Rabbit corneal epithelial wound repair: tight junction reformation

Previous studies have shown that the corneal epithelium will close a limbus to limbus scrape wound in four to five days, but requires 10 days to become firmly attached to the stroma. In order to determine if the restoration of the corneal epithelial barrier follows a similar sequence, we have used freeze-fracture to study tight junction reformation in a rabbit epithelial wound model. Dutch-belted rabbit corneal epithelium was removed with a limbus to limbus scrape wound and sampled from 0 to 30 days post-wounding. A minimum of 3 animals from each time point were processed for electron microscopy and freeze-fracture. Freeze-fracture showed that the cells at the wound margin had a reduction in the number of intramembrane particles on their apical surface. In areas adjacent to the wound edge, fragments of tight junctions were first observed on two days post-wounding specimens. The junctions became progressively more complex in the area behind the wound edge until wound closure at four days when the junctions were also present in the central region of the cornea. The maturation of the junctions continued and at five days after surgery they resembled control junctions. This sequence suggests that the the establishment of morphologically mature tight junctions may be necessary before the corneal epithelium can firmly reattach to the stroma.