Introduction: Our aim was to evaluate the effects of 36 months of treatment with citicoline and vitamin B 12 eye drops on macular function in patients with type 1 diabetes (DM1) with mild signs of non-proliferative diabetic retinopathy (NPDR). Methods: A prospective, randomized, interventional, monocentric, double-masked study was conducted. Twenty patients with DM1 were enrolled and randomly divided into two groups: the DC group (10 patients; mean age ± standard deviation 46.86 ± 8.78 years) in which one eye of each patient was treated with citicoline and vitamin B 12 eye drops (OMK2 Ò , Omikron Italia srl, Italy, one drop thrice daily) for a period of 36 months; the DP group (10 patients; mean age ± standard deviation 47.89 ± 7.74 years) in which one eye of each patient was treated with placebo (eye drops containing hypromellose 0.3%, one drop thrice daily) for a period of 36 months. A total of 18 eyes (10 from the DP and 8 from the DC group, respectively) completed the study. In both groups, multifocal electroretinogram (mfERG) recordings were assessed at baseline and after 36 months. In mfERG analysis, the N1-P1 response amplitude density (RAD) evaluated in the 0-2.5°(ring 1), in the 2.5-5°(ring 2), in the 5-10°(ring 3), and in the 0-10°( ring 1 ? ring 2 ? ring 3) were considered. Results: With respect to baseline, after 36 months of follow-up, the mfERG RADs recorded in R1, R2, R3, and R1 ? R2 ? R3 were significantly increased (i.e., R1 ? R2 ? R3 RAD from 21.552 ± 2.522 nV/degree 2 at baseline to 26.912 ± 2.850 nV/degree 2 at 36 months) in DC eyes, whereas in DP eyes they were significantly reduced (i.e., R1 ? R2 ? R3 RAD from 21.033 ± 3.574 nV/degree 2 at baseline to 16.151 ± 3.571 nV/degree 2 at 36 months). Conclusions: This study indicates that patients with NPDR treated with citicoline and vitamin B 12 eye drops for a 36-month period achieved an improvement of the macular bioelectrical responses detectable by mfERG recordings. By contrast, during the same period of follow-up, patients with NPDR treated with placebo showed a worsening of the macular function.