再治疗根管粪肠球菌生物膜形成与临床表现相关性分析

目的检测粪肠球菌形成生物膜的能力,探讨其生物膜形成能力与临床表现之间的关系。方法采用96孔板法形成生物膜,结合结晶紫染色,检测临床样本中分离的53株粪肠球菌形成生物膜的能力,分析其生物膜形成能力与患牙临床表现之间的关系。结果53株粪肠球菌中,40株（75.47%）具有生物膜形成能力;在患牙的多种临床表现中,瘘道与再治疗根管粪肠球菌生物膜形成具有相关性,结果具有统计学意义（P＜0.05）。结论再治疗根管中,无瘘道的患牙分离出来的粪肠球菌生物膜形成能力强于有瘘道的患牙,临床治疗中应予以注意。     

再感染根管内粪肠球菌生物膜的研究进展

粪肠球菌是顽固性和继发性根管感染中最易分离到的细菌,其主要致病机制之一是形成生物膜.笔者下面就再感染根管内粪肠球菌的分离与鉴定、影响粪肠球菌生物膜形成的相关因素等作一综述.     

粪肠球菌与复发性根尖周炎的相关性及其机制

粪肠球菌可在恶劣的环境中持续生长和繁殖,而在生物膜中具有的极强的生存和致病能力,使其成为根尖周炎复发的重要的致病因素。粪肠球菌的毒力因子,可引起健康组织损伤,增强细菌的黏附能力。粪肠球菌的检出率,在原发性感染根管内为7.5%,但在治疗失败的感染根管中可达到70%以上,即粪肠球菌在原发性根尖周炎患牙的根管内并非主要致病菌。粪肠球菌在根管冠1/3段的感染较重,在根中1/3和根尖1/3段的感染较轻。粪肠球菌对常规的根管消毒和抗菌药物有极强的耐药性,很难用传统的方法将其于根管内彻底清除,因此研发可以有效地抑制或杀灭粪肠球菌药物十分必要。     

根管治疗后疾病相关性粪肠球菌生物膜的形成机制

粪肠球菌生物膜持续向牙根尖释放致病因子,导致根管治疗后相关疾病发生.有效控制粪肠球菌生物膜的形成,是提高根管治疗成功率的关键.本文就根管内粪肠球菌生物膜的结构、生物学特性以及影响其生物膜形成的黏附因子和环境因子等作一综述.     

根管治疗后疾病中粪肠球菌的致病性和检测及清除

粪肠球菌是根管治疗后疾病最常见的细菌,在再感染根管内的检出率为24%~77%。粪肠球菌的致病性与其形成的生物膜高度相关,而其耐药性也与其致病性密不可分,是引发根管内慢性感染的关键。粪肠球菌的检测以细菌培养和聚合酶链反应（PCR）为主,而PCR用于检测感染根管内粪肠球菌较细菌培养更为敏感。根管治疗后疾病中粪肠球菌的清除方法多种多样,结果各不相同。有研究显示抑菌率,质量分数2%的氯己定为100%,10%的盐酸氯丙嗪为88.8%,4%的利多卡因凝胶为76.4%和5%的盐酸阿米洛利为71.4%;有研究则显示,混合物-四环素-异构体-酸-去污剂、QMiX和次氯酸钠皆较2%氯己定更有效。还有研究显示,铒：钇-铝石榴石激光对粪肠球菌生物膜有清除作用。     

根管内粪肠球菌感染的研究进展

粪肠球菌（E.faecalis）在治疗失败根管内的检出率较高,是根管持续感染和再感染的重要微生物之一。粪肠球菌在治疗前后根管内的感染特点不同,并且对抗菌药物有较强耐药性。目前的根管清理和消毒方法难以将定植于根管中的粪肠球菌彻底清除。本文就有关根管内粪肠球菌的感染特点及其对抗菌剂的敏感性研究进展作一综述。     

