| The clinical indications of implant dentures are extensive,in recent ten years,with the rapid development of implant denture,more and more patients with tooth defect began to fix their missing teeth with the implant denture..Implantology has become one of the most rapid development of the two disciplines of Oral Science,it is no exaggeration to say that Implantology has changed the dental implant model.Peri-implant inflammation(also known as peri-implant mucositis)is a reversible inflammation of the soft tissue around the implant.The underlying cause is bacterial microorganisms on the implant.Due to the poor oral hygiene,the accumulation of plaque are gathering to the implants,stimulates the body to produce inflammatory response.The clinical manifestations show as the swelling,bleeding or bleeding of gingival and mucous.It is the chronic inflammation of the tissue around the implant after the implantation of the implant or the function,which results in the bone resorption around the implant,and the final outcome of the development is the loss of the implant.The good quality of osseointegration and the "cuff" closure of the soft tissue attached to the implant neck are the biological basis for maintaining the long-term stability of the implant."Cuff" structure,can play a role in the biological barrier,can effectively prevent bacteria and other inflammatory factors invasion,prevention of peri implantitis,reduce bone resorption,improve the clinical success rate of implant.In this study,we aimed to explore a method of healing abutment surface which is more beneficial to the attachment of gingival fibroblasts,and to improve the design of the abutment surface.The implant can be stable for a long time and has a good effect on the oral environment.Although the implant and natural teeth with epithelial junctional epithelium around similar to hemidesmosome and basal plate attached on the surface of implant;but the combination of implant and connective tissue has been controversial.The gingival connective tissue in gingival fibroblasts are the most important position,so we study Ti6A14V alloy micro arc oxidation and micro arc oxidation of silver treatment on gingival fibroblasts and its biological behavior influence in guiding us to explore a method for surface treatment of healing abutment is conducive to the formation of the cuff to deposit a certain clinical significance.Chapter ?:Primary culture and identification of Human gingival FibroblastsObjective:To isolate and culture Human gingival Fibroblasts(HGFs)in vitro,and identify their morphological,histological and growth characteristics.Method:Selection of young patients in South Hospital of Southern Medical University Department of Orthodontics who needs extraction treatment,age in 12 to 20 years,patients with healthy gingival and without dental caries and taking drugs which make gingival hyperplasia.With the consent of local anesthesia,the gingival tissue was removed and the tissue was cultured by tissue explant method.Results:1.The first results of microscope show that primary cells with cultured about 8-10d swim out from the black organization.Around to the block edge,cells are adhenting to the bottles,the cells' morphology likes long fusiform,uniform cytoplasm,we can see the single nuclears in the cytoplasm of central oral,2-3 nuclears get in the center of it.With the increase of the number of days by cultureing,the cells extend from the center of the tissue block to the periphery,and the tissue block is in the center.The cells covered the bottom of Petridish can be seen around the area of about 90%HGFs cells closely arranged elongated spindle cells arranged in whorls and braided,cultured human gingival fibroblast growth,epithelial cells gradually disappear,traits tend to be stable.2.Cells which under the light microscope by immunohistochemical staining exhibited anti vimentin staining was positive in the cytoplasm of positive particles,uniform distribution,clear nucleus,no staining;anti keratin staining was negative,that the cells derived from the mesoderm,without skin derived cells mixed.There was no positive staining in negative control group.The results showed that the cultured cells derived from mesenchymal stem cells and epithelial cells.Conclusion:In this study,we can see epithelial cells can be culturesd from the tissue,and be identified by immunohistochemistry.Although the proliferation of these cells is weak,but they can last longer in the vitroChapter ?:Study on physical and chemical properties of Ti6A14V titanium alloy by micro arc oxidationObjective:Using scanning electron microscope(SEM),energy spectrum analyzer(EDS),surface roughness measurement