Incorporation of functionalized reduced graphene oxide/magnesium nanohybrid to enhance the osteoinductivity capability of 3D printed calcium phosphate-based scaffolds

Improving bone regeneration is one of the most pressing problems facing bone tissue engineering (BTE) which can be tackled by incorporating different biomaterials into the fabrication of the scaffolds. The present study aims to apply the 3D-printing and freeze-drying methods to design an ideal scaffold for improving the osteogenic capacity of Dental pulp stem cells (DPSCs). To achieve this purpose, hybrid constructs consisted of 3D-printed Beta-tricalcium phosphate (β-TCP)-based scaffolds filled with freeze-dried gelatin/reduced graphene oxide-Magnesium-Arginine (GRMA) matrix were fabricated through a novel green method. The effect of different concentrations of Reduced graphene oxide-Magnesium-Arginine (RMA) (0, 0.25% and 0.75%wt) on the morphology, mechanical properties, and biological activity of the 3D scaffolds were completely evaluated. Our findings show that the incorporation of RMA hybrid into the scaffold can remarkably enhance its mechanical features and improve cell proliferation and differentiation simultaneously. Of all scaffolds, β-TCP/0.25GRMA showed not only the highest ALP activity and cell proliferation after 14 days but it up-regulated bone-related genes and proteins (COL-I, RUNX2, OCN). Hence, the fabricated 3D printed β-TCP/0.25GRMA porous scaffolds can be considered as a high-potential candidate for BTE.