Purpose To evaluate the effect of intravitreal anti-vascular endothelial growth factor (anti-VEGF) injection on central choroidal thickness (CCT), central macular thickness (CMT) and best-corrected visual acuity (BCVA) in diabetic macular edema (DME). Methods Retrospective, cohort analysis of 90 eyes of 90 patients receiving anti-VEGF therapy for DME. In patients' records, measurements of CCT, CMT, and BCVA before treatment and at 2 years after treatment were recorded. Using enhanced-depth imaging optical coherence tomography (EDI-OCT) images, choroidal thickness and macular thickness measurements were recorded in the subfoveal area and 1 mm nasal to 1 mm temporal to the central foveal area. The baseline and final CMT and CCT values measured from all three quadrants were analyzed statistically. Results Mean age of the patients was 59.60 +/- 9.78 (range, 40-77) years. Mean baseline nasal-CT 226.4 +/- 52.5 mu m, central-CT 243.2 +/- 51.1 mu m and temporal-CT 224.6 +/- 47.9 mu m. Mean final nasal-CT 220.0 +/- 50.2 mu m, central-CT 235.3 +/- 53.6 mu m, temporal-CT 220.5 +/- 48.1 mu m (p = 0.122, p = 0.056, p = 0.184, respectively). Mean baseline nasal- MT 385.3 +/- 67.7, central-MT 345.5 +/- 119.7 mu m and temporal-MT 365.0 +/- 64.9 mu m. Mean final nasal-MT 359.6 +/- 59.2 mu m, central-MT 306.2 +/- 98.4 mu m and temporal-MT 353.4 +/- 63.3 mu m (p = 0.001, p = 0.002, p = 0.234, respectively). The BCVA improved from 0.52 +/- 0.44 logMAR at baseline to 0.38 +/- 0.33 at final (p = 0.002). Conclusion After treatment of diabetic macular edema with intravitreal anti-VEGF injection, CMT and BCVA improved significantly, but CCT did not decrease significantly.