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In this study, graphene oxide (GO) and reduced-thiolated GO (rGOSH) were used as 2D substrate to fabricate nanocomposites with nanoparticles of gold nanospheres (AuNS) or nanorods (AuNR), via in situ reduction of the metal salt precursor and seed-mediated growth processes. The plasmonic sensing capability of the gold-decorated nanosheets were scrutinized by UV-visible (UV-VIS) spectroscopy. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), thermogravimetric analyses (TGA), and atomic force microscopy (AFM) were performed in order to prove the actual reduction that occurred concomitantly with the thiolation of GO, the increase in the hydrophobic character as well as the size, and preferential gathering of the gold nanoparticles onto the nanosheet substrates, respectively. Moreover, the theoretical electronic and infrared absorption (UV-VIS and IR) spectra were calculated within a time-dependent approach of density functional theory (DFT). Eventually, in vitro cellular experiments on human neuroblastoma cells (SH-SY5Y line) were carried out in order to evaluate the nanotoxicity of the nanocomposites by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide tetrazolium reduction (MTT) colorimetric assay. Results pointed out the promising potential of these hybrids as plasmonic theranostic platforms with different hydrophilic or hydrophobic features as well as cytotoxic effects against cancer cells.