Suspended graphene oxide nanoparticle for accelerated multilayer osteoblast attachment.

Mimicking bone tissues having layered structures is still a significant challenge because of the lack of technologies to assemble osteoblast cell types into bone structures. One of the promising and attractive materials in biomedical and different engineering fields is graphene and graphene-based nanostructures such as graphene oxide (GO) because of their unique properties. In most studies, GO was synthesized using chemical vapor deposition method, and was coated on the substrate. In this study, we proposed a simple technique for assembly of cells that facilitates the construction of osteoblast-like structures using suspended GO synthesized by graphite powder, H2 SO4 , and KMnO4 .Toxicity effects of GO on human mesenchymal stem cells (hMSCs) derived from bone marrow were analyzed. In addition to normal MSCs, toxicity effects of GO on human cancer cell line saos-2 as an abnormal cell line that possess several osteoblastic features, was examined. The attachment and expression of osteoblast cells genes were evaluated after differentiation of MSCs to osteoblast cells in presence of suspended GO by scanning electron microscopy and real time PCR. We found that the toxicity effects of GO are dose dependent and in oseogenic medium containing suspended GO the expression level of osteoblast genes osteopontin and osteocalcin and cell adhesion markers connexin were higher than control group. Interestingly, through this method GO was found to induce multilayer osteoblast cell morphology and enhance the number of cell layer. We expect that the presented method would become a highly useful approach for bone tissue engineering.