茶多酚、MTAD和次氯酸钠液抑制粪肠球菌及其生物膜的作用比较

目的:检测中药茶多酚、MTAD、52.5 g/L次氯酸钠抑制根管壁粪肠球菌生物膜的能力,并分析中药茶多酚作为根管冲洗液的可行性。方法:采用微孔板滴定法检测茶多酚,MTAD和52.5 g/L次氯酸钠液对粪肠球菌的最小抑菌浓度(MIC)和最小杀菌浓度(MBC),采用琼脂扩散实验检测3种抗菌剂对粪肠球菌的抑菌圈。通过建立粪肠球菌感染根管3周和6周的模型,分别用茶多酚、MTAD和52.5 g/L次氯酸钠液对粪肠球菌感染后的根管进行处理,然后对根管壁生物膜中的细菌生存情况进行定性定量分析,比较这3种根管冲洗液抗粪肠球菌生物膜的能力。结果:MIC、MBC和琼脂扩散实验显示:3组冲洗剂对粪肠球菌均有抑制作用。在粪肠球菌感染根管3周的生物膜中,MTAD和次氯酸钠液可以完全抑制粪肠球菌生物膜,茶多酚处理后虽仍有粪肠球菌生长,但明显低于生理盐水处理组的细菌生存数,差异有统计学意义(P<0.05)。在粪肠球菌感染根管6周的生物膜中,次氯酸钠液可以完全抑制细菌生长,茶多酚和MTAD作用后仍有活菌生长,但与对照组相比,细菌生存数均显示了8个log值的下降,差异有统计学意义(P<0.05)。结论:52.5 g/L次氯酸钠液具有很强的抑制根管内粪肠球菌生物膜作用,而茶多酚和MTAD也具有一定的抑制粪肠球菌生物膜的能力。     

PIPS联合纳米银对根管粪肠球菌抗菌效果研究

目的 探究激光引发光声流系统(PIPS)和纳米银(AgNPs)联合使用对根管内粪肠球菌生物膜的抗菌效果。方法 收集36颗单根管离体牙,建立粪肠球菌感染根管实验模型,将样本随机分成6组,采用0.9%NaCl、2%NaClO、0.1%AgNPs溶液分别联合传统手动冲洗(CNI)或PIPS对根管进行冲洗,使用菌落计数法测定治疗前后根管内粪肠球菌生物膜菌落数,并计算菌落计数减少的百分比。结果 所有实验组粪肠球菌生物膜的抑制效果均强于对照组(P<0.05),使用PIPS辅助0.9%NaCl、2%NaClO、0.1%AgNPs冲洗组的降幅均分别大于CNI辅助0.9%NaCl、2%NaClO、0.1%AgNPs冲洗组(P<0.05)。PIPS辅助0.1%AgNPs冲洗组的降幅明显大于PIPS辅助2%NaClO冲洗组(P<0.05)。结论 PIPS辅助AgNPs溶液冲洗可以显著提高根管内粪肠球菌生物膜的清除效果。     

再治疗根管内粪肠球菌检出及其与临床表征关系的分析研究

目的检测临床再治疗根管内的粪肠球菌,分析粪肠球菌检出率与临床症状及体征之间的关系。方法临床收集需根管再治疗的患牙108颗,记录症状和体征,根管内采集细菌样本,提取细菌基因组DNA,用聚合酶链反应定性检测粪肠球菌。结果根管内粪肠球菌的检出率为47.2%。在有症状病例、有体征病例、既有症状又有体征病例中,根管内粪肠球菌的检出率分别为52.6%、57.9%、62.5%,其中,有体征病例与无体征病例粪肠球菌的检验率差异有统计学意义（P＜0.05）,既有症状又有体征病例与无体征病例组粪肠球菌的检出率差异有统计学意义（P＜0.05）;在有症状的病例组中,有咬合痛的病例粪肠球菌的检出率为66.7%,与无咬合痛病例组相比差异有统计学意义（P＜0.05）。结论再治疗根管内粪肠球菌的存在与临床症状或体征密切相关。     

