侵袭性牙周炎和慢性牙周炎的龈下优势菌分析

目的 :分析侵袭性牙周炎 (aggressiveperiodontitis ,AgP)与慢性牙周炎 (chronicperiodontitis ,CP)的龈下优势菌群 ,为探讨牙周炎分类、病因和诊断提供实验依据。方法 :将中学生流调筛选 (16例 )及牙周病专科就诊(2 4例 )的AgP和CP患者 ,采集龈下菌斑样本 ,在厌氧菌基础培养基 (CDC)和选择性培养基 (TSBV)上培养分析。结果 :局限型AgP患者的伴放线放线杆菌 (Actinobacillusactinomycetemcomitans ,Aa)及兼性厌氧菌的检出率显著高于中度CP患者 (P <0 .0 5 ,P <0 .0 1) ,而广泛型AgP和重度CP患者的厌氧菌总数较局限型AgP和中度CP患者显著增加 (P <0 .0 5 )。结论 :局限型AgP和中度CP的龈下优势菌有明显差别 ,Aa是一个重要的危险因子。     

牙周炎患者唾液中伴放线放线杆菌的检出状况分析

目的 检测不同类型牙周炎患者唾液中的伴放线放线杆菌(Actinobacillusactinomycetemcomitans,Aa),探讨唾液和集合龈下菌斑中Aa检出率的差异以及唾液中Aa的存在状况与牙周临床指标的关系. 方法 收集50例侵袭性牙周炎(aggressive periodontitis,AgP)患者、48例慢性牙周炎(chronic periedontitis,CP)患者和25例非牙周炎者的非刺激性全唾液和集合龈下菌斑,应用聚合酶链反应(PcR)技术检测两种样本中的Aa. 结果 Aa在AgP患者唾液中的检出率(32%)显著高于非牙周炎者(4%)和CP患者(15%),差异均有统计学意义(P<0.01,P<0.05),同时Aa在AgP患者唾液中的检出率也显著高于集合龈下菌斑样本(16%),差异亦有统计学意义(P<0.05).年龄≤30岁是唾液中存在Aa的危险指征(OR=3.23,P<0.05);出血指数≥3的位点超过70%与唾液中存在Aa有关(OR=19.21,P<0.01). 结论 AgP患者唾液样本中Aa的检出率明显高于集合龈下菌斑样本,亦高于CP患者和非牙周炎者,提示Aa可能参与AgP的发生和发展.     

实时荧光定量PCR检测牙周炎患者龈下菌斑的分布

目的 应用实时荧光定量PCR技术探索侵袭性牙周炎（aggressive periodontitis,AgP）、慢性牙周炎（chronic periodontitis,CP）患者龈下菌斑中伴放线聚集杆菌(A. actinomycetemcomitans,Aa)、牙龈卟啉单胞菌（P. gingivalis,Pg）的分布规律。方法 采集32例AgP、33例CP、32例牙周健康者的龈下菌斑,构建含有2种待测细菌基因片段的重组质粒,建立定量标准,采用TaqManMGB探针实时荧光定量PCR方法检测样本中细菌数量。结果 本实验构建的引物及TaqManMGB探针特异性及敏感性较好。AgP组龈下菌斑Aa的检出率高于CP组（P＜0.01）,但2种细菌数量在组间无显著差异,两组内Pg的检出率及数量都明显高于Aa（P＜0.001）,另外AgP组Aa的数量、CP组Pg数量与牙周探诊深度密切相关（P＜0.01及P＜0.001）。结论 龈下菌斑Aa的检出率可能与牙周炎类型存在一定关联,Aa可能并不是中国人群样本AgP患者龈下菌斑的优势菌,实时荧光定量PCR对牙周病学研究有广泛应用前景。     

