Microorganisms from canals of root-filled teeth with periapical lesions

AIM: The objective of the present study was to identify the microbial flora within root canals of teeth with failed root-canal treatment and to determine the association of the various species with clinical features. METHODOLOGY: Sixty root-filled teeth with persisting periapical lesions were selected for this study. During nonsurgical endodontic re-treatment, the root-filling material was removed and the canals were sampled. Microbial sampling, isolation and species determination were performed using advanced microbiological techniques for anaerobic species. The association of microbiological findings with clinical features was investigated. RESULTS: Microorganisms were recovered from 51 teeth. In most cases, one or two strains per canal were found. Of the microbial species isolated, 57.4% were facultative anaerobic species and 83.3% Gram-positive microorganisms. Enterococcus faecalis was the most frequently recovered bacterial species. Obligate anaerobes accounted for 42.6% of the species and the most frequently isolated genera was Peptostreptococcus. which was associated with clinical symptoms (P < 0.01). Significant associations were also observed between: (a) pain or history of pain and polymicrobial infections or anaerobes (P < 0.05): (b) tenderness to percussion and Prevotella intermedia/P. nigrescens (P < 0.05); (c) sinus and Streptococcus spp. (P < 0.001) or Actinomyces spp. (P < 0.01); (d) coronally unsealed teeth and Streptococcus spp. or Candida spp. (both with P < 0.01). CONCLUSION: The microbial flora in canals after failure of root-canal treatment were limited to a small number of predominantly Gram-positive microbial species. Facultative anaerobes, especially E. faecalis, were the most commonly isolated microorganisms, however, polymicrobial infections and obligate anaerobes were frequently found in canals of symptomatic root-filled teeth.     

Evaluation of root canal microorganisms isolated from teeth with endodontic failure and their antimicrobial susceptibility

Studies of the microbiota from the canals of teeth with failure of endodontic therapy have revealed that it differs markedly from that of untreated necrotic dental pulps. This study aimed to evaluate the microbiota of 30 root-filled teeth with persisting periapical lesions and to test the antibiotic susceptibility of the most prevalent species. Microbial samples, isolation and speciation were done using advanced microbiologic techniques for anaerobic species. A total of 55 bacterial species were isolated, 80% were gram-positives and 58% facultative anaerobic microorganisms. The bacterial genera most frequently recovered were Enterococcus, Streptococcus, Peptostreptococcus and Actinomyces. Antibiotic sensitivity of Enterococcus faecalis and Peptostreptococcus spp. was accomplished with the E-test system. All species studied were susceptible to benzylpenicillin, amoxicillin, amoxicillin combined with clavulanate. However, 20% of the E.faecalis strains were resistant to erythromycin and 60% to azithromycin. It was concluded that microbial flora in canals after endodontic failure comprised predominantly facultative anaerobes and gram-positive species. E.faecalis was the species most frequently isolated and showed erythromycin and azithromycin resistance among the isolates.     

根管治疗失败病例根管内微生物的分子生物学检测

目的:通过对根管治疗失败病例根管内微生物进行分子生物学检测,研究失败病例根管内各种细菌的检出率及优势菌;分析临床症状和体征与特殊微生物的关系.方法:选择40 例根管治疗失败病例共40 颗患牙,根据症状分为疼痛组、窦道组和无症状组,去除根管内充填物,根管内细菌取样,PCR检测鉴定.结果: 根管治疗失败病例根管主要呈混合感染,共检出6 种细菌,粪肠球菌是最常检出的细菌,产黑普氏菌与疼痛、衣氏放线菌与瘘管有一定的相关性.结论:根管治疗失败的主要原因是根管内的微生物感染持续存在,根管治疗失败病例根管内微生物组成有其特殊性.     

