牙龈卟啉单胞菌牙龈素在牙周炎症过程中的作用及其机制

牙周炎是口腔临床实践中最为常见的疾病之一。牙龈卟啉单胞菌与牙周炎密切相关,牙龈卟啉单胞菌表面的牙龈素(gingipains)、菌毛和脂多糖被认为是其最重要的毒力因子,是近年来牙周炎机制研究的热点,引起研究者的广泛关注。本文结合近年来的研究进展,就牙龈卟啉单胞菌gingipains的结构、引起牙周炎症的机制和所涉及的信号通路作一综述。     

牙龈卟啉单胞菌脂多糖引起牙周炎症的机制和特点

革兰阴性厌氧菌牙龈卟啉单胞菌和牙周炎密切相关，脂多糖是革兰阴性菌外膜中的主要结构成分，具有广泛的生物学活性，被认为是革兰阴性菌的主要毒力因子之一。本文就牙龈卟啉单胞菌脂多糖的化学组成和结构，所启动炎症信号通路及其起始受体，引起牙周炎症的机制和特点作一综述。     

大鼠实验性牙周炎中牙龈卟啉单胞菌rgpB和kgp变化的研究

目的:通过建立大鼠实验性牙周炎模型,检测不同时段大鼠上颌第一磨牙龈下人工定植的牙龈卟啉单胞菌、牙龈素rgpB和kgp相对含量的改变,动态观察牙周炎发展不同阶段牙龈卟啉单胞菌致病基因的变化。方法:选择6周的成年雄性大鼠13只,应用钢丝结扎法和细菌种植法建立大鼠实验性牙周炎模型。分别在4周和8周取上颌第一磨牙龈下菌斑,提取细菌DNA,应用PCR方法进行牙龈卟啉单胞菌、牙龈素基因rgpB和kgp特异引物扩增,应用SPSS13.0统计软件,分析大鼠牙周炎不同时段rgpB和kgp的相对变化。结果:大鼠实验性牙周炎模型,龈下菌斑中牙龈卟啉单胞菌的相对含量实验组明显高于对照组,8周组高于4周组;实验组牙龈卟啉单胞菌牙龈素rgpB和kgp明显高于对照组,4周组和8周组rgpB和kgp相对含量无显著性差异。结论:rgpB基因和kgp基因与牙周炎的致病性有关而与牙周炎的严重程度可能无直接相关。     

牙龈卟啉单胞菌脂多糖引起牙周炎症的机制和特点

革兰阴性厌氧菌牙龈卟啉单胞菌和牙周炎密切相关,脂多糖是革兰阴性菌外膜中的主要结构成分,具有广泛的生物学活性,被认为是革兰阴性菌的主要毒力因子之一。本文就牙龈卟啉单胞菌脂多糖的化学组成和结构,所启动炎症信号通路及其起始受体,引起牙周炎症的机制和特点作一综述。     

牙龈卟啉单胞菌研究进展

牙周炎是常见的口腔疾病之一,是中国成年人失牙的主要原因.牙龈卟啉单胞菌在引发牙周组织破坏过程中发挥着重要的作用,成为国内外学者研究的热点之一.本文就牙龈卟啉单胞菌与口腔疾病和其他全身性疾病的关系、牙龈卟啉单胞菌全基因组,牙龈卟啉单胞菌与牙龈上皮细胞的相互作用等研究作一综述,以拓宽牙龈卟啉单胞菌的研究思路,进血为牙周炎的...     

牙龈素促进牙龈卟啉单胞菌免疫逃逸的机制

有关牙周炎病因的关键致病菌假说近年来引起学者们的关注,该假说认为牙龈卟啉单胞菌在牙周炎的发病过程中发挥着重要作用,是牙周炎的关键致病菌.牙龈素是牙龈卟啉单胞菌产生的重要致病因子之一,它可以协助牙龈卟啉单胞菌逃逸巨噬细胞、中性粒细胞及补体系统的杀伤作用.本文就牙龈素促进牙龈卟啉单胞菌免疫逃逸的机制相关研究进展作一综述,对这一机制的深入了解有助于进一步理解牙龈素的致病机制,同时也可以为牙周炎的防治探索新的途径.     

牙龈卟啉单胞菌牙龈蛋白酶的研究进展

牙周炎是一种牙齿支持组织的慢性炎症性疾病,能引起结缔组织和牙槽骨的破坏而导致渐进性附着丧失,最终可能导致牙齿脱落.革兰氏阴性厌氧菌牙龈卟啉单胞菌是慢性牙周炎的重要致病菌,具有包括脂多糖、菌毛、血细胞凝集素和胞外蛋白酶在内的多种毒力因子,其中牙龈蛋白酶提供的蛋白水解活性占该生物体产生的一般蛋白水解活性的大部分(85％),...     

