title: Corneal Regeneration by Deep Anterior Lamellar Keratoplasty (DALK) Using Decellularized Corneal Matrix.
abstract: The purpose of this study is to demonstrate the feasibility of DALK using a decellularized corneal matrix obtained by HHP methodology. Porcine corneas were hydrostatically pressurized at 980 MPa at 10°C for 10 minutes to destroy the cells, followed by washing with EGM-2 medium to remove the cell debris. The HHP-treated corneas were stained with H-E to assess the efficacy of decellularization. The decellularized corneal matrix of 300 μm thickness and 6.0 mm diameter was transplanted onto a 6.0 mm diameter keratectomy wound. The time course of regeneration on the decellularized corneal matrix was evaluated by haze grading score, fluorescein staining, and immunohistochemistry. H-E staining revealed that no cell nuclei were observed in the decellularized corneal matrix. The decellularized corneal matrices were opaque immediately after transplantation, but became completely transparent after 4 months. Fluorescein staining revealed that initial migration of epithelial cells over the grafts was slow, taking 3 months to completely cover the implant. Histological sections revealed that the implanted decellularized corneal matrix was completely integrated with the receptive rabbit cornea, and keratocytes infiltrated into the decellularized corneal matrix 6 months after transplantation. No inflammatory cells such as macrophages, or neovascularization, were observed during the implantation period. The decellularized corneal matrix improved corneal transparency, and remodelled the graft after being transplanted, demonstrating that the matrix obtained by HHP was a useful graft for corneal tissue regeneration.
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Hashimoto
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Funamoto
authors.affiliation: Department of Ophthalmology, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Shuji
authors.initials: S
authors.lastname: Sasaki
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Negishi
authors.affiliation: International Center for Materials Nanoarchitectonics, National Institute for Materials Science, Tsukuba, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Honda
authors.affiliation: International Center for Materials Nanoarchitectonics, National Institute for Materials Science, Tsukuba, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Hattori
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Kwangwoo
authors.initials: K
authors.lastname: Nam
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Kimura
authors.affiliation: Department of Ophthalmology, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Mochizuki
authors.affiliation: International Center for Materials Nanoarchitectonics, National Institute for Materials Science, Tsukuba, Japan.
authors.forename: Hisatoshi
authors.initials: H
authors.lastname: Kobayashi
authors.affiliation: Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Akio
authors.initials: A
authors.lastname: Kishida
mesh_terms: D000818:Animals; D003315:Cornea / Q000473:pathology / Q000502:physiology*; D003316:Corneal Diseases / Q000601:surgery*; D016039:Corneal Transplantation; D008297:Male; D011817:Rabbits; D012038:Regeneration; D044968:Regenerative Medicine; D034421:Sus scrofa