title: The Alteration of the Epidermal Basement Membrane Complex of Human Nevus Tissue and Keratinocyte Attachment after High Hydrostatic Pressurization.
abstract: We previously reported that human nevus tissue was inactivated after high hydrostatic pressure (HHP) higher than 200 MPa and that human cultured epidermis (hCE) engrafted on the pressurized nevus at 200 MPa but not at 1000 MPa. In this study, we explore the changes to the epidermal basement membrane in detail and elucidate the cause of the difference in hCE engraftment. Nevus specimens of 8 mm in diameter were divided into five groups (control and 100, 200, 500, and 1000 MPa). Immediately after HHP, immunohistochemical staining was performed to detect the presence of laminin-332 and type VII collagen, and the specimens were observed by transmission electron microscopy (TEM). hCE was placed on the pressurized nevus specimens in the 200, 500, and 1000 MPa groups and implanted into the subcutis of nude mice; the specimens were harvested at 14 days after implantation. Then, human keratinocytes were seeded on the pressurized nevus and the attachment was evaluated. The immunohistochemical staining results revealed that the control and 100 MPa, 200 MPa, and 500 MPa groups were positive for type VII collagen and laminin-332 immediately after HHP. TEM showed that, in all of the groups, the lamina densa existed; however, anchoring fibrils were not clearly observed in the 500 or 1000 MPa groups. Although the hCE took in the 200 and 500 MPa groups, keratinocyte attachment was only confirmed in the 200 MPa group. This result indicates that HHP at 200 MPa is preferable for inactivating nevus tissue to allow its reuse for skin reconstruction in the clinical setting.
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kansai Medical University, Hirakata, Japan.
authors.forename: Naoki
authors.identifier: 0000-0002-0712-3172
authors.initials: N
authors.lastname: Morimoto
authors.affiliation: 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: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, Suita, Japan.
authors.forename: Atsushi
authors.identifier: 0000-0002-4075-6019
authors.initials: A
authors.lastname: Mahara
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kansai Medical University, Hirakata, Japan.
authors.forename: Michiharu
authors.initials: M
authors.lastname: Sakamoto
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kansai Medical University, Hirakata, Japan.
authors.forename: Natsuko
authors.initials: N
authors.lastname: Kakudo
authors.affiliation: Japan Tissue Engineering Co., Ltd., Gamagori, Japan.
authors.forename: Masukazu
authors.initials: M
authors.lastname: Inoie
authors.affiliation: Department of Biomedical Engineering, Osaka Institute of Technology, Osaka, Japan.
authors.forename: Toshia
authors.initials: T
authors.lastname: Fujisato
authors.affiliation: 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: Department of Plastic and Reconstructive Surgery, Kansai Medical University, Hirakata, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kusumoto
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, Suita, Japan.
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Yamaoka
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D001485:Basement Membrane / Q000378:metabolism / Q000473:pathology*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002648:Child; D002675:Child, Preschool; D024143:Collagen Type VII / Q000378:metabolism; D004817:Epidermis / Q000378:metabolism / Q000473:pathology*; D005260:Female; D006801:Humans; D006874:Hydrostatic Pressure; D007223:Infant; D015603:Keratinocytes / Q000378:metabolism / Q000473:pathology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009506:Nevus / Q000378:metabolism / Q000473:pathology*; D055815:Young Adult
chemical_list: D015815:Cell Adhesion Molecules; D024143:Collagen Type VII