牙周炎患者牙周袋内硫化物水平与牙龈卟啉单胞菌的关系

目的分析牙周袋内硫化物的检出率、浓度与牙龈卟啉单胞菌的关系。方法选取慢性牙周炎初诊患者20名,采集龈下菌斑,用定量PCR方法检测菌斑中牙龈卟啉单胞菌,同时用金刚探针牙周诊断仪测量同一取样位点的硫化物水平,并记录临床探诊深度、出血指数。结果牙龈卟啉单胞菌的检出率（92％）明显高于硫化物的检出率（34％）（P〈0．05）;在深牙周袋、出血的位点,硫化物的检出率、浓度均明显高于浅牙周袋、不出血的位点（P〈0．05）;而在深牙周袋、出血的位点,牙龈卟啉单胞菌的检出率和浓度无显著性增高;在硫化物检测阳性的位点牙龈卟啉单胞菌的数量高于硫化物检测阴性的位点,但差异无统计学意义。结论牙周袋内硫化物的水平不是反映牙龈卟啉单胞菌数量的敏感指征,但可在一定程度上反映牙周组织的破坏程度和炎症的存在。     

牙龈卟啉单胞菌对慢性牙周炎致病作用研究进展

牙龈卟啉单胞菌是公认的牙周主要致病菌之一,是与慢性牙周炎密切相关的红色复合体的重要成员,在引发牙周组织破坏的过程中发挥重要作用。其中牙龈卟啉单胞菌的毒力因子及牙龈卟啉单胞菌与牙龈上皮细胞的相互作用在这一过程中发挥关键作用。本文对牙龈卟啉单胞菌在慢性牙周炎中发挥的作用及其自身在此过程中的改变做一综述。     

维吾尔族慢性牙周炎中牙龈卟啉单胞菌菌fimA毒力基因型的分布

目的：分析维吾尔族慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌菌毛fimA毒力基因型的分布情况。方法：收集52例维吾尔族慢性牙周炎患者的龈下菌斑，采用16SrRNA PCR法检测牙龈卟啉单胞菌，并根据菌毛fima毒力基因型的特异引物，用聚合酶链反应（PCR）检测Ⅱ型fimA和Ⅳ型fima菌株的分布。结果：16SrRNA PCR法检测牙龈卟啉单胞菌在龈下菌斑中阳性检出率是76．9％。牙周袋PPD〉6mm位点龈下菌斑标本的P．gingivalis检出率高于4〈PPD≤6mm采样的位点，2组差异有统计学意义（P〈0．05）。牙龈卟啉单胞菌菌毛．fimA毒力基因型在牙龈卟啉单胞菌感染者的检出率分别是：Ⅱ fimA型为37．5％，ⅣfimA型为22．5％。结论：维吾尔族慢性牙周炎患者龈下菌斑中牙龈卟啉单胞菌有较高的检出率。牙龈卟啉单胞菌存在fima毒力基因多态性。     

牙龈卟啉单胞菌在牙周病防治中的应用

牙周病是口腔两大类主要疾病之一,具有较高的发病率,因此,探索预防牙周病的有效途径十分重要。本文介绍了国外学者以牙龈卟啉单胞菌不同形式的抗原进行免疫学防治牙周炎的试验,其中包括对牙龈卟啉单胞菌菌毛、血凝素、牙龈素和外膜蛋白的研究。     

Immunodominant antigens of Porphyromonas gingivalis in patients with rapidly progressive periodontitis

