3D printed reduced graphene oxide-GelMA hybrid hydrogel scaffolds for potential neuralized bone regeneration

Peripheral nerves participate in bone growth and repair by secreting neurotransmitters, and enable new bone to possess physiological bone-sensing capability. However, it is difficult to achieve synchronized nerve regeneration during the healing process of large bone defects at present. As a bioactive nanomaterial, reduced graphene oxide (rGO) can promote neuronal differentiation and myelination of Schwann cells (SCs), while enhancing the adhesion and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) through its strong non-covalent binding ability. In this study, 3D printing-based rGO/GelMA hydrogels with enhanced osteogenic and neurogenic dual differentiation were used to simultaneously load SCs and BMSCs. By changing the concentration of rGO(0.03%/0.05%/0.1%), the compressive strength, rheological properties and aperture of the hydrogel can be improved. In vitro, cell live/death staining, phalloidin staining and SEM showed that cells loaded on the hydrogel had a high survival rate (85%) and good adhesion ability. In vivo, we found that the rGO/GelMA hydrogel exhibited the same low inflammatory response compared to the pure-GelMA group and the cell-only group, but surrounded by collagen fibers. Meanwhile, the osteogenic and neural proteins in the rGO/GelMA group were found to be highly expressed in immunohistochemistry and immunofluorescence. In this study, a scaffold material containing double cells was used to promote synergistic regeneration of nerves and bone, providing a promising strategy for the preparation of personalized and functionalized biomimetic bone material.