title: Regeneration of corneal epithelium utilizing a collagen vitrigel membrane in rabbit models for corneal stromal wound and limbal stem cell deficiency.
abstract: OBJECTIVE
  This study was performed to evaluate the potential of a collagen-based membrane, collagen vitrigel (CV), for reconstructing corneal epithelium in the stromal wound and limbal stem cell deficiency (LSCD) models.
  
  
  METHODS
  Three groups of rabbits were used in the stromal wound model: CV affixed using fibrin glue (CV + FG group, n = 9), fibrin glue only (FG group, n = 3) and an untreated control group (n = 3). In the LSCD model, one group received CV containing human limbal epithelial cells (CV + hLEC group, n = 2) and the other was an untreated control (n = 1). Gross observation, including fluorescent staining, pathological examination, immunohistochemistry and electron microscopy, was used to evaluate the effect of CV on the corneal epithelium.
  
  
  RESULTS
  In the stromal wound model, fluorescent staining showed that epithelial reconstruction occurred as rapidly in the CV + FG group as it did in the control group. The pathological examination proved that the CV supported a healthy corneal epithelium in the CV + FG group, whereas FG led to hypertrophy and inappropriate differentiation of corneal epithelium in the FG group. In the LSCD model, the corneas in the CV + hLEC group showed sustained tissue transparency with good epithelialization, low inflammatory response and reduced neovascularization. However, the control cornea was translucent and showed high amounts of inflammation and neovascularization.
  
  
  CONCLUSIONS
  We have demonstrated that CV supports corneal epithelial differentiation and prevents epithelial hypertrophy, in addition to serving as a scaffold for hLEC transplantation, without complications.
authors.affiliation: Translational Tissue Engineering Center, Wilmer Eye Institute and Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: J Jeremy
authors.initials: JJ
authors.lastname: Chae
authors.forename: Winnette McIntosh
authors.initials: WM
authors.lastname: Ambrose
authors.forename: Freddy A
authors.initials: FA
authors.lastname: Espinoza
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Mulreany
authors.forename: Shengyong
authors.initials: S
authors.lastname: Ng
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Takezawa
authors.forename: Morgana M
authors.initials: MM
authors.lastname: Trexler
authors.forename: Oliver D
authors.initials: OD
authors.lastname: Schein
authors.forename: Roy S
authors.initials: RS
authors.lastname: Chuck
authors.forename: Jennifer H
authors.initials: JH
authors.lastname: Elisseeff
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D018929:Cell Culture Techniques; D017690:Cell Transplantation; D024042:Collagen Type I; D003316:Corneal Diseases / Q000628:therapy*; D003319:Corneal Stroma / Q000293:injuries; D004195:Disease Models, Animal; D019573:Epithelium, Corneal / Q000166:cytology / Q000502:physiology*; D007150:Immunohistochemistry; D016850:Limbus Corneae / Q000473:pathology*; D008567:Membranes, Artificial; D008854:Microscopy, Electron; D011817:Rabbits; D012038:Regeneration / Q000502:physiology*; D013234:Stem Cells / Q000473:pathology; D054457:Tissue Scaffolds; D014945:Wound Healing
chemical_list: D001672:Biocompatible Materials; D024042:Collagen Type I; D008567:Membranes, Artificial
keywords: cell transplantation; collagen vitrigel; corneal epithelium; corneal wound; fibrin glue; limbal stem cell deficiency