We studied 4 isolates of Porphorymonas gingivalis , ATCC 33277, 381, A7A1-28, and W50, to identify major cell surface antigens and select the best strain from which to obtain antigen for a test vaccine. Immunoglobulin G (IgG) titers measured by enzyme-linked immunosorbent assay using whole-cell sonicates as antigen were significantly elevated for the sera of 64 rapidly progressive periodontitis patients relative to sera of 30 normal control subjects for each of the 4 strains studied. Western blots were prepared for all 4 strains and developed using sera from 22 patients and 20 control subjects to identify and determine the frequency of antibody-binding components. The intensity of binding by patient sera was greatest for the 75-kDa and 55-kDa components. The 43-kDa component was also widely recognized. Strains ATCC 33277 and 381 appeared to be antigenically similar. Because of the higher serum antibody titers, the larger proportion of seropositive patients and higher frequency of binding to specific protein components in Western blots, our efforts were focused on strain ATCC 33277. Whole-cell sonicates, proteinase K-digested sonicate, lipopolysaccharide, capsular polysaccharide, and whole-cell protein fractions were prepared and evaluated for anti-genie activity. By dot immunoblot, most of the antibody binding activity was found in the whole-cell protein fraction, with much lesser amounts in lipopolysaccharide and none in capsular polysaccharide. The antibody-binding activity was accessible on the cell surface, since 98.9% of P. gingivalis -specific antibody, including antibody binding to the 43-kDa, 55-kDa and 75-kDa components on Western blot, was removed by whole-cell adsorption. Furthermore, the 43-kDa and 55-kDa but not the 75-kDa component on intact cells were accessible for labeling with 125_I, confirming their cell surface location and accessibility.     

Immunization of Macaca fascicularis against experimental periodontitis using a vaccine containing cysteine proteases purified from Porphyromonas gingivalis

INTRODUCTION: Periodontitis is a common infectious disease to which Porphyromonas gingivalis has been closely linked, in which the attachment tissues of the teeth and their alveolar bone housing are destroyed. We conducted a study to determine if immunization using a purified antigen could alter the onset and progression of the disease. METHODS: Using the ligature-induced model of periodontitis in Macaca fascicularis, we immunized five animals with cysteine protease purified from P. gingivalis and used an additional five animals as controls. Alveolar bone loss was measured by digital subtraction radiography. RESULTS: Immunization induced high titers of specific immunoglobuin G serum antibodies that were opsonic. Total bacterial load, levels of P. gingivalis in subgingival plaque and levels of prostaglandin E(2) in gingival crevicular fluid were significantly reduced. Onset and progression of alveolar bone loss was inhibited by approximately 50%. No manifestations of toxicity were observed. CONCLUSIONS: Immunization using a purified protein antigen from P. gingivalis inhibits alveolar bone destruction in a ligature-induced periodontitis model in M. fascicularis.     

牙龈卟啉单胞菌脂蛋白基因在慢性牙周炎及牙周健康者中的检测

目的 应用基因芯片技术检测3种脂蛋白基因PG0717、PG0183、PG2135在慢性牙周炎患者和牙周健康者牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)中的分布,探讨这些基因与牙周临床指数之间的关系,为研究脂蛋白在Pg致病过程中的作用提供依据.方法 选取41例慢性牙周炎患者(牙周炎组)及76例牙周健康者(健康对照组),记录探诊深度、附着丧失、探诊出血及牙齿松动度,取龈下菌斑进行细菌分离培养,以临床采集的样本提取的DNA为探针,以抑制消减杂交技术获得的PgW83特异基因片段PG0717、PG0183、PG2135为目标序列,采用Cy5荧光标记目标序列.应用基因芯片技术检测PG0717、PG0183、PG2135基因在牙周炎组病变部位、非病变部位和健康对照组Pg中的分布.结果在牙周炎组病变部位PG0717、PG0183、PG2135基因的检出率分别为90%(18/20)、70%(14/20)、70%(14/20),非病变部位的检出率分别为60%(12/20)、45%(9/20)、40%(8/20),而在健康对照组PG0717、PG0183、PG2135的检出率分别为55%(11/20)、25%(5/20)、30%(6/20).3种脂蛋白基因在牙周炎组病变部位和健康对照组中的检出率差异均有统计学意义(P＜0.05),并且与牙周探诊深度、临床附着丧失、探诊出血及牙齿松动度相关.结论 带有PG0717、PG0183、PG2135基因的Pg菌株致病力强.     

牙龈卟啉单胞菌PG1055基因在临床菌株中的表达

目的应用基因芯片技术检测PG1055基因在不同人群的牙龈卟啉单胞菌(P.gingivalis)中分布,探讨这些基因与牙周临床指数之间的关系。方法取龈下菌斑进行细菌分离培养,以临床采集样本提取的DNA为探针,以抑制消减杂交技术获得P.gingivalisW83的特异基因片段PG1055为目标序列,采用Cy5荧光标记目标序列。应用基因芯片技术检测PG1055基因在牙周病患者及健康人群的牙龈卟啉单胞菌中的分布。结果PG1055基因在牙周病患者及健康人群中的检出率有统计学差异,并且与牙周临床指数相关。结论PG1055基因与P.gingivalis的致病性有关。     

