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Alex Cohen, Timothy Boyce, Xiaowu Gu, Michael Elliott; Novel Therapeutic Strategies for Corneal Trauma. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3874.
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To evaluate the role of caveolin-1 (Cav-1) in corneal wound healing.
Immunofluorescence microscopy was performed upon human and mouse corneas to identify the pattern of Cav-1 expression. Next, primary corneal epithelial cells were prepared from mouse corneas and in vivo wound healing assays were performed. In addition, Wild-type (WT) and Cav-1 knockout (KO) mice were subjected to in vivo corneal epithelial and stromal wounds. Some corneas were prepared for immunofluorescence microscopy while the rate and pattern of wound healing was observed in others.
Cav-1 is preferentially expressed in the basal epithelium of the limbus of human and mouse corneas. This location is where the corneal stem cells (CSC) reside. The next experiments showed that Cav-1 immuno-reactivity is increased at the leading edge of the cells responsible for closing an epithelial wound both in vitro and in vivo. Next, a 100% epithelial defect was created in the eyes of WT and Cav-1 KO mice. Here we found that the Cav-1 KO mouse corneas healed at a faster rate than their WT counterparts. At 96h following the wound, the Cav-1 KO corneas were almost completely healed, while the WT mice showed a 50% or greater epithelial defect. This was confirmed in vitro using primary WT and Cav-1 KO cultured corneal epithelial cells. At 24 hours after the initial wound, the Cav-1 KO wound was approximately 80% closed while the WT wound was only about 30% closed. Next, mice were subjected to a deep, stromal wound with a diamond burr. The mice were allowed to heal and monitored for a period of 4 weeks. At the end of this time period the corneas were imaged using a digital slit lamp camera and we found that the corneas of WT mice showed dense central scar formation while those of Cav-1 KO were essentially clear.
Cav-1 is necessary for proper corneal epithelial wound healing in vivo and there is compelling evidence for a role in stromal scar formation as well.
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