Nanoprecipitation was used for producing CAI-PLGA nanoparticles that were below 220 nm and 0.2 polydispersion index (PDI) representing a reproducible method to generate nanoparticles of acceptable size and particle size distribution (
Figs. 1A,
1B). Wild type mice underwent laser injury and immediately post laser were injected with polymeric CAI-PLGA nanoemulsion (0.5 µg, 1.0 µg, or 2 µg) or aflibercept (10 µg) or the combination of aflibercept and CAI 1 µg in a 2 µL total volume (
Fig. 2A). Fluorescein angiography (FA) was used to confirm the success of laser rupture and location of the lesions (
Fig. 2B). The volume of the laser burn was quantified using OCT (
Fig. 2C,
Fig. 3). Seven days post intravitreal injection of the polymeric CAI-PLGA nanoemulsion, a significant decrease in choroidal neovascular volume was observed with a dose-dependent effect for CAI at 0.5 and 1.0 µg doses when compared to saline injected mice (saline = 100 ± 8.93,
n = 5 vs. CAI 0.5 µg = 49.27 ± 2.89,
n = 4,
P = 0.0066, saline versus CAI 1.0 µg = 24.87 ± 2.49,
n = 5,
P = 0.0001). However, a higher dose of CAI did not show any significant effect (saline versus CAI 2.0 µg = 70.43 ± 9.08,
n = 3,
P = 0.295). Aflibercept (saline versus aflibercept = 80.29 ± 7.93,
n = 4,
P = 0.388) or the combination (saline versus aflibercept/CAI 1.0 µg = 57.09 ± 5.08,
n = 3,
P = 0.127) did not show efficacy at day 7 suggesting no additive or synergistic effects. On day 14 post treatment, CAI at the 1.0 µg dose showed reduced neovascularization compared to saline (saline = 100 ± 16.72,
n = 5 versus CAI = 30.52 ± 7.01,
n = 3,
P = 0.030). This response compared favorably with the effect of 10 µg intravitreal aflibercept at week 2 (saline versus aflibercept = 41.29 ± 6.95,
n = 3,
P = 0.052; see
Fig. 2C). There did not appear to be a greater anti-angiogenic effect with increased amounts of CAI-nanoparticles (see
Fig. 2C). Combination treatment with 1.0 µg CAI-nanoparticle and 10 µg aflibercept was no more effective than polymeric CAI-PLGA nanoemulsion alone at week 2 (CAI 1.0 µg = 30.52 ± 7.01,
n = 3, versus CAI/aflibercept = 40.06 ± 2.86,
n = 3,
P = 0.356; see
Fig. 2C).