New wound-healing model using cultured corneal endothelial cells. 1. Quantitative study of healing process

A new in vitro model to evaluate the corneal endothelial wound-healing process was developed. Confluent monolayer cell sheets of either rabbit or bovine corneal endothelial cells were cultured on cover slips, the surface of which had been precoated with hydroxyethyl methacrylate (HEMA) except for the 8 mm-diameter area for cell culture. Wounds were made by scraping off cells in the center of the cell sheets using a rotating silicone tip. The areas of these wounds were measured by a method of computed planimetry. The average wound area was 3.00 +/- 0.21 mm2 in rabbit cell cultures and 3.18 +/- 0.28 mm2 in bovine. The healing process after the wounding was observed using an inverted phase contrast microscope and wound-healing rates (healed area per hour) were calculated. Wounded areas were recovered by 60 hours in rabbit experiments, and by 84 hours in bovine experiments. Wound-healing rates were different among the observation periods and showed differences between the two species. The healing rates in the rabbit were higher than those in the bovine in the early phase. This in vitro model can be used for quantitative evaluation of the wound-healing process of the corneal endothelium.