降钙素和1，25维生素D拮抗牙周炎大鼠牙槽骨丧失的疗效观察

发布时间：2018-06-07 00:02

本文选题：牙周炎 + 降钙素　；参考：《天津医科大学》2014年硕士论文

【摘要】：目的：本实验通过建立大鼠牙周炎模型并对其应用1,25维生素D及降钙素后,观察牙周探诊深度(probing depth, PD)、牙槽骨丧失水平,检测血清中的骨碱性磷酸酶(bone alkaline phosphatase, BALP)及骨钙素(osteocalcin, OC)的浓度的变化情况。以探讨降钙素与1,25维生素D对牙周炎导致大鼠牙槽骨丧失的影响,从而为牙周炎的辅助治疗探索一种新的可行方法。方法：清洁级健康成年雄性Wistar大鼠125只,随机分为A、B、C、D、E5组(n=25只／组),即A组为空白对照组,B组为牙周炎对照组,C组为降钙素治疗组,D组为1,25维生素D治疗组,E组为降钙素和1,25维生素D联合治疗组。B、C、D、E组用正畸结扎丝结扎双侧上颌第一磨牙牙颈部加高糖软食的方法建立牙周炎大鼠模型。A组麻醉后不作处理。用正畸结扎丝结扎上颌磨牙牙颈部加高糖软食的方法建立牙周炎大鼠模型后,给予药物治疗,降钙素治疗组(C组,降钙素溶于生理盐水,2μg/kg,皮下注射)、1,25维生素D治疗组(D组,维生素D溶于花生油中,2μg/kg,灌胃)、降钙素和1,25维生素D联合治疗组(E组)。观察各组大鼠的一般指标：体质量变化,活动情况,毛色等。在第2周、4周、6周和8周末各组处死5只大鼠,测量双侧上颌第一磨牙牙周探诊深度,观察牙槽骨丧失水平；第8周末各组采取10只大鼠腹部降主动脉血,用酶联免疫吸附试验法(Enzyme-linked immunosorbent assay, ELISA)检测血清中的骨碱性磷酸酶(Bone alkaline phosphatase, BALP)以及骨钙素(Osteocalcin,OC)的浓度。留取大鼠上颌骨,经10%甲醛固定后,15%的EDTA脱钙,常规石蜡包埋切片HE染色观察牙周组织的病理变化,包括牙周膜纤维排列和炎细胞浸润程度以及牙槽骨情况。应用SPSS17.0统计软件包分析实验数据,符合正态分布和方差齐性计量资料采用均数±标准差(x±s)表示。多组资料间均数比较采用单因素方差分析,组间多重比较采用LSD-t检验。P0.05为差异有统计学意义。结果：A组大鼠活泼好动,形体均匀,反应敏捷,每日饮量少且尿量少,毛色光滑,体质量随着周龄的增长而稳步的增长。B、C、D、E组大鼠于结扎术3天后,表现为食欲下降,唾液量增多,生长基本良好,每日饮量少且尿量少,随后食欲逐渐如常,体质量随着周龄的增长而稳步的增长。通过对A组和B组大鼠双侧上颌第一磨牙PD的比较可以发现,用药2周,B组大鼠的PD高于A组大鼠(P0.05)。B组与C、D、E组PD差异无统计学意义(P0.05)；用药后4周、6周和8周,B组PD均高于C、D、E组(P0.05),且E组PD低于C组和D组。通过对A组和B组大鼠双侧上颌第一磨牙牙槽骨丧失水平的比较可以发现,用药2周,B组大鼠牙槽骨丧失水平高于A组(P0.05)。B组大鼠与C、D、E组大鼠牙槽骨丧失水平差异无统计学意义(P0.05)；用药4周时,B组与C、D、E组牙槽骨丧失水平差异无统计学意义(P0.05)；用药6周和8周时,B组牙槽骨丧失水平高于C、D、E组(P0.05),且E组低于C组和D组；在第8周末进行血清中BALP及OC的检测。8周时,A组BALP、OC均高于B组(P0.05)；且B、C、D、E组间BALP和OC的水平差异均有统计学意义(P0.05)。8wk时组织病理学观察发现差异,HE染色显示A组牙龈上皮完整,结合上皮位于釉牙骨质界,牙槽嵴顶无吸收。B组牙周袋形成,牙龈乳头丧失,牙龈上皮表面糜烂,牙周膜纤维排列紊乱炎症细胞浸润牙槽骨吸收严重。E组大鼠牙周膜纤维排列较整齐,牙槽峭顶有新骨及新生牙骨质形成。结论： 1.维生素D缺乏,钙、磷的缺乏或不平衡可引起牙槽骨成骨能力的降低。 2.降钙素和1,25维生素D均能部分拮抗牙周炎导致的牙槽骨丧失,二者联合应用效果更佳。
[Abstract]:Objective: To observe the changes of the concentration of bone alkaline phosphatase (bone alkaline phosphatase, BALP) and Osteocalcin (osteocalcin, OC) in the serum by establishing a rat model of periodontitis and using 1,25 vitamin D and calcitonin, and observing the level of probing depth (PD), the loss of alveolar bone, and the changes of the concentration of the serum alkaline phosphatase (bone alkaline phosphatase, BALP) and Osteocalcin (osteocalcin, OC) in the serum. The effects of calcium and 1,25 vitamin D on alveolar bone loss in rats with periodontitis, so as to explore a new feasible method for adjuvant treatment of periodontitis.
Methods: 125 healthy adult male Wistar rats were randomly divided into A, B, C, D, E5 group (n=25 only / group), that is, group A as the blank control group, the B group was the control group of periodontitis, the C group was the calcitonin treatment group, the D group was the 1,25 vitamin treatment group, and the group was ligated with orthodontic ligature and ligature. The.A group of the rat model of periodontitis was not treated with the method of high sugar soft food in the neck of the first molar of the maxillary first molar to establish a rat model of periodontitis with orthodontic ligation of the maxillary molar and high sugar and soft food. The treatment group (group C, calcitonin soluble in physiological saline, 2 u g/kg, subcutaneous injection), 1,25 Vitamin D group (group D, vitamin D dissolved in peanut oil, 2 mu g/kg, gavage), calcitonin and 1,25 vitamin D combined treatment group (group E). Observe the general indexes of rats in each group: body mass change, activity condition, hair color and so on. 5 rats were killed in each group at 4 weeks, 6 weeks and 8 weekends in each group, and the depth of