Sulforaphane (SFN) is a bioactive component found in broccoli that has neuroprotective and anti-cancer effects. SFN has been encapsulated in various drug carriers to improve its stability and effectiveness. Specifically, this study presents an SFN delivery system based on human Amniotic Fluid-EVs (hAFEVs) for the treatment of neurological disorders. The binding affinity between hAFEVs (<200 nm) and SFN was investigated by spectroscopic analysis, revealing a temperature-dependent binding mechanism. The binding constants (ksv, kq, and kb) were also assessed, indicating a static quenching mechanism. Additionally, the docking studies of SFN and CD9, CD81, HSP70, and TSG101 were performed using AutoDock 4.2.6 software's Lamarckian genetics algorithm to investigate the type of interaction and their effects on the targetability of hAFEVs. The obtained results showed that SFN-loaded hAFEVs had a favorable effect on increasing the survival and proliferation of human PC12 cells (137%). The results of gene expression studies showed that this release system can upregulate the expression of Nrf2 and decrease the expression of IL-6, which shows the support of nerve cells against oxidative stress. In the In-Ovo study, treatment with SFN-hAFEVs resulted in improved grey and white matter cohesion and the improved orientation of bipolar neurons. The results indicate a significant increase in synaptophysin expression and the number of doublecortin-positive cells improved neuronal development and synapse formation. The findings of this study suggest that SFN-hAFEVs have great potential as a treatment for neurological disorders.