牙周炎患者基础治疗后牙龈卟啉单胞菌的定植研究

目的 通过对牙周炎患者龈下菌斑中牙龈卟啉单胞菌(Porphyromonas gingivalu,Pg)的检测,探讨慢性牙周炎(chronic periodontitis,CP)和侵袭性牙周炎(aggressive periodontitis,AgP)患者牙周基础治疗后Pg的定植规律.方法 选取90例CP患者和90例AgP患者,在牙周基础治疗前、治疗后6周、12周共采集龈下菌斑样本1620个,运用AmpliFluor终末点定量聚合酶链反应方法 检测Pg含量.结果 治疗后6周CP和AgP组Pg活动位点分别为61(22.6%)和66(24.4%)个,两者差异无统计学意义(P>0.05);治疗后6周Pg活动位点在治疗前检测的牙周临床指数高于Pg静止位点.治疗后12周两组Pg活动位点分别为96(35.6%)和18(6.7%)个,差异有统计学意义(P<0.05);治疗后12周Pg活动位点在治疗后6周时检测的牙周临床指数高于Pg静止位点.结论 在牙周基础治疗后6周时,CP和AgP患者Pg定植均已开始,仉是两组定植规律存在一定差异.在牙周基础治疗后,治疗前牙周组织炎性反应严重的龈下位点Pg易于定植.     

伴放线放线杆菌在龈下菌斑和颊黏膜分布研究

目的:比较伴放线放线杆菌(actinobac illus actinomycetem com itans,A.a)在不同类型牙周炎患者龈下菌斑和颊黏膜中的分布。方法:通过聚合酶链反应(polym erase chain reaction,PCR)对侵袭性牙周炎患者(AgP)、慢性牙周炎患者(CP)、牙周健康者口腔龈下菌斑和颊黏膜中的A.a进行检测,分析该菌分别在两部位的相对含量。结果:AgP组菌斑和颊黏膜样本中A.a阳性检出率均为41.7%,分别高于CP组(菌斑16.7%、颊黏膜10.0%)和牙周健康组(菌斑和颊黏膜均为0%)。AgP组A.a在菌斑和颊黏膜的相对含量分别为38.5%和22.2%,高于CP组(菌斑19%、颊黏膜12.75%)。结论:A.a不仅存在于龈下菌斑中,也能够粘附于颊黏膜;A.a是AgP的主要优势菌也参与了CP的菌群组成。     

侵袭性牙周炎及慢性牙周炎患者血清Dickkopf-1水平的影响因素研究

目的 观察牙周炎与血清Dickkopf-1(DKK-1)水平的关系,探讨影响血清DKK-1水平的可能因素.方法 收集侵袭性牙周炎(aggressive periodontitis,AgP)、慢性牙周炎(chronic periodontitis,CP)患者各20例(分别为AgP组、CP组)及分别与AgP、CP患者年龄匹配的健康对照者各20名(分别为H1组、H2组),采用酶联免疫吸附测定法检测血清DKK-1水平.采用t检验比较各组血清中DKK-1水平的差异,并采用相关分析和多元线性回归方法分析血清DKK-1水平与牙周临床参数及年龄等因素的关系,检测水准为双侧α =0.05.结果 AgP组血清DKK-1水平[(12.36±3.19) μg/L]显著高于CP组[(8.90 ±2.73) μg/L] (P=0.001),但与其对照组H1组[(12.37±2.74) μg/L]相比差异无统计学意义(P=0.99),CP组与H2组[(8.85 ±2.56) μg/L]相比差异也无统计学意义(P =0.896);AgP与CP组血清DKK-1水平与牙周探诊深度、附着丧失、出血指数、探诊出血阳性位点百分比间均未见显著相关(AgP组r值分别为-0.029、-0.223、0.062、-0.412;CP组r值分别为-0.156、0.185、-0.379、0.051);总体样本血清DKK-1水平与年龄呈显著负相关(r=-0.453,P =0.000);多元线性回归分析显示年龄对血清DKK-1水平有影响(β=-0.391,t=-3.626,P=0.001).结论 未发现牙周炎对血清DKK-1水平产生影响;年龄是影响血清DKK-1水平的重要因素,血清DKK-1水平随年龄增加而降低.     

