| The titanium implants have become a conventional method which can replace the missing tooth and repair the facture now. As a substitute of the natural tooth the dental implant will have implant-bone interface with jaw and have implant-soft interface with periodontium when it is implanted in the jaw. If the implant want to stay and have the function in a complex physical oral environment for a long time, a solid bony union between the planting and alveolar is very important. The implant neck soft tissue sealing barrier is the same implant. The barrier mainly is made up by gingival epithelium and collagen fiber which bonding on the implant.The connective around the implant which is made up by the collagen fiber which is secreted by the gingival fibroblasts and the gingival fibroblasts is an important part for the soft tissue healing of the implant wear transparent part. The healing good structure no only provide solid surface epithelial basal tissue, prevent epithelial root migration, strengthen the edge sealing reduce bacteria invasion, reduce the implant surrounding soft tissue inflammation caused by gum back beautiful front teeth problems, but also protect the deep tissue healing. Studies have shown that chemical properties on the surface of the implant neck and the character of implant surrounding area, biocompatibility materials properties and geometric structure can affect the cell’s adhesion; the research on the surface of the implant neck geometric structure has become a hot spot at present.The micron grade groove design on the implant wear transparent part which has good hydrophilic and biocompatibility can increase implant wear transparent part surface area, provide more binding site for adhesion protein such as fiber connection protein(FN), is good to promote fibroblasts secrete more cellular FN, adsorb and active more platelet at the same time that will promote the follow-up more gingival fibroblasts adhere to gather. Studies have shown that T60/10(60 microns wide 10 microns deep) groove morphology’s hydrophilic and biocompatibility is best than any other groove morphology, but the plasma fibronectin(FN) connection adhesive ability of T60/10 is limited. As a kind of macromolecular extra cellular matrix protein FN has many biological function. For example it promote fibroblasts go to wound which can accelerate wound healing. If there is not a good soft tissue closed on the implant neck in the early time, it will cause infection easily. It is crucial to make up the lack of T60/10 consequently. Objective:The purpose of the topic is to improve the early soft tissue healing ability of T60/10 groove collar. The plasma fibronectin(FN) will be modified on the surface of micron level titanium groove, and it must improve adsorption rate of plasma FN. When the plasma fibronectin(FN) was modified on the surface of T60/10, we will evaluation the influence to biological behavior of gingival fibroblasts such as proliferation, adhesion and secretion, etc. The relationships between the modified material and the early soft tissue attachment will be studied then, and the study could provide experimental basis for the future development of new materials. Methods:This subject adopts the method of silicon oxidation to modify the plasma fibronectin(FN) on the surface of the microgroove titanium surfaces. The content and distribution of FN will be detected by X-ray diffraction spectroscopy analysis, FN protein immunofluorescence technology, ELISA.Field emission scanning electron microscopy(SEM), atomic force microscopy and contact angle meter will be use to test o the surface topography, roughness and surface hydrophilic of the surface of FN modified material.Human gingival fibroblasts are cultured on the modified FN T60/10 groove titanium surface and smooth titanium surface respectively. The proliferation of HGF is detected by CCK8. Field emission scanning electron microscopy(SEM) and cell immunofluorescence technology test are used to the adhesion of the HGF. Coomassie brilliant blue staining techniques, type I collagen fiber ELISA experiment and Real- time PCR technique are used to test the secretion of HGF which come from the materials. Results:(1)When plasma FN was modified on the surface of micron grade titanium groove by the method of silicon oxide, XPS analysis that there were chemical bonds which strength was higher than the direct immersion group between FN and silicon oxide of titanium plate. The elements which come from the surface of the Silicon oxide materials was tested by XPS and the OD value of ELISA detection showed that FN content was significantly more than the direct immersion group, with statistical significance(P < 0.05). When we used fluorescence microscope observe the fluorescent dye FN, It was found FN distributed evenly on the material by,the fluorescence pixels were significantly higher than the control group, with statistically significant(P < 0.05).(2)Field emission scanning electron microscopy(SEM) discover that the surface of the silicon oxide T60/10 had not been etched by chemicals which were used or produced in the process of silicon oxidation reaction, and the size remains the same. It was found by Atomic force microscope that the modified FN material’s roughness had increased, with statistical significance(P < 0.05). It was also found by contact Angle meter that he modified FN material’s wetting Angle was less than the control group, with statistical significance(P < 0.05).(3)The cell proliferation and adhesion: DAPI staining was used to detect the early adhesion number of HGF which is vaccinated in the surface of the modified FN groove and smooth, undecorated FN group and smooth titanium plates after 2h, 4h, and 6h. The Fluorescent number which are on the surface of four group materials is less, almost equal. It is discovered that the number of the cell which adhered on the four groups materials increased obviously from 4 h, and the number of cells on the two silicon oxide materials groups are greater than the blank group. The numbers of the silicon oxidation groove group were largest in the four groups. The numbers of cells on the silicon oxide smooth material were larger than the blank group, and the cell number of silicon oxide smooth group is as same as the blank groove group. The cell on the blank groove material is larger than smooth group. At 6h the cell number on four materials groups increased more obviously at the same time, the number of cell on the silicon oxidation groove group increased more significantly than other three groups, the silicon oxide smooth more than blank groove group also. The result of DAPI staining experiment on different time after HGF was vaccinated; we discovered the materials which were modified FN through silicon oxide methods can effectively promote more HGF to adhere on the surface of materials on the early time. CCK8 was used to test the number of HGF were vaccinated on the modified FN groove and smooth, undecorated FN groove and smooth titanium plates at 6h, 12 h, 1d, 3d, 5d, 7d. It is found that the number of cells on four material groups increased with time. The number of cells on the surface of the silicon oxide groove materials was lager than other three groups of materials in the six period, and there are statistically significant(P < 0.05); The number of cells which was on the surface of silicon oxide smooth group were below the undecorated groove group all time except 1d(P < 0.05); The number of cells which was on the surface of the blank smooth group was still lower than the other three groups in each time(P < 0.05). According to the observation come from Fluorescence microscope at 1d, 3d, 5d, the number of cells on different materials were consistent with CCK8 results. The cells were elongated and regular array along with groove on silicon oxidation groove titanium group and blank groove titanium group. The cells were polygonal and irregular array on silicon oxidation smooth titanium and blank titanium group, cell arrangement is irregular, cells are polygonal. The cells which were on two silicon oxidation material groups spread larger, the pseudopodium of the cells were longer than the other.(4)According to the results of coomassie brilliant blue staining at 1d, 3d, 5d and type I collagen fiber ELISA experiments at 5d, the number of extra cellular total protein and extra cellular type I collagen fiber from two silicon oxide material groups were larger than the number of blank material. According to Real-time PCR analysis, type I collagen m RNA expression level of HGF which were on two silicon oxide material groups were higher than blank group, the above differences were statistically significant(P<0.05). Conclusions:The plasma fibronectin(FN) was modified on T60/10 by silicon oxidation would not change the surface of the original groove shape, but also increased the hydrophilic of material and make up for the lack of adsorption capability for plasma fibronectin at the same time. The FN modification materials which have more biocompatibility than blank group could promote the proliferation and early adhesion of HGF effectively. HGF which come from the FN modification materials synthetic more protein and type I collagen fiber than the blank group, and type I collagen m RNA expression level were higher than the blank group at the same time. Type I collagen fiber which is synthetic by HGF is an important part of the "collar", so the FN modification materials will be in implant to the neck soft tissue healing, and have a positive effect to prevent infection at the same time. |
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