Red bacterial complex is associated with the severity of chronic periodontitis in a Thai population

Background:  The distribution of periodontal pathogens differs in various geographic locations and racial/ethnic groups. This study investigated the microbiological features of chronic periodontitis (CP) patients in Thailand.
Methods:  Subgingival plaque samples from 20 non-periodontitis subjects, 20 patients with mild CP, and 20 patients with moderate to severe CP were examined using polymerase chain reaction (PCR) to identify Porphyromonas gingivalis , Tannerella forsythia , Treponema denticola , and Aggregatibacter actinomycetemcomitans .
Results:  In the moderate to severe CP patients, there was high prevalence of P. gingivalis (95%), T. forsythia (95%), T. denticola (80%), as well as the red complex (coexistence of all three species at the same lesion) (75%). A. actinomycetemcomitans was detected in only 35% of the patients in this study group. P. gingivalis was detected in as high as 45% of the non-periodontitis controls. CP and disease severity were significantly related to the presence of T. forsythia together with T. denticola and the red complex. The red complex was not found in any non-periodontitis site.
Conclusion:  Red complex bacteria were predominant periodontal pathogens of the moderate to severe form of CP in this Thai population. The presence of T. forsythia together with T. denticola , and the red complex species at the same site were significantly associated with the disease severity.     

Microbial comparison of smoker and non-smoker adult and early-onset periodontitis patients by polymerase chain reaction

A number of bacterial species are involved in the aetiology of periodontitis and include Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus and Treponema denticola. Several studies have shown differences in the microflora between the various forms of periodontal disease. It is recognised that smoking is a risk factor for periodontal disease, but there are conflicting reports on whether or not smoking has an effect on the periodontal microflora. We utilised the polymerase chain reaction to determine the presence of A. actinomycetemcomitans, P. gingivalis, P. intermedia, B. forsythus and T. denticola in subgingival plaque samples in 33 adult periodontitis (AP) patients and 24 generalized early-onset periodontitis (GEOP) patients prior to treatment. When GEOP and AP patients were compared there were significant differences in the number of positive patients and sites for both A. actinomycetemcomitans and B. forsythus (p=0.0023 and 0.00001, respectively). No statistically significant differences in the prevalence of these organisms were found between smoker and non-smoker groups. These results confirm that AP and GEOP sites harbour varied microflora, but show that B. forsythus and A. actinomycetemcomitans were detected to a significantly greater extent in this group of GEOP than in the AP patients investigated. Our findings do not support the hypothesis that smokers have significant differences in the prevalence of periodontal pathogens from non-smokers.     

吸烟与非吸烟者慢性牙周炎龈下福赛类杆菌的检测

目的：利用聚合酶链反应（PCR）检测吸烟与非吸烟者龈下福赛类杆菌（Bf）水平,通过Bf的检出率进一步探讨吸烟与牙周炎的关系。方法：健康者与慢性牙周炎伴吸烟与非吸烟患者共75例,用纸捻法提取每位观察者龈沟液,PCR检测。结果：慢性牙周炎组Bf检出率（59．5％）高于健康组（24．2％）,具有显著性差异（P〈0．01）;吸烟者慢性牙周炎组Bf检出率（72．0％）高于非吸烟者健康组、吸烟者健康组、非吸烟者慢性牙周炎组（15．8％、35．7％、41．2％）,P〈0．05。结论;Bf与吸烟及慢性牙周炎有密切关系。     

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。     

Clinical and microbiological evaluation of ligature-induced peri-implantitis and periodontitis in dogs

The purpose of this study was to evaluate the attachment loss around teeth and implants by clinical and microbiological analysis. The mandibular premolars were extracted in 5 mongrel dogs and, 3 months later, two titanium implants were installed on each side of the mandible and, after another 3 months, abutment connection was performed. Plaque control in the implants and maxillary premolars was maintained for two weeks prior to the start of the main experiment. On day 0 and 30 days after ligature placement, microbiological samples were obtained and relative attachment level was measured for the teeth and implants. The presence of Porphyromonas gingivalis, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, Prevotella intermedia and Prevotella nigrescens was evaluated by polymerase chain reaction technique on day 0 and 30 days after ligature placement. None of the above bacteria were detected on day 0. Thirty days after ligature placement, P. gingivalis was present in 95% and 85% and B. forsythus was present in 80% and 85% of the implants and teeth sites, respectively. Statistical analysis (one-way RM-ANOVA) showed a significant difference (P<0.01) between pre- and post-induction measurements around teeth and implants. However, there was no significant difference (P=0.41) in the rate of attachment loss, between periodontitis and peri-implantitis. It can be concluded that: (1) P. gingivalis and B. forsythus were strongly associated with induced peri-implantitis and periodontitis, and (2) induced peri-implantitis and periodontitis presented a similar rate of attachment loss.     

