Human Corneal Wound Healing

The ability of the cornea to heal and maintain its clarity has paramount importance in preserving the eyesight. In the ocular surface, a pathologic wound healing process, along with local inflammation and neovascularization, can induce failure in the functional recovery of the ocular surface tissues, which can lead to corneal blindness. In particular, corneal healing process under inflammatory conditions is not fully understood.

In this model, confluent monolayers of the human corneal epithelial cell line are consistently wounded, creating a cell-free area, based on the technique described by Liang et al. (Liang et al., 2007). Cultures are gently washed with DMEM/F12 to remove loose cells. Cells are then exposed to different cytokines in culture medium. Cells for control conditions are also scratched, washed, and maintained in culture medium after the scratch. Immediately after the scratch and/or at different time points, at least four images of the scraped area are captured using phase contrast microscopy. The remaining wounded area and the scratch width at six different points per image are measured. The same scratched area is selected for the measurements at each time of study.

Using this model, the influence of inflammatory molecules or treatments in corneal wound closure can be studied, and relative contribution of cell proliferation and cell migration processes analyzed.