instrument and contact angle measurement instrument to take test of microarc oxidation and microarc oxidation with Ag treated Ti6A14V titanium alloy surface mor:phology characteristics,elemental composition,crystal structure,surface roughness and surface level contact angle.Method:Make Ti6A14V alloys in 8mm diameter,0.8mm thickness;take Ti6A14V titanium alloy as anode and platinum electrode as the cathode,set the voltage at the level of 350V,frequency at 50Hz,duty cycle at 50%,cool the electrolyte with ice to keep the temperature at 40? for 5mins.Samples with micro arc oxidation should be washed by deionized water after ultrasonic by 15min,drying plastic sealing packaging stand.Make the silver nitrate solution strength of 0.006 mol/L.Take The platinum electrode as anode,the sample after micro arc oxidation treatment as cathode,silver nitrate solution as the electrolyte,set the voltage at 2V,deal with samples for 30S(storage of electrolyte,wash samples in ultrasonic cleaning and drying,obtained silver microarc oxidation samples.Using scanning electron microscopy(SEM)to observe the surface morphology and structure;calculateing the surface roughness,measuring contact angle and liquid surface materials the surface free energy calculation.Results:Scanning electron microscope was used to observle the surface of titanium alloy after micro arc oxidation,the surface of porous ceramic was observed,and the pore size was about 2-5?m.Detection of Ti6A14V-MAO and Ti6Al4V-MAO-Ag surface roughness show roughness of 1.59+-0.056m.The contact angle of Ti6Al4V-MAO and Ti6A14V-MAO-Ag group were less than that of smooth group.Conclusion:By means of scanning electron microscopy(SEM),energy dispersive spectroscopy,surface roughness and contact angle measurement,it was proved that Ti6A14V was successfully treated by micro arc oxidation and micro arc oxidation.Chapter ?:Effects of silVer coating on Ti6A14V titanium alloy with micro arc oxidation on the adhesion,cell morphology and proliferation of human gingival fibroblastsObject:Study on the effect of silver coating on Ti6A14V titanium alloy by micro arc oxidation and micro arc oxidation on the early adhesion,cell morphology and proliferation of human gingival fibroblasts.Method:In Ti6A14V titanium alloy surface smooth,Ti6A14V-MAO and Ti6A14V-MAO-Ag were inoculated in primary cultured human gingival fibroblast cells by scanning electron microscopy,fluorescence microscope,CCK-8 was detected to observe the HGFs early adhesion,proliferation and morphology of cells.Results:1.Cells in the early adhesion of smooth Ti6A14V titanium alloy,Ti6Al4V-MAO and Ti6Al4V-MAO-Ag surface were inoculated with human gingival fibroblast cells,4h cells were inoculated with DAPI,12h dye on the surface of the nuclears and cytoskeleton were dyed with the number of cells was calculated under fluorescence microscope.6h,12h group and Ti6A14V-MAO-Ag group,the number of surface cells was significantly more than the smooth group,there was a significant difference(P<0.05)(Ti6A14V-MAO).2.Cell morphology:treatment by fluorescent staining of HGFs were inoculated on smooth Ti6A14V titanium alloy and Ti6Al4V-MAO surface after 24 h of cytoskeleton fluorescence was observed under the microscope,found that gingival fibroblasts arranged along the lines on the polished specimen with smooth surface of titanium alloy,a spindle type line,cells showed polar arrangement.While the Ti6Al4V-MAO and Ti6A14V-MAO-Ag groups were spindle type surface growth,cell spreading area.Scanning electron microscope obserVation on gingival fibroblasts and sample temperature after 6h co cultured cells not fully extended,round and oral.While the MAO and MAO-Ag group co cultured gingival fibroblasts and filopodia extending pseudo wire.After continuing to culture to 12h,the smooth group cells were found to be extended,and the MAO and MAO-Ag groups had significantly increased gingival fibroblasts.3.Cell proliferation:using CCK8 method to detect HGFs inoculated in the control group,MAO group and MAO-Ag group,the smooth surface of 1d,3d,5d group,7d after the number of cell proliferation,found in culture to Id,proliferation number did not change significantly.But after 3d,5d and 7d,the number of cells on the surface of the MAO-Ag group was significantly higher than that of the smooth group,and there was statistical difference.Conclusion:Microarc oxidation and micro arc oxidation treatment of Ag Ti6A14V could promote the growth of adhesion on gingival fibroblast proliferation,silver treatment did not inhibit the proliferation of Ti6A14V alloy by micro arc oxidation on cell growth and adhesion of fiber gum. |
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