Distribution of enzymatically pathogenic bacteria from periodontal pocket in advancing periodontitis

Production of 9 enzymatic activities of 527 strains freshly isolated from periodontal pockets in advancing periodontitis were investigated. Of these isolates, two strains showed lecithinase activity on egg yolk agar plate. Collagenase, plasmin and lipase were produced by 28 strains, 26 strains and 22 strains, respectively. Two lecithinase-producing strains were identified as Bacteroides intermedius. Nineteen strains of B. intermedius and 1 strain of Fusobacterium species produced lipase on egg yolk agar plate. All of the 28 collagenase-producing strains were B. gingivalis. B. gingivalis (20 strains) and non black-pigmented Bacteroides (6 strains) showed plasmin activity. These results indicate that Bacteroides species, mainly B. gingivalis and B. intermedius may exert an important influence on the exacerbation of the disease.     

Sulfate-reducing bacteria in the periodontal pocket

This report is the first to describe the occurrence of sulfate-reducing bacteria in the human mouth. Samples of subgingival dental plaque were examined for the presence of sulfate-reducing bacteria. Using enrichment cultures, sulfate-reducing bacteria were detected in 25 (58%) of 43 individuals, and in 39 (48%) of the 82 samples. Pure isolates of sulfate-reducing bacteria, obtained from a limited number of enrichment cultures, belonged to the genera Desulfobacter and Desulfovibrio. These genera are also the predominant sulfate-reducing bacteria in the human large intestine. The sulfate-reducing bacteria use sulfate as terminal electron acceptor to oxidize low-molecular-weight organic compounds, mainly products of microbial fermentation such as acetate, lactate etc. The numbers of sulfate-reducing bacteria in the mouth are assumed to be limited by sulfate. Potential sources of sulfate in the subgingival area include free sulfate in pocket fluid and glycosaminoglycans from periodontal tissues.     

A study of the bacteria associated with advancing periodontitis in man

Samples of apical plaque were taken by means of an anaerobic gas-flushed syringe from 21 sites in eight patients. The samples were anaerobically dispersed, diluted and plated and incubated in an atmosphere of 80% N2, 10% H2 and 10% CO2 for 7-21 days. All colonies on plates containing 20-50 isolates were picked, repeatedly restreaked, characterized and identified where possible by a probabilistic computer identification program. The sites were divided into four groups on the basis of clinical features. The significance of differences between bacterial populations in the groups was determined by the Kruskal Wallis and Mann-Whitney U tests, while the Spearman rank correlation coefficient was used to determine the rank correlation of clinical features of diseases and microbial species. The subgingival microbiota in advanced destructive sites was predominated by Gram-negative rods. The microbiota of two young adult patients with generalized extensive bone loss, extensive clinical inflammation and suppuration was dominated by Bacteroides asaccharolyticus and an organism with characteristics consistent with Actinobacillus actinomycetemcomitans. The predominant cultivable microbiota in two patients with extensive bone loss but minimal clinical inflammation was predominated by Bacteroides melaninogenicus ss intermedius and Eikenella corrodens in one patient and E. corrodens and a slow growing fusiform-shaped Bacteroides in a second patient. A third group of four patients demonstrated moderate levels of clinical inflammation and evidence of continued bone loss in the last year. Predominant organisms in this group were more heterogeneous and included B. asaccharolyticus, Fusobacterium nucleatum, the "fusiform" Bacteroides and anaerobic vibrios. Sites with minimal disease in the patients revealed higher proportions of Gram-positive organisms including Rothia dentocariosa, Actinomyces naeslundii and Actinomyces viscosus. A positive rank correlation could be detected between clinical inflammation including suppuration and B. asaccharolyticus and a negative rank correlation between inflammation and E. corrodens.     

