| Objective: Periodontitis is an diseases which cause four kinds ofperiodontal support organization inflammation and defect, it’s major clinicalmanifestations is periodontal pocket formation, periodontal attachment lossand alveolar bone absorption. Along with the age increased the prevalence andseverity of periodontitis increases. Oral epidemiological survey results showedthat periodontitis has become the first reason to adult’s tooth lost in China,which seriously impacts the patients’ chewing function and periodontal beauty.Medical research indicate that periodontitis is closely related to varioussystemic disease, and effects of periodontitis on blood lipid and blood sugarlevel could be the important reasons which increased probability ofcardiovascular disease, diabetes etc system disease. In the clinical treatment ofperiodontitis, the traditional local and systemic drugs and periodontalfoundation treatment can control gum inflammation and improve theperiodontal status, but with little effect for the reconstruction of theperiodontal tissue. Therefore, it becomes urgent clinical problems how tomake periodontal tissue damaged and lost repair and regenerate to restoreperiodontal beautiful appearance and normal chewing function. Theemergence of periodontal tissue engineering technology provides forperiodontal tissue regeneration a new train of thought.Periodontal tissue engineering achieve repair and regeneration throughusing the principle, method and technical of life science and biologicalmaterials science to reconstruct periodontal tissue with new original form andfunction, and it includes three fundamental aspects: seed cell, support materialand growth factor. In this research shuanghuangbu as growth factor，withgingival fibroblasts as seed cells and zein support material combination werecombined and applied in animal bone defect model to study the effect and mechanism of in the periodontal tissue repair and regeneration, in order toprovide theory for gingival fibroblast-zein-shuanghuangbu complex inperiodontal tissue engineering development and application.Methods:20teeth of two beagle dogs were chosen and divided into4groups with5teeth in each group. Bone defect models were made and donethe following treatment respectively. Experimental groupⅠ:flap operation+zein; experimental groupⅡ:flap surgery+gingival fibroblast-zein;experimental group Ⅲ: flap surgery+gingival fibroblast-zein-shuanghuangbucomplex; control group: flap surgery. The specimens were separately obtainedin three months after operation, according to the Cone beam CT inspection,histological measurement(experimental defect,new alveolar bone formation,new cementum formation,the lenghth of junctional epithelium) and immuno-histochemical staining quantitative analysis (bone morphogenetic protein-2,bone sialoprotein),to research the effect and mechanism of gingival fibroblast-zein-shuanghuangbu complex on the regeneration of the experimentalperiodontium.Results:1Histologic observationAt the3month after operation, inflammatory cells infiltration wasevident in gingival and periodontium of experimental group Ⅰand Ⅱ, butthere wasn’t obvious inflammatory cells in the experimental group Ⅲ. bonedefect area of experimental groupⅠ showed the new bone formation, withbone trabecular slender around by lots of fiber organization; there was sheetnew bone, osteoblast arranged on edge of bone defect and more new bloodvessels in periodontal membrane in experimental Ⅱ; There were somegradually mature new bone tissue in experimental group Ⅲ,and haversiansystem were observed; In defect area of control group,there was majorfibrosis bone, with lesser new bone trabecular and osteoblast, cementoblast.Four groups showed evident junctional epithelium shift to root.2Histology measurement results Height measurement of the new alveolar bone in the experimental groupⅠ, Ⅱ, Ⅲ separately were1.02±0.09mm,1.03±0.06mm and1.14±0.07mm, higher than those in the control group±0.06mm (P<0.05). The newcementum height of experimental Ⅲ was1.00±0.08mm, higher than that0.91±0.10mm in control group (P <0.05); And there was not a significantdifference of the lenghth of junctional epithelium in four groups (P>0.05).3Cone beam CT inspection resultIn bone defect area of group experimental there was new indistinct boneformation, blending with surrounding bone tissue gradually; Point and platesareas of high density were observed in experimental group Ⅱ; Theexperimental Ⅲ displayed vast new bone formation in bone defect area, andcortical bone of defect edge was continuous; The bone defect area in controlgroup was lost, but there was the most low density shadow in four groups.4Bone mineral density of alveolar bone measurement resultsThe percentage of bone mineral density difference between normal areaand operative area in the experimental group Ⅱ and Ⅲ was lower than that incontrol group on three months postoperatively (P<0.05),and percentage ofbone mineral density difference in experimental group Ⅲ was lower than thatin experimental group Ⅱ,but there was no significant difference between theexperimental group Ⅰand the control group (P>0.05).5Immunohistochemical resultsBone morphogenetic proteins-2positive expression was in cytoplasmwith brownish-yellow granules or briquette. There were bone morphogeneticproteins-2positive expression in four groups. Osteoblasts around new alveolarbone and cementoblast were positive staining. In experimental group Ⅱ, Ⅲpositive expression rate of bone morphogenetic proteins-2were higher thanthat in the control group (P<0.05), there was no significant difference betweenexperimental group Ⅰand control group (P>0.05).Bone sialoprotein positive expression was in cytoplasm with browncoloring.There were bone sialoprotein positive expression in four groups,brownish-yellow granules or mass was visible in cytoplasm of cementoblast and osteoblast. Compared with the control group, bone sialoprotein positiveexpression rate in experimental group Ⅱ, Ⅲ obviously increased (P <0.05),there was no significant difference between experimental group Ⅰand controlgroup (P>0.05).Conclusion:1Gingival fibroblast-zein-shuanghuangbu complex may be speed up theformation and maturity of bone matrix through promoting proliferation ofgingival fibroblasts and osteoblast differentiation,to accelerate theregeneration of the periodontal tissue.2Gingival fibroblast-zein-shuanghuangbu complex can increase bonemor-phogenetic proteins-2and bone sialoprotein expression in the periodontaltissue, and accelerate the new bone and cementum formation.3Shuanghuangbu plays an important part in inhibiting bacterial growthand controlling periodontal inflammation. |
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