慢性根尖周炎感染根管内粪肠球菌和白色念珠菌的检测

目的检测未经治疗和根管再治疗的慢性根尖周炎感染根管内粪肠球菌和白色念珠菌,探讨粪肠球菌和白色念珠菌与慢性根尖周炎发病的关系。方法选取未经根管治疗和需根管再治疗的慢性根尖周炎病例各30例,采集根管内细菌样本,提取DNA,分别设计粪肠球菌和白色念珠菌的引物进行PCR检测。结果所有根管内均可检测到细菌。粪肠球菌在慢性根尖周炎根管再治疗病例中检出率为53.3%(16/30),慢性根尖周炎未经根管治疗病例的根管为10%(3/30)。白色念珠菌仅检出2例,均在慢性根尖周炎根管再治疗病例中检出(6.67%),而未经根管治疗病例的检出率为0。结论粪肠球菌和白色念珠菌在原发的慢性根尖周炎感染根管中检出率较低,在再治疗根管中检出率增高,可能与根管治疗时再感染有关。     

A preliminary study investigating the survival of tetracycline resistant Enterococcus faecalis after root canal irrigation with high concentrations of tetracycline

AIM: To compare the ability of two Enterococcus faecalis strains to survive exposure to an irrigation solution containing a high concentration of tetracycline in the root canals of bovine teeth. METHODOLOGY: The root canals of twelve bovine incisor root sections were chemo-mechanically prepared using commercially available drills, sodium hypochlorite and ethylenediamine tetra-acetic acid. The root sections were divided into two groups and inoculated with either a tetracycline sensitive or resistant strain of E. faecalis. The strains are isogenic, however one contains a conjugative transposon related to Tn916 which confers resistance to tetracycline, and the other strain is sensitive to the antibiotic. After 26 days of incubation the root canals were irrigated using one of three solutions (sterile distilled water, 50% ethanol or tetracycline at a concentration of 30 mg mL(-1)). The roots were sampled by grinding dentine and canal contents and the debris collected were incubated in broth to assess growth. RESULTS: Irrigation with sterile distilled water or 50% ethanol did not remove all of the cells present. The tetracycline containing solution was efficient in preventing any growth of sensitive E. faecalis, however the resistant strain was able to survive a 5 min exposure at 30 mg mL(-1). CONCLUSIONS: The presence of the Tn916-like conjugative transposon containing the tetracycline resistance gene tet(M) allowed an E. faecalis strain to survive irrigation using a solution containing an extremely high concentration of tetracycline in a root canal model.     

Isolation of Enterococcus faecalis in previously root-filled canals in a Lithuanian population

The occurrence of Enterococcus faecalis in root canals of previously root filled teeth with apical periodontitis requiring retreatment was studied in Lithuanian patients. Twenty-five asymptomatic teeth were included in the study. Avoiding contamination microbiological samples were taken from the canals before and after preparation and irrigation with sodium hypochlorite and EDTA. Microbes were isolated from 20 of 25 teeth. E. faecalis was isolated from 14 of those 20 culture positive teeth, usually in pure culture or as a major component of the flora. Second samples taken after preparation revealed growth in 7 of the 20 teeth. Five of the seven cases were E. faecalis in pure culture. Isolation of E. faecalis was not related to the use of any particular root filling material in the original root filling. The results indicate that, rather than previous chemical treatment, it is the ecological conditions present in the incompletely filled root canal that are important for the presence of E. faecalis in these teeth.     

Biofilm formation in medicated root canals

The hypothesis that Enterococcus faecalis resists common intracanal medications by forming biofilms was tested. E. faecalis colonization of 46 extracted, medicated roots was observed with scanning electron microscopy (SEM) and scanning confocal laser microscopy. SEM detected colonization of root canals medicated with calcium hydroxide points and the positive control within 2 days. SEM detected biofilms in canals medicated with calcium hydroxide paste in an average of 77 days. Scanning confocal laser microscopy analysis of two calcium hydroxide paste medicated roots showed viable colonies forming in a root canal infected for 86 days, whereas in a canal infected for 160 days, a mushroom-shape typical of a biofilm was observed. Analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis showed no differences between the protein profiles of bacteria in free-floating (planktonic) and inoculum cultures. Analysis of biofilm bacteria was inconclusive. These observations support potential E. faecalis biofilm formation in vivo in medicated root canals.     