Comparative analysis of putative periodontopathic bacteria by multiplex polymerase chain reaction

Background and Objective:  The polymerase chain reaction (PCR) has been applied for the rapid and specific detection of periodontopathic bacteria in subgingival plaque and is potentially of clinical benefit in the diagnosis and treatment of periodontitis subjects. However, several technical points need to be modified before the conventional PCR detection system can be used by clinicians.
Material and Methods:  To develop a PCR-based technique more applicable for clinical use than conventional PCR, we established a multiplex PCR for five putative periodontopathic ( Treponema denticola , Porphyromonas gingivalis , Aggregatibacter actinomycetemcomitans , Prevotella intermedia and Tannerella forsythia ) and two nonperiodontopathic ( Streptococcus sanguinis and Streptococcus salivarius ) species of bacteria using whole-plaque suspension as templates, and detected bacteria in subgingival plaque taken from 85 subjects at the supportive periodontal therapy stage after active periodontal treatments.
Results:  Among putative periodontopathic bacteria, the detection frequency of T. denticola and P. gingivalis was elevated in parallel with higher probing pocket depth and clinical attachment loss, and had 4.2–14.1 times increasing odds of the clinical parameters tested. Detection of any of the five species of putative periodontopathic bacteria markedly increased the odds ratio of a higher probing pocket depth, clinical attachment loss and bleeding on probing.
Conclusion:  The multiplex PCR system developed in this study enabled the detection of all the bacteria under investigation in one reaction tube in a less time- and labor-intensive manner than conventional PCR. These results support the potential clinical use of multiplex PCR for detecting periodontopathic bacteria and for evaluating therapeutic strategies and predicting the prognosis for each subject.     

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。     

Clinical and microbiological effect of scaling and root planing in smoker and non-smoker chronic and aggressive periodontitis patients

OBJECTIVES: To compare the effects of scaling and root planing (SRP) on clinical and microbiological parameters at selected sites in smoker and non-smoker chronic and generalized aggressive periodontitis patients. MATERIALS AND METHODS: Clinical parameters including probing depth (PD), relative attachment level (RAL), and bleeding upon probing (BOP), and subgingival plaque samples were taken from four sites in 28 chronic periodontitis (CP) and 17 generalized aggressive periodontitis (GAgP) patients before and after SRP. Polymerase chain reaction assays were used to determine the presence of A. actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia and Treponema denticola. RESULTS: Both CP and GAgP non-smokers had significantly greater reduction in pocket depth (1.0+/-1.3 mm in CP smokers versus 1.7+/-1.4 mm in non-smokers, p=0.007 and 1.3+/-1.0 in GAgP smokers versus 2.4+/-1.2 mm in GAgP non-smokers, p<0.001) than respective non-smokers, with a significant decrease in Tannerella forsythensis in CP sites (smokers 25% increase and non-smokers 36.3% decrease, p<0.001) and Prevotella intermedia at GAgP sites (smokers 25% reduction versus 46.9% in non-smokers, p=0.028). CONCLUSION: SRP was effective in reducing clinical parameters in both groups. The inferior improvement in PD following therapy for smokers may reflect the systemic effects of smoking on the host response and the healing process. The lesser reduction in microflora and greater post-therapy prevalence of organisms may reflect the deeper pockets seen in smokers and poorer clearance of the organisms. These detrimental consequences for smokers appear consistent in both aggressive and CP.     

慢性牙周炎与口腔幽门螺杆菌的相关性研究

目的：探讨慢性牙周炎患者与其口腔中的幽门螺杆菌（helicobacter pylori,HP）的相关性。方法：依据HP特异的尿素酶C和cagA基因设计引物,建立PCR方法。检测37例牙周炎患者口腔菌斑和含漱液中的HP,同时设立牙周健康对照组,比较两组的检测结果。结果：尿素酶C基因在牙周炎组的阳性率为64.8%,健康组为38.5%;尿素酶C基因和cagA基因共同阳性率在牙周炎组为32.4%,健康组为7.7%;两组差异有统计学意义;且轻型牙周炎患者口腔HP阳性率低于中、重型患者。龈下菌班HP阳性率高于龈上菌斑。结论：HP参与了慢性牙周炎的致病过程,抑或是加强了其它因素的致病过程,HP与牙周炎有相关性。     