Circulating antibodies after experimental chronic infection in the root canal of teeth in monkeys

Abstract – Root canals of 49 teeth with necrotic pulp tissue in five monkeys were infected with Streptococcus faecalis ss liquefaciens, Actinornyces bovis, Fusobacteriam nucleatum, Peptostreptococcus anaerobiuS; and Bacteroidcs oralis in various combinations. After 6 months the root canals and the periapical tissues were subjected to radiographic., foacteriologic, and histologic examinations. Signs of periapical inflammation were radiographically registered in 41 teeth, most frequently in teeth inoculated with a mixed flora. In 11 out of 16 teeth infected separately with Strep, faecalis ss iiquefaciens, apical periodontitis was observed. Sera from the monkeys prior to and after the experimental inoculation were analyzed by means of gel diffusion, hemagghitination (HA), and complement binding test for antibodies against different antigen preparations of the homologous bacterial strains used for the inoculation. Detectable antibodies were seen with antigens of B. oralis in all five monkeys. Agglutinating antibodies were demonstrated with lipopolysaccharide-antigen of B. oralis in titers between 1:40 and 1:320. A marked reduction of the antibody level against lipopolysaccharide (LPS) antigen after mercaptoethanol treatment indicated that a main part of the antibodies registered was of IgM-class. The study shows that certain antigens of bacterial origin from infected root canals, while affecting the periapical tissues, also stimulate the production of circulating antibodies.     

Bacterial coronal leakage after obturation with three root canal sealers

The purpose of this study was to compare the bacterial leakage of root canals obturated with three root canal sealers, using Endodontalis faecalis as a microbial tracer to determine the length of time for bacteria to penetrate through the obturated root canal to the root apex. Seventy-five, single-rooted teeth with straight root canals had the crown cut off at the cementoenamel junction. Root canals were instrumented by a step-back technique. The prepared teeth were randomly divided into 3 groups of 19 teeth each and another 2 groups as positive and negative controls (9 teeth each). The experimental groups were dependent on the sealer used: AH-Plus, Apexit, and Ketac-Endo. The root canals were obturated using a lateral condensation technique. After 24 h the teeth were attached to microcentrifuge tubes with 2 mm of the root apex submerged in Brain Heart Infusion broth in glass test tubes. The coronal portions of the root canal filling materials were placed in contact with E. faecalis. The teeth were observed for bacterial leakage daily for 30 and 60 days. With the chi2 test for comparing pairs of groups at the 0.05 level (p < 0.05), there was no statistical difference between Ketac-Endo and AH-Plus (p > 0.06), but Apexit had significantly higher leakage (p < 0.05) at 30 days. After 60 days there was no statistical difference between Ketac-Endo and Apexit (p > 0.05), but Apexit leaked more than AH-Plus. The conclusion drawn from this experiment was that epoxy resin root canal sealer was found to be more adaptable to the root canal wall and filling material than a calcium hydroxide sealer when bacterial coronal leakage was studied.     

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.     

根管治疗失败病例根管内微生物检测及药敏试验

目的通过对根管治疗失败病例根管内微生物进行培养检测，了解失败病例根管内菌群特征，同时对检测出的细菌进行药敏试验，为临床根管治疗提供参考。方法选择86例根管治疗失败病例共86颗惠牙，去除根管内充填物后，对根管内细菌取样，进行培养和鉴定，用琼脂稀释法对根管内分离的优势菌进行药敏试验。结果根管治疗失败病例根管中微生物菌群组成以兼性厌氧菌为优势菌，主要以1-2种革兰阳性菌为主，肠球菌是最常检出的细菌。失败病例根管内检出的兼性及专性厌氧菌对青霉素G、甲醛甲酚、甲硝唑敏感。结论细菌与根管治疗失败密切相关，根管治疗失败病例根管内微生物组成有其独特性。临床应选择敏感性药物进行治疗。     

An in vitro comparison of bacterial leakage of three common restorative materials used as an intracoronal barrier

The purpose of this in vitro study was to evaluate quantitatively the effectiveness of three different restorative materials used as an intracoronal barrier to prevent microleakage of endodontically treated teeth. Fifty-five extracted human single-canal teeth were used in this study. The teeth were endodontically prepared and obturated. Forty-five teeth were randomly assigned to three experimental groups: group 1: sealed with Ketac-Cem (3M ESPE, St Paul, MN) (n = 15), group 2: sealed with Clearfil Protect Bond/Clearfil AP-X (Kuraray, New York, NY) (n = 15), and group 3: sealed with Maxcem (Kerr, Orange, CA) (n = 15). Ten teeth were also randomly assigned to a negative control group (n = 5) and a positive control group (n = 5). Microleakage was tested by using a sterile two-chamber bacterial method and Enterococcus faecalis was used as a microbial marker. Samples were incubated aerobically at 37 degrees C for 120 days. Bacterial leakage was determined by change in turbidity in the medium. Statistical analysis was performed using a Wald chi-square test. No significant difference (p > 0.05) in bacterial leakage was found between the three experimental groups tested. All positive controls leaked within 60 days and broth of the negative control group remained clear throughout the entire experimental period.     

