Construction Of Naringin-loaded Multilayered Films On Titanium Surfaces And Their Effects On Osteoblasts And Osteoclasts

Titanium-based materials have been widely used as bone implant and dental implant materials in clinical field because of their good biocompatibility and mechanical properties.However,due to the existence of biological inertia on the surface of titanium materials,the osteointegration of titanium materials is poor at the early stage of implantation,especially for the pathological fractures caused by osteoporosis.At present,how to improve the biological activity of titanium implant surface,enhance the ability of titanium implant osseointegration,and increase the success rate of implant surgery is the principal direction of research.Therefore,to resolve the fractures in patients with osteoporosis,in this study the titanium surfaces were treated by acid etching and further the naringin(NA)-loaded multilayered films were constructed on titanium surfaces by layer by layer self-assembly technology(LBL),the NA-loaded multilayered films modified titanium surfaces can promote osteoblast differentiation,inhibiting osteoclast formation.The research contents and results of this paper are as follows:(1)Construction and characterization of naringin-loaded multilayered films on titanium surfaceThe microstructed titanium(Micro-Ti)surfaces were prepared by acid etching,and a layer of polydopamine was added on the surface to increase the binding property for further modification.Then,the multilayered films and naringin-loaded multilayered films were constructed on titanium surfaces by LBL technology.Scanning electron microscopy(SEM),atomic force microscopy(AFM),X-ray photoelectron spectroscopy(XPS)and contact angle measuring instrument were used to characterize the materials,and the release rate of NA was detected.It was proved that the naringin-loaded multilayered films was successfully constructed on the surface of titanium material,and the structure had a certain sustained-release effect.(2)The effect of naringin-loaded multilayered films modified titanium surfaces on the morphology,proliferation and differentiation of osteoblasts Fluorescence staining,CCK8 detection,ALP detection,mineralization detection and q RT-PCR were conducted in this study after the osteoblasts were inoculated on the surface of different titanium materials.The results were as follows:compared with the other groups,the osteoblasts grown on naringin-loaded multilayered films modified titanium surfaces showed better spreading ability,no significant difference in cell proliferation,and showed the higher differentiation ability at early and late osteogenic stages.The q RT-PCR results showed that compared with the other groups,the expressions of the osteoblastic differentiation markers Runx2,ALP,COL I,OCN,OPN and OPG were higher in naringin-loaded multilayered films modified titanium group.These results indicated that naringinloaded multilayered films modified titanium surfaces could promote osteoblast differentiation.(3)The effect of naringin-loaded multilayered films modified titanium surfaces on the morphology and differentiation of osteoclastsAfter macrophages were inoculated on the surfaces of titanium materials,the osteoclasts were cultured in osteoclast culture medium and detected by fluorescence staining,ACP kit and q RT-PCR.The results showed that there were obvious multinucleated osteoclasts in the other groups,but there was almost no osteoclast observed in naringin-loaded multilayered films modified titanium group,the ACP activity and the expression levels of osteoclastic differentiation related genes CTSK,NFAT,TRAP and VATP also were the lowest in this group.It is proved that naringin-loaded multilayered films modified titanium surfaces can inhibit the formation of osteoclasts.In conclusion,this study demonstrated that the naringin-loaded multilayered films modified titanium surfaces can regulate the balance of bone formation and bone resorption,and accelerate bone remodeling.It provides a new idea for the surface modification and optimization of titanium implants,and also provides theoretical support for the study of titanium implants.