ObjectiveRestoring large boney defects using bone grafts alone is an unpredictable procedure. Biodegradable polymeric scaffolds suffer rapid biodegradation and lack sufficient osteo-conductivity. The aim of this study was to histomorphometrically evaluate three-dimensional printed graphene oxide-enriched poly(ε-caprolactone) (PCL) scaffolds for bone regeneration in a rabbit defect model using two different concentrations of graphene oxide. Basic characteristic properties and mount of new bone regeneration formation were evaluated.Methodstwo concentrations of graphene oxide (1 and 3 wt%) were added to PCL scaffolds using hot blind technique while pure PCL scaffolds served as a control. Laboratory characterization included scanning electron microscopy (SEM), x-ray diffraction analysis (XRD), contact angle, internal porosity, in addition to density measurements. All scaffolds were subjected to biodegradation evaluation and cell cytotoxicity test. In vivo bone regeneration was evaluated in the tibia defect of a rabbit model by measuring the amount of new bone formation (n = 15, ά = 0.05).ResultsSEM images showed slight reduction in pore size and increase in filament width of scaffolds with increasing GO contents. However, the printed scaffolds matched well with the dimensions of the original design. XRD patterns revealed characteristic peaks identifying microstructure of scaffolds. Addition of GO increased crystallinity of the scaffolds. The contact angle and porosity readings indicated reduction in measurements with increased content of GO indicating improved wetting properties while the density followed an opposing pattern. Higher biodegradability values were associated with higher GO content resulting in acceleration of observed biodegradation. The results of cytotoxicity test showed reduction in cell viability with higher GO content. Bone regeneration was significantly enhanced for 1 wt% GO scaffolds compared to other groups as was evident by higher bone density observed in x-ray images and higher amount of new bone formation observed at different time intervals.SignificanceGraphene oxide improved the physical and biological properties of PCL scaffolds and significantly enhanced new bone regeneration.