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RANKL和OPG在大鼠牙移动性根吸收及早期修复中的表达研究

【作者】 冯慧

【导师】 陈文静;

【作者基本信息】 南京医科大学 , 口腔正畸学, 2008, 硕士

【摘要】 第一部分RANKL和OPG在大鼠牙移动性根吸收中的表达研究目的初步研究大鼠正畸牙移动性根吸收中破骨细胞和骨保护因子(osteoprotegerin,OPG)、破骨细胞分化因子(receptor activator ntlclearfactor kappa B ligand,RANKL)的变化和规律,探讨破骨细胞在正畸导致的炎性牙根吸收中的作用。方法选用6-8周龄Wistar雄性大鼠39只,随机分为实验组及对照组,对照组3只,实验组每组6只。实验组建立大鼠正畸牙移动性根吸收模型。分别在0d、1d、3d、5d、7d、10d及14d全麻下处死动物。HE染色观察细胞形态反应,TRAP染色观察破骨细胞的变化,免疫组化检测OPG、RANKL的表达。结果正畸牙移动性根吸收模型建立的第7天起可见显著的牙根吸收现象,牙槽骨边缘及牙骨质出现骨吸收陷窝。实验第1、3、5、7天,实验组TRAP阳性单核破骨细胞增加;实验第3、5、7、10天,实验组TRAP阳性多核破骨细胞增加,实验组与对照组有显著性差异。实验第3、5、7、10天,实验组RANKL强阳性表达;实验第5、7、10天,实验组OPG强阳性表达,实验组与对照组有显著性差异结论正畸牙移动性根吸收过程中,破骨细胞起关键作用,并随作用时间,破骨细胞数量和部位发生变化。OPG、RANKL的表达水平与破骨细胞的分化成熟变化过程密切相关,发挥重要作用。第二部分RANKL和OPG在大鼠牙移动性根吸收修复早期中的表达研究目的初步研究大鼠正畸牙移动性根吸收修复早期中破骨细胞和骨保护因子(osteoprotegerin,OPG)、破骨细胞分化因子(receptor activatornuclear factor kappa B ligand,RANKL)的变化和规律,探讨破骨细胞在正畸导致的炎性牙根吸收修复中的作用。方法建立大鼠正畸牙移动性根吸收修复早期模型,TRAP染色观察破骨细胞的变化,免疫组化检测OPG、RANKL的表达。结果实验第1天起,TRAP染色阳性多核破骨细胞细胞逐渐减少,实验第7、10、14天,TRAP染色阳性多核破骨细胞数量已逐渐恢复至生理水平。实验第1天,OPG、RANKL表达量下降,第3天以后逐渐恢复至生理水平。结论施力终止后,大鼠牙根即停止吸收,并于1-3天后开始牙根修复,OPG、RANKL的表达水平与此过程密切相关。

【Abstract】 Part one Expression of OPG and RANKL in the process of orthodontic induced root resorption in rats.Objective To investigate the role of OPG and RANKL in induction of osteoclasts during the process of orthodontic induced tooth root resorption of rats.Methods The process of orthodontic induced tooth resorption of upper molars was performed in 39 Wistar rats(6-8 weeks).The rats were divided into control group(3 rats)and experimental group(6 rats each group).The model of orthodontic induced tooth resorption was peformed in the experimental group.The animals were sacrisfied in 0d,1d,3d,5d, 7d,10d,and 14d respectively.The morphologic change was observed. The expression of OPG and RANKL protein at the compression sites was detected by immunohistochemical staining technique.Furthermore,we observed the TRAP positive cells in this process.Results After 7 days of the establish of orthodontic induced root resorption model,the obvious root resorption was observed,and absorption lacunas were detected at the edge of alveolar bone and in the cementum.The TRAP positive mononucleated osteoclast were increased in 1d,3d,5d and 7d;while the TRAP positive multinucleated osteoclast were increased in 3d,5d,7d and 10d.Immunohistochemical staining revealed that amount of OPG protein was increased in 3d,5d,7d,10d; that amount of RANKL protein was increased in 5d,7d,10d.Conclusion It indicates that the osteoclast plays key role in the process of orthodontic induced root resorption,and the amount and location of osteoclasts changed with time.The expression of OPG、RANKL is related to this process.Part two Expression of OPG and RANKL in the repair process of orthodontic induced root resorption in rats.Objective To investigate the role of OPG and RANKL in induction of osteoclasts during the repair process of tooth root resorption.Methods The repair of orthodontic induced tooth resorption of upper molars was performed in Wistar rats.The expression of OPG protein at the compression sites was detected by immunohistochemical staining technique.Furthermore,we observed the TRAP positive cells in this process.Results The TRAP positive cells were decreased from the first day after removing the force,and the amount of the TRAP positive cells was almost the same as normal after 7 days.Immunohistochemical staining revealed that amount of OPG、RANKL protein was decreased from the first day after removing the force,and from the third day on,the amount of the OPG、RANKL protein was almost the same as normal.Conclusion It indicates that the root of rats stopped resorption immediately after the the removal of the force,and started to repair after 1-3 days.The expression of OPG、RANKL is related to this process.

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