牙周炎患者的血脂、血糖水平分析

目的分析牙周炎对患者血糖和血脂水平的影响。方法对117例侵袭性牙周炎(AgP)患者、40例慢性牙周炎(CP)患者及37名牙周健康者,空腹抽取静脉血,检测血脂、血糖水平,比较3组之间的差异。结果AgP患者平均甘油三酯水平[(1．09±0．79)mmol／L]显著高于健康对照者[(0．94±0．28)mmol／L,P<0．05];CP和AgP患者的血糖水平[分别为(5．40±1．01)mmol／L和(5．07±0．66)mmoL／L]均高于健康对照者(4．62±0．64 mmol／L),P< 0．01;CP组的总胆固醇高于健康对照组,P<0．05;AgP组的重度位点百分比与总胆固醇呈正相关(r=0．25,P<0．01)。结论牙周炎可能会影响血糖和血脂的水平。     

牙周袋内挥发性硫化物与牙周炎症程度的关系

目的初步探讨牙周袋内挥发性硫化物与侵袭性牙周炎(aggressive periodontitis,AgP)和慢性牙周炎(chronic periodontitis,CP)炎症程度的关系。方法用金刚牙周探针诊断仪检查探诊深度、附着丧失、探诊出血等临床指标的同时,检测牙周袋内硫化物水平。共检查15例AgP和16例CP患者870个患牙的5 220个位点。结果无论是AgP还是CP患者,硫化物阳性位点的各项临床指标均明显高于阴性位点(P<0.001);硫化物水平与各项临床指标间都有明显的正相关关系(P<0.001);中、深袋组的硫化物水平和阳性位点率均明显高于浅袋位点(P<0.001);有附着丧失位点的硫化物水平和阳性位点率均高于无附着丧失位点,在浅袋组其差异有显著性。结论牙周袋内挥发性硫化物的检测可以反映侵袭性牙周炎和慢性牙周炎患者的牙周炎症程度。     

牙周炎患者TLR2,TLR4基因多态性研究

目的：研究TLR2、TLR4基因多态性分布,探讨其与牙周炎的发病和严重程度之间的相关性。方法：选择40位侵袭性牙周炎（AgP）患者及50名慢性牙周炎（CP）患者为实验对象,以100名牙周健康者作为对照组。从全血样本中提取基因组DNA,采用PCR-RFLP方法进行TLR2、TLR4基因多态性分析,使用凝胶电泳和溴乙锭检测等位基因。结果：正常对照组和（AgP）、（CP）患者均为TLR2 Arg677Trp基因的杂合型。在A妒组与CP组患者中未发现TLR2 Arg753Gln的突变型,而对照组中该基因频率为6％。在所有研究对象中都未发现TLR4 Asp299Gly和Thr399Ile的基因多态性。结论：TLR2、TLR4基因的多态性与牙周炎之间无相关性。     

不同牙周状态下龈下菌斑中人类巨细胞病毒的检测

目的 :研究人类巨细胞病毒 (HCMV)与慢性牙周炎及其活动性的相关性。方法 :采集 62例慢性牙周炎患者 (男性 2 7例 ,女性 3 5例 )的牙周炎活动部位 ,牙周炎静止部位 ,以及轻度龈炎部位的龈下菌斑 ,使用SeekVi ralDNAkit试剂盒提取DNA ,采用巢式PCR法检测HCMV ,比较同一患者不同牙周状态的检出率并加以分析。结果 :慢性牙周炎患者牙周炎活动部位、牙周炎静止部位、轻度龈炎部位的HCMV检出率为分别为 3 8.7%、14 .5 %、12 .9% ;HCMV在牙周炎活动部位的检出率高于牙周炎静止期部位 (P <0 .0 5 ) ,以及轻度龈炎部位 (P <0 .0 5 ) ;牙周炎静止部位与轻度龈炎部位的HCMV检出率的差异无统计学意义 (P >0 .0 5 )。结论 :提示HCMV感染与慢性牙周炎及其活动性的有一定的相关性。     

Real-time PCR quantification of cytomegalovirus in aggressive periodontitis lesions using TaqMan technology

