伴2型糖尿病的慢性牙周炎牙周可疑致病菌的检测

目的 检测伴2型糖尿病(diabetes mellitus,DM)的慢性牙周炎(chronic periodontitis,CP)患者龈下菌斑中牙周可疑致病菌的种类和构成,从微生物学角度探讨牙周炎与DM的相互作用机制.方法 采集伴2型DM的CP患者154例(DM组)、不伴DM的单纯CP患者120例(CP组)及40名全身及牙周健康者(N组)的龈下集合菌斑,传统酚-氯仿法提取菌斑DNA,以牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg),伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa),具核梭杆菌(Fusobacterium nucleatum,Fn),中间普氏菌(Prevotella intermedia,Pi),福塞坦氏菌(Tannerella forsythia,Tf),齿垢密螺旋体(Treponema denticola,Td)为目标菌,应用以16SrRNA为基础的聚合酶链反应(PCR)技术对龈下菌群进行检测.结果 Pg、Aa、Fn、Pi、Tf、Td在DM组中均可检出;与CP组相比,在性别、年龄、牙周状况基本一致的情况下,轻度牙周炎者DM组Pi的检出率为35%(8/23),CP组为65%(13/20),两组间差异有统计学意义(P<0.05);重度牙周炎者DM组Pg、Aa、Tf的检出率分别为78%(72/92)、27%(25/92)、67%(62/92),CP组分别为58%(35/60)、17%(10/60)、43%(26/60),DM组均显著高于CP组,差异有统计学意义(P<0.05).同时,DM组Aa、Tf PCR产物的平均灰度值(average gradation,AVG)比值显著高于CP组,Pi的AVG比值明显低于CP组,P<0.05.结论 与单纯CP相比,伴2型DM的牙周炎患者龈下菌斑中Pg、Aa、Tf的数量增多,Pi的数量减少.     

伴2型糖尿病的慢性牙周炎牙周可疑致病菌的检测

目的 检测伴2型糖尿病(diabetes mellitus,DM)的慢性牙周炎(chronic periodontitis,CP)患者龈下菌斑中牙周可疑致病菌的种类和构成,从微生物学角度探讨牙周炎与DM的相互作用机制.方法 采集伴2型DM的CP患者154例(DM组)、不伴DM的单纯CP患者120例(CP组)及40名全身及牙周健康者(N组)的龈下集合菌斑,传统酚-氯仿法提取菌斑DNA,以牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg),伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa),具核梭杆菌(Fusobacterium nucleatum,Fn),中间普氏菌(Prevotella intermedia,Pi),福塞坦氏菌(Tannerella forsythia,Tf),齿垢密螺旋体(Treponema denticola,Td)为目标菌,应用以16SrRNA为基础的聚合酶链反应(PCR)技术对龈下菌群进行检测.结果 Pg、Aa、Fn、Pi、Tf、Td在DM组中均可检出;与CP组相比,在性别、年龄、牙周状况基本一致的情况下,轻度牙周炎者DM组Pi的检出率为35%(8/23),CP组为65%(13/20),两组间差异有统计学意义(P<0.05);重度牙周炎者DM组Pg、Aa、Tf的检出率分别为78%(72/92)、27%(25/92)、67%(62/92),CP组分别为58%(35/60)、17%(10/60)、43%(26/60),DM组均显著高于CP组,差异有统计学意义(P<0.05).同时,DM组Aa、Tf PCR产物的平均灰度值(average gradation,AVG)比值显著高于CP组,Pi的AVG比值明显低于CP组,P<0.05.结论 与单纯CP相比,伴2型DM的牙周炎患者龈下菌斑中Pg、Aa、Tf的数量增多,Pi的数量减少.     

