牙周致病菌内毒素耐受的研究进展

内毒素耐受(endotoxin tolerance)是指先前的脂多糖(lipopolysaccharide,LPS)刺激使机体或体外培养的细胞对后续刺激反应性降低的一种现象,可能导致炎症因子释放的减少,进而在牙周炎发生发展过程中发挥重要作用。本文就牙周致病菌内毒素耐受的作用和机制的研究进展作一综述。     

慢性牙周炎主要牙周致病菌和口臭的相关性分析

目的：分析慢性牙周炎主要牙周致病菌与口臭程度的相关性。方法：选取慢性牙周炎伴口臭患者43例作为研究对象,检测其主要牙周致病菌VSCs含量和OS值,并对OS值和VSCs水平与临床指标以及VSCs水平与Pg、Pi、Tf、Td进行相关分析。结果：治疗前研究组的PD、BI、CAL、OS值和VSCs水平高于对照组,治疗后研究组PD、BI、CAL、OS值和VSCs水平与治疗前相比明显降低,差异均有统计学意义（P＜0．05）。治疗后研究组OS值和VSCs水平与PD、BI、CAL呈正相关,且VSCs水平与舌背、龈下菌斑中Pg、Pi、Tf、Td也呈正相关。结论：慢性牙周炎主要牙周致病菌与口臭程度存在着高度相关性,有效降低牙周致病菌水平,可以改善口臭值。     

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。     

从牙周致病菌检出探讨冠修复体与牙周炎的相关性

目的:探讨不同材质冠修复体对基牙牙周状况的影响。方法:选取冠修复患者120例,分别用镍铬合金(Ni-Cr)、钴铬合金(Co-Cr)、全瓷冠组(All ceramic)进行全冠修复(n=40),修复前后检查记录临床牙周指数(牙龈出血指数、菌斑指数和牙周探诊深度);采集龈下菌斑,PCR技术检测标本中的牙龈卟啉单胞菌(Pg)、伴放线嗜血杆菌(Aa)、中间普氏菌(Pi)、具核酸杆菌(Fn)。结果:修复后6~24月全瓷冠组牙周指数与修复前无显著差异(P>0.05),Ni-Cr组和Co-Cr组牙周指数均升高(P<0.05)。Ni-Cr组合金烤瓷冠在修复后12个月牙周致病菌Pg、Pi、Fn检出率显著高于Co-Cr组和氧化锆全瓷组(P<0.05);全瓷组牙周致病菌的检出率最低。结论:牙冠修复中全瓷对牙周组织的损害小于镍铬合金、钴铬合金。     

动脉粥样硬化斑块中牙周致病菌的检测

目的:探讨牙周炎的程度是否影响动脉粥样硬化斑块中牙周致病菌的检出率.方法:29例患有不同程度牙周炎患者,行冠状动脉搭桥手术时,取动脉粥样硬化斑块及术前取龄下菌斑,采用chelex-100法提取细菌DNA,以聚合酶链反应法检测牙龄卟啉单胞菌、伴放线放线杆菌、福赛斯坦氏菌3种牙周炎相关致病菌.29人分为2组,重度牙周炎组1...     

动脉粥样硬化斑块中牙周致病菌DNA的检测

目的研究动脉粥样硬化斑块中的主要牙周致病菌。方法选择需要行动脉重建外科手术的病例,收集术中分离的动脉粥样硬化斑块标本10例,提取斑块基因组DNA,用PCR扩增细菌16SrDNA片段方法检测标本中3种牙周炎特异致病菌包括伴放线放线杆菌、牙龈卟啉单胞菌和福赛坦氏菌的DNA并测序。结果10例患者动脉粥样硬化斑块中,伴放线放线杆菌检出率为0%,牙龈卟啉单胞菌为20%,福赛坦氏菌为100%。扩增之目的片段经测序与参考菌株序列一致。结论牙周致病菌在动脉粥样硬化的发生、发展中可能起着重要的作用。     

牙周致病菌免疫逃逸机制的研究进展

牙周致病菌可引起牙周组织慢性炎症,由其引发的宿主过度的免疫炎症反应会进一步加重牙周组织的损伤。本文就龈沟的天然免疫防御机制,微生物对龈沟防御机制的敏感性,中性粒细胞和牙龈上皮细胞对牙周致病菌的免疫反应进行系统阐述,旨在表明真正的牙周致病菌因不能很好地激发或者抑制宿主的免疫反应,从而逃避宿主的免疫防御,导致牙周炎的发生,为牙周病的治疗和早期预防提供参考依据。     