Relationship between some subgingival bacteria and periodontal pocket depth and gain or loss of periodontal attachment after treatment of adult periodontitis

We studied the association between post-treatment periodontal disease activity and subgingival Bacteroides gingivalis, Bacteroides intermedius, spirochetes and motile rods. 20 adults, 22-62 years, with moderate-to-severe periodontitis participated in a split-mouth treatment study. All individual quadrants received supragingival cleaning and in addition, subgingival scaling and a NaHCO3-NaCl-H2O2 slurry, subgingival scaling alone, slurry alone, or no subgingival treatment. Post-treatment periodontal disease status was determined over a period of 12 months by changes in probing periodontal pocket depth and probing periodontal attachment level. Subgingival specimens obtained by paper point-sampling were evaluated for B. gingivalis and B. intermedius using indirect immunofluorescence and for spirochetes and motile rods using bright light phase contrast microscopy. A total of 142 periodontitis lesions representing all 4 quadrants of the 20 subjects were studied. The relationship between clinical data and bacteria was analyzed using logistic regression. The probability of the study organisms being present in subgingival sites at 3 to 6 months after treatment increased with increased residual pocket depth. The presence of B. gingivalis showed a strong positive association (p less than 0.004) with loss of periodontal attachment. A significant association was also found for spirochetes (p less than 0.008) but not for motile rods (p greater than 0.35) or B. intermedius (p greater than 0.13). Similar results were obtained at 12 months after therapy, except that the presence of motile rods was significantly associated with loss of periodontal attachment (p less than 0.03). Caution must be exercised when using B. gingivalis or spirochetes to evaluate treatment efficacy. If the presence of these organisms was utilized to indicate progressing periodontitis, many active lesions could be identified, and only 1 to 17% and 13 to 43% of sites in remission at 3-6 months after therapy would be expected to harbor B. gingivalis and spirochetes, respectively. The consequences of treating periodontal sites in remission would mainly be limited to cost and inconvenience. However, since several active periodontitis lesions did not reveal the organisms, treatment decisions based solely on the absence of the organisms may result in the omission of needed therapy. As a practical consideration, periodontal treatment should be continued as long as B. gingivalis and maybe spirochetes are detectable in the subgingival microflora. In the absence of these organisms, and until additional periodontal pathogens have become known, the decision to continue or conclude periodontal therapy must b     

牙周炎患者牙周袋内硫化物水平与牙周临床指标的相关分析

目的评价牙周炎患者牙周袋内硫化物（pS）与口气挥发性硫化物（VSC）及牙周临床指标的相关性。方法全口洁治后1周，对20例慢性牙周炎伴口臭的患者龈下刮治和根面平整（SRP）前后用Halimeter检测口气中VSC，用金刚探针检测pS，并对牙周袋内pS及菌斑指数、出血指数、探诊深度、临床附着水平纵向观察6个月。结果口内pS〉0的位点数（r=0．592，P〈0．001）和ps值（r=0．506，P〈0．001）与口气VSC均呈显著正相关；治疗后6周、3个月、6个月，pS与出血指数呈低度相关，r值分别为0．183、0．411、0．350（P〈0．05）；SRP后6周，pS及各临床指标较基线水平显著改善（P〈0．001），并在第3、6个月时进一步改善或保持稳定。结论pS值可以特异性地反映牙周袋内硫化物的水平，可作为牙周疗效判断的客观指标，反映和监测维护期的牙周组织炎症状况。     

牙周牙髓病的髓腔与牙周袋内细菌相关性研究

目的：对牙髓牙周综合症患牙进行细菌学分析，以探讨牙髓炎、牙周炎在病原学上的相互关系。方法：对牙髓腔、牙周袋内的细菌进行需氧，厌氧培养，并用自动微生物鉴定系统对其进行分类鉴定。结果：髓腔和牙周袋内的细菌组成有相关性，主要为厌氧的产黑色素杆菌。结论：牙髓炎、牙周炎可通过根尖孔互为感染源，而导致牙髓、牙周综合征     

牙周洁治对人牙周炎袋底微生物和龈沟液量的影响

目的:探讨牙周洁治对人牙周炎袋底微生物和龈沟液量的影响,为研究牙周洁治机制提供依据。方法:选60例牙周炎患者,随机分为2组,每组30例,洁治组作牙周洁治,对照组用常规漱口液漱口,分别在治疗前、治疗后1周和治疗后1月用刚果红负性染色法观察袋底微生物的变化,并测定牙周探诊深度(PD)、龈指数(GI)和龈沟液量(GCF)的变化。结果:洁治组在洁治后1周时牙周袋底的球菌比率明显增加,杆菌和螺旋体比率及PD和GCF明显降低;对照组GI轻度降低,但PD和GCF以及袋底的球菌、杆菌、螺旋体比率无明显变化;洁治组和对照组的球菌、杆菌和螺旋体比率及PD、GI和GCF均有显著性差异。洁治后1月时PD、GI和GCF量及袋底的微生物比率与洁治1周时无显著性差异。结论:牙周洁治对人牙周炎袋底微生物有明显的改善作用,可通过减少袋底致病菌比率和降低PD以减轻牙周炎症和减少龈沟液的分泌。     