伴2型糖尿病的慢性牙周炎牙周可疑致病菌的检测

目的 检测伴2型糖尿病(diabetes mellitus,DM)的慢性牙周炎(chronic periodontitis,CP)患者龈下菌斑中牙周可疑致病菌的种类和构成,从微生物学角度探讨牙周炎与DM的相互作用机制.方法 采集伴2型DM的CP患者154例(DM组)、不伴DM的单纯CP患者120例(CP组)及40名全身及牙周健康者(N组)的龈下集合菌斑,传统酚-氯仿法提取菌斑DNA,以牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg),伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa),具核梭杆菌(Fusobacterium nucleatum,Fn),中间普氏菌(Prevotella intermedia,Pi),福塞坦氏菌(Tannerella forsythia,Tf),齿垢密螺旋体(Treponema denticola,Td)为目标菌,应用以16SrRNA为基础的聚合酶链反应(PCR)技术对龈下菌群进行检测.结果 Pg、Aa、Fn、Pi、Tf、Td在DM组中均可检出;与CP组相比,在性别、年龄、牙周状况基本一致的情况下,轻度牙周炎者DM组Pi的检出率为35%(8/23),CP组为65%(13/20),两组间差异有统计学意义(P<0.05);重度牙周炎者DM组Pg、Aa、Tf的检出率分别为78%(72/92)、27%(25/92)、67%(62/92),CP组分别为58%(35/60)、17%(10/60)、43%(26/60),DM组均显著高于CP组,差异有统计学意义(P<0.05).同时,DM组Aa、Tf PCR产物的平均灰度值(average gradation,AVG)比值显著高于CP组,Pi的AVG比值明显低于CP组,P<0.05.结论 与单纯CP相比,伴2型DM的牙周炎患者龈下菌斑中Pg、Aa、Tf的数量增多,Pi的数量减少.     

牙周可疑致病菌的传播性研究进展

现代分子生物学的快速进展为微生物的分类学研究提供了多种高敏感度的方法,使研究各类细菌的传播途径成为可能。近年来研究表明牙周可疑致病菌可交互传播,本文着重就其中与牙周疾病关系密切的相关细菌在人群中传播的可能性及途径的研究进展进行综述。     

未获培养微生物与牙周可疑致病菌的牙周共生状态

目的：探索未获培养微生物与牙周可疑致病菌的共生关系。方法：使用基于16SrRNA基因检测的PCR法，对慢性牙周炎患者龈下菌斑中牙周可疑致病微生物和代表性未获培养微生物进行检测，通过KAPPA检验分析任意两种细菌对间的关系。结果：在56个细菌对中发现6对细菌的KAPPA值具有显著性。结论：Pg／T.d、B．f/T．d、B．f／F.n细菌组合的共生关系与文献报道一致，AU126／P．n、AU126／F．n、AU126／X112细菌对间的共生关系显示了未获培养微生物在龈下菌群中扮演的角色。     

臭氧水溶液对牙周可疑致病菌作用的研究

目的：评价臭氧水溶液对常见牙周可疑致病菌的杀菌效果。方法：采用细菌悬液定量实验法,测定不同浓度的臭氧水溶液对牙龈卟啉单胞菌（P．g）ATCC33277、伴放线菌嗜血菌（H．a）ATCC29522、具核梭杆菌（F．n）ATCC10957和牙龈卟啉单胞菌临床分离株作用30、60、90、120 s 的杀菌率,阳性对照组采用不同浓度的过氧化氢溶液,阴性对照组采用蒸馏水。实验样本在厌氧培养72 h 后进行菌落计数并计算杀菌率。结果：臭氧水溶液对几种牙周可疑致病菌均有杀灭效果,且杀菌率随浓度的增加而提高（P ＜0．01）。对 P．g 标准株和临床分离株的杀菌率差异无统计学意义（P ＞0．05）。线性回归分析臭氧水溶液对3种细菌杀灭作用的浓度影响因素β值均大于0．95,时间影响因素β值均小于0．11。结论：臭氧水溶液对牙龈卟啉单胞菌、伴放线菌嗜血菌和具核梭杆菌具有浓度依赖性杀灭效果。     

牙周炎可疑致病菌对慢性肾脏病的影响及可能机制

慢性肾脏病发病率高、病死率高,已逐步成为严重威胁人类身体健康的全球公共卫生问题。牙周炎作为常见的口腔慢性感染性疾病,与慢性肾脏病具有相关性。牙周可疑致病菌是牙周炎的始动因子,在慢性肾脏病的发生、发展中可能起到重要作用。文章就牙周可疑致病菌对慢性肾脏病的影响及可能机制做一综述。     