牙周致病菌在动脉粥样硬化发生发展中的作用研究

牙周炎与全身健康特别是心血管疾病关系密切,牙周致病菌积极参与了动脉粥样硬化的形成与发展。目前已有研究证据表明,牙周致病菌感染是心血管事件的独立危险因素,其通过多种免疫炎症以及代谢相关分子机制促进动脉粥样硬化病变的发生发展。本文从牙周炎及牙周致病菌与心血管疾病的相关性、牙周致病菌影响动脉粥样硬化的机制等方面,对这一主题进...     

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.     

儿童口腔唾液中牙周病病原菌的检测

目的:确立儿童唾液中牙周可疑病原菌检测方法,了解牙周病原菌在儿童口腔中的分布。方法:利用细菌16SRNA设计10种牙周病原菌PCR引物,采用PCR技术检测291名儿童唾液中牙周病原菌。结果:唾液中共有9种病原菌被检出。检出率分别为:生痰二氧化碳噬纤维菌(C.s)75.9%,黄褐二氧化碳噬纤维菌(C.o)72.8%,伴放线放线杆菌(A.a)64.6%,啮蚀艾肯氏菌(E.c)48.5%,直肠弯曲菌(C.r)45.0%,变黑普氏菌(P.n)29.0%,福赛拟杆菌(B.f)22.0%,中间普氏菌(P.i)5.2%,牙龈卟啉单胞菌(P.g)5.2%,未检出齿垢密螺旋体(T.d)。细菌检出率与性别无关,不同年龄组总体检出率无统计学差异。P.g只在8岁以上儿童中检出。结论:通过PCR方法可以简单、快速地检测牙周病原菌在唾液中的分布,牙周病原菌的检出为研究儿童牙周病发病提供了重要的理论依据。     

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。     

Identification of periodontopathic bacteria in gingival tissue of Japanese periodontitis patients

INTRODUCTION: The identification of invading periodontopathic bacteria in tissues is important to determine their role in the pathogenesis of periodontal disease. The objective of this study was to identify periodontopathic bacteria in diseased gingival tissue of periodontitis patients. METHODS: Subgingival plaque and gingival tissue were collected from 32 generalized chronic periodontitis (CP), 16 generalized aggressive periodontitis (GAgP) and eight localized aggressive periodontitis (LAgP) patients. Detection frequencies and quantities of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Tannerella forsythensis were investigated by polymerase chain reaction. The prevalences of Streptococcus oralis and Streptococcus sobrinus were also examined and the distribution of A. actinomycetemcomitans serotypes was observed. RESULTS: P. gingivalis and T. forsythensis were detected in approximately 70% of tissue samples and 50% of plaque samples in the three periodontitis groups. Prevalence of A. actinomycetemcomitans in tissue samples was higher in the LAgP (63%) group than in either the CP (16%) or the GAgP (38%) group. A. actinomycetemcomitans serotype c was detected in 50% of LAgP patients. Detection frequencies of S. oralis and S. sobrinus were markedly low in both plaque and tissue samples from all three periodontitis groups. Amounts of P. gingivalis, A. actinomycetemcomitans and T. forsythensis in the tissue samples were not different among the three periodontitis groups. CONCLUSION: P. gingivalis, A. actinomycetemcomitans and T. forsythensis can localize in diseased gingival tissue and may be involved in periodontal tissue destruction. Serotype c is the predominant serotype of A. actinomycetemcomitans in Japanese LAgP patients.     

