The influence of stromal contraction in a wound model system on corneal epithelial stratification.
OBJECTIVE
The healing process of some corneal wounds involves closure by stromal contraction and the renewal of the stratified epithelium. In wound gape injury such stromal contraction occurs with epithelial stratification. In previous in vitro studies of noncontracted and contracted corneal fibroblast-seeded collagen gels (FSCGs) it was shown that initiation of wound contraction by the myofibroblast phenotype (present within the wounded stroma) was dependent on vitronectin and/or fibronectin. This study considers one aspect of the epithelial-stromal interaction that occurs during wounding. The stratification of corneal epithelial cells on noncontracted and contracted corneal FSCGs was compared.


METHODS
Dissociated bovine corneal epithelial cells were seeded on noncontracted and contracted corneal FSCGs, and these assemblies were cultured for 7 days. The epithelium that formed was evaluated using laser confocal microscopy and immunohistochemical markers directed against cytokeratin 3, desmoplakin I and II, integrin alpha-6 subunit, laminin, and collagen VII. The characteristics of the epithelium were compared with stromal carriers comprised of dissociated bovine corneal epithelial cells seeded on intact stroma and basement membrane (stromal carrier biopsies).


RESULTS
The stratified epithelium that developed on contracted corneal fibroblast-seeded collagen gels was similar to that formed on stromal carriers, whereas nonstratified epithelium formed on noncontracted FSCGs.


CONCLUSIONS
These studies showed that the contracted state of fibroblast-seeded gels enhanced the development of well-organized, stratified corneal epithelium.
Taliana
L
L
Cooperative Research Centre for Eye Research and Technology, Sydney/CSIRO Division of Molecular Science, Sydney/University of New South Wales, Sydney, North Ryde, NSW, Australia. talia101@doc.mssm.edu
Evans
M D
MD
Dimitrijevich
S D
SD
Steele
J G
JG
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001485:Basement Membrane; D002417:Cattle; D002450:Cell Communication; D002452:Cell Count; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D003319:Corneal Stroma / Q000166:cytology* / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism; D051180:Desmoplakins; D019573:Epithelium, Corneal / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D039503:Integrin alpha6; D007633:Keratins / Q000378:metabolism; D007797:Laminin / Q000378:metabolism; D018613:Microscopy, Confocal; D008954:Models, Biological; D014945:Wound Healing
D015703:Antigens, CD; D003598:Cytoskeletal Proteins; D051180:Desmoplakins; D039503:Integrin alpha6; D007797:Laminin; D007633:Keratins; D003094:Collagen