Intravitreal Delivery of VEGF-A165-loaded PLGA Microparticles Reduces Retinal Vaso-Obliteration in anIn VivoMouse Model of Retinopathy of Prematurity

PURPOSE: Retinopathy of prematurity (ROP) is a condition of abnormal retinal vascularization with reduced levels of vascular endothelial growth factor (VEGF) causing vaso-obliteration (Phase I), followed abnormal neovascularization from increased VEGF (Phase II). We hypothesized that intravitreal pro-angiogenic VEGF-A in microparticle form would promote earlier retinal revascularization in an oxygen-induced ischemic retinopathy (OIR) mouse model. MATERIALS AND METHODS: Wildtype mice (39) were exposed to 77% oxygen from postnatal day 7 (P7) to P12. VEGF-A(165)-loaded poly(lactic-co-glycolic acid) (PLGA) (n=15) or empty PLGA (n=14) microparticles were fabricated using a water-in-oil-in-water double emulsion method, and injected intravitreally at P13 into mice right eyes (RE). Left eyes (LE) were untreated. At P20, after retinal fluorescein angiography, vascular parameters were quantified. Retinal VEGF levels at P13 and flatmounts at P20 were performed separately. RESULTS: VEGF-A(165)-loaded microparticles had a mean diameter of 4.2 μm. with a loading level of 8.6 weight.%. Retinal avascular area was reduced in VEGF-treated RE (39.5 ± 9.0%) compared to untreated LE (52.6 ± 6.1%, P< 0.0001) or empty microparticle-treated RE (P< 0.001) and untreated LEs (P= 0.001). Retinal arteries in VEGF-treated RE were less tortuous than untreated LE (1.08 ± 0.05 vs. 1.18 ± 0.08, P< 0.001) or empty-microparticles-treated RE (P= 0.02). Retinal arterial tortuosity was similar in the LE of VEGF and empty microparticle-treated mice (P> 0.05). Retinal vein width was similar in VEGF-treated and empty microparticle-treated RE (P> 0.9), which were each less dilated than their contralateral LE (P< 0.01). VEGF levels were higher in P13 OIR mice than RA mice (P< 0.0001). Retinal flatmounts showed vaso-obliteration and neovascularization. CONCLUSIONS: Endogenous retinal VEGF is suppressed in OIR mice. Exogenous intravitreal VEGF-A(165)-loaded microparticles in OIR mice reduced retinal vaso-obliteration and accelerated recovery from vein dilation and arterial tortuosity. This may be beneficial in preventing Phase II ROP without systemic effects.