Biocompatibility and potential efficacy in biological applications rely on the bio-interactions of graphene nanoparticles with biological tissues. Analyzing and modulating cellular and device-level activity requires non-invasive electrical stimulation of cells. To address these needs, G-optrodes, bio-interfaces based on graphene, have been developed. These devices use light to stimulate cells without modifying their genetic code. Optoelectronic capabilities, in particular the capacity to transform light energy into electrical energy, will be maintained throughout the procedures of neural stimulation. G-optrodes have also been studied as thin films on a range of substrates, and they have been designed to function at a very small scale. This study examines the impact of G-optrode-based substrate designs on the optical stimulation of pheochromocytoma (PC-12). Graphene electrodes, known as G-optrodes, are responsible for converting light into electrical pulses with stimulating effects. G-optrode bio-interfaces provide a stimulus that is independent of wavelength range but is sensitive to changes in illuminance. The authors have performed a comprehensive investigation based on the correct effects of the medication in vitro, employing substrate-based G-optrode biointerfaces. In substrate-based systems, the authors have proven that graphene is biocompatible. PC-12 cells were cultured on graphene for 7 days. Based on the findings, 20-nm and 50-nm thick G-optrodes are being studied for possible use in biological and artificial retinal applications. The findings of this study highlight the significance of biocompatibility in the selection and use of G-optrodes for biomedical purposes.