In this study, non-enzymatic biosensor based on nickel nanoparticles (Ni-) /Graphene: GelMA hybrids were synthesized. Physical, chemical and electrochemical behavior of hybrid in the application of glucose sensing were evaluated. We applied a facile polyol approach to decorate Ni- on the reduced graphene oxide (rGO) nanosheets (GNs) for the nonenzymatic electrochemical detection of glucose. Furthermore, the effects of Ni-NP content (0.14, 0.28, 0.42 and 0.56 wt.%) on the electrochemical properties of the hybrids were evaluated. Nickel nanoparticles examined by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), dynamic light scattering (DLS), cyclic voltammetry and electrochemical impedance spectroscopy (EIS). Results showed that spherical nickel nanoparticles with an average size 20 ± 2 nm with high purity were synthesized. Electrochemical results show the electrocatalytic behavior of nickel nanoparticles for sensing glucose. In addition, Ni-/rGO hybrids containing varying amounts of Ni-NP were synthesized by facile procedures and one-step in situ reduction approach. Results revealed that GO is almost completely reduced to rGO and functionalized Ni- hosted on the surface of graphene layer. Modified glassy carbon electrode with Ni-/rGO due to simultaneous effect of Ni- and rGO had better electrochemical behavior in comparison with Nickel nanoparticles and graphene. Results showed that in the presence of ethylene glycol, spherical-like Ni- were deposited on rGO nanosheets. As prepared Ni-/rGO: glassy carbon electrode (GCE) revealed significantly improved electrochemical performance to the oxidation of glucose in alkaline solution compared to rGO: GCE, depending on Ni-NP content. Specifically, hybrid 0.28 wt.% Ni-/ rGO: GCE revealed the linear range for glucose (0.25 µM to 1200 mM) with low detection limit of 0.01 µM and sensitivity of 2511.276 µA.µM -1 .cm -2 . After synthesizing of methacrylate gelatin (GelMA), the ternary hybrid hydrogels of Ni-/rGO: GelMA were prepared by the combination of different ratios of Ni-/rGO hybrid with GelMA (95: 5 and 90:10) and evaluated. According to the results, GelMA with the highest degree of Methacrylation (81.4 ± 0.4%) was synthesized. Electrochemical behavior of GelMA showed that although there was not any redox peak in the absence of glucose, well-defined redox peak was revealed in the presence of glucose. Moreover, electrochemical results showed the presence of glucose oxidation peaks in the modified electrode with low concentration of Ni-/rGO hybrid (Ni-/rGO: GelMA = 95: 5). With the increase of Ni-/rGO hybrid (Ni-/rGO: GelMA = 90: 10), the electrochemical behavior of modified glassy carbon electrode with triple hybrid hydrogel was similar to the hybrid of Ni-/rGO. Finally, after optimization of triple hybrid hydrogel of Ni-/rGO :GelMA in the ratio of 90:10, other parameters of sensory were calculated. According to the results, linear range and sensitivity were 100-10000 ?M 3658.278 µA.µM -1 .cm -2 , which compared to other non-enzymatic glucose biosensors is prominent. Keywords : Biosensor, Graphene, Nickel nanoparticles, Gelatin methacrylate (GelMA).