title: Development of a bioengineered corneal endothelial cell sheet to fit the corneal curvature.
abstract: OBJECTIVE
  To evaluate a novel bioengineered corneal endothelial cell sheet that fits the curvature of the posterior corneal surface.
  
  
  METHODS
  A spherically curved gelatin hydrogel sheet (SCGS) was prepared by the dehydrothermal cross-linking method, and its permeability to water and protein was tested. Monkey corneal endothelial cells (MCECs) were seeded onto these hydrogel sheets, and the cells were examined by immunohistochemistry. Then MCEC-SCGS constructs were transplanted in monkeys with bullous keratopathy to assess the efficacy of the hydrogel sheets as a scaffold.
  
  
  RESULTS
  The hydrogel sheets showed similar permeability to water and protein as that of atelocollagen and vitrigel sheets. After transplantation, the SCGS did not show wrinkling and adhered tightly to the posterior corneal surface, whereas the flat sheets developed wrinkles that inhibited tight adhesion. Monkey corneal endothelial cells grown on hydrogel sheets expressed anti-zonula occludens-1 (ZO-1), N-cadherin, and sodium, potassium, and adenosine triphosphatase (Na,K-ATPase) along the plasma membrane. In a monkey model of bullous keratopathy, transplanted MCEC-SCGS constructs showed good adhesion to the posterior corneal surface, with subsequent improvement of corneal edema and transparency.
  
  
  CONCLUSIONS
  A novel MCEC-SCGS construct was effective in a monkey model of bullous keratopathy. The SCGS achieves close adhesion to the posterior corneal surface without wrinkling and may contribute to clinical transplantation of corneal endothelial cell sheets.
authors.affiliation: Corneal Regeneration Research Team, Foundation for Biomedical Research and Innovation, Kobe, Japan.
authors.forename: Miwa
authors.initials: M
authors.lastname: Kimoto
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Shima
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Hiraoka
authors.forename: Shiro
authors.initials: S
authors.lastname: Amano
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamagami
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D003315:Cornea / Q000601:surgery*; D016039:Corneal Transplantation / Q000379:methods*; D004195:Disease Models, Animal; D004728:Endothelium, Corneal / Q000166:cytology*; D000882:Haplorhini; D006801:Humans; D007150:Immunohistochemistry; D007634:Keratitis / Q000473:pathology / Q000601:surgery*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
keywords: corneal endothelium; curved gelatin hydrogel sheet; transplantation