To compare postoperative outcomes of 27-gauge (G) and 25-G vitrectomy conducted as day surgery for proliferative diabetic retinopathy (PDR). One hundred eighty-five consecutive PDR patients (185 eyes) who underwent primary vitrectomy (27-G in 64 eyes, 25-G in 121 eyes) were analyzed. The 27-G and 25-G groups did not differ significantly in preoperative Early Treatment Diabetic Retinopathy Study (ETDRS) score, age, or preoperative intraocular pressure. The proportions of simultaneous cataract surgery (27-G vs. 25-G: 59.4% vs. 62.4%) and air-filled eyes (76.6% vs. 85.1%) were not significantly different between two groups. Both groups showed significant improvement in ETDRS score at postoperative 1, 3, and 6months (all, P < 0.0001). Mean gain in ETDRS score from baseline was apparently better in 27-G group than in 25-G group at 1, 3, and 6months, but there were no significant differences (1month: 20.3 vs. 13.1 letters, P = 0.0703; 3months: 22.9 vs. 17.5 letters, P = 0.1561; 6months: 24.3 vs. 19.3 letters, P = 0.3313). Operation time was apparently longer for 27-G vitrectomy, but there was no significant difference (54.0 vs. 51.1min, P = 0.3676). The same was observed for postoperative intraocular pressure at postoperative day 1 (19.7 vs. 18.1mmHg, P = 0.1353). Incidence of postoperative retinal detachment (1.6% vs. 0.8%) and reoperation due to vitreous hemorrhage (6.3% vs. 6.6%) was not different between two groups. The 27G system is as safe and as usefulas the 25G system when used for PDR and can be expected to achieve earlier recovery of postoperative visual acuity.