BACKGROUND: Herpesviruses are implicated in the pathogenesis of human periodontitis. However, the quantity of herpesviruses in periodontal sites remains unknown. OBJECTIVE: The aim of this study was to compare levels of subgingival human cytomegalovirus (HCMV) in aggressive periodontitis patients and in periodontally healthy subjects. METHODS: A total of 16 consecutive subjects with aggressive periodontitis and 15 healthy control subjects were included in the study. Subgingival specimens were collected by a periodontal curette. TaqMan real-time polymerase chain reaction (PCR) assay was used to quantify HCMV. RESULTS: HCMV was detected in 68.8% of aggressive periodontitis lesions but not in any of the periodontally healthy study sites. HCMV viral load in positive subgingival specimens ranged from 5 x 10(2) to 7.4 x 10(3) copies/ml. CONCLUSIONS: The TaqMan real-time PCR technology seems to provide a rapid and sensitive method for quantifying HCMV in periodontal pockets. The present findings confirm the frequent presence of HCMV in aggressive periodontitis lesions. Determining HCMV levels in different types of periodontitis may help elucidate the periodontopathic role of the virus and advance our understanding of the disease pathogenesis.     

Real-time polymerase chain reaction quantification of human cytomegalovirus and Epstein-Barr virus in periodontal pockets and the adjacent gingiva of periodontitis lesions

BACKGROUND: Genomic sequences of human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV), two herpesviruses, can frequently be detected in periodontal pockets of progressive periodontitis lesions, but the prevalence and load of the two viruses in gingival tissue are unknown. This study determined levels of HCMV and EBV DNA in the periodontal pocket and in the adjacent gingiva of periodontitis lesions using a real-time polymerase chain reaction (PCR) assay. MATERIAL AND METHODS: A total of 20 systemically healthy periodontitis patients participated in the study. Nine patients below 35 years of age were tentatively diagnosed as having aggressive (early onset) periodontitis, and 11 patients 36-56 years of age as having chronic (adult) periodontitis. Clinical parameters were evaluated using established methods. Using periodontal curettes, specimens were harvested from 6-10 mm periodontal pockets and from the adjacent inflamed periodontal pocket wall. A 5'-nuclease (TaqMan) real-time PCR assay was used to identify and quantify genomic copies of periodontal HCMV and EBV. RESULTS: HCMV DNA was detected in 78% of subgingival and 33% of gingival tissue samples from aggressive periodontitis lesions, but only in 46% of subgingival and 9% of gingival tissue samples from chronic periodontitis lesions. In aggressive periodontitis, HCMV subgingival and gingival tissue counts were positively correlated with periodontal pocket depth and probing attachment loss at sample sites (p6 mm, but none of 14 patients having mean pocket depth at sample teeth相似文献   

Low prevalence of subgingival viruses in periodontitis patients

Background: Viruses such as Human Cytomegalovirus (HCMV) and Epstein–Barr virus (EBV) have been proposed to be periodontal pathogens. The aim of this study was to analyse the presence of herpesvirus DNA in subgingival plaque samples of patients with different forms of periodontitis and in healthy periodontia.
Materials and Methods: A total of 140 ethnically mixed (prevalently Caucasian) subjects took part in the study. Sixteen were affected by localized aggressive periodontitis (LAgP), 64 by generalized aggressive periodontitis (GAgP), 20 by chronic periodontitis (CP) and 40 were periodontally healthy. Polymerase chain reaction (PCR) analyses were performed to detect HCMV and EBV. Sera were tested for anti-HCMV and EBV IgG antibodies. PCRs for herpes simplex (HSV) and varicella zoster virus (VZV) were performed in subgingival samples from a subset of 20 AgP subjects.
Results: HCMV DNA was not detected in any plaque samples. EBV DNA was detected in four LAgP (25%), two GAgP (3%) subjects and four healthy individuals (10%). HSV DNA and VZV DNA were not detected in the subset of studied individuals.
Conclusions: This study challenges the previously reported high prevalence of herpesvirus DNA in subgingival samples from periodontitis patients and so questions whether they act as pathogens in such patients.     

