根面处理牙周炎牙骨质后内毒素水平变化及对牙周膜成纤维细胞影响的研究

发布时间：2018-05-05 12:55

  本文选题：牙周炎 + 内毒素　； 参考：《中国人民解放军医学院》2017年硕士论文

【摘要】：牙周炎是最常见的口腔疾病之一,是导致牙齿缺失的主要疾病,其病因及发病机制仍是牙周病学研究热点。牙周炎患牙的部分牙根表面牙骨质暴露于牙周袋内,感染后会发生结构性和理化性变化,严重影响牙周功能性细胞在牙根表面的生长和附着,阻碍牙周组织再生修复。人牙周膜成纤维细胞(human periodontal ligament fibroblasts,hPDLFs)是构成牙周韧带的重要细胞,在牙周致病菌及其毒性产物作用下会发生直接损伤或引发免疫应答反应,导致牙周组织破坏。本研究主要观察内毒素对hPDLFs增殖及IL-8分泌的影响;不同方法处理牙周炎患牙根面牙骨质后内毒素水平变化及其根面处理后的牙周炎牙骨质片对牙周膜成纤维细胞分泌细胞因子水平变化,为选择临床牙周炎治疗方法提供依据。本研究主要分为三部分实验其目的、方法和结果如下:实验一:内毒素对人牙周膜成纤维细胞增殖及IL-8分泌的影响目的:观察不同浓度的内毒素(Lipopolysaccharides ,LPS )对PDLFs的影响及IL-8的分泌情况。方法:本实验分为五组,PDLFs组(阴性对照组)、11μg/mlLPS组、10μg/mlLPS 组、100μg/mlLPS组、200μg/mlLPS 组。各组细胞进行培养,MTT 法观察细胞增殖变化。24h, 96h ELISA法检测IL-8分泌情况。结果:与PDLFs组比较,10μg/ml LPS 促进 PDLFs 增殖;200μg/ml LPS 抑制 PDLFs 增殖;24h: 200μg/ml LPS组促进IL-8分泌,有显著性统计学差异。96h: 1OOμg/mlLPS、200μg/mlLPS组促进IL-8分泌,有显著性统计学差异。结论:不同浓度的内毒素对PDLFs增殖、IL-8分泌合成释放有调节作用。实验二:根面处理牙周炎患牙根面牙骨质后内毒素水平变化研究目的:观察不同方法处置牙周炎患牙根面牙骨质后内毒素水平变化。方法:选择临床上正畴拔除的健康的前磨牙6例,重度牙周炎拔除的后牙36例。每颗牙进行编号,依分组根面处理后于釉牙骨质界下2 mm制成2片4 mmx4 mmx1 mm的牙骨质片。健康牙为阴性对照组,36颗牙周炎患牙每颗牙中1个牙片不进行根面处理,为牙周炎组。其余36片牙周炎牙片随机平均分成6组:龈下刮治及根面平整(scaling and root planning ,SRP)组、SRP+ 抗菌肽 A 组、SRP+ 抗菌肽 B 组、SRP+EDTA 组、SRP+ Nd:YAG激光组和SRP +Er: YAG激光组。显色基质鲎试剂检测每个牙片内毒素浓度。按照编号记录每颗牙齿内毒素浓度,计算每组每个牙周炎患牙处理前后内毒素浓度变化。采用SPSS17.0软件包对数据进行统计学分析。结果:与牙周炎组相比,各处理组内毒素浓度有不同程度降低,均有显著性差异(P0.01 )。与SRP组相比,SRP+抗菌肽A组、SRP+抗菌肽B组、SRP +Er: YAG激光组,内毒素浓度显著下降(P0.01 );SRP+EDTA组、SRP+Nd: YAG激光组均无显著性差异(P0.05)。结论:不同方法处理牙周炎患牙根面均可降低内毒素含量,抗菌肽A+SRP处理牙周炎根面降低内毒素含量最多。实验三:根面处理牙周炎患牙根面牙骨质对牙周膜成纤维细胞分泌细胞因子的影响目的:观察不同方法处理牙周炎患牙根面对PDLFs分泌细胞因子的影响。方法:实验二根面处置的牙周炎牙骨质片(6组)及健康牙牙骨质片与PDLFs共培养,ELISA检测24h时上清液IL-6、IL-8、TNF-α水平。结果:(1)与SRP组比较:其余组IL-8、TNF-α含量均低于SRP组,有显著性差异(P0.01)。(2)抗菌肽A+SRP组IL-6含量与健康牙组无显著性差异(P0.05)。(3)除SRP组,其余根面处理组IL-8含量与健康牙组比较无显著性差异(P0.05 )。( 4 ) Nd: YAG激光+SRP组与Er: YAG激光+SRP的TNF-α含量与健康牙组无显著性差异(P0.05)。结论:抗菌肽(A、B)、Er: YAG激光、Nd: YAG激光、EDTA联合SRP处理组IL-8、TNF-α含量均少于SRP组,其IL-8含量组间无差异。抗菌肽A+SRP在处理组中IL-6含量最低。Nd: YAG激光+SRP在处理组中TNF-α含量最低。
[Abstract]:Periodontitis is one of the most common oral diseases and is the main disease causing tooth loss. Its etiology and pathogenesis is still a hot spot in periodontitis. The surface of the root surface of periodontitis teeth is exposed to the periodontal pocket, and the structural and physical changes will occur after infection, which seriously affects the surface of periodontal functional cells on the root surface. The growth and attachment of human periodontal ligament fibroblasts (hPDLFs) is an important cell for the periodontal ligament, and the periodontal pathogens and their toxic products will be directly damaged or triggered by the immune response, leading to the destruction of periodontal tissue. This study is mainly observed in this study. The effects of endotoxin on the proliferation of hPDLFs and the secretion of IL-8; the changes in endotoxin level after treatment of periodontitis and the root surface of periodontitis and the level of the secretory cytokines in periodontal ligament fibroblasts after root surface treatment are the basis for the treatment of clinical periodontitis. This study is mainly divided into three parts. The purpose, methods and results are as follows: Experiment 1: the effect of endotoxin on the proliferation and IL-8 secretion of human periodontal ligament fibroblasts: the effect of Lipopolysaccharides (LPS) on PDLFs and the secretion of IL-8. Methods: this experiment was divided into five groups, PDLFs group (negative control group), 11 g/mlLPS group, 10 u g/m. Group lLPS, 100 mu g/mlLPS group and 200 g/mlLPS group. Cells were cultured