發(fā)布時間：2018-11-07 20:16

【摘要】：背景：臨床常見由于過大正畸力或咬合干擾引起的咬合創(chuàng)傷。正常情況下的(?)力對牙周組織可以產(chǎn)生良性刺激,而異常的(?)力則會造成牙周組織的病理性損傷。臨床中常見(?)創(chuàng)傷或不適當?shù)恼υ斐傻难乐芨街鴨适�、牙根吸收以及牙齒松動等。在牙周膜中膠原纖維不斷改建,已有文獻報道,如果對成纖維細胞功能的破壞均能夠引起牙周支持組織的喪失,在牙周組織的改建過程中,胰島素樣生長因子(IGF)以及破骨細胞與改建過程密切相關(guān)。目的：本研究有效建立了實驗性大鼠磨牙咬合創(chuàng)傷的動物模型,通過觀察在咬合創(chuàng)傷及創(chuàng)傷刺激因素去除前后牙周組織中膠原纖維及成纖維細胞形態(tài)學(xué)變化,胰島素樣生長因子(IGF-I)的表達變化及破骨細胞數(shù)量活性變化等情況,探討咬合創(chuàng)傷性牙周改變的發(fā)生發(fā)展以及修復(fù)過程,為相應(yīng)的臨床治療提供實驗依據(jù)。方法：選取Wistar大鼠作為實驗動物,雄性,三月齡,體重250-280g,無齲病、牙周病,共25只。隨機分為對照組和實驗組,對照組5只大鼠；實驗組分為1周組、3周組、4周組及咬合創(chuàng)傷2周后去除(?)創(chuàng)傷2周組,共五組,每組5只大鼠。實驗組大鼠于下頜右側(cè)第一磨牙粘3/4鈷鉻金屬冠,金屬冠高出(?)面0.8mm,對照組不做任何處理。將大鼠在各自的觀察時間點麻醉后心臟灌注并處死,切取右側(cè)下頜第一磨牙區(qū)軟硬組織,固定,脫鈣,包埋,正中矢狀面連續(xù)切片,切片厚度為5μm。首先對切片進行HE染色,觀察大鼠磨牙區(qū)牙周支持組織的形態(tài)學(xué)改變；對切片進行免疫組織化學(xué)染色,觀察IGF-I表達變化；對切片進行TRACP染色以及CK免疫組化染色,觀察破骨細胞數(shù)量及活性變化。使用ImagePro-Plus6.0軟件對IGF-I和CK免疫組化染色的平均光密度值計算及隨機視野內(nèi)TRACP陽性細胞計數(shù),用Graphpad Prism6. x. C統(tǒng)計軟件進行數(shù)據(jù)分析。結(jié)果：1建立咬合創(chuàng)傷動物實驗?zāi)Ｐ碗S著咬合創(chuàng)傷的時間延長,大鼠逐漸性情急躁,被毛雜亂無光澤,進食減少,后不喜運動,反應(yīng)較慢。咬合創(chuàng)傷四周組大鼠平均體質(zhì)量增長較對照組減少30g,而對照組大鼠行為正常,被毛順滑有光澤。2 HE染色結(jié)果對照組：牙周膜纖維結(jié)構(gòu)致密,排列有序,成纖維細胞沿纖維長軸排列。實驗組：1w組牙周纖維排列稍紊亂,牙周膜血管擴張出血；3w組牙周膜增寬,牙周纖維排列紊亂,牙槽骨的破壞更加明顯；4w組病損的程度最重,骨破壞最明顯；去除咬合創(chuàng)傷組牙周組織接近正常,可見組織修復(fù)3牙周膜中IGF-I表達變化對照組：IGF-I在牙周膜中呈陰性表達或弱陽性表達。實驗組：牙周支持組織中IGF-I濃度逐漸增加,在4w組時表達最多；去除合創(chuàng)傷組IGF-I陽性細胞的數(shù)量減少。主要陽性表達部位為成牙骨質(zhì)細胞和成骨細胞。各實驗組統(tǒng)計學(xué)結(jié)果與對照組相比均P0.05,有統(tǒng)計學(xué)意義。4牙周組織破骨細胞中TRACP變化對照組：破骨細胞中TRACP呈弱陽性表達,少量表達于牙周膜與牙槽骨交界處。實驗組：1w組TRACP在牙周組織中呈弱陽性表達,主要表達于牙槽骨邊界的破骨細胞(p0.01)；3w組牙周組織染色陽性反應(yīng)增強,紅色陽性個數(shù)增多(p0.01)；4w組牙槽骨中可見大量破骨細胞,破骨細胞中出現(xiàn)染色陽性反應(yīng),在骨吸收陷窩內(nèi)有強陽性表達(p0.01)；去除創(chuàng)傷組與3w、4w組相比,TRACP陽性表達略減弱,骨陷窩內(nèi)可見成骨細胞修復(fù),部分破骨細胞中無陽性反應(yīng)(p0.01)。3w組與1w、4w組p0.05,無統(tǒng)計學(xué)意義,其他各實驗組間均p0.01,有統(tǒng)計學(xué)意義。5牙周組織中破骨細胞CK變化對照組：偶可見CK表達陽性的破骨細胞。實驗組：1w組可見少量CK表達陽性的破骨細胞；3w組破骨細胞活性增強,主要位于臨近牙周膜的牙槽骨；4w組破骨細胞活性最強,骨吸收明顯,牙槽骨中可見大量陽性破骨細胞；去除創(chuàng)傷組破骨細胞陽性減弱,可見骨組織修復(fù)。1w、3w、4w組與對照組統(tǒng)計學(xué)結(jié)果均P0.05,有統(tǒng)計學(xué)意義。結(jié)論：1咬合創(chuàng)傷可引起大鼠牙周組織的病理改變,牙周膜增寬,牙周纖維紊亂,牙槽骨吸收。創(chuàng)傷時間越久,病理改變越明顯。2去除合創(chuàng)傷及局部刺激因素有利于牙周組織的愈合。3咬合創(chuàng)傷去除前后,IGF-I可能通過促進成纖維細胞的分化與增殖的機制參與了組織修復(fù)。4咬合創(chuàng)傷可促使破骨細胞分化,引起牙槽骨破壞。
[Abstract]:Background: The clinical common is due to overlarge orthodontic force or occlusal trauma. Under normal conditions (?) The force can cause a benign stimulus to the periodontal tissue, and the abnormal (?) the force may cause pathological damage to the periodontal tissue. What is common in the clinical (?) Periodontal attachment loss, root resorption, and tooth loosening due to trauma or inappropriate orthodontic force. In the course of the remodeling of the periodontal tissue, the insulin-like growth factor (IGF) and the osteoclast are closely related to the remodeling process. Objective: In this study, an animal model of experimental rat molar occlusion was established, and the morphological changes of the collagen fibers and fibroblasts in the periodontal tissues were observed by observing the factors of occlusion and trauma. The changes of the expression of insulin-like growth factor (IGF-I) and the change of the number of osteoclasts were discussed. Methods: Wistar rats were selected as experimental animals, male, three-year-old, body weight of 250-280g, caries-free and periodontal disease. The control group and the experimental group were randomly divided into control group and experimental group. The experimental group was divided into 1 week group, 3 week group, 4 week group and 2 weeks after occlusion.) The 2-week-old group was divided into five groups, each group of 5 rats. The first molar of the experimental group was 3/ 4 cobalt-chromium metal crown on the right side of the lower jaw, and the metal crown was higher (?) The surface was 0.8mm, and the control group did not do any treatment. After the rats are anesthetized with each observation time point, the heart is perfused and executed, the soft and hard tissue of the first molar area of the right lower jaw is cut, the soft and hard tissue of the right mandibular first molar area is fixed, decalcified, embedded, the median sagittal plane is continuously cut, and the thickness of the slice is 5. m The morphological changes of the periodontal supporting tissues in the molar areas of the rats were observed, and the expression of IGF-I was observed by immunohistochemical