Relationship of Actinobacillus actinomycetemcomitans serotypes to periodontal condition: prevalence and proportions in subgingival plaque

No study available has utilized the new classification scheme (the consensus report of the American Academy of Periodontology 1999) to determine the prevalence of Actinobacillus actinomycetemcomitans in different periodontal conditions. The purpose of this study was to investigate prevalence and proportions of A. actinomycetemcomitans serotypes in subgingival plaque samples from a young Taiwanese population with aggressive periodontitis, chronic periodontitis and no periodontal disease. A total of 221 subgingival plaque samples from 171 diseased subjects (70 had aggressive periodontitis, and 101 had chronic periodontitis) (mean age 25.0 +/- 8.2 yr) and 50 periodontally healthy subjects (mean age 18.4 +/- 9.5 yr) were screened for A. actinomycetemcomitans. Serotypes of A. actinomycetemcomitans were determined by an indirect immunofluorescence assay using serotype-specific polyclonal antisera to A. actinomycetemcomitans strains ATCC 29523 (serotype a), ATCC 43728 (serotype b) and ATCC 33384 (serotype c). Prevalence (% of positive samples) of A. actinomycetemcomitans was 84.3% in aggressive periodontitis, 60.4% in chronic periodontitis, and 64.0% in periodontally healthy subjects. Proportions of A. actinomycetemcomitans (mean percentage per total bacteria) in periodontally healthy subjects were significantly lower than in aggressive periodontitis subjects. The proportion of serotype b in subjects with aggressive periodontitis and subjects with chronic periodontitis were significantly greater than that in periodontally healthy subjects. The proportion of serotype c in periodontally healthy subjects was much higher than that in chronic periodontitis subjects. The results of this study suggest that prevalence and proportions of A. actinomycetemcomitans are significantly greater in patients with aggressive periodontitis than in those with chronic periodontitis. Serotype b is the predominant serotype of A. actinomycetemcomitans in patients with diseased periodontal conditions. Serotype c is a more common serotype detected in periodontally healthy subjects.     

Real-time polymerase chain reaction quantification of Epstein--Barr virus in chronic periodontitis patients

BACKGROUND: Although herpes viruses have been implicated in the pathogenesis of chronic and aggressive periodontitis, few data in the literature refer to quantification of these viruses in periodontal sites, especially in relation to serological findings. OBJECTIVE: The aim of the present study was to compare Epstein--Barr virus (EBV) DNA load in subgingival specimens from chronic periodontitis patients and in periodontally healthy subjects, in relation to serologic testing of IgM and IgG antibodies to EBV. METHODS: A total of 22 chronic periodontitis patients and 13 controls participated in the present study. Seventy-nine subgingival specimens (one pooled, one from a deep and one from a shallow site), sampled with paper points, were analysed with real-time polymerase chain reaction for EBV. Subjects were also examined for anti-EBV IgG and IgM levels in serum, using an enzyme-linked immunosorbent assay. RESULTS: One subject was seronegative for EBV. Three subjects (one patient and two controls) displayed anti-EBV IgM. Their data were excluded from further analysis. All three displayed EBV in their subgingival samples. Nine out of the remaining 20 chronic periodontitis patients and 10 out of 11 controls did not display EBV subgingivally. A statistically significant difference in viral load was observed between pooled and shallow-pocket samples from periodontitis patients but not between samples from deep and shallow pockets (Kruskall--Wallis anova, Dunn's multiple comparisons test). CONCLUSIONS: Data from the present study do not strongly support the pathogenetic significance of EBV in chronic periodontitis lesions. The data do, however, suggest that parallel serological assessments provide a useful insight into the association of viruses with periodontal disease.     

Gene expression signatures in chronic and aggressive periodontitis: a pilot study

This pilot study examined gene expression signatures in pathological gingival tissues of subjects with chronic or aggressive periodontitis, and explored whether new subclasses of periodontitis can be identified based on gene expression profiles. A total of 14 patients, seven with chronic and seven with aggressive periodontitis, were examined with respect to clinical periodontal status, composition of subgingival bacterial plaque assessed by checkerboard hybridizations, and levels of serum IgG antibodies to periodontal bacteria assayed by checkerboard immunoblotting. In addition, at least two pathological pockets/patient were biopsied, processed for RNA extraction, amplification and labeling, and used to study gene expression using Affymetrix U-133 A arrays. Based on a total of 35 microarrays, no significantly different gene expression profiles appeared to emerge between chronic and aggressive periodontitis. However, a de novo grouping of the 14 subjects into two fairly robust clusters was possible based on similarities in gene expression. These two groups had similar clinical periodontal status and subgingival bacterial profiles, but differed significantly with respect to serum IgG levels against the important periodontal pathogens Porphyromonas gingivalis, Tannerella forsythensis and Campylobacter rectus. These early data point to the usefulness of gene expression profiling techniques in the identification of subclasses of periodontitis with common pathobiology.     

