Sustained release of sulforaphane by bioactive extracellular vesicles for neuroprotective effect on chick model.

Abstract

Novel studies have shown neurological treatment possibilities with extracellular vesicles (EVs) as natural particles with a special composition that are produced by different cell types. Their stability, natural structure, composition, and bioavailability make them good candidates as drug vehicles. Here, EVs were isolated from amniotic fluid (AF) through differential centrifugation, and characterized for size (<200 nm), structure, and composition, their effectiveness on the human PC12 cell line, and brain of chick embryos exposed to sodium valproate (animal autistic model). Sulforaphane (SFN) was employed as a bioactive compound and then encapsulated into Evs using three methods including passive (incubation), active (sonication), and active-passive (sonication-incubation). Further, the loading and in vitro releases of SFN fitted the Korsmeyer-Peppas (R²  = 0.99) kinetic model by non-Fickian diffusion case II (n = 0.44, passive loading) and Fickian diffusion case I (n = 0.41, active and active-passive loading). SFN-loaded EVs (SFN@EVs; 11 μM: 103 nM) stimulated hPC-12 cell proliferation. The gene expression analysis revealed that SFN@EVs could upregulate Nrf2 and reduce IL-6 expression. Eventually, histopathological results of the coronal cross-section of the chick embryos brain showed treatment with SFN@EVs. This treatment illustrated normality in the gray and white matter and the orientation of the bipolar neurons. Our findings showed EVs' potentially acting as a gene expression regulator in autism spectrum disorder.