Green nanotechnology applied to neuroscience offers innovative solutions for the treatment of central nervous system disorders. This study reports on the biosynthesis of gold nanoparticles (AuNPs) using extracts from the psilocybin-producing mushroom Psilocybe cubensis and the evaluation of their neurostimulatory potential. The AuNPs were synthesized via the biological reduction of HAuCl₄ and characterized using UV-Vis spectroscopy, TEM, DLS, and FTIR. The physicochemical results confirmed the production of monodisperse spheroidal nanoparticles with an average diameter of 18.94 nm and high colloidal stability (zeta potential: −31.0 mV). FTIR analysis suggested that fungal metabolites constitute the stabilizing organic corona of the nanomaterial. Bioactivity was evaluated in the Neuro2a (N2a) neuronal model using the IncuCyte® real-time monitoring platform and 3D holotomographic microscopy. A dose-dependent hormetic response was identified: concentrations between 22.91 and 183.25 µg/mL induced a significant increase in cell proliferation and viability without morphological compromise, while doses ≥ 733.0 µg/mL resulted in acute cytotoxicity. 3D holotomography confirmed that the biostimulatory treatment (91.63 µg/mL) promotes a phenotype with active cytoplasmic extensions, consistent with early stages of neurite outgrowth. In conclusion, P. cubensis AuNPs constitute a promising nanotherapeutic system with neuroprotective and pro-proliferative capabilities, laying the groundwork for future applications in regenerative medicine and neurorehabilitation.

Gold nanoparticles, Green synthesis, Holotomography, Neuroregeneration, Psilocybe cubensis.