Bacteria isolated after unsuccessful endodontic treatment in a North American population

OBJECTIVE: The purpose of this study was to determine the composition of the microbial flora present in teeth after the failure of root canal therapy in a North American population. These results were then compared with those of the previous Scandinavian studies. STUDY DESIGN: Fifty-four root-filled teeth with persistent periapical radiolucencies were selected for retreatment. After removal of the root-filling material, the canals were sampled with paper points, and by reaming of the apical dentin. Both samples were grown under aerobic and strict anaerobic conditions. Then the bacterial growth was analyzed. RESULTS: The microbial flora was mainly of 1 to 2 strains of predominantly gram-positive organisms. Enterococcus faecalis was the most commonly recovered bacterial species. CONCLUSIONS: Bacteria were cultivated in 34 of the 54 teeth examined in the study. E faecalis was identified in 30% of the teeth with a positive culture.     

利用粪肠球菌感染根管模型评价不同根管封闭剂的抗菌效果

目的 体外建立粪肠球菌根管感染模型,比较3种生物陶瓷类根管封闭剂的抗菌性能.方法 选择人单直根管的离体前牙48颗,接种粪肠球菌并孵育4周以构建体外根管感染模型.完成根管成形和清理后,随机将样本分为3个实验组和2个对照组,每组按照如下方法进行根管充填:A组,Biodentine+牙胶;B组,iRoot BP+牙胶;C组,...     

氯化镧对根管内粪肠球菌抗菌效果的体外实验

目的:研究氯化镧、FC、CP、氢氧化钙丙二醇糊剂4种根管消毒剂对离体牙根管内粪肠球菌的抗菌性能。方法:选取72个新鲜拔除的单根管前磨牙,随机分为4个实验组(氯化镧组、FC组、CP组、氢氧化钙丙二醇糊剂组)、1个阴性对照组和1个阳性对照组,每组12个牙。所有牙清理根管后灭菌,除阴性对照组不感染细菌外,其余各组均建立粪肠球菌感染根管模型。建模后,4个实验组分别在根管内放置氯化镧药液、FC、CP、氢氧化钙丙二醇糊剂;阴性和阳性对照组放置生理盐水,37℃、50 mL/L CO2培养箱内培养。分别于培养后3 d和7 d时,取各组根管内壁牙本质粉末用BHI培养基继续培养72 h,比浊仪检测各样本的浊度。结果:①药物处理第3天时,氯化镧组牙本质小管中残留菌量较阳性对照组少,但高于阴性对照组和FC组、氢氧化钙丙二醇糊剂组(P<0.05),与CP组无显著性差异(P>0.05);②药物处理第7天时,FC组、CP组、氢氧化钙丙二醇糊剂组和氯化镧组4种消毒剂牙本质小管内残留细菌量较阳性对照组有明显减少(P<0.05),与阴性对照组相比,差异均无统计学意义(P>0.05)。结论:根管内放置氯化镧药液7 d可有效抑制粪肠球菌生长。     

Apical periodontitis development and bacterial response to endodontic treatment. Experimental root canal infections in monkeys with selected bacterial strains

