Purpose. Povidone iodine has been proven to be a valuable antiseptic solution in preparing the eye for surgery and is an alternative to postoperative topical antibiotics. No study has addressed the intraocular toxicity of povidone iodine after injection into the anterior chamber. We investigated the potential toxicity of povidone iodine on the corneal endothelium after injections into the anterior chamber in a rabbit model. Methods. In this study we used 24 eyes of 12 albino rabbits. The eyes were divided into the following three groups according to the drugs tested: group A, 5% povidone iodine; group B, 10% povidone iodine; group C, balanced salt solution. The injected eyes were evaluated by biomicroscopy, specular microscopy, corneal pachymetry, and transmission and scanning electron microscopy. Results. Corneal edema was observed in all eyes of groups A and B. In groups A and C, the endothelial cell morphology was not significantly changed and the mean endothelial cell count of the eyes did not change significantly (p = 0.5054). There was no significant difference in corneal thickness between groups A and C (p = 0.3823), but there was a significant difference between groups B and C (p = 0.0002). Transmission and scanning electron microscopy results were normal in group C but not in groups A and B. Conclusion. Povidone iodine in both 5% and 10% concentrations demonstrates severe toxicity when one drop of either concentration is placed directly in the anterior chamber. When povidone iodine is used in preparing the eye for intraocular surgery and as an alternative to postoperative antibiotics, the inadvertent leakage of povidone iodine into the anterior chamber must definitely be prevented.