牙龈卟啉菌、中间普氏菌的分离、培养和鉴定

目的：采用厌氧培养和鉴定技术，分离牙周炎患者龈下菌斑中的牙龈卟啉菌和中间普氏菌。方法：采集牙周炎患者的龈下菌斑，厌氧培养，分离产黑色素菌。经长波长紫外灯观察，各种生化分析和间接免疫荧光染色，分离和鉴定牙龈卟啉菌，中间普氏菌。结果：在受检的33例牙周炎患者中，24例患者检出了牙龈卟啉菌，总检出率为72．77％，共分离了79株。18例患者检出了中间普氏菌，总检出率为54．54％。共分离了32株。结论：厌氧培养，生化反应鉴定技术的发展，使牙龈卟啉菌和中间普氏菌的分离与培养准确可靠，为今后在分子水平上了解各菌株的致病机理打下了基础。     

Intracellular localization of Porphyromonas gingivalis thiol proteinase in periodontal tissues of chronic periodontitis patients

OBJECTIVES: Porphyromonas gingivalis is a significant periodontal pathogen that has been shown in vitro to be able to invade gingival epithelial cells and grow intracellularly. The aim of the present study was to detect P. gingivalis in gingival tissues from chronic periodontitis (CP) patients. MATERIALS AND METHODS: Monoclonal antibodies specific to a cell membrane-bound thiol proteinase of P. gingivalis were used to detect the microbe in gingival tissues of CP patients (n = 13) by immunohistochemistry. The presence of P. gingivalis was also analysed by polymerase chain reaction (PCR). RESULTS: Immunohistochemical analysis of the periodontal tissues revealed positive staining for P. gingivalis thiol proteinase in 11 of the 13 patients. Positive staining was mainly located intracellularly in the perinuclear region of the cytoplasm in the periodontal epithelial cells and it could be detected throughout the whole depth of both pocket and oral epithelium. The sensitivity of immunohistochemistry was found to be comparable with that of PCR. CONCLUSIONS: Our results provide in vivo evidence of the ability of P. gingivalis to enter human gingival epithelial cells. Intracellular localization of P. gingivalis contributes to its evasion of the host immune surveillance and eventually increases its resistance to conventional treatments of periodontal diseases.     

Antibody reactive with Porphyromonas gingivalis hemagglutinin in chronic and generalized aggressive periodontitis

BACKGROUND: Porphyromonas gingivalis, a black-pigmented, gram-negative anaerobe, is found in periodontitis lesions and its presence in subgingival plaque significantly increases the risk for periodontitis. We have previously shown that patients with aggressive forms of periodontitis that are seropositive for P. gingivalis have less attachment loss than those that are seronegative. This suggests that antibody reactive with antigens of P. gingivalis may be protective and decrease disease severity and extent. Recent studies in the murine abscess model and in the host antibody response in chronic periodontitis patients suggest that antibody reactive with P. gingivalis hemagglutinin may be an important protective antibody response. OBJECTIVES: In this study, we tested the hypothesis that there was a significant relationship between antibody reactive with P. gingivalis hemagglutinin and measures of periodontal attachment loss. METHODS: We determined the immunoglobulin G (IgG) antibody concentration reactive with recombinant P. gingivalis hemagglutinin in 117 chronic periodontitis and 90 generalized aggressive periodontitis patients. We also determined the IgG subclass distribution for antibody reactive with P. gingivalis hemagglutinin. RESULTS AND CONCLUSIONS: We found IgG reactive with P. gingivalis hemagglutinin in both chronic periodontitis and generalized aggressive periodontitis patients. Most of this IgG antibody was of the IgG1 and IgG3 subclasses. Antibody reactive with P. gingivalis hemagglutinin, however, did not have a significant relationship with measures of periodontal attachment loss.     

Trypsin-like activity levels of Treponema denticola and Porphyromonas gingivalis in adults with periodontitis

Abstract Treponema denticola (Td) and Porphyromonas gingivalis (Pg) are associated with human moderate and severe adult periodontal diseases. This study Quantifies these two anaerobes and their trypsin-like (TL) activities in subgingival plaque collected from both clinically healthy and periodontally diseased sites of human periodontitis patients. Antigen levels of the microorganisms were determined by monoclonal antibodies and TL activities were measured by the fluorescent substrate Z-gly-gly-arg-AFC in a disc format. Significant positive correlations were observed between the antigen levels and the TL activities when the data were subjected to statistical analyses both on a site-specific and on a patient basis. Anaerobe synergism was found between Td and Pg in a continental US population, and positive correlations were found between anaerobe levels (individually and total) and clinical indicators of adult periodontitis.