Changes of factors on disease activity in advancing periodontitis

Two test teeth, anteriors with greater than or equal to 6 mm deep periodontal pockets from each of 10 patients with advancing periodontitis were included in this study. The clinical signs of advancing periodontitis, generalized moderate to deep pockets and to severe loss of alveolar bone, were observed in young adult. There have been several reports on factors, which reflect the conversion clinically from infection by highly pathogenic plaque bacteria to a from of periodontitis displaying relatively rapid loss of clinical attachment. The purpose of this investigation was to detect parameters in fluid, which could leak from the underlying inflamed connective tissue into the gingival crevice, and which could shown correlatively the progressive variations of periodontal disease by recurrent acute stage. In order to determine active disease sites and to monitor guantitatively response to therapy or to measure degree to susceptibility of future breakdown. Examinations on following parameters at pre- and post- periodontal treatment stages were carried out. Endotoxin, collagenase, alkaline phosphatase, beta-glucuronidase, interleukin-1 alpha, IgG antibody levels to Bacteroides gingivalis, Bacteroides intermedius were measured in gingival exudate samples, which were collected by the microtips technique from periodontal pockets. The following results were obtained: 1) Considering the effect of periodontal therapy, pathogenic responses on total colony forming unit (CFU), interleukin-1 alpha and changes of endotoxin and beta-glucuronidase levels after the treatment have indicated that specific changes in humoral responses. 2) There was not significant relation between alkaline phosphatase, collagenase, IgG antibodies level to Bacteroides gingivalis, Bacteroides intermedius and responses in active and also inactive disease sites. 3) This study has been resulted in the development of diagnostic techniques which requires strict criteria on the disease activity of the periodontal disease very specific in order to permit a more scientific approach to the care of periodontitis patients and to speculate the prognosis of the patients after the treatment.     

慢性牙周炎栖牙密螺旋体和牙周袋内硫化物水平的相关性研究

目的 研究慢性牙周炎病人牙周袋内栖牙密螺旋体和牙周袋内硫化物水平的关系。方法 临床上选取17例诊断为慢性牙周炎的病人,采用金刚探针/牙周诊断仪检测牙周袋内硫化物水平,记录牙周袋探诊深度、临床附着丧失以及探诊出血相关牙周指标。同时,采用16S rRNA PCR检测相同位点的栖牙密螺旋体。结果 慢性牙周炎病人栖牙密螺旋体检出率为88.2%,硫化物阳性位点和硫化物阴性位点中栖牙密螺旋体的检出率分别为68.5%和43.2%。硫化物阳性位点与阴性位点中牙周附着丧失（clinical attachment loss, CAL）平均值分别为（2.84±2.33）mm和（1.83±1.60）mm,两者差异有统计学意义（P<0.01）。硫化物阳性位点与阴性位点牙周探诊深度（probing depth, PD）平均值分别为（4.20±1.57）mm和（3.83±1.30）mm,两者差异无统计学意义(P＞0.05)。硫化物阳性位点中牙周探诊出血（bleeding on probing, BOP）阳性检出为率92.5%,大于硫化物阴性位点（75.8%）,两者差异有统计学意义（P<0.01）。结论 慢性牙周炎病人牙周袋内的硫化物水平能反映牙周栖牙密螺旋体分布情况,与牙周附着丧失存在相关性。     

Effects of different blood groups on the reproduction of periodontal pocket bacteria