the periodontal detection of the bilateral maxillary first molar was measured. The level of alveolar bone loss was observed, and the blood of abdominal descending aorta in 10 rats was taken at the end of eighth weeks. The concentrations of bone alkaline phosphatase (Bone alkaline phosphatase, BALP) and Osteocalcin (Osteocalcin, OC) in the serum were detected by Enzyme-linked immunosorbent assay (ELISA) in each group. The maxillofacial bone was retained in the rat and 10% formaldehyde was obtained. After fixation, 15% of EDTA decalcification and routine paraffin embedded HE staining were used to observe the pathological changes of periodontal tissue, including periodontal ligament fiber arrangement, infiltration degree of inflammatory cells and alveolar bone. The SPSS17.0 statistical package was used to analyze the experimental data, which accorded with the normal distribution and variance homogeneity measurement data with mean mean + standard deviation (x + s). Comparison of data between groups was conducted by one-way ANOVA. Multiple comparisons between groups were performed by LSD-t test,.P0.05 was statistically significant.
Results: the rats in group A were active and active, with a uniform body, quick reaction, less daily drink and less urine and smooth hair color. The body mass increased steadily with the age of.B, C, D, E, 3 days after ligation, showing a decline in appetite, increased saliva, good base, less daily drink and less urine, and the appetite gradually as usual, body and body gradually as usual, body gradually, body gradually as usual, body then gradually as usual, body appetite gradually as usual, body After 2 weeks of drug use, the PD of group B rats was higher than that of A group (P0.05).B group and C, D, E group, and there was no significant difference between the A group and the C group (P0.05).B group, and the 4 weeks, 6 weeks and 8 weeks after the drug use were higher than those of the group of A and group A. In group A and group D, by comparing the loss of alveolar bone in the bilateral maxillary first molar of rats in group A and B, the loss of alveolar bone in group B rats was higher than that of group A (P0.05).B group and C, and there was no significant difference in the loss level of alveolar bone in the group of D and E group (P0.05). The difference of the loss of alveolar bone in the B group was different from that of the E group (P0.05). There was no statistical significance (P0.05). At 6 and 8 weeks, the loss of alveolar bone in group B was higher than that of C, D, E group (P0.05), and the E group was lower than the C and D groups. The HE staining showed that the gingival epithelium was intact in group A, the epithelium was located in the glazed cementum boundary, the top of the alveolar ridge was not absorbed by the.B group, the papilla was lost, the surface of the gingiva was erosive, the periodontal ligament was arranged in disorder, and the inflammatory cells infiltrated the alveolar bone, and the periodontal ligament of the alveolar bone was neatly arranged and the alveolar crest was on the top. The formation of new bone and new cementum. Conclusion:
1. vitamin D deficiency, calcium or phosphorus deficiency or imbalance can cause osteogenesis of alveolar bone.
2. calcitonin and 1,25 vitamin D can partly antagonize the alveolar bone loss caused by periodontitis, and the combined application of the two is better.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.42

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