牙周基础治疗对2型糖尿病相关性牙周炎患者血糖控制的Meta分析

目的系统评价牙周基础治疗对2型糖尿病相关性牙周炎患者血糖控制的影响,探讨牙周基础治疗在糖尿病治疗中的具体作用。方法计算机检索Cochrane图书馆对照试验注册中心、Medline、EMbase、SIGLE、GreyNet、NTIS、中国生物医学文献数据库、中文科技期刊全文数据库、中国期刊全文数据库和万方数据库,查找有关牙周基础治疗对糖尿病血糖控制影响的研究。检索时限均为1991-2011年4月31日。均由2名评价者独立选择试验、提取资料和评估方法学质量,然后采用RevMan 5．1软件对资料进行Meta分析。纳入7个研究,共计471例受试患者。结果牙周基础治疗能明显降低2型糖尿病相关性牙周炎患者糖化血红蛋白的水平,组间差异有统计学意义（95％CI：-0．94—0．22,P=0．001）。治疗组牙周袋探诊深度低于对照组,组间差异也有统计学意义（95％CI：-2．26～0．69,P=0．0002）。结论牙周基础治疗有利于2型糖尿病相关性牙周炎患者总体血糖水平的控制。     

2型糖尿病伴慢性牙周炎牙周基础治疗的疗效观察

目的观察牙周基础治疗对2型糖尿病伴慢性牙周炎患者在临床指标及血糖控制方面的影响。方法选取52例2型糖尿病伴慢性牙周炎患者并随机分为2组。实验组行牙周基础治疗和药物治疗，对照组仅给予药物治疗。治疗前后分别记录患者各项临床指标和糖化血红蛋白水平。结果治疗后16周实验组菌斑指数（PLI）及出血指数显著减少，且PLI与对照组差异有统计学意义（P〈0.001）：两组患者其他各项临床指标和糖化血红蛋白水平在治疗后有一定改善，但差异无统计学意义。结论牙周基础治疗有利于2型糖尿病伴慢性牙周炎患者减轻牙周炎症及在一定程度上改善代谢状况。     

牙周基础治疗对Ⅱ型糖尿病伴慢性牙周炎患者血糖水平的影响

目的:观察牙周基础治疗对Ⅱ型糖尿病伴慢性牙周炎患者牙周炎症的控制和糖化血红蛋白的影响。方法:选取62例患者分为牙周治疗组(31例)和非牙周治疗组(31例),牙周治疗组给予牙周基础治疗和药物治疗,记录术前和术后8周龈沟出血指数(SBI)、牙周探诊深度(PD)、附着水平丧失(AL)、糖化血红蛋白(HbAIc)。非牙周治疗组仅给予药物治疗并记录治疗前及治疗后8周的PD、AL、HbAIc。结果:牙周治疗组PD、AL、HbAIc均有明显下降(P<0.01)。非牙周治疗组无显著性差异。结论:牙周基础治疗对Ⅱ型糖尿病伴慢性牙周炎患者具有良好的治疗反应。     

Antibiotic susceptibility of putative periodontal pathogens in advanced periodontitis patients

In the present study, the antibiotic susceptibility of most prevalent micro-organisms in advanced periodontitis patients was evaluated. In 56 patients, pooled subgingival plaque samples were taken from the deepest site of each quadrant and were cultivated anaerobically. From each patient, the 4 most frequently encountered types of bacterial colonies were subcultured and identified (Rapid ID 32 A). From all bacterial species identified in the 224 subcultures, the 4 most prevalent were used for susceptibility testing to tetracycline, metronidazole and amoxicillin/clavulanate using the E Test. The most prevalent microorganisms were Fusobacterium nucleatum (38/214), Peptostreptococcus micros (33/214), Prevotella oralis (33/214) and Porphyromonas gingivalis (32/214). Regarding antibiotic susceptibility it could be shown that minimal inhibitory concentration (MIC) in all cases was below antibiotic concentrations achievable in gingival crevicular fluid. However, antibiotic resistance was seen in 3 to 29% of the investigated microorganisms.     