Background : Many studies have been conducted proving the relation between different blood groups and certain diseases. It was claimed in these studies that H pylori showed different distribution according to different blood groups. In dentistry, the relation between blood groups and dental diseases was investigated in a limited number of studies and it was concluded that there might be a relation between them. The aim of the present study was to determine whether the bacteria isolated from the periodontal pockets of individuals with periodontal diseases indicated differences in CFU amounts to form colonies in different ABO blood groups. Methods : Bacterial samples obtained from the individuals with periodontal diseases from the worst affect sites were inoculated into culture media formed by blood taken from 32 individuals who were systemically and periodontally healthy and who had different blood groups. The colony numbers of these bacteria were observed. Results : Although periodontal pocket bacteria formed colonies in different numbers in different ABO blood groups (p<0.05), no statistically significant difference was determined in the reproduction of these bacteria in different Rh blood groups and different sexes (p>0.05). Conclusion : Different ABO blood groups may show differences in significant rates in the colonisation numbers of the bacteria that are the main cause of periodontal diseases.     

Enzymatic degradation of collagen-guided tissue regeneration membranes by periodontal bacteria

Bacterial infection in the vicinity of guided tissue regeneration barrier membranes was shown to have a negative effect on the clinical outcomes of this increasingly used technique. Several oral and specifically periodontal bacteria were shown to adhere to such membranes in vivo and in vitro with a higher affinity to membranes constructed from collagen. The present study examined the role of periodontal bacteria and their enzymes in the degradation of commercially used collagen membranes. Degradation of two collagen membranes [Biomend (Calcitek, Colla-Tec Inc., Plainsboro, NJ) and Bio-Gide (Geistlich Biomaterials, Wolhousen, Switzerland)] labeled by fluorescein isothiocyanate was examined by measuring soluble fluorescence. Porphyromonas gingivalis, Treponema denticola and Actinobacillus actinomycetemcomitans and their enzymes were evaluated. Collagenase from Clostridium hystolyticum was used as a positive control. While whole cells of P. gingivalis were able to degrade both types of membranes, T. denticola could degrade Bio-Gide membranes only and A. actinomycetemcomitans whole cells could degrade none of the membranes. Fractionation of P. gingivalis cells revealed that cell membrane associated proteases were responsible for the degradation of the two collagen membranes. In T. denticola, the purified major phenylalanine protease was found to be responsible for the degradation of Bio-Gide membranes. These results suggest that proteolytic bacterial enzymes may take part in the degradation of collagen barrier membranes used for guided tissue regeneration.     

慢性牙周炎患者牙周袋内臭氧油存留时间及有效浓度研究

目的 研究慢性牙周炎患者牙周袋内臭氧油有效浓度及存留时间。方法 选取2017年1月至2018年1月廊坊市人民医院口腔科就诊的慢性牙周炎患者200例,将其随机分为A组（84例）与B组（116例）,疗效考察指标为：菌斑指数（PLI）、牙龈指数（GI）、探诊深度（PD）、探诊出血（BOP）和附着丧失指标（AL）;将A组患者随机分为6组,每组14例患者,设对照组、臭氧浓度10 g/L、40 g/L、60 g/L、80 g/L、110 g/L进行单因素试验,对臭氧油浓度与治疗效果进行对比研究;将B组患者随机分为6组,设定臭氧油存留时间30 s、10 min、20 min、30 min、40 min及60 min进行单因素试验,对臭氧油存留时间与治疗效果进行对比研究。结果 与治疗前相比,A组患者通过不同浓度的臭氧油治疗后,各组AL、PD、PLI、GI、BOP指标均显著降低（P<0.05）,且当臭氧油浓度大于60 g/L时,治疗后的PLI、GI、AL均显著低于对照组（P<0.05）;与治疗前相比,B组患者治疗过程中保持臭氧浓度为60 g/L,且在牙周袋内存留时间大于等于30 min时,AL、PD、PLI均显著降低（P<0.05）。结论 慢性牙周炎患者牙周袋内臭氧油有效浓度及存留时间对于患者的病情改善具有重要的意义,值得临床重点关注。     

Stimulatory response of neutrophils from periodontitis patients with periodontal pathogens

