We examined the protective properties of purpurogallin, a naturally occurring phenol, in delaying necrosis of cultured corneal endothelial cells caused by oxygen free radicals. Endothelial cell cultures were prepared from New Zealand white rabbits using microcarrier cell culture techniques. Corneal endothelial cells were treated with hypoxanthine (2 mM) and xanthine oxidase (67 IU/L) to generate free radicals. The criteria for cell necrosis were cytoplasmic shrinkage, dissolution of plasma membranes and presence of "haloes" around the cells on phase contrast microscopy, confirmed by transmission electron microscopy. More than 95% of second-generation cells exhibited morphologic evidence of necrosis within 4.62 +/- 0.82 minutes after exposure to oxyradicals. The addition of purpurogallin (0.25 or 1.0 mM) significantly increased time to cell necrosis to 8.18 +/- 0.83 and 11.59 +/- 1.71 minutes respectively (p < 0.05). Further studies are under way to determine whether purpurogallin may be useful in preventing endothelial cell damage in corneas preserved for corneal transplantation.