Release of neuronotrophic factor from rabbit corneal epithelium during wound healing and nerve regeneration

Epithelial neuronotropic factor (ENF) is secreted by cultured epithelial cells of rabbit cornea and conjunctiva, and is active in promoting survival and inducing neurite outgrowth of cultured trigeminal neurons. This study evaluated the relation of ENF to corneal nerve regeneration utilizing a model of heptanol-induced epithelial wounding. The organ culture technique was used to collect ENF from the intact corneal epithelium, and a neuronal bioassay was utilized to quantify ENF. The results revealed no change in ENF secretion either during initial wound closure or after 1 week, when the epithelium had regenerated. However, ENF secretion was elevated 2.4 times in 2 weeks after wounding. Morphometric analysis of corneal nerves stained by gold chloride impregnation showed that the first sign of regeneration of intraepithelial nerves was observed after 2 weeks, and the normal pattern of epithelial neural density was re-established after 3 weeks. However, the neural density was still subnormal (35-47% less than the control) in the wounded epithelium up to 4 weeks after wounding. Thus it appears that a surge in ENF secretion occurred after epithelial regeneration but before nerve regeneration. The results suggest that ENF may mediate corneal nerve regeneration.