OBJECTIVE: Neutrophils play a crucial role in the defense of invading bacteria by releasing biologically active molecules. The response of peripheral blood neutrophils was studied in periodontitis-affected patients and in healthy controls towards stimulation to Porphyromonas gingivalis (Pg) and Actinobacillus actinomycetemcomitans (Aa) extracts. MATERIALS AND METHODS: Peripheral venous blood was drawn from 23 adult patients with moderate to advanced chronic periodontitis (probing depth >or=5 mm, attachment loss >or=3 mm), and 30 healthy volunteers. Neutrophil response followed by metalloproteinase-9 (MMP-9) and interleukin-8 (IL-8) secretion was assayed by zymography and enzyme-linked immunosorbent assay, respectively, on both whole blood and purified neutrophils. In addition to periodontal pathogen extracts, known stimulating agents were tested, such as Escherichia coli-lipopolysaccharide (LPS), phytohemagglutinin, and zymosan A. RESULTS: Neutrophil response, expressed as a secretion ratio under stimulated and non-stimulated conditions, measured in whole blood, showed no differences between periodontitis and healthy controls. Instead, in purified neutrophils from patients, MMP-9 exhibited a significantly higher secretion ratio with LPS and Pg (1.5- to 2-fold), whereas IL-8 showed a larger increase in secretion ratio (3- to 7-fold) in the presence of Pg, Aa, LPS, and zymosan A. CONCLUSION: Peripheral neutrophils of periodontitis-affected patients are more reactive as suggested by their significantly higher response toward periodontal pathogen extracts and other stimulating agents.     

脉冲Nd：YAG激光照射对实验性大鼠慢性牙周炎的作用

目的:探讨脉冲Nd:YAG激光照射对大鼠实验性牙周炎牙周临床指标及牙周组织中一氧化氮(NO)含量的影响.方法:健康Sprague-Dawley大鼠共64只,随机分为8组,正常对照组(N组),药物对照治疗组(D组),牙周炎阳性对照组(P组),其余5组按照给予不同剂量激光照射治疗.末次治疗15 d后测量牙周临床指标牙龈指数(GI),牙槽骨吸收值ABL(Alver bone loss),菌斑指数(plaque index,PLI),采用分光光度仪测定大鼠牙周组织中NO的含量.结果:各剂量组ABL、NO值较末照射组(P组)显著降低(P＜0.05).1.5～2.0 W 20 pps剂量组激光照射后牙周指数均较P组显著降低(P＜0.05),与D组无显著差异.结论:在1.5～2.0 W 20pps剂量范围内,采用脉冲Nd:YAG激光牙周袋内照射能有效改善牙周指数,降低实验大鼠牙周组织中NO含量,起到治疗牙周炎的作用.     

In vitro activity of taurolidine gel on bacteria associated with periodontitis

Objectives

The purpose of this in vitro study was to determine the antimicrobial activity of two different taurolidine gel formulations in comparison with minocycline microspheres.

Methods

Three percent taurolidine gel (TLG3) and 2 % taurolidine gel (TLG2) were compared to minocycline microspheres (MINO) against single bacterial species and a 12-species-mixture. The antimicrobial activity was proven by determination of minimal inhibitory concentrations (MICs), killing assays, after exposure of the antimicrobials as well as within a biofilm.

Results

The MICs against the single species were between 0.5 and 2 mg/ml of taurolidine. MICs of the used mixed microbiota were 1.5 mg/ml (TLG3) and 4 mg/ml (TLG2). Fusobacterium nucleatum and Porphyromonas gingivalis were completely killed by 10 % TLG3 and TLG2 (equivalent to 3 and 2 mg/ml taurolidine) after 6 h. The mixture of 12 species was not completely killed by any of the test substances. Taurolidine gels showed a post-antimicrobial activity, however being less than that of MINO. On biofilms, taurolidine gels reduced concentration dependently the colony forming unit (CFU) counts (multi-species biofilms by 3.63 log10 after 100 % (30 mg/ml) of TLG3), reductions were 2.12 log10 after MINO (1000 μg/ml minocycline).

Conclusions

Taurolidine gel formulations exert antimicrobial activity against bacteria associated with periodontal disease. Nevertheless, a complete elimination of biofilms seems to be impossible and underlines the importance of mechanical removal of biofilms prior to application of the antimicrobial.

Clinical relevance

Taurolidine gels may represent a potential alternative for adjunctive topical antimicrobial treatment in periodontitis and infectious peri-implant diseases.