in each group. MTT method was used to observe cell proliferation and.24h, 96h ELISA method was used to detect IL-8 secretion. Results: compared with group PDLFs, 10 mu g/ml LPS promoted PDLFs proliferation, and 200 micron inhibited the proliferation of IL-8. H: 1OO mu g/mlLPS, 200 g/mlLPS group promote IL-8 secretion, there are significant statistical differences. Conclusion: different concentrations of endotoxin can regulate the proliferation of PDLFs and the release of IL-8 secretion. Experiment two: root surface treatment of periodontitis with dental root surface endoendotoxin level changes: Observation of different methods of treatment of periodontitis and tooth root teeth Methods: 6 healthy premolars, 36 cases of severe periodontitis and 36 cases of posterior teeth extracted from severe periodontitis were selected, and each tooth was numbered. 2 slices 4 mmx4 mmx1 mm of cementum were made from 2 mm in the enamel cementum boundary. The healthy teeth were negative control group, 36 periodontitis and each tooth in 1 teeth. The other 36 slices of periodontitis were divided into 6 groups randomly: subgingival scaling and root surface leveling (scaling and root planning, SRP), SRP+ antibacterial peptide A group, SRP+ antibacterial peptide B group, SRP+EDTA group, SRP+ Nd:YAG laser group and laser group. Chromogenic matrix limulus test was used to detect each tooth The concentration of endotoxin in each tooth was recorded by serial number and the endotoxin concentration in each group of periodontitis and teeth was calculated before and after treatment. The data were statistically analyzed by SPSS17.0 software package. Results: compared with the periodontitis group, the concentration of endotoxin in each treatment group decreased in varying degrees (P0.01) and SRP. Group SRP+ antibacterial peptide A, SRP+ antibacterial peptide B group, SRP +Er: YAG laser group, the concentration of endotoxin decreased significantly (P0.01), SRP+EDTA group, SRP+Nd: YAG laser group had no significant difference (P0.05). Conclusion: different methods to treat periodontitis with root surface can reduce endotoxin content, antibacterial peptide treatment of periodontitis root surface to reduce endotoxin content of the most Experiment three: the effect of root surface treatment on periodontitis with root surface cementum on the secretory cytokines of periodontal ligament fibroblasts in order to observe the effects of different methods to treat PDLFs secretory cytokines in periodontitis and tooth roots. Methods: the treatment of periodontitis with two surfaces (6 groups) and healthy teeth bone slices were co cultured with PDLFs, E LISA detected the level of IL-6, IL-8, and TNF- alpha in the supernatant of 24h. Results: (1) the content of IL-8 and TNF- in the rest group was lower than that of the SRP group, and there was a significant difference (P0.01). (2) there was no significant difference between the IL-6 content of the antibacterial peptide A+SRP group and the healthy teeth group. (3) there was no significant difference in the content of the remaining root surface treatment group compared with the healthy tooth group. (.05) (4) there was no significant difference in the TNF- alpha content between the Nd: YAG laser +SRP group and the Er: YAG laser +SRP in the healthy teeth group (P0.05). The content of TNF- alpha in the treated group was the lowest in the light +SRP.

【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R781.4

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