staining of the slices, and the number of osteoclasts and the changes of the activity were observed by TRACP staining and immunohistochemical staining of the slices. The average optical density values of IGF-I and CK immunohistochemical staining were calculated using ImagePro-Plus6.0 software and the TRACP positive cells in the random field of view were counted and Graphpad Prim6 was used. The x. C statistical software is used for data analysis. Results: 1 The experimental model of the animal model of the occlusion of the wound was prolonged with the time of the occlusion, and the rats were impetuous, the hair was mixed with the hair, the food was reduced, the exercise was not exhilarated, and the response was slow. The average mass of the rats in the peripheral group increased by 30g in the control group, while the rats in the control group were normal, and the hair was smooth and smooth. The results of HE staining showed that the fibrous structure of the periodontal ligament was dense, the order was ordered, and the fibroblasts were arranged along the long axis of the fiber. In the experimental group, the periodontal fibers in the 1w group were arranged slightly, and the periodontal ligament was expanded and bleeding; the periodontal ligament of the 3w group was widened, the arrangement of the periodontal fibers was disordered and the destruction of the alveolar bone was more obvious; the degree of the lesion in the 4w group was the most and the bone destruction was the most obvious; the periodontal tissue in the group of the bite wound group was close to normal, The expression of IGF-I in the periodontal ligament was observed by the visible tissue. The expression of IGF-I was negative or weakly positive in the periodontal ligament. In the experimental group, the concentration of IGF-I in the periodontal supporting tissues increased gradually, and the expression of IGF-I was the most in the 4w group, and the number of IGF-I positive cells in the treated group was decreased. The main positive expression site is cementum and osteoblast. The results showed that the TRACP in the osteoclast of the periodontal tissues showed a weak positive expression, and a small amount of the TRACP in the osteoclasts was expressed in the junction of the periodontal ligament and the alveolar bone. The expression of TRACP in the experimental group was weakly positive in the periodontal tissues, and the expression of TRACP in the alveolar bone was mainly expressed in the osteoclasts (p0.01). In the 3w group, the staining positive reaction of the periodontal tissues was enhanced, the number of red-positive cells increased (p0.01), and a large amount of osteoclasts were found in the alveolar bone of the 4w group. The positive expression of TRACP was positive in osteoclasts (p0.01). Compared with the 3w and 4w group, the positive expression of TRACP was slightly decreased, and the osteoblast in the bone was repaired, and no positive reaction was found in some osteoclasts (p0.01). 3w group and 1w and 4w group p0.05, There was no statistical significance. There was a statistical significance between the other experimental groups and the control group of osteoclast CK in the periodontal tissues. In the experimental group, a small amount of CK-expressing osteoclasts was found in the 1w group, and the osteoclast activity in the 3w group was enhanced mainly in the alveolar bone near the periodontal ligament; the osteoclast activity of the 4w group was the strongest, the bone resorption was clear, and a large number of positive osteoclasts were found in the alveolar bone; The results showed that there was no significant difference between the group and the control group. Conclusion: 1. The occlusal trauma can cause the pathological changes of the periodontal tissue of the rat, the widening of the periodontal ligament, the disorder of the periodontal fiber and the absorption of the alveolar bone. the longer the wound time, the more obvious the pathological change. The removal of the wound and local stimulation is beneficial to the healing of the periodontal tissue. Before and after the occlusion, IGF-I may be involved in the tissue repair by the mechanism of promoting the differentiation and proliferation of fibroblasts. causing alveolar bone destruction.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R783.5

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