慢性牙周炎龈下菌斑中福赛斯坦纳菌的检测

目的检测慢性牙周炎患者和牙周健康者龈下菌斑中福赛斯坦纳菌的数量和所占比例,探讨福赛斯坦纳菌与牙周炎发生发展的关系。方法采集经常规聚合酶链反应（PCR）法检测福赛斯坦纳菌为阳性的61例慢性牙周炎患者和12例牙周健康者的龈下菌斑,应用TaqMan实时荧光定量PCR法对菌斑的细菌总量和福赛斯坦纳菌的数量进行定量检测,构建含有福赛斯坦纳菌和真细菌靶基因的重组质粒,建立定量标准。结果本研究设计的引物和探针具有良好的特异性和敏感性;慢性牙周炎患者病变位点的福赛斯坦纳菌数量和细菌总量均高于牙周健康者的健康位点,且福赛斯坦纳菌在龈下菌斑中的比例也比健康位点高（P<0.05）;龈下菌斑的细菌数量与探诊深度呈正相关（P<0.001）;龈下菌斑中福赛斯坦纳菌所占比例在不同的探诊深度间无统计学差异（P>0.05）。结论龈下菌斑中福赛斯坦纳菌的数量与牙周状况有密切关系,实时荧光定量PCR法是研究牙周病病因及治疗方法的有效手段。     

Viruses in periodontal disease - a review

The purpose of this review was to evaluate the evidence supporting the hypothesis that viral infection plays a role in the development of periodontitis. An involvement in periodontal diseases has been suspected specifically for human immunodeficiency virus (HIV) and herpes viruses. An association has been demonstrated between HIV infection and some distinct forms of periodontal infection, i.e. necrotizing lesions. Furthermore, reports of increased prevalence and severity of chronic periodontitis in HIV-positive subjects suggests that HIV infection predispose to chronic periodontitis. Several studies, most of them from the same research group, have demonstrated an association of herpesviruses with periodontal disease. Viral DNA have been detected in gingival tissue, gingival cervicular fluid (GCF) and subgingival plaque from periodontaly diseased sites. In addition markers of herpesviral activation have been demonstrated in the GCF from periodontal lesions. Active human cytomegalovirus (HCMV) replication in periodontal sites may suggest that HCMV re-activation triggers periodontal disease activity. Concerns regarding sampling, methods and interpretation cast doubts on the role of viruses as causes of periodontal disease.     

Prevalence of human herpesviruses in patients with aggressive periodontitis

BACKGROUND: Recent studies have demonstrated that various human viruses, especially cytomegalovirus (HCMV) and Epstein-Barr virus type-1 (EBV-1), seem to play a part in the pathogenesis of human periodontitis. The aim of this investigation was to evaluate the subgingival presence of HCMV and EBV in patients with aggressive periodontitis (AgP) and healthy subjects and to examine the effect of treatment on the incidence of these viruses 3 months following surgery. METHODS: A polymerase chain reaction (PCR) method determined the presence of HCMV and EBV-1. Subgingival plaque samples from 17 consecutive AgP patients and 16 healthy controls were collected. The following indices were measured: plaque index (PI), gingival index (GI), probing depths (PD), and clinical attachment loss (CAL). Clinical parameters were assessed pretherapy and at 3 months following surgical and antimicrobial therapy. RESULTS: HCMV was detected in 64.7% of AgP patients but not detected in healthy subjects (P < 0.001) and EBV-1 in 70.6% of AgP patients and 6.3% of the healthy controls (P < 0.001). HCMV and EBV-1 coinfection was detected in 41.7% of AgP patients. A statistically significant decrease was found in all clinical parameters 3 months after treatment. There was a statistically significant decrease in HCMV and EBV-1 following therapy (P < 0.001; no HCMV; 1 patient with EBV-1). CONCLUSIONS: These findings indicate that subgingival presence of EBV-1 HCMV is strongly associated with aggressive periodontitis, and coinfection with HCMV and EBV-1 appears to be particularly deleterious to periodontal health.