title: Development and evaluation of porcine atelocollagen vitrigel membrane with a spherical curve and transplantable artificial corneal endothelial grafts.
abstract: OBJECTIVE
  To develop a collagen vitrigel (CV) optimized as a corneal endothelial cell (CEC) carrier and create an artificial corneal endothelial graft.
  
  
  METHODS
  We first developed a flat-shaped collagen vitrigel for regenerative medicine (CV-RM) using porcine atelocollagen and ultraviolet (UV) irradiation. The optimal UV amount was determined by measuring the CV-RM transparency under various irradiating conditions. The collagen vitrigel for corneal endothelial regenerative treatment (CV-CERT), a transparent porcine atelocollagen with a curved shape, was made using spherically curved molds and UV irradiation. The membrane permeability of the CV-CERT was tested in vitro. The biocompatibility, transparency, and adhesiveness of the CV-CERT were evaluated in rabbit eyes. We also developed a culture technique for distributing human CECs on the curved CV-CERT.
  
  
  RESULTS
  The optimal amount of UV irradiation for CV-RM transparency was 2400 mJ/cm(2). Membrane permeability of CV-CERT at day 5 was higher than that of commercially available CV (P = 0.032). The CV-CERT was transparent and biocompatible in rabbit corneas for up to 4 months. The CV-CERT remained attached to the rabbit corneal posterior surface, whereas the flat-shaped CV-RM, differing only in shape from the CV-CERT, dislocated soon after surgery. Human CECs seeded on the CV-CERT using our technique were evenly distributed with a single layer structure and a mean cell density of 2650 ± 100 cells/mm(2).
  
  
  CONCLUSIONS
  We developed a transparent and biocompatible porcine-derived atelocollagen vitrigel membrane with a spherical curvature. A transplantable artificial endothelial graft was created by combining cultured human CECs and the CV-CERT.
authors.affiliation: Department of Ophthalmology, University of Tokyo Graduate School of Medicine, Tokyo, Japan.
authors.forename: Junko
authors.initials: J
authors.lastname: Yoshida
authors.affiliation: Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki, Japan.
authors.forename: Ayumi
authors.initials: A
authors.lastname: Oshikata-Miyazaki
authors.affiliation: Department of Ophthalmology, University of Tokyo Graduate School of Medicine, Tokyo, Japan.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Yokoo
authors.affiliation: Department of Ophthalmology, University of Tokyo Graduate School of Medicine, Tokyo, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamagami
authors.affiliation: Division of Animal Sciences, National Institute of Agrobiological Sciences, Ibaraki, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Takezawa
authors.affiliation: Department of Ophthalmology, University of Tokyo Graduate School of Medicine, Tokyo, Japan.
authors.forename: Shiro
authors.initials: S
authors.lastname: Amano
mesh_terms: D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D003315:Cornea / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D003318:Corneal Opacity / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D016039:Corneal Transplantation / Q000379:methods*; D004195:Disease Models, Animal; D004728:Endothelium, Corneal / Q000637:transplantation*; D006801:Humans; D008297:Male; D008567:Membranes, Artificial; D011817:Rabbits; D013552:Swine; D016896:Treatment Outcome
chemical_list: D008567:Membranes, Artificial; C059821:atelocollagen; D003094:Collagen
keywords: collagen vitrigel; corneal endothelial cell transplantation; regenerative medicine