Detection and Quantification of Periodontal Pathogens in Smokers and Never‐Smokers With Chronic Periodontitis by Real‐Time Polymerase Chain Reaction

Background: The purpose of the present investigation is to compare the presence and number of periodontal pathogens in the subgingival microbiota of smokers versus never‐smokers with chronic periodontitis and matched probing depths (PDs) using real‐time polymerase chain reaction (RT‐PCR). Methods: Forty current smokers and 40 never‐smokers, matched for age, sex, and mean PD of sampling site, were included in this investigation. A full‐mouth periodontal examination was performed, and a pooled subgingival plaque sample was collected from the deepest site in each quadrant of each participant. To confirm smoking status, expired carbon monoxide (CO) concentrations were measured with a CO monitor. The presence and quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were determined using RT‐PCR. Results: Smokers had greater overall mean PD (P = 0.001) and attachment loss (P = 0.006) and fewer bleeding on probing sites (P = 0.001). An association was observed between smoking status and the presence of A. actinomycetemcomitans (P <0.001). The counts of A. actinomycetemcomitans (P <0.001), P. gingivalis (P = 0.042), and T. forsythia (P <0.001) were significantly higher in smokers. Conclusions: Smokers showed significantly greater amounts of P. gingivalis, A. actinomycetemcomitans, and T. forsythia than never‐smokers. There was a significant association between smoking and the presence of A. actinomycetemcomitans.     

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。     

慢性牙周炎致病菌与冠状动脉粥样硬化相关性研究

目的 探讨慢性牙周炎与冠状动脉粥样硬化性心脏病(简称冠心病)发病的相关性机制。方法 收集36例行冠状动脉搭桥手术并伴有慢性牙周炎患者的冠状动脉粥样硬化斑块和龈下菌斑,50例慢性牙周炎患者的龈下菌斑,采用Chelex-100法提取细菌DNA,以聚合酶链反应(PCR)法检测冠状动脉粥样硬化斑块和龈下菌斑中牙龈卟啉单胞菌(Porphyromonasgingivalis,Pg)、伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa)、具核梭杆菌(Fusobacterium nucleatum,Fn)、中间普氏菌(Preuotella intermedia,Pi)、福赛斯坦纳菌(Tannerella forsythensis,Tf)5种慢性牙周炎相关致病菌。结果 36例患者冠状动脉粥样硬化斑块中,各种牙周炎相关致病菌的检出率分别为:Pg 36%,Aa 3%,Pi 14%,Fn 25%,Tf 47%;同一患者冠状动脉粥样硬化斑块及龈下菌斑中同时检出5种牙周炎相关致病菌的一致的例数:Tf 15例,Pg 13例,Fn 5例,Pi 3例,Aa 0例;冠心病组的Pi检出率高于对照组,二者差异具有统计学意义(P<0.05)。结论 在冠状动脉粥样硬化斑块中慢性牙周炎相关致病菌DNA的检出,提示慢性牙周炎相关致病菌在冠心病的发生、发展中可能起着重要的作用;中间普氏菌可能是冠心病的危险因素。     

Detection frequency of periodontitis-associated bacteria by polymerase chain reaction in subgingival and supragingival plaque of periodontitis and healthy subjects

The aim of this study was to compare the detection frequencies of 25 bacterial species in subgingival and supragingival plaque of 18 untreated periodontitis subjects and 12 periodontally healthy subjects. Genomic DNA was extracted from subgingival and supragingival plaque samples, and bacterial detection was performed by polymerase chain reaction of the 16S rRNA genes. Fourteen bacteria showed no relationship with periodontitis, and 11 of these 14 species were frequently detected (≥50%) in subgingival plaque in both periodontitis and healthy subjects. Nine bacteria such as Eubacterium saphenum, Prevotella intermedia, and Treponema denticola seemed to be related to periodontitis; their detection frequencies in subgingival plaque samples were higher in periodontitis than in healthy subjects, but these differences were not statistically significant by multiple comparisons (0.002≤ P< 0.05). Two species ( Mogibacterium timidum and Porphyromonas gingivalis ) were detected significantly more frequently in subgingival plaque of periodontitis subjects than of healthy subjects ( P< 0.002), with P. gingivalis being detected only in periodontitis subjects, suggesting that these two species are closely related to periodontitis. There were no significant differences in the detection frequencies of the 25 bacteria between subgingival and supragingival plaque, suggesting that the bacterial flora of supragingival plaque reflects that of subgingival plaque.     

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.