Purpose: This study compared clinical outcomes and complications between 23-gauge (23g) and 25-gauge (25g) transconjunctival sutureless vitrectomy in patients with proliferative diabetic retinopathy. Study Design: It was a retrospective study using data prospectively defined and collected. 80 eyes underwent 23g transconjunctival sutureless vitrectomy, and 80 eyes underwent 25g surgery using the same vitrectomy system by one surgeon. Primary outcome measures were best-corrected visual acuity, intraocular pressure (IOP), and incidence of intraoperative and postoperative complications. Results: Vision was significantly improved after intervention in both groups (p ≥ 0.0001). There was no significant difference in visual outcomes between the groups (p = 0.43) or in the type and frequency of retinal breaks occurring during surgery (p = 0.63). The 23g group had significantly more patients with a day 1 IOP of <6 mm Hg (p = 0.034) and significantly more patients requiring a sclerostomy suture postoperatively (p = 0.014). Conclusion and Message: Both gauges are equally effective for the treatment of proliferative diabetic retinopathy.