Regeneration of corneal epithelium utilizing a collagen vitrigel membrane in rabbit models for corneal stromal wound and limbal stem cell deficiency

Acta Ophthalmol. 2015 Feb;93(1):e57-66. doi: 10.1111/aos.12503. Epub 2014 Dec 11.

Abstract

Purpose: 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.

Conclusion: 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.

Keywords: cell transplantation; collagen vitrigel; corneal epithelium; corneal wound; fibrin glue; limbal stem cell deficiency.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Biocompatible Materials
  • Cell Culture Techniques
  • Cell Transplantation*
  • Collagen Type I*
  • Corneal Diseases / therapy*
  • Corneal Stroma / injuries
  • Disease Models, Animal
  • Epithelium, Corneal / cytology
  • Epithelium, Corneal / physiology*
  • Immunohistochemistry
  • Limbus Corneae / pathology*
  • Membranes, Artificial*
  • Microscopy, Electron
  • Rabbits
  • Regeneration / physiology*
  • Stem Cells / pathology
  • Tissue Scaffolds
  • Wound Healing

Substances

  • Biocompatible Materials
  • Collagen Type I
  • Membranes, Artificial