Abstract Background To evaluate the efficacy and outcomes of one-way surgical technique for the treatment of vitreous hemorrhage post vitrectomy on proliferative diabetic retinopathy (PDR) patients. Methods This retrospective case series include 47 PDR patients who had vitrectomy with balanced saline solution tamponade and have developed vitreous hemorrhage without significant absorption. The one-way air-fluid exchange procedure which involves the application of a 0.22-μm pore size filter to exchange about 4.5–5.5 ml of fluid with a 10 ml syringe was performed on 47 patients (47 eyes). Post procedure, additional treatments were administered when needed. Best corrected visual acuity (BCVA), occurrence of intra-procedural and post-procedural complications were recorded and analyzed. Results A total of 47 eyes of 47 PDR patients with a mean age of 50.8 ± 12.0 years were reviewed. Because of vitreous hemorrhage or tractional retinal detachment of PDR, all 47 eyes underwent vitrectomy with balanced saline solution tamponade prior to the exchange procedure. Four patients (8.51%) and 43 patients (91.5%) were diagnosed with type 1 diabetes mellitus (T1DM), or type 2 diabetes mellitus (T2DM), respectively. All 47 eyes were given the one-way air-fluid exchange procedure in the treatment room. Forty-two cases (89.4%) needed the air-fluid exchange procedure only once, 4 cases (8.51%) underwent the procedure twice, and 1 case (2.13%) was given the procedure three times, followed by additional retinal photocoagulation and one intravitreal injection of Conbercept. In addition to the procedure, no further treatment was needed for 5 eyes (10.6%) while additional retinal laser treatment was provided for 41 eyes (87.2%). The BCVA at the final follow-up was significantly improved from the initial acuity baseline in all cases. No complications were observed during the follow-ups. Conclusion This one-way air-fluid exchange procedure can effectively exchange the vitreous hemorrhage and improve visual acuity of PDR patients who develop vitreous rehemorrhage post vitrectomy without obvious complications.