cop><pub>Public Library of Science</pub><pmid>32822395</pmid><doi>10.1371/journal.pone.0237985</doi><tpages>e0237985</tpages><orcidid>https://orcid.org/0000-0002-0712-3172</orcidid><orcidid>https://orcid.org/0000-0002-0024-6807</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Biology and Life Sciences Burns Care and treatment Cell viability Cryopreservation Diabetes Diabetes mellitus Electron microscopy Epidermis Freeze-thawing Health aspects Keratinocytes Medicine Medicine and Health Sciences Methods Microscopy Polydactyly Reconstructive surgery Research and Analysis Methods Room temperature Skin Skin grafts Tissue culture Tissue engineering Transmission electron microscopy Ulcers Wound care Wound healing |
| title | Human cultured epidermis accelerates wound healing regardless of its viability in a diabetic mouse model |
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