Purpose - The purpose of the study was to examine the effect of topical application of substance P (SP), insulin-like growth factor-1 (IGF-1) and vasoactive intestinal polypeptide (VIP) on corneal re-epithelialization in galactosemic rats. Methods - Experimental galactosemia was induced by feeding a diet containing 30% galactose for 4–6 months. The corneal epithelium was debrided bi-laterally by scraping with a blunted scalpel blade. One eye (control) received only a saline solution whilst the other eye received a solution of SP and/or IGF-1 or VIP. A single drop of control or test solution was administered 4 times daily until wound closure. Corneas were stained with fluorescein and videotaped under ultraviolet illumination at regular time intervals after debridement. After digitizing the video image, the wound area was calculated using an image analysis program (NIH Image). Results - Corneal re-epithelialization was significantly delayed in galactosemic compared to normal animals. Rates of healing were 1.3 ± 0.07 mm 2 /hour and 1.02 ± 0.02 mm 2 /hour for normal and galactosemic animals, respectively. Topical application of SP in concentrations ranging from 25 pg/ml up to 250 µg/ml had no significant effect on the rate of corneal re-epithelialization. Similarly, IGF-1 (1 µg/ml) or VIP (1 µg/ml) when applied alone did not affect re-epithelialization. Furthermore, resurfacing of the debrided area was not affected by co-application of SP (250 µg/ml) and IGF-1 or VIP. Conclusion - Independent or combined topical application of SP, VIP or IGF-1 at the concentrations tested, does not modulate corneal epithelial wound healing in rats with galactosemia induced by 30% galactose.