title: Inactivation of Human Nevus Tissue Using High Hydrostatic Pressure for Autologous Skin Reconstruction: A Novel Treatment for Giant Congenital Melanocytic Nevi.
abstract: Giant congenital melanocytic nevi are intractable lesions associated with a risk of melanoma. High hydrostatic pressure (HHP) technology is a safe physical method for producing decellularized tissues without chemicals. We have reported that HHP can inactivate cells present in various tissues without damaging the native extracellular matrix (ECM). The objectives of this study were to inactivate human nevus tissue using HHP and to explore the possibility of reconstructing skin using inactivated nevus in combination with cultured epidermis (CE). Human nevus specimens 8 mm in diameter were pressurized by HHP at 100, 200, 500, and 1000 MPa for 10 min. The viability of specimens just after HHP, outgrowth of cells, and viability after cultivation were evaluated to confirm the inactivation by HHP. Histological evaluation using hematoxylin-eosin staining and immunohistochemical staining for type IV collagen was performed to detect damage to the ECM of the nevus. The pressurized nevus was implanted into the subcutis of nude mice for 6 months to evaluate the retention of human cells. Then, human CE was applied on the pressurized nevus and implanted into the subcutis of nude mice. The viability of pressurized nevus was not detected just after HHP and after cultivation, and outgrowth of fibroblasts was not observed in the 200, 500, and 1000 MPa groups. Human cells were not observed after 6 months of implantation in these groups. No apparent damage to the ECM was detected in all groups; however, CE took on nevus in the 200 and 500 MPa groups, but not in the 1000 MPa group. These results indicate that human nevus tissue was inactivated by HHP at more than 200 MPa; however, HHP at 1000 MPa might cause damage that prevents the take of CE. In conclusion, all cells in nevus specimens were inactivated after HHP at more than 200 MPa and this inactivated nevus could be used as autologous dermis for covering full-thickness skin defects after nevus removal. HHP between 200 and 500 MPa will be optimal to reconstruct skin in combination with cultured epidermal autograft without damage to the ECM.
authors.affiliation: 1 Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan .
authors.forename: Chizuru
authors.initials: C
authors.lastname: Jinno
authors.affiliation: 2 Department of Plastic and Reconstructive Surgery, Kansai Medical University, Hirakata, Japan .
authors.forename: Naoki
authors.initials: N
authors.lastname: Morimoto
authors.affiliation: 3 Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, Suita, Japan .
authors.forename: Atsushi
authors.initials: A
authors.lastname: Mahara
authors.affiliation: 1 Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan .
authors.forename: Pham Hieu
authors.initials: PH
authors.lastname: Liem
authors.affiliation: 1 Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan .
authors.forename: Michiharu
authors.initials: M
authors.lastname: Sakamoto
authors.affiliation: 1 Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan .
authors.forename: Shuichi
authors.initials: S
authors.lastname: Ogino
authors.affiliation: 2 Department of Plastic and Reconstructive Surgery, Kansai Medical University, Hirakata, Japan .
authors.forename: Natsuko
authors.initials: N
authors.lastname: Kakudo
authors.affiliation: 4 Japan Tissue Engineering Co., Ltd. , Gamagori, Japan .
authors.forename: Masukazu
authors.initials: M
authors.lastname: Inoie
authors.affiliation: 5 Department of Biomedical Engineering, Osaka Institute of Technology, Osaka, Japan .
authors.forename: Toshia
authors.initials: T
authors.lastname: Fujisato
authors.affiliation: 2 Department of Plastic and Reconstructive Surgery, Kansai Medical University, Hirakata, Japan .
authors.forename: Kenji
authors.initials: K
authors.lastname: Kusumoto
authors.affiliation: 1 Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan .
authors.forename: Shigehiko
authors.initials: S
authors.lastname: Suzuki
authors.affiliation: 3 Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, Suita, Japan .
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Yamaoka
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000378:metabolism; D004817:Epidermis / Q000473:pathology; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D006874:Hydrostatic Pressure; D007150:Immunohistochemistry; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009508:Nevus, Pigmented / Q000473:pathology / Q000628:therapy*; D019919:Prosthesis Implantation; D012038:Regeneration; D012867:Skin / Q000473:pathology*; D012878:Skin Neoplasms / Q000473:pathology / Q000628:therapy*; D014022:Tissue Survival; D014182:Transplantation, Autologous
chemical_list: D024141:Collagen Type IV