Toxicity issues and biocompatibility concerns with traditional classical chemical cross-linking processes prevent them from being universal approaches for hydrogel fabrication for tissue engineering. Physical cross-linking methods are non-toxic and widely used to obtain cross-linked polymers in a tunable manner. Therefore, in the current study, argon micro-plasma was introduced as a neutral energy source for cross-linking in fabrication of the desired gelatin-graphene oxide (gel-GO) nanocomposite hydrogel scaffolds. Argon microplasma was used to treat purified gelatin (8% w/v) containing 0.1∼1 wt% of high-functionality nano-graphene oxide (GO). Optimized plasma conditions (2,500 V and 8.7 mA) for 15 min with a gas flow rate of 100 standard cm3/min was found to be most suitable for producing the gel-GO nanocomposite hydrogels. The developed hydrogel was characterized by the degree of cross-linking, FTIR spectroscopy, SEM, confocal microscopy, swelling behavior, contact angle measurement, and rheology. The cell viability was examined by an MTT assay and a live/dead assay. The pore size of the hydrogel was found to be 287 ± 27 µm with a contact angle of 78° ± 3.7°. Rheological data revealed improved storage as well as a loss modulus of up to 50% with tunable viscoelasticity, gel strength, and mechanical properties at 37 °C temperature in the microplasma-treated groups. The swelling behavior demonstrated a better water-holding capacity of the gel-GO hydrogels for cell growth and proliferation. Results of the MTT assay, microscopy, and live/dead assay exhibited better cell viability at 1% (w/w) of high-functionality GO in gelatin. The highlight of the present study is the first successful attempt of microplasma-assisted gelatin-GO nano composite hydrogel fabrication that offers great promise and optimism for further biomedical tissue engineering applications.