The present work investigates the influence of graphene oxide incorporation on the physio-chemical properties and biocompatibility of chitosan-guar gum based porous 3D scaffolds. In this study, a porous 3-dimensional chitosan/guar gum-GO scaffolds was fabricated by freeze drying method with different concentrations of GO (0, 0.5, 1 and 1.5 wt%). To improve their stability in the aqueous medium, the scaffolds was crosslinked using sodium tetraborate (Borax) resulting in borax crosslinked chitosan/guar gum-GO scaffolds (CGB) and characterized using FT-IR, XRD, Raman and TGA analysis. Crosslinking of the polymer matrix by borax was confirmed by FT-IR analysis, and XRD spectra shows that GO and borax incorporation did not affect the crystallinity of the polymer matrix. Increase in thermal stability was observed as the concentration of GO increased in the scaffolds. Porous morphology of the scaffold can be observed under scanning electron microscopy and the scaffolds exhibited around 60 % porosity. Irrespective of GO concentration, all the scaffolds exhibited good surface wettability and the degradation of the scaffolds slowed as the concentration of GO increased. In vitro cytocompatibility studies with Vero cells showed that both the CG and CGB scaffolds exhibited more than 90 % cell viability irrespective of GO concentration and the cell attached well on the scaffolds with GO when compared to control scaffolds. Overall, the prepared scaffold proved to be highly suitable for biological applications such as drug delivery and tissue engineering.