Prevalence of periodontal pathogens in Brazilians with aggressive or chronic periodontitis

OBJECTIVES: Previous studies suggest differences between geographically and racially distinct populations in the prevalence of periodontopathic bacteria as well as greater periodontal destruction associated with infection by highly leucotoxic Actinobacillus actinomycetemcomitans. The present study examined these hypotheses in Brazilians with aggressive or chronic periodontitis. MATERIALS AND METHODS: Clinical, radiographical, and microbiological assessments were performed on 25 aggressive periodontitis and 178 chronic periodontitis patients including 71 males and 132 females, 15-69 years of age. RESULTS: The prevalence of Porphyromonas gingivalis was similar to that of other South American populations. The prevalence of A. actinomycetemcomitans and its highly leucotoxic subgroup was higher in Brazilians. Highly leucotoxic A. actinomycetemcomitans was more prevalent in aggressive periodontitis (chi2=27.83) and positively associated with deep pockets (>6 mm, chi2=18.26) and young age (<29 years, chi2=18.68). Greater mean attachment loss was found in subjects with highly leucotoxic A. actinomycetemcomitans than in subjects with minimally leucotoxic (p=0.0029) or subjects not infected (p=0.0001). CONCLUSION: These data support the hypothesis of differences between populations in the prevalence of periodontopathic bacteria and of greater attachment loss in sites infected with highly leucotoxic A. actinomycetemcomitans. Detection of highly leucotoxic A. actinomycetemcomitans in children and adolescents may be a useful marker for aggressive periodontitis.     

Investigation and quantification of key periodontal pathogens in patients with type 2 diabetes

Field CA, Gidley MD, Preshaw PM, Jakubovics N. Investigation and quantification of key periodontal pathogens in patients with type 2 diabetes. J Periodont Res 2012; 47: 470–478. © 2012 John Wiley & Sons A/S Background and Objective: Diabetes is a recognized risk factor for periodontitis. There are conflicting data regarding whether healthy diabetic patients or diabetic patients with chronic periodontitis have an altered subgingival microbiota compared with nondiabetic individuals. The aim of the present study was to detect quantitative differences in selected periodontopathogens in the subgingival plaque of diabetic patients using TaqMan quantitative PCR. Material and Methods:  Type 2 diabetes mellitus patients with (n = 9) or without chronic periodontal disease (n = 15) were recruited and matched to nondiabetic control subjects (n = 12 periodontally healthy, n = 12 chronic periodontitis). Subgingival plaque samples were collected from deep (> 4 mm probing depth) and shallow sites (≤ 3 mm probing depth) using paper points, and Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum and Porphyromonas gingivalis were quantified. Results: Forty‐eight subjects (69 samples) were recruited. Marked differences were seen in the levels of all three bacterial species, relative to the total bacterial population, according to periodontal health status. Using real‐time quantitative PCR, bacterial counts for P. gingivalis were significantly higher in deep pockets of diabetic and nondiabetic subjects compared with periodontally healthy subjects (p < 0.05) but did not differ significantly between diabetics and nondiabetics. A. actinomycetemcomitans was detected in all groups in low quantities, and counts did not differ significantly between groups (p > 0.05). F. nucleatum was abundant in all groups, with no clear significant differences between groups. P. gingivalis was found in higher quantities in periodontitis than in periodontally healthy subjects (p < 0.05). Statistically significant positive correlations were identified between pocket depth and counts for all three species tested (p < 0.05). Conclusion: A. actinomycetemcomitans, F. nucleatum and P. gingivalis were present in significantly different quantities and proportions in subgingival plaque, according to periodontal disease status. No significant differences were identified between the subgingival microbiota of type 2 diabetes mellitus patients compared with nondiabetic subjects.     