Herpes viruses and periodontopathic bacteria in early-onset periodontitis

OBJECTIVES: This study examined the occurrence of human herpes viruses and suspected periodontopathic bacteria in early-onset periodontitis patients who experienced progressive disease in at least 2 periodontal sites during the maintenance phase of therapy. MATERIAL AND METHODS: In each of 16 individuals (9 male and 7 female; mean age 33.1+/-2.6 years), subgingival plaque samples were collected from 2 deteriorating and 2 stable periodontitis sites. A nested polymerase chain reaction method determined the presence of human cytomegalovirus (HCMV), Epstein-Barr virus type 1 (EBV-1) and herpes simplex virus (HSV). A 16s rRNA polymerase chain reaction method identified Porphyromonas gingivalis, Dialister pneumosintes, Bacteroides forsythus and Actinobacillus actinomycetemcomitans. RESULTS: HCMV was detected in 59.4% of active and in 12.5% of stable sites (p<0.001), EBV-1 in 43.8% of active and in 12.5 % of stable sites (p=0.01), HSV in 34.5% of active and in 9.4% of stable sites (p=0.03), and co-infection with any of the 3 test herpesviruses in 43.8% of active and in 3.1% of stable sites (p<0.001). P. gingivalis was detected in 71.9% of active and in 37.5% of stable sites (p=0.01), D. pneumosintes in 62.5% of active and in 18.8% of stable sites (p=0.04), co-infection with P. gingivalis and D. pneumosintes in 50% of active and in 0% of stable sites (p<0.001), and co-infection with any 3 or 4 of the test bacteria in 40.6% of active and in 0% of stable sites (p=0.001). All periodontitis sites showing herpesvirus co-infection and all but one site showing P. gingivalis and D. pneumosintes co-infection revealed bleeding upon probing. CONCLUSIONS: HCMV, EBV-1, HSV and herpesvirus co-infection, as well as P. gingivalis, D. pneumosintes and P. gingivalis-D. pneumosintes co-infection were statistically associated with active periodontitis. Herpesviruses are immunosuppressive and may set the stage for overgrowth of subgingival P. gingivalis, D. pneumosintes and other periodontopathic bacteria. Understanding the significance of herpesviruses in human periodontitis may allow for improved diagnosis, more specific therapy and, ultimately, disease prevention.     

In vitro activity of tetracyclines, macrolides, quinolones, clindamycin and metronidazole against periodontopathic bacteria

We re-evaluated several antibiotics including newer ones, for their in vitro killing activity, as well as their inhibitory activity, against clinical isolates of periodontopathic bacteria. Tetracyclines were active against Porphyromonas gingivalis , and were highly active against Prevotella intermedia , but demonstrated only a low killing activity against Actinobacillus actinomycetemcomitans . Rokitamycin. a new macrolide, and clindamycin were highly active against P. gingivalis and P. intermedia , but showed very weak killing activity against A. actinomycetemcomitans . Quinolones demonstrated excellent bactericidal activity against A. actinomycetemcomitans , and good inhibitory and bactericidal activity against P. gingivalis and P. intermedia . Metronidazole had an activity almost equivalent to quinolones against P. gingivalis and P. intermedia ; but it was the least active against A. actinomycetemcomitans .     

Inhibitory effects of macrocarpals on the biological activity of Porphyromonas gingivalis and other periodontopathic bacteria

BACKGROUND/AIMS: Macrocarpals, which are phloroglucinol derivatives contained in eucalyptus leaves, exhibit antimicrobial activity against a variety of bacteria including oral bacteria. This study examined effects of macrocarpals A, B, and C on periodontopathic bacteria, especially Porphyromonas gingivalis. METHODS: Macrocarpals A, B, and C were purified from a 60% ethanol-extract of Eucalyptus globules leaves. To investigate antibacterial activity, representative periodontopathic bacteria were cultured in media with or without various amounts of macrocarpals; subsequently, the optical density at 660 nm was measured. Macrocarpal inhibition of P. gingivalis Arg- and Lys-specific proteinases was assessed by spectrofluorophotometric assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The effect of macrocarpals on P. gingivalis binding to saliva-coated hydroxyapatite beads was examined with (3)H-labeled P. gingivalis. RESULTS: Growth of P. gingivalis was inhibited more strongly than growth of Prevotella intermedia or Prevotella nigrescens and Treponema denticola by macrocarpals, however, Actinobacillus actinomycetemcomitans and Fusobacterium nucleatum were much more resistant. Macrocarpals inhibited P. gingivalis Arg- and Lys-specific proteinases in a dose-dependent manner. The enzyme-inhibitory effect of macrocarpals was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis in which hemoglobin degradation by P. gingivalis proteinase was inhibited by macrocarpals. P. gingivalis binding to saliva-coated hydroxyapatite beads was also strongly attenuated by macrocarpals. CONCLUSIONS: Macrocarpals A, B and C demonstrated antibacterial activity against periodontopathic bacteria. Among tested bacteria, P. gingivalis displayed the greatest sensitivity to macrocarpals; additionally, its trypsin-like proteinase activity and binding to saliva-coated hydroxyapatite beads were inhibited by macrocarpals. These results indicate that eucalyptus leaf extracts may be useful as a potent preventative of periodontal disease.     