Isolation of yeasts and enteric bacteria in root-filled teeth with chronic apical periodontitis

AIMS: The aim of this study was to determine the occurrence and role of yeasts, enteric gram-negative rods and Enterococcus species in root-filled teeth with chronic apical periodontitis, and the antimicrobial effect of iodine potassium iodide (IKI) irrigation. METHODOLOGY: Forty symptom-free root-filled teeth with chronic apical periodontitis were included in the study. The patients were divided into two groups. In group A the canals were filled with calcium hydroxide for 10-14 days after cleaning and shaping; in group B the canals were irrigated with IKI for 5 min after cleaning and shaping followed by a permanent root filling. Microbiological samples were taken from the canals before and after the chemomechanical preparation and after iodine irrigation (group B). RESULTS: Microbes were isolated from 33 of 40 teeth in the initial sampling. Yeasts were isolated from six teeth, three of them together with E. faecalis. Enteric rods (Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis) were present in three teeth and E. faecalis was isolated from 21 of the 33 culture positive teeth, 11 in pure culture. Growth was detected in 10 teeth of the second samples. Six of the 10 cases were E. faecalis, with five being a pure culture. All third samples (after IKI) except one were negative. The number of microbial cells per sample did not correlate with lesion size. Two flare-ups were recorded, both in teeth with a mixed infection. CONCLUSION: The high prevalence of enteric bacteria and yeasts in root-filled teeth with chronic apical periodontitis was established. IKI improved the antimicrobial effect of the treatment.     

Microbial analysis of canals of root-filled teeth with periapical lesions using polymerase chain reaction

The objective of the present study was to investigate the presence of nine bacterial species in root-filled teeth associated with periapical lesions using a polymerase chain reaction analysis and to correlate these species with clinical features of the cases. DNA was extracted from 45 canal samples of root-filled teeth with periapical lesions. A PCR assay using species-specific primers of 16S rDNA and the downstream intergenic spacer region was used for microbial detection. Enterococcus faecalis was the most prevalent species, detected in 77.8% of the study teeth, followed by Peptostreptococcus micros, detected in 51.1%. Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia, and Prevotella nigrescens were detected in 35.6%, 22.2%, 11.1%, and 11.1% of the sampled teeth, respectively. Moreover, PCR detected Filifactor alocis in 26.7%, Treponema denticola in 24.4%, and Tannerella forsythia in 4.4% of the samples. T. denticola and P. micros were statistically associated with tenderness to percussion (p < 0.05). P. nigrescens was associated with the presence of spontaneous pain and abscess (p < 0.05). P. endodontalis and P. nigrescens were associated with purulent exudates (p < 0.05). Synergistic relationship was also observed between some species. The results of this study indicated that E. faecalis was the most frequently identified test species by PCR in teeth with failing endodontic treatment.     

Identification of Enterococcus faecalis in root-filled teeth with or without periradicular lesions by culture-dependent and-independent approaches

Enterococcus faecalis is the most commonly found species in root-filled teeth evincing recalcitrant periradicular lesions and as a consequence, a role in causation of endodontic treatment failure has been suggested. The purpose of this study was to evaluate the prevalence of this bacterial species in root-filled teeth with or without periradicular lesions. Identification of E. faecalis was carried out by polymerase chain reaction (PCR) or conventional culture procedures. Overall, E. faecalis was detected by species-specific 16S rRNA gene-based PCR in 40/50 teeth (80%), while culture revealed the occurrence of this species in 8/50 teeth (16%). PCR was significantly more effective than culture in detecting this bacterial species (p < 0.001). Of 27 root-filled teeth with no periradicular lesions, E. faecalis was found in 22 cases (81.5%) by PCR and in five cases (18.5%) by culture. Of 23 root-filled teeth with periradicular lesions, E. faecalis was identified in 18 cases (78%) by PCR and in three cases (13%) by culture. Regardless of the identification technique used, no significant difference was observed when comparing the occurrence of E. faecalis in root-filled teeth with and without periradicular lesions (p > 0.05). Although these findings apparently put into question the status of E. faecalis as the main species causing endodontic treatment failure, other related factors still need to be clarified before this assumption turns into certainty.     

