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Aims To investigate the effect of intravitreal injection (IVI) on the optic nerve in infants with retinopathy of prematurity (ROP) during long-term follow-up.Methods The preterm infants included were divided into control, untreated and treated groups. The treatment group was further divided into the plus disease group and the no plus disease group. The vertical cup-to-disc ratio (C/D) was measured and compared in fundus photography at 36 weeks to 6 months of postmenstrual age (PMA) in the three groups of infants, respectively.Results A total of 381 eyes were included in this study. As the PMA passed from 36 weeks to 6 months, the C/Ds of the infants in all three groups increased to varying degrees, in which those of the treated group were significantly larger than those of the untreated group and the control group at all follow-up periods except at 36 weeks of PMA. In addition, in the treatment group, the C/D was significantly lower in the plus disease group than in the no-plus disease group, except at 6 months of PMA.Conclusions The present study found that IVI for ROP would result in an enlargement of the C/D in prematurity. The presence of plus disease was associated with an underestimation of the C/D. And the C/D in preterm infants continued to increase to varying degrees in at least 6 months of PMA.