Background: The use of topical mitomycin C in ophthalmology is increasing in all sub-specialties, but the risk-benefit ratio should be considered, taking into account its complications. The dose and duration of application of MMC depends on the surgery in which it is used but is still controversial. Objective: To evaluate the changes in the normal histology of the anterior surface structures of the rabbit eye “conjunctiva, cornea and sclera" after subconjunctival injection of different doses of Mitomycin C (0.1, 0.2 and 0.4 mg/ml) after different periods (1, 7 and 14 days). Material and Methods: A comparative prospective case-control animal study included eighteen eyes of eighteen healthy New Zealand rabbits conducted at Faculty of Medicine, Al-Azhar University. Results: Group B and C rabbits' eyes clinically throughout the study revealed significant findings in the form of severe congestion, severe chemosis, lid edema and lacerations while group A showed non-significant clinical findings. Histopathological results showed significant decrease of the conjunctival goblet cells and scleral elastic tissues with MMC administration at different doses even the low dose from the 1st to the 3rd sacrifice in dose and time-dependent manner. Scleral thickness showed significant decrease after 2 weeks of MMC administration. Corneal thickness showed non-significant decrease within the different sacrifices. Conclusion: Preoperative subconjunctival injection of MMC by 0.1 mg/ml concentration one day before, is mainly do not affects the cornea and have the least effects on the conjunctiva and the sclera.