Microbiological analysis of infected root canals from symptomatic and asymptomatic teeth with periapical periodontitis and the antimicrobial susceptibility of some isolated anaerobic bacteria

The purpose of the present study was to investigate the correlation between the composition of the bacterial flora isolated from infected root canals of teeth with apical periodontitis with the presence of clinical signs and symptoms, and to test the antibiotic susceptibility of five anaerobic bacteria mostly commonly found in the root canals of symptomatic teeth against various substances using the E-test. Microbial samples were taken from 48 root canals, 29 symptomatic and 19 asymptomatic, using adequate techniques. A total of 218 cultivable isolates were recovered from 48 different microbial species and 19 different genera. Root canals from symptomatic teeth harbored more obligate anaerobes and a bigger number of bacterial species than the asymptomatic teeth. More than 70% of the bacterial isolates were strict anaerobes. Statistical analysis used a Pearson Chi-squared test or a one-sided Fisher's Exact test as appropriate. Suggested relationships were found between specific microorganisms, especially gram-negative anaerobes, and the presence of spontaneous or previous pain, tenderness to percussion, pain on palpation and swelling amoxicillin, amoxicillin + clavulanate and cephaclor were effective against all the strains tested. The lowest susceptibility rate was presented by Prevotella intermedia/nigrescens against Penicillin G. Our results suggested that specific bacteria are associated with endodontic symptoms of infected teeth with periapical periodontitis and the majority of the anaerobic bacterial species tested were susceptible to all antibiotics studied.     

Microbiologic analysis of teeth with failed endodontic treatment and the outcome of conservative re-treatment

Objective. The purposes of this study were to determine what microbial flora were present in teeth after failed root canal therapy and to establish the outcome of conservative re-treatment.Study design. Fifty-four root-filled teeth with persisting periapical lesions were selected for re-treatment. After removal of the root filling, canals were sampled by means of advanced microbiologic techniques. The teeth were then re-treated and followed for up to 5 years.Results. The microbial flora was mainly single species of predominantly gram-positive organisms. The isolates most commonly recovered were bacteria of the species Enterococcus faecalis. The overall success rate of re-treatment was 74%.Conclusions. The microbial flora in canals after failed endodontic therapy differed markedly from the flora in untreated teeth. Infection at the time of root filling and size of the periapical lesion were factors that had a negative influence on the prognosis. Three of four endodontic failures were successfully managed by re-treatment.     

Microbial leakage of Enterococcus faecalis after post space preparation in teeth filled in vivo with RealSeal versus Gutta-percha

The purpose of this study was to compare microbial leakage of Enterococcus faecalis after post space preparation in teeth filled in vivo with RealSeal (SybronEndo, Orange, CA) versus gutta percha. Twenty-six premolars in which extraction was indicated for orthodontic reasons were endodontically treated before extraction to simulate normal clinical conditions. Ten of these bicuspids were obturated following the RealSeal protocol; the other 10 were filled with gutta percha and Grossman-based formula cement. Post space was prepared immediately after filling leaving 5 mm of apical obturation material. The rest of the teeth were equally divided into the positive and negative control groups. Forty-eight hours later teeth were extracted and submitted to a dual-chamber leakage model using E faecalis. All of the RealSeal-filled teeth showed leakage during the first week, having a mean leakage of 3.5 days (+/-2.32 days). Gutta-percha-filled teeth showed a mean leakage of 10 days (+/-11.53 days). Mann-Whitney U comparison between groups revealed a p = 0.09. Under the conditions of the present study, there is no statistically significant difference in the microleakage of teeth filled with RealSeal compared with gutta percha when post space is prepared.     

