Effect of zoledronic acid and graphene oxide on the physical and in vitro properties of injectable bone substitutes

The aim of this work was to develop injectable bone substitutes (IBS) consisting of zoledronic acid (ZOL) and graphene oxide (GO) for the treatment of osteoporosis and metastasis. The powder phase was consisting of tetra calcium phosphate (TTCP), dicalcium phosphate dihyrate (DCPD) and calcium sulfate dihyrate (CSD), while the liquid phase comprised of methylcellulose (MC), gelatin and sodium citrate dihyrate (SC), ZOL and GO. The structural analysis of IBS samples was performed by Fourier Transform Infrared Spectroscopy (FTIR). Injectability, setting time and mechanical strength were investigated. Additionally, in vitro properties of synthesized IBS were analyzed by means of bioactivity, ZOL release, degradation, pH variation, PO43- ion release and cell studies. Overall, all IBS exhibited excellent injectability results with no phase separation. The setting time of the IBS was prolonged with ZOL incorporation while the prolonging effect decreased with GO incorporation. The mechanical properties decreased with ZOL addition and increased with the incorporation of GO. The maximum compressive strength was found as 25.73 MPa for 1.5GO0ZOL incorporated IBS. In vitro results showed that ZOL and GO loaded IBS also revealed clinically suitable properties with controlled release of ZOL, pH value and PO43- ions. In in vitro cell studies, both the inhibitory effect of ZOL and GO loaded IBS on MCF-7 cells and proliferative effect on osteoblast cells were observed. Moreover, the prepared IBS led to proliferation, differentiation and mineralization of osteoblasts. The results are encouraging and support the conclusion that developed IBS have promising physical and in vitro properties which needs to be further validated by gene expression and in vivo studies.