Purpose:To investigate Wnt3a and vascular endothelial growth factor (VEGF) levels in the vitreous fluid of patients with proliferative diabetic retinopathy (PDR) and to examine their correlation with PDR activity.Methods:Vitreous samples from 45 eyes with PDR and 28 eyes with nondiabetic macular disease were collected. Active PDR was present in 24 patients and inactive PDR in 21 patients, according to retinal neovascularization. The Wnt3a and VEGF level of vitreous fluid samples were measured by enzyme-linked immunosorbent assay.Results:Comparison revealed that mean intravitreal levels of Wnt3a increased significantly in PDR eyes compared with control eyes (13.55 ng/mL vs. 1.57 ng/mL, P < 0.001). The mean VEGF concentrations in the vitreous fluid of patients with PDR were also higher than those in nondiabetic controls, with the values being 723.21 pg/mL and 20.81 pg/mL, respectively (P < 0.001). In addition, vitreous concentrations of Wnt3a and VEGF were significantly higher in active PDR than in eyes with inactive PDR (P = 0.016 and P = 0.008, respectively). Furthermore, a significant positive correlation was detected between Wnt3a and VEGF levels in the vitreous.Conclusion:Intravitreous levels of Wnt3a and VEGF in patients with PDR are increased and correlated mutually. Wnt3a may be an important player in the development of diabetic retinopathy and its activity in vitreous fluid can be biomarker of PDR.