A microbiological profile of symptomatic teeth with primary endodontic infections

The aim of this study was to evaluate the composition of the microbiota of primary endodontic infections associated with symptomatic teeth. Samples were collected by means of a #15 H-type file and 2 sterile paper points from 60 symptomatic (n = 30) or asymptomatic (n = 30) single-rooted teeth with necrotic pulp. The presence of 40 bacterial species was determined by the checkerboard DNA-DNA hybridization method. The species found in higher counts (x10(5)) in symptomatic cases were Fusobacterium nucleatum ssp. vincentii, Veillonella parvula, Treponema socranskii, Enterococcus faecalis, and Campylobacter gracilis and in asymptomatic cases were F. nucleatum ssp. vincentii, Fusobacterium nucleatum ssp. nucleatum, E. faecalis, Eubacterium saburreum, and Neisseria mucosa. Total bacterial counts and counts of Tannerella forsythia were significant higher in symptomatic cases (p < 0.05), whereas levels of Propionibacterium acnes were reduced in this group of teeth. The data of the present investigation suggested an association between higher total bacterial counts and levels of T. forsythia and the presence of pain.     

Detection of bacteria in endodontic samples by polymerase chain reaction assays and association with defined clinical signs in Italian patients

BACKGROUND/AIMS: The presence of selected bacteria (Enterococcus faecalis, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, Treponema denticola) in infected root canals was studied using polymerase chain reaction (PCR) assays, and the association of bacteria with clinical signs of endodontic disease was assessed. The null hypothesis, that no difference could be observed between clinical signs of apical periodontitis and a specific bacterial strain, was tested. METHODS: Microbial samples were obtained from 62 teeth in 54 patients with endodontic disease. For each tooth, clinical data including patient symptoms were collected. Teeth were categorized by diagnosis as having acute apical periodontitis (AAP, teeth with clinical symptoms but no periapical radiolucency, n=22), chronic apical periodontitis (CAP, teeth with radiolucency but no clinical symptoms, n=15) or exacerbated apical periodontitis (EAP, teeth with symptoms and radiolucency, n=25). Seventy-one percent of cases were primary endodontic infections, and 29% were recurrent ('secondary') endodontic infections (failing cases). PCR assays were used to detect the presence of the selected bacteria. RESULTS: T. denticola and E. faecalis were each detected in 15 of 62 samples (24%), P. gingivalis in 8 samples (13%), P. intermedia in 5 samples (8%), and T. forsythensis in 4 samples (7%). T. denticola was detected in 56% of teeth with EAP. E. faecalis was found in 60% of teeth with CAP and in 72% of teeth with secondary infection. Statistical analysis demonstrated an association of CAP and secondary endodontic infection with the presence of E. faecalis. (P<0.01). EAP was associated with the presence of T. denticola (P<0.01). CONCLUSION: T. denticola was associated with symptomatic endodontic disease in the presence of apical bone resorption. E. faecalis was associated with treatment failures. We suggest that these species may play critical roles in endodontic pathology.     

Microbiota of periapical lesions refractory to endodontic therapy

The periapical microbiota of 36 teeth with refractory apical periodontitis was investigated. None of the teeth had responded to conventional endodontic or long-term (> 6 months), calcium-hydroxide treatment. Eight patients had received antibiotics systemically. After anaerobic culture, a total of 148 microbial strains were detected among 67 microbial species. One of the 36 lesions was culture-negative. Approximately half (51.0%) of the bacterial strains were anaerobic. Gram-positive species constituted 79.5% of the flora. Facultative organisms, such as Staphylococcus, Enterococcus, Enterobacter, Pseudomonas, Stenotrophomonas, Sphingomonas, Bacillus, or Candida species were recovered from 27 of the lesions (75%). Sulfur granules were found in 9 lesions (25%). In these granules Actinomyces israelii, A. viscosus, A. naeslundii, and A. meyeri were identified. Other bacterial species, both gram-positive and gram-negative, were detected in the granules as well. Two sulfur granules did not contain Actinomyces. Scanning electron microscopy demonstrated rod- and spirochete-like cells in the granules, and transmission electron microscopy revealed organisms with copious amounts of extracellular material. Outer membrane vesicles were also seen. Some of the granules were calcified. This study demonstrated a wide variety of microorganisms, particularly gram-positive ones, in the periapical lesions of teeth with refractory apical periodontitis.     

