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7059915 
Journal Article 
Surface Functionalization of Titanium with BMP-7/RGD/Hyaluronic Acid for Promoting Osteoblast Functions 
Zhou, L; Wu, J; Wu, D; Yu, J; , 
2019 
1 
Journal of Biomaterials and Tissue Engineering
ISSN: 2157-9083
EISSN: 2157-9091 
AMER SCIENTIFIC PUBLISHERS 
VALENCIA 
32-39 
WOS:000468536600004 
Titanium (Ti) is widely used in dental implants because of its good mechanical properties. It is very important to modify the surface of Ti properly to improve the osteogenesis efficiency. In this study, 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and N-Hydroxysuccinimide (NHS) were used to activate Arg-Gly-Asp (RGD), and then RGD was connected with Hyaluronate acid (HA) to obtain HA-RGD. Element content of the product was analyzed by Vario EL III elemental analyzer. The gene sequence of BMP-7 was amplified by PCR and linked to the pIRES2-EGFP overexpression vector. The pIRES2-EGFP-BMP-7 was wrapped with HA-RGD and then the HA-RGD-BMP-7 was obtained. The envelope of HA-RGD-BMP-7 was detected by gel electrophoresis block test. The shape of HA-RGD-BMP-7 complex was observed by atomic force microscopy (AFM). The surface morphology of titanium sheet was observed with field emission scanning electron microscopy (SEM). The HA-RGD-BMP-7 composite was grafted onto the surface of the treated titanium sheet by electrostatic adsorption method. Golden View staining was performed and the surface of the titanium sheet was photographed by ultraviolet imaging system. Human bone marrow mesenchymal stem cells (MC3T3 E1) were cultured on the surface of titanium sheet. Cell morphology was detected by SEM. BMP-7 expression efficiency was detected by immunofluorescence. Osteogenic status was detected by Alizarin Red-S. BMP-7, osteocalcin (OC), bone sialoprotein (BSP) and osteopontin (OPN) protein levels were detected by Western Blotting. The results showed that RGD was successfully linked to HA and pIRES2-EGFP-BMP-7 vector was obtained by gene cloning. AFM results showed that the HA-RGD-BMP-7 complex was granular in shape and its particle size was 198.2 nm. Golden View staining showed that HA-RGD-BMP-7 complex adsorbed stably on the surface of titanium sheet. SEM results showed that MC3T3 E1 cells could nest in the pits on the surface of titanium sheet. Immunofluorescence and Western Blotting assay showed that BMP-7 could stably express on MC3T3 E1 cells.