Spontaneous chronic corneal epithelial defects (SCCEDs) are characteristic ulcers in dogs that are refractory to healing. The aim of the study was to evaluate the use of a topical regenerative agent to promote healing of SCCEDs. Nineteen dogs (20 eyes) were randomized to receive either regenerative agent (10 eyes) or placebo (10 eyes) every 48 h following corneal debridement, which was repeated 1 week later if the SCCED had not yet healed. The mean ± standard deviation time to re-epithelialization was 17.3 ± 12.8 days for the group treated with a topical regenerative agent and 19.3 ± 11.7 days for the group treated with a placebo; the cumulative healing rates were not statistically different (P > 0.650). A positive association was found between the initial size of the ulcer and the time to re-epithelialization (r = 0.555, P = 0.011). Although well tolerated by dogs, there was no therapeutic advantage in using a topical regenerative agent for re-epithelialization of SCCEDs.