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Change in epithelial keratin expression during healing of rabbit corneal wounds
Abstract:
Corneal epithelial wound healing following full-thickness trephination and transcorneal freeze injury was studied by electron microscopy and immunofluorescent microscopy using monoclonal antibodies AE1, AE2, and AE3 to human epithelial keratin. Wounds were evaluated at various time intervals between 4 hr and 2 mo after injury. By scanning and transmission electron microscopy, epithelial migration was evident 4 hr after injury and was characterized by thinning of the epithelium and extension of filopodial processes. AE1 monoclonal antibody, which stains specifically the superficial cells of normal corneal epithelium, reacted to cells at the leading edge of the migrating epithelium. By 24 hr, all cells migrating over the wound displayed positive fluorescence with AE1 while the epithelium over the undamaged cornea exhibited normal fluorescence limited to the superficial epithelial cells. In full-thickness corneal wounds, reepithelialization was complete by 1-2 wk; however, all epithelial cells covering the wound remained positive for the AE1 antikeratin antibody. By 2 mo, the AE1 fluorescence returned to normal. In transcorneal freeze injuries, reepithelialization was complete by 4 to 7 days after injury, with all cells overlying the wound reacting with the AE1 antibody. By 2 wk after freeze injury, all epithelial cells appeared to express a normal AE1 staining pattern. No change was noted in the fluorescent distribution of either AE2 antibody, which did not react with the corneal epithelium, or AE3, which reacts with all corneal epithelial cells. These results suggest that healing of corneal epithelial wounds involves changes in keratin expression of the corneal epithelium.
Insights
Corneal wound healing involves changes in keratin expression, specifically with the AE1 antibody, indicating altered cell behavior during the repair process in both full-thickness and freeze injuries.
Area of Science:
- Ophthalmology
- Cell Biology
- Wound Healing Research
Background:
- Corneal epithelial wound healing is crucial for maintaining ocular surface integrity.
- Understanding the molecular changes during healing can inform therapeutic strategies.
Purpose of the Study:
- To investigate the changes in keratin expression during corneal epithelial wound healing.
- To characterize the role of specific keratins in the repair process following injury.
Main Methods:
- Utilized electron microscopy (scanning and transmission) for ultrastructural analysis.
- Employed immunofluorescent microscopy with monoclonal antibodies (AE1, AE2, AE3) targeting human epithelial keratin.
- Evaluated corneal wounds at various time points post-injury (4 hr to 2 mo).
Main Results:
- Epithelial migration observed as early as 4 hr post-injury, with filopodial extension.
- AE1 antibody, normally staining superficial cells, marked migrating cells and all cells in healing wounds.
- Reepithelialization completed within 1-2 weeks for full-thickness wounds and 4-7 days for freeze injuries.
- AE1 staining normalized by 2 months in full-thickness wounds and 2 weeks in freeze injuries.
- AE2 and AE3 antibodies showed no significant changes in distribution during healing.
Conclusions:
- Corneal epithelial wound healing is associated with dynamic changes in keratin expression.
- The AE1 antibody highlights altered keratin profiles in migrating and regenerating corneal epithelial cells.
- These findings suggest keratin expression changes are integral to the corneal wound repair mechanism.