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

J. Jeremy Chae, Winnette McIntosh Ambrose, Freddy A. Espinoza, Daniel G. Mulreany, Shengyong Ng, Toshiaki Takezawa, Morgana M. Trexler, Oliver D Schein, Roy S. Chuck, Jennifer Hartt Elisseeff

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)e57-e66
JournalActa Ophthalmologica
Volume93
Issue number1
DOIs
StatePublished - Feb 1 2015

Fingerprint

Corneal Epithelium
Regeneration
Collagen
Stem Cells
Rabbits
Membranes
Wounds and Injuries
Fibrin Tissue Adhesive
Cornea
Hypertrophy
Staining and Labeling
Control Groups
Electron Microscopy
Transplantation
Epithelial Cells
Immunohistochemistry

Keywords

  • cell transplantation
  • collagen vitrigel
  • corneal epithelium
  • corneal wound
  • fibrin glue
  • limbal stem cell deficiency

ASJC Scopus subject areas

  • Ophthalmology
  • Medicine(all)

Cite this

Regeneration of corneal epithelium utilizing a collagen vitrigel membrane in rabbit models for corneal stromal wound and limbal stem cell deficiency. / Chae, J. Jeremy; McIntosh Ambrose, Winnette; Espinoza, Freddy A.; Mulreany, Daniel G.; Ng, Shengyong; Takezawa, Toshiaki; Trexler, Morgana M.; Schein, Oliver D; Chuck, Roy S.; Elisseeff, Jennifer Hartt.

In: Acta Ophthalmologica, Vol. 93, No. 1, 01.02.2015, p. e57-e66.

Research output: Contribution to journalArticle

Chae, J. Jeremy ; McIntosh Ambrose, Winnette ; Espinoza, Freddy A. ; Mulreany, Daniel G. ; Ng, Shengyong ; Takezawa, Toshiaki ; Trexler, Morgana M. ; Schein, Oliver D ; Chuck, Roy S. ; Elisseeff, Jennifer Hartt. / Regeneration of corneal epithelium utilizing a collagen vitrigel membrane in rabbit models for corneal stromal wound and limbal stem cell deficiency. In: Acta Ophthalmologica. 2015 ; Vol. 93, No. 1. pp. e57-e66.
@article{2852bb77dd524fb88441fc1d46ee1126,
title = "Regeneration of corneal epithelium utilizing a collagen vitrigel membrane in rabbit models for corneal stromal wound and limbal stem cell deficiency",
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",
author = "Chae, {J. Jeremy} and {McIntosh Ambrose}, Winnette and Espinoza, {Freddy A.} and Mulreany, {Daniel G.} and Shengyong Ng and Toshiaki Takezawa and Trexler, {Morgana M.} and Schein, {Oliver D} and Chuck, {Roy S.} and Elisseeff, {Jennifer Hartt}",
year = "2015",
month = "2",
day = "1",
doi = "10.1111/aos.12503",
language = "English (US)",
volume = "93",
pages = "e57--e66",
journal = "Acta Ophthalmologica",
issn = "0001-639X",
publisher = "Copenhagen Scriptor",
number = "1",

}

TY - JOUR

T1 - Regeneration of corneal epithelium utilizing a collagen vitrigel membrane in rabbit models for corneal stromal wound and limbal stem cell deficiency

AU - Chae, J. Jeremy

AU - McIntosh Ambrose, Winnette

AU - Espinoza, Freddy A.

AU - Mulreany, Daniel G.

AU - Ng, Shengyong

AU - Takezawa, Toshiaki

AU - Trexler, Morgana M.

AU - Schein, Oliver D

AU - Chuck, Roy S.

AU - Elisseeff, Jennifer Hartt

PY - 2015/2/1

Y1 - 2015/2/1

N2 - 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.

AB - 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.

KW - cell transplantation

KW - collagen vitrigel

KW - corneal epithelium

KW - corneal wound

KW - fibrin glue

KW - limbal stem cell deficiency

UR - http://www.scopus.com/inward/record.url?scp=84921606894&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84921606894&partnerID=8YFLogxK

U2 - 10.1111/aos.12503

DO - 10.1111/aos.12503

M3 - Article

VL - 93

SP - e57-e66

JO - Acta Ophthalmologica

JF - Acta Ophthalmologica

SN - 0001-639X

IS - 1

ER -