In six monkeys, 160 root canals were inoculated with a combination of four bacterial strains belonging to species Streptococcus milleri, Peptostreptococcus anaerobius, Prevotella oralis, and Fusobacterium nucleatum. In two other monkeys, 24 root canals were inoculated with a five-strain combination consisting of these strains and a strain of Enterococcus faecalis. All strains were previously isolated from an infected monkey root canal. After 8-12 months, survival of the strains was recorded bacteriologically, and the reaction in the periapical region was radiographed. From 180 of 184 root canals, one or more of the bacterial strains were reisolated. The two facultative strains were more frequently reisolated than the anaerobic strains. Apical periodontitis was registered in the periapical region of more than 96% of root canals with reisolated bacteria but in none of those without reisolated bacteria. Endodontic treatment was carried out in two sessions with an interval of 14 d without interappointment dressings, and the effect was evaluated bacteriologically before and after each treatment. The chemo-mechanical treatment reduced significantly the number of strains and bacterial cells. The facultative bacteria were more resistant to the treatment than the anaerobic bacteria. The five-strain combination had a higher survival rate than the four-strain combination.     

Recovery of Enterococcus faecalis after single- or multiple-visit root canal treatments carried out in infected teeth ex vivo

AIM: To assess the presence of Enterococcus faecalis after root canal treatment in single or multiple visits in an ex vivo model. METHODOLOGY: Forty-five premolar teeth were infected ex vivo with E. faecalis for 60 days. The canals were then prepared using a crowndown technique with System GT and Gates-Glidden burs and irrigated with 2% chlorhexidine gel. The specimens were divided into five groups (G1, G2, G3, G4 and G5) according to the time elapsed between chemical-mechanical preparation and root canal filling, the irrigant solution used and the use or nonuse of a calcium hydroxide intra-canal medicament. The teeth were then root-filled and incubated for 60 days at 37 degrees C. Dentine chips were removed from the canal walls with sequential sterile round burs at low speed. The samples obtained with each bur were immediately collected in separate test tubes containing Brain-Heart Infusion broth. These samples were placed onto agar plates and colony forming units were counted after 24 h at 37 degrees C. Data were ranked and analysed using the Kruskal-Wallis statistical test. RESULTS: Enterococcus faecalis was recovered from 20% (three of 15 specimens) of G1 (chlorhexidine irrigation and immediate root filling in a single visit), 25% (four of 15 specimens) of G2 (chlorhexidine irrigation and filling after 14 days use of a calcium hydroxide dressing in multiple visits), 40% (two of five specimens) of G3 (chlorhexidine irrigation and filling after 7 days), 60% (three of five specimens) of G4 (saline irrigation and filling after 7 days) and from 100% (five of five specimens) of G5 (saline irrigation and immediate filling without sealer). CONCLUSIONS: Neither single- nor multiple-visit root canal treatment ex vivo, eliminated E. faecalis completely from dentinal tubules. Up to 60 days after root filling, E. faecalis remained viable inside dentinal tubules. When no sealer was used, E. faecalis presented a higher growth rate.     

Phenotypic and genotypic identification of enterococci isolated from canals of root-filled teeth with periapical lesions

The objectives of the present study were to identify enterococcal species isolated from the canals of root-filled teeth with periapical lesions using biochemical and molecular techniques, and to investigate the genetic diversity of the isolates. Twenty-two Enterococcus strains, isolated from the canals of root-filled teeth with persisting periapical lesions, were identified to species level using rapid ID 32 STREP galleries and partial 16S rDNA sequencing. To subtype the strains, genomic DNA from the isolates was analyzed by pulsed field gel electrophoresis (PFGE) after digestion with SmaI. Intragenic regions of two genes, ace and salA, were sequenced for further differentiation of the isolates. All strains were identified as Enterococcus faecalis by both commercial kit and partial 16S rDNA sequencing. PFGE with SmaI of 22 isolates demonstrated 18 macrorestriction profiles, whereas 13 distinct genotypes were identified after analysis of the ace and salA composite sequences. Most of the isolates from distinct patients had different PFGE profiles. Moreover, in two cases, different E. faecalis strains were found in different root-filled teeth from the same mouth. E. faecalis was the only enterococcal species isolated from the canals of root-filled teeth with periapical lesions. Genetic heterogeneity was observed among the E. faecalis isolates following PFGE and sequence-based typing method. Furthermore, the genetic diversity within root canal strains was similar to previous reports regarding E. faecalis isolates from different clinical and geographic origins.