咬合创伤影响大鼠牙周组织改建的机制初探
发布时间：2018-11-07 20:16
【摘要】：背景：临床常见由于过大正畸力或咬合干扰引起的咬合创伤。正常情况下的(?)力对牙周组织可以产生良性刺激,而异常的(?)力则会造成牙周组织的病理性损伤。临床中常见(?)创伤或不适当的正畸力造成的牙周附着丧失、牙根吸收以及牙齿松动等。在牙周膜中胶原纤维不断改建,已有文献报道,如果对成纤维细胞功能的破坏均能够引起牙周支持组织的丧失,在牙周组织的改建过程中,胰岛素样生长因子(IGF)以及破骨细胞与改建过程密切相关。目的：本研究有效建立了实验性大鼠磨牙咬合创伤的动物模型,通过观察在咬合创伤及创伤刺激因素去除前后牙周组织中胶原纤维及成纤维细胞形态学变化,胰岛素样生长因子(IGF-I)的表达变化及破骨细胞数量活性变化等情况,探讨咬合创伤性牙周改变的发生发展以及修复过程,为相应的临床治疗提供实验依据。方法：选取Wistar大鼠作为实验动物,雄性,三月龄,体重250-280g,无龋病、牙周病,共25只。随机分为对照组和实验组,对照组5只大鼠；实验组分为1周组、3周组、4周组及咬合创伤2周后去除(?)创伤2周组,共五组,每组5只大鼠。实验组大鼠于下颌右侧第一磨牙粘3/4钴铬金属冠,金属冠高出(?)面0.8mm,对照组不做任何处理。将大鼠在各自的观察时间点麻醉后心脏灌注并处死,切取右侧下颌第一磨牙区软硬组织,固定,脱钙,包埋,正中矢状面连续切片,切片厚度为5μm。首先对切片进行HE染色,观察大鼠磨牙区牙周支持组织的形态学改变；对切片进行免疫组织化学染色,观察IGF-I表达变化；对切片进行TRACP染色以及CK免疫组化染色,观察破骨细胞数量及活性变化。使用ImagePro-Plus6.0软件对IGF-I和CK免疫组化染色的平均光密度值计算及随机视野内TRACP阳性细胞计数,用Graphpad Prism6. x. C统计软件进行数据分析。结果：1建立咬合创伤动物实验模型随着咬合创伤的时间延长,大鼠逐渐性情急躁,被毛杂乱无光泽,进食减少,后不喜运动,反应较慢。咬合创伤四周组大鼠平均体质量增长较对照组减少30g,而对照组大鼠行为正常,被毛顺滑有光泽。2 HE染色结果对照组：牙周膜纤维结构致密,排列有序,成纤维细胞沿纤维长轴排列。实验组：1w组牙周纤维排列稍紊乱,牙周膜血管扩张出血；3w组牙周膜增宽,牙周纤维排列紊乱,牙槽骨的破坏更加明显；4w组病损的程度最重,骨破坏最明显；去除咬合创伤组牙周组织接近正常,可见组织修复3牙周膜中IGF-I表达变化对照组：IGF-I在牙周膜中呈阴性表达或弱阳性表达。实验组：牙周支持组织中IGF-I浓度逐渐增加,在4w组时表达最多；去除合创伤组IGF-I阳性细胞的数量减少。主要阳性表达部位为成牙骨质细胞和成骨细胞。各实验组统计学结果与对照组相比均P0.05,有统计学意义。4牙周组织破骨细胞中TRACP变化对照组：破骨细胞中TRACP呈弱阳性表达,少量表达于牙周膜与牙槽骨交界处。实验组：1w组TRACP在牙周组织中呈弱阳性表达,主要表达于牙槽骨边界的破骨细胞(p0.01)；3w组牙周组织染色阳性反应增强,红色阳性个数增多(p0.01)；4w组牙槽骨中可见大量破骨细胞,破骨细胞中出现染色阳性反应,在骨吸收陷窝内有强阳性表达(p0.01)；去除创伤组与3w、4w组相比,TRACP阳性表达略减弱,骨陷窝内可见成骨细胞修复,部分破骨细胞中无阳性反应(p0.01)。3w组与1w、4w组p0.05,无统计学意义,其他各实验组间均p0.01,有统计学意义。5牙周组织中破骨细胞CK变化对照组：偶可见CK表达阳性的破骨细胞。实验组：1w组可见少量CK表达阳性的破骨细胞；3w组破骨细胞活性增强,主要位于临近牙周膜的牙槽骨；4w组破骨细胞活性最强,骨吸收明显,牙槽骨中可见大量阳性破骨细胞；去除创伤组破骨细胞阳性减弱,可见骨组织修复。1w、3w、4w组与对照组统计学结果均P0.05,有统计学意义。结论：1咬合创伤可引起大鼠牙周组织的病理改变,牙周膜增宽,牙周纤维紊乱,牙槽骨吸收。创伤时间越久,病理改变越明显。2去除合创伤及局部刺激因素有利于牙周组织的愈合。3咬合创伤去除前后,IGF-I可能通过促进成纤维细胞的分化与增殖的机制参与了组织修复。4咬合创伤可促使破骨细胞分化,引起牙槽骨破坏。
[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.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R783.5
本文编号：2317456
[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.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R783.5
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