Polymerase chain reaction identification of microorganisms in previously root-filled teeth in a South Korean population

A large body of evidence indicates that microorganisms are the primary causative agents of endodontic treatment failures. This study intended to assess the occurrence of nine putative endodontic pathogens in root-filled teeth associated with periradicular lesions in a South Korean population using a culture-independent molecular approach. Fourteen root-filled teeth with persistent periradicular diseases were selected for retreatment. After removal of the root canal filling, the canals were sampled, and a polymerase chain reaction assay using taxon-specific oligonucleotide primers was used for microbial detection. Bacteria were present in all cases, as revealed by amplification using ubiquitous 16S rDNA primers. The most frequently detected taxon was Enterococcus faecalis (64%), followed by Streptococcus spp. (21%) and Tannerella forsythensis (14%). The results of this study using a highly sensitive identification method are concurrent with those from other geographical locations using diverse identification methods in that E. faecalis is the main species found in cases of root-filled teeth associated with periradicular lesions.     

Antibiotic resistance and production of inhibitory substances among bacterial strains isolated from the oral cavities of BALB/c mice

In the oral cavities of BALB/c mice, microbial population levels are regulated by multifactorial processes. Factors include the production of inhibitory substances and the exchange of genetic material. In this work, 371 isolates from different sites (saliva, tongue, teeth, and mucosa) of the oral cavities of BALB/c mice were screened for resistance to antibiotics and antimicrobial activity. Antibiotic-resistant strains represented 25% of the total flora. Among the predominant species, all the S. faecalis isolates showed multiresistance, and 23% of the Lactobacillus murinus isolates and 15% of the Staphylococcus aureus were resistant to at least one antibiotic. Resistance to aminoglycosides (neomycin, streptomycin, kanamycin, and gentamicin) was most frequently encountered. In S. faecalis, high levels of resistance were recorded to neomycin and streptomycin but not to gentamicin or kanamycin. Macrolides (M), lincosamides (L), streptogramin B (S), tetracycline (Tc), and chloramphenicol (Cm) resistance was also present in multiresistance patterns, especially among S. faecalis isolates. Hemolytic (Hly+) streptococci were less resistant to MLS, Tc, and Cm than were non-hemolytic (Hly-) isolates. Resistance to beta-lactam antibiotics was detected only among staphylococci and with a low prevalence (4%). The frequencies of strains producing antimicrobial substances against the indicator strains (S. mutans LG-1, S. sanguis Ny 101, and A. viscosus Ny 1) were high for L. murinus (76%) and S. faecalis (57% for Hly- and 90% for Hly+), but low for S. aureus (7%). These results indicate that the indigenous oral flora could interfere with colonization by allochthonous micro-organisms and that resistance patterns should be taken into account for the elimination of the oral indigenous flora by antibiotic treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Resistance of experimental glass ionomer cement sealers to bacterial penetration in vitro

This study evaluated penetration of Enterococcus faecalis in canals filled with glass ionomer cement sealers. Canals of 40 extracted teeth in four equal experimental groups were prepared and filled with KT-308 (experimental sealer) and a single gutta-percha cone (SC), ZUT (KT-308 with an antibacterial agent) and SC, Kerr sealer and vertically compacted gutta-percha (VC), or Kerr sealer with the antibacterial agent and VC. Eight additional teeth served as positive controls and four as negative controls. Pulp chambers were inoculated with E. faecalis, and bacterial penetration through the filled canals was assessed over a period of 90 days. Incidence of bacterial penetration was significantly higher for ZUT/SC than for Kerr sealer/VC (p < 0.05). KT-308/SC did not differ significantly from any other material. The positive controls showed significantly faster penetration than all the other groups (p < 0.05). It was concluded that KT-308 effectively prevented penetration of E. faecalis, whereas ZUT required further development.