Effect Of Gradient Nanostructured Pure Ti On Proliferation And Adhesion Of MG63 Cells And Its Preliminary Mechanism

Objective: The effect of gradient nano-metal pure titanium on proliferation and adhesion of MG63 cells was investigated by this experiment.It was investigated and verified whether the gradient nano-metal pure titanium promoted the proliferation and adhesion of MG63 cells by changing the surface perormance.Theoretical and experimental basis for the application of gradient nano-metal pure titanium in oral cavity.Methods:1.Material preparation: After the SMAT treatment,the medical metal pure titanium plate was electrically spark-cut into a gradient nanometer pure titanium titanium piece with a diameter of 11 mm and a height of 2.5 mm.In order to eliminate the influence of roughness on the experimental results,the control group was subjected to SMAT treatment and then passed Vacuum annealing at 680°C,on the basis of the material grain size remains basically unchanged,a pure titanium material with similar surface roughness is obtained.2.Material treatment: 36%-38% dilute hydrochloric acid,acetone,absolute ethanol,distilled water was ultrasonically washed for 15 minutes,and then stored under high temperature and high pressure.3.The growth curve of MG63 cells was detected by MTS assay.4.MTS method was used to detect the proliferation of MG63 on gradient nano-metal pure titanium and pure titanium materials.5.Scanning electron microscopy(SEM)was used to detect the adhesion of MG63 on gradient nano-metal pure titanium and pure titanium materials.6.The surface roughness of the experimental material was calculated using a surface roughness meter and a CLSM.7.The topological structure of the surface samples of the gradient nano-metal pure titanium and pure titanium was analyzed by Atomic Force Microscope(AFM).8.The gradient nano-metal pure titanium and pure titanium after high temperature and high pressure sterilization were respectively immersed in the simulated body fluid,and the morphology of the material was observed by scanning electron microscopy(SEM).The elemental composition of the mineralized product after soaking was analyzed by X-ray energy spectrum(EDX).9.The protein content of the gradient nano-metal pure titanium and pure titanium surface was detected by a microplate reader.10.The hydrophilicity of the surface of the gradient nano-metal pure titanium and pure titanium materials was measured using a contact angle measuring instrument.11.Using SPSS17.0 statistical analysis software to process experimental data.Results: 1.Growth curve results: MG63 cell growth curve is similar to "S" shape,0-1days is the incubation period,2-5 days is the logarithmic growth phase,the cell number reaches the peak at 5 days,and then enters the plateau.2.Proliferation results: MG63 cells were co-cultured with the two groups of materials for 1,3,5,and 7 days.The MTS results showed that the OD values of the two groups increased gradually on the 1st,3rd,and 5th day,and the OD values of the two groups decreased slightly at 7 days,and the OD values of the nano-groups Both were higher than the pure titanium group,and the differences were statistically significant.3.SEM observation of adhesion results: After MG63 cells were co-cultured with the two groups of materials for 2 days,the surface of the nano-materials adhered to a wider range of cells,and a large number of pseudo-footes were attached to make the cells and materials closely connected,and the number of cells adhering to the nano-group was higher than that of the pure titanium group.4.Roughness results: the surface roughness of the gradient nano-metal pure titanium is1.73±0.11μm(N=10),and the surface roughness of the coarse-grain titanium is2.54±0.06μm(N=10).5.Topological structure test results: the surface of the two groups of materials showed a peak-like undulating topography.The nano-group peak-like topology has small undulations and the undulation range is +243.4 nm to-276.3 nm,which is characterized by a rough surface nano-scale surface.The pure titanium group has a large peak-like topology with an undulation ranging from +2.6 μm to-2.4 μm,which is characterized by a coarse-grained surface with a rough surface.6.Biomimetic mineralization results: Gradient nano-metal pure titanium surface induces the formation of new compounds,the surface forms a spherical coating,and the surface of pure titanium is snow-like deposited.EDX detection of the deposits on the surface of the two materials showed that the nano-composites mainly contained calcium and phosphorus,and the pure titanium compounds did not contain calcium and phosphorus.7.Protein adsorption results: The protein adsorption amount on the surface of the two groups increased gradually with time and the nano titanium protein adsorption amount exceeded the pure titanium,the protein adsorption increased rapidly in the first four hours,and the protein adsorption amount reached saturation in 4-24 hours.At 24 hours,the protein adsorption capacity of the two groups decreased,and the difference was statistically significant.8.Hydrophilic test results: The hydrophilicity test results showed that the contact angle of nano titanium water was 53.717 degrees,the contact angle of pure titanium water was96.883 degrees,the surface of water contacted with nano titanium was completely spread for 0.5 seconds,and the water contacted the surface of pure titanium.1.5 seconds completely spread out.Conclusions: Gradient nano-metal pure titanium promotes the proliferation and adhesion of MG63 cells,and its mechanism is closely related to the change of surface perormance of the material.