Nanohybrid composed of graphene oxide functionalized with sodium hyaluronate accelerates bone healing in the tibia of rats

Abstract

This study synthesized and characterized a nanohybrid composed of graphene oxide (GO) functionalized with sodium hyaluronate (HY) (GO-HY), evaluated its effect in vitro and determined its osteogenic potential in vivo. The synthesized nanohybrid was analyzed by Scanning electron microscopy (SEM), Raman spectrometry, Thermogravimetry, Fourier-transform infrared (FTIR) spectroscopy and X-ray diffraction. MC3T3-E1 cell viability was assessed by MTT assay in 48 and 72 h. Bone defects were created in tibia of 40 Wistar rats and filled with blood clot (control), 1% HY, GO (50, 100 and 200 μg/mL) and the nanohybrid (50, 100 and 200 μg/mL). After 7 and 14 days, histomorphometric analysis was carried out to assess osteogenic potential of the nanohybrid. Immunohistochemical analysis evaluated the expression of vascular endothelial growth factor (VEGF) in bone defects. Thermogravimetric analysis, Raman and FTIR spectrometry confirmed the functionalization of GO with HY by covalent bonds. Five μg/mL concentrations of the nanohybrid did not alter the viability of the MC3T3-E1 cells. Histomorphometric analysis demonstrated that the nanohybrid at 100 μg/mL significantly accelerated the bone repair in tibia of rats when compared to controls (p < 0.01). Immunohistochemical analysis showed a significantly less intense VEGF expression in tibia treated with the nanohybrid when compared to controls (p < 0.05). The nanohybrid composed of GO functionalized with HY was able to induce the acceleration of the tissue regeneration process in bone defects created in the tibia of rats. This novel nanohybrid is a promising material for the field of bone tissue engineering.