Production of Soybean Oil Nanoemulsion (SONE) and Evaluation of Angiogenic and Embryotoxic Activity

Abstract

The SONE oil/water was prepared using the pseudo-ternary phase diagram, using a low energy method with phase inversion by changing the composition. In order to optimize the preparation of SONE, three speeds were used in the agitator arm and two different rods. The particle size and polydispersity index were determined by Dynamic Light Scattering (DLS) and the stability test by the freeze-thaw cycle. Angiogenesis in chicken embryo egg chorioallantoic membrane and zebrafish (Danio rerio) embryotoxicity was performed. The lower stirring speed and propeller shaft induced smaller particle size (550.2 nm). Regarding angiogenesis, there was a statistically significant difference for all the evaluated parameters (length, caliber, junctions, and number of blood vessel complexes) and the result was higher in SONE when compared to the inhibition control (Dexamethasone), but lower than the induction control (Regederm®) and there was no statistically significant difference between SONE and distilled water. It was observed that the exposure of the zebrafish embryos to SONE caused an increase in the mortality rate dependent on time and concentration. The LC50 for SONE decreased statistically with increasing exposure (p-value = 0.046). Heart rate decreased significantly with increasing concentration at all exposure times (p-value < 0.05), as a result of progressive embryo mortality. The hatching rate was late until the concentration of 0.0193 mg/mL and no hatching rate was verified from that concentration. Exposure of zebrafish embryos to different concentrations of SONE induced malformations such as spinal changes, pericardial edema and yolk sac edema, but there was no significant difference in the malformation rate of embryos exposed to SONE when compared to the control group. The SONE produced remained stable in the freeze-thaw cycle, with changes only in pH. Despite the low results for embryotoxicity, further studies are needed, aiming at the ideal formulation for angiogenesis purposes.