慢性牙周炎龈下菌斑中五种牙周可疑致病微生物的分布

目的研究五种牙周可疑致病微生物在慢性牙周炎患者龈下菌斑的分布。方法选择27例慢性牙周炎患者,每位患者选取牙周袋最深的两个位点作为观察位点,采集龈下微生物样本,采用多重聚合酶链反应和反杂交的方法对伴放线菌嗜血菌、牙龈卟啉单胞菌、福赛斯坦纳菌、中间普雷沃菌和齿垢密螺旋体五种微生物进行半定量检测。结果在所检测的54个位点中,牙龈卟啉单胞菌、中间普雷沃菌、福赛斯坦纳菌和齿垢密螺旋体均有较高的检出率,分别为98.15%、92.59%、100%和98.15%;伴放线菌嗜血菌检出率较低,为20.37%。牙龈卟啉单胞菌和福赛斯坦纳菌的检出量明显高于其他三种微生物,其差异有统计学意义（P<0.05）。结论慢性牙周炎患者多存在牙龈卟啉单胞菌、福赛斯坦纳菌、中间普雷沃菌和齿垢密螺旋体的同时感染,且牙龈卟啉单胞菌和福赛斯坦纳菌的感染量较高。     

牙周基础治疗对侵袭性牙周炎患者唾液和龈沟液中牙周致病菌的作用

目的 分析侵袭性牙周炎(aggressive periodontitis,AgP)患者牙周基础治疗前后唾液和龈沟液中6种牙周致病菌的变化及差异,为评价AgP的治疗效果和预后提供一个微生物学手段.方法 16例AgP患者在牙周基础治疗前、治疗后2周及2、4、6个月采集唾液和龈沟液,聚合酶链反应技术检测伴放线放线杆菌(Aggregatibacter actinomycetemcomitans,Aa)、牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)、福赛坦菌(Tannerella forsythia,Tf)、齿垢密螺旋体(Treponema denticola,Td)、中间普氏菌(Prevotella intermedia,Pi)和具核梭杆菌(Fusobacterium nucleatum,Fn)的分布.结果 16例AgP患者唾液中Aa、Pg、Tf、Td、Pi和Fn在治疗前均检出较高,分别为7、16、13、14、9和14例患者检山;治疗后2周降至最低,分别为3、9、5、4、4和7例患者检出;治疗后2个月开始升高,治疗后6个月时唾液中牙周致病菌检出基本回到治疗前水平.龈沟液中各位点Aa、Pg、Tf、Td、Pi和Fn的检出率在治疗前分别为17％ (11/64)、95％ (61/64)、84％(54/64)、72％(46/64)、41％ (26/64)、56％(36/64);治疗后2周显著降低,在治疗后6个月时仍保持相对较低的水平,分别为13％ (8/64)、73％(47/64)、42％(27/64)、23％(15/64)、8％ (5/64)和38％(24/64),除Aa外其余5种牙周致病菌的检出率均显著低于治疗前(P＜0.05).唾液和龈沟液中6种牙周致病菌检出率存在差异.结论 AgP患者牙周致病菌的变化早于临床指标的变化;定期每2个月的牙周维护在短期内可使龈沟液中致病菌维持在低水平;唾液中牙周致病菌的检测不能完全代替位点检测.     

5种牙周龈下可疑致病微生物与慢性牙周炎局部不同牙周状态间的关系

目的 观察5种龈下微生物检出水平与慢性牙周炎局部牙周状态的关系。方法 选择20例慢性牙周炎患者的80个位点及10例牙周健康者的20个位点为观察位点,采集龈下微生物样本,记录牙周探诊深度（PD）,根据所测位点的PD进行分组。PD≤4 mm为A组,4 mm＜PD≤6 mm为B组,PD>6 mm为C组,健康对照组为H组。通过聚合酶链反应（PCR）和DNA探针反杂交技术半定量检测各组伴放线菌嗜血菌、牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌的检出率和检出水平。结果 B、C组牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌的检出率和检出水平均高于H组,A组牙龈卟啉单胞菌的检出率和检出水平也高于H组,C组福赛斯坦纳菌和齿垢密螺旋体检出水平高于B组,以上差异均有统计学意义（P<0.05）;伴放线菌嗜血菌在各组间的检出率及检出水平都无明显差异。结论 随着牙周袋的加深,牙龈卟啉单胞菌、福赛斯坦纳菌、齿垢密螺旋体和中间普氏菌体的阳性检出率和检出水平都有随之增加的趋势;牙龈卟啉单胞菌与慢性牙周炎的早期炎症关系较为密切,而福赛斯坦纳菌和齿垢密螺旋体与中重度慢性牙周炎炎症位点的严重程度有关。