The distribution of periodontopathic bacteria among Japanese children and their parents

OBJECTIVE AND BACKGROUND: It is not well known how periodontopathic bacteria colonize in the oral cavity during childhood. The purpose of this study was to investigate the distribution of periodontopathic bacteria in oral cavities of children and their parents and the relationship between the bacterial findings and clinical parameters. METHODS: Fifty-six children (mean age: 8.3 +/- 3.5, range: 1-15 years), including 15 with deciduous dentition, 26 with mixed dentition and 15 with permanent dentition, and their parents participated in this study. Whole saliva and dental plaque of the children and whole saliva of their parents were collected for detection of seven species of periodontopathic bacteria (Actinobacillus actinomycetemcomitans, Tannerella forsythensis (Bacteroides forsythus), Campylobacter rectus, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens and Treponema denticola) using the polymerase chain reaction method. Clinical parameters including simplified Oral Hygiene Index and Papillary-Marginal-Attachment Index were recorded for the children and their accompanied parents. RESULTS: The detection frequencies of T. forsythensis, C. rectus, P. nigrescens, T. denticola, A. actinomycetemcomitans and P. gingivalis in the oral cavities of children were 42.9%, 94.6%, 42.9%, 48.2%, 1.8% and 8.9%, respectively. T. forsythensis, P. gingivalis and T. denticola were detected more frequently in the saliva of parents (54.8%, 54.8%, 88.1%, respectively) than in the saliva of children (25.5%, 7.3%, 41.8%, respectively). Different detection frequencies of P. nigrescens were found among the oral cavities of children with deciduous, mixed and permanent dentitions. In mixed dentition, females harbored T. forsythensis more frequently than males did. Children who harbored T. forsythensis, P. intermedia, P. nigrescens and T. denticola showed high scores for oral debris measurement by simplified Oral Hygiene Index. T. forsythensis, P. intermedia and P. nigrescens were detected more frequently in children whose parents were positive for these pathogens than in children whose parents were negative. CONCLUSIONS: High plaque retention seems to promote the colonization of periodontal pathogens in the oral cavities of children. T. forsythensis, P. intermedia and P. nigrescens were detected more frequently in the oral cavities of children whose parents already harbored these bacteria. Familial transmission of these bacteria is suggested.     

Effect of periodontal treatments on serum IgG antibody titers against periodontopathic bacteria

Abstract. Serum IgG antibody titers to 7 periodontopathic bacteria in periodontitis patients were measured at the Is1 visit and after various periodontal treatments with clinically successful improvement, in order to evaluate what kind of factors are associated with changes of serum antibody titers. 20 patients (10 male and 10 female from 23 to 61 years old) with adult, rapidly progressive periodontitis were enrolled in this study. All patients received initial preparation and most of them also underwent surgical procedure. After the treatments, the mean probing pocket depths decreased from 3.72 mm to 1.56 mm. Serum samples were collected from patients at the initial and final examinations. Serum IgG antibody titers against sonicated antigens of Porphyromonas gingivalis FDC 381, Prevotella intermedia ATCC 25611. Prevotella loescheii ATCC 15930. Fusobacterium nucleatum subspecies nucleatum ATCC 25586. Actinobacillus actinomycetemcomitans FDC Y4. Eikenella corrodens FDC 1073 and Capnocylophaga ochracea #M 12 were determined by enzyme-linked immunosorbent assay. The mean antibody titers to P. gingivalis and P. intermedia decreased significantly after the treatment as compared to their pretreatment levels. The antibody titer to P. gingivalis, especially, decreased in all of the patients examined. A significant relationship was found between the decreased antibody titer to P. gingivalis and the number of teeth which received periodontal surgery, as well as treatment length, and the relationship between the decreased antibody titer to P. intermedia and the number of extracted teeth was also significant. These results suggest that the changes of serum IgG titers against P. gingivalis and P. intermedia are related to the suppression of such pathogens in subgingival plaque.