Developing smart scaffolds with drug release capability is one of the main approaches to bone tissue engineering. The current study involves the fabrication of novel gelatin (G)-hydroxyapatite (HA)-/vitamin D (VD)-loaded graphene oxide (GO) scaffolds with different concentrations through solvent-casting method. Characterizations confirmed the successful synthesis of HA and GO, and VD was loaded in GO with 36.87 ± 4.87% encapsulation efficiency. Physicochemical characterizations showed that the scaffold containing 1% VD-loaded GO had the best mechanical properties and its porosity percentage and density was in the range of natural spongy bone. All scaffolds were degraded after 1-month, subjecting to phosphate buffer saline. The release profile of VD did not match any mathematical kinetics model, porosities and the degradation rate of the scaffolds were dominant controlling factors of release behavior. Studies on the bioactivity of scaffolds immersed in simulated body fluid indicated that VD and HA could encourage the formation of secondary apatite crystals in vitro. Buccal fat pad-derived stem cells (BFPSCs) were seeded on the scaffolds, MTT assay, alkaline phosphatase activity as an indicator of osteoconductivity, and cell adhesion were conducted in order to evaluate in vitro biological responses. All scaffolds highly supported cell adhesion, MTT assay indicated better cell viability in 0.5% VD-loaded GO containing scaffold, and the scaffold enriched with 2% VD-loaded GO performed the most ALP activity. The results demonstrated the potential of these scaffolds to induce bone regeneration. Developing smart scaffolds with drug release capability is one of the main approaches to bone tissue engineering. The current study involves the fabrication of novel gelatin (G)-hydroxyapatite (HA)-/vitamin D (VD)-loaded graphene oxide (GO) scaffolds with different concentrations through solvent-casting method. Characterizations confirmed the successful synthesis of HA and GO, and VD was loaded in GO with 36.87 ± 4.87% encapsulation efficiency. Physicochemical characterizations showed that the scaffold containing 1% VD-loaded GO had the best mechanical properties and its porosity percentage and density was in the range of natural spongy bone. All scaffolds were degraded after 1-month, subjecting to phosphate buffer saline. The release profile of VD did not match any mathematical kinetics model, porosities and the degradation rate of the scaffolds were dominant controlling factors of release behavior. Studies on the bioactivity of scaffolds immersed in simulated body fluid indicated that VD and HA could encourage the formation of secondary apatite crystals in vitro. Buccal fat pad-derived stem cells (BFPSCs) were seeded on the scaffolds, MTT assay, alkaline phosphatase activity as an indicator of osteoconductivity, and cell adhesion were conducted in order to evaluate in vitro biological responses. All scaffolds highly supported cell adhesion, MTT assay indicated better cell viability in 0.5% VD-loaded GO containing scaffold, and the scaffold enriched with 2% VD-loaded GO performed the most ALP activity. The results demonstrated the potential of these scaffolds to induce bone regeneration.