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PubMed2002

Comparison of 16S rDNA-based PCR and checkerboard DNA-DNA hybridisation for detection of selected endodontic pathogens.

Molecular methods have been used recently to investigate the bacteria encountered in human endodontic infections. The aim of the present study was to compare the ability of a 16S rDNA-based PCR assay and checkerboard DNA-DNA hybridisation in detecting Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Peptostreptococcus micros, Porphyromonas endodontalis, Por. gingivalis and Treponema denticola directly from clinical samples. Specimens were obtained from 50 cases of endodontic infections and the presence of the target species was investigated by whole genomic DNA probes and checkerboard DNA-DNA hybridisation or taxon-specific oligonucleotides with PCR assay. Prevalence of the target species was based on data obtained by each method. The sensitivity and specificity of each molecular method was compared with the data generated by the other method as the reference--a value of 1.0 representing total agreement with the chosen standard. The methods were also compared with regard to the prevalence values for each target species. Regardless of the detection method used, T. denticola, Por. gingivalis, Por. endodontalis and B. forsythus were the most prevalent species. If the checkerboard data for these four species were used as the reference, PCR detection sensitivities ranged from 0.53 to 1.0, and specificities from 0.5 to 0.88, depending on the target bacterial species. When PCR data for the same species were used as the reference, the detection sensitivities for the checkerboard method ranged from 0.17 to 0.73, and specificities from 0.75 to 1.0. Accuracy values ranged from 0.6 to 0.74. On the whole, matching results between the two molecular methods ranged from 60% to 97.5%, depending on the target species. The major discrepancies between the methods comprised a number of PCR-positive but checkerboard-negative results. Significantly higher prevalence figures for Por. endodontalis and T. denticola were observed after PCR assessment. There was no further significant difference between the methods with regard to detection of the other target species.

Aggregatibacter actinomycetemcomitans↗
PubMed2006

Identification of bacteria in endodontic infections by sequence analysis of 16S rDNA clone libraries.

A significant proportion of oral bacteria are unable to undergo cultivation by existing techniques. In this regard, the microbiota from root canals still requires complementary characterization. The present study aimed at the identification of bacteria by sequence analysis of 16S rDNA clone libraries from seven endodontically infected teeth. Samples were collected from the root canals, subjected to the PCR with universal 16S rDNA primers, cloned and partially sequenced. Clones were clustered into groups of closely related sequences (phylotypes) and identification to the species level was performed by comparative analysis with the GenBank, EMBL and DDBJ databases, according to a 98% minimum identity. All samples were positive for bacteria and the number of phylotypes detected per subject varied from two to 14. The majority of taxa (65.2%) belonged to the phylum Firmicutes of the Gram-positive bacteria, followed by Proteobacteria (10.9%), Spirochaetes (4.3%), Bacteroidetes (6.5%), Actinobacteria (2.2%) and Deferribacteres (2.2%). A total of 46 distinct taxonomic units was identified. Four clones with low similarity to sequences previously deposited in the databases were sequenced to nearly full extent and were classified taxonomically as novel representatives of the order Clostridiales, including a putative novel species of Mogibacterium. The identification of novel phylotypes associated with endodontic infections suggests that the endodontium may still harbour a relevant proportion of uncharacterized taxa.

Adolescent↗
PubMed2007

Phylogenetic analysis of bacterial and archaeal species in symptomatic and asymptomatic endodontic infections.

Phylogenetic analysis of bacterial and archaeal 16S rRNA was used to examine polymicrobial communities within infected root canals of 20 symptomatic and 14 asymptomatic patients. Nucleotide sequences from approximately 750 clones amplified from each patient group with universal bacterial primers were matched to the Ribosomal Database Project II database. Phylotypes from 37 genera representing Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria and Proteobacteria were identified. Results were compared to those obtained with species-specific primers designed to detect Prevotella intermedia, Porphyromonas gingivalis, Porphyromonas endodontalis, Peptostreptococcus micros, Enterococcus sp., Streptococcus sp., Fusobacterium nucleatum, Tannerella forsythensis and Treponema denticola. Since members of the domain Archaea have been implicated in the severity of periodontal disease, and a recent report confirms that archaea are present in endodontic infections, 16S archaeal primers were also used to detect which patients carried these prokaryotes, to determine if their presence correlated with severity of the clinical symptoms. A Methanobrevibacter oralis-like species was detected in one asymptomatic and one symptomatic patient. DNA from root canals of these two patients was further analysed using species-specific primers to determine bacterial cohabitants. Trep. denticola was detected in the asymptomatic but not the symptomatic patient. Conversely, Porph. endodontalis was found in the symptomatic but not the asymptomatic patient. All other species except enterococci were detected with the species-specific primers in both patients. These results confirm the presence of archaea in root canals and provide additional insights into the polymicrobial communities in endodontic infections associated with clinical symptoms.

Archaea↗
PubMed2004

Endodontic treatment of primary teeth using a combination of antibacterial drugs.

AIM: To evaluate the clinical outcome of 'Lesion Sterilization and Tissue Repair' (LSTR) therapy for endodontic treatment of primary teeth. METHODOLOGY: A mixture of metronidazole, ciprofloxacin, and minocycline (3Mix) in ointment (macrogol mixed with propylene glycol: MP) or in a root canal sealer was used to disinfect infected root canals of 56 patients aged 4-18 years. Out of a total of 87 primary teeth, 81 cases had physiologic root resorption. A total of 54 radiolucent periradicular lesions were present. The root canals were not prepared before or after disinfection. 3Mix medicament (3Mix-MP/3Mix-sealer) was placed at orifices of root canals or on the bottom of pulp chambers, and then sealed with glass-ionomer cement and further reinforced by a composite resin inlay prepared using a direct method and cemented with resin. RESULTS: In all cases, clinical symptoms such as gingival swelling (52 cases), sinus tracts (22 cases), induced dull pain (3 cases), spontaneous dull pain (26 cases), and pain on biting (46 cases) disappeared after treatment, although in four cases clinical signs and symptoms were finally resolved only after retreatment using the same procedures. Thus, gingival abscesses and fistulae, if present, disappeared after a few days. Successor permanent teeth erupted without any disorders, or were found radiographically to be normal and in the process of eruption. All the cases were evaluated as successful. The mean function time of the primary teeth was 680 days (range: 68-2390 days), except for one case in which the successor permanent tooth was congenitally missing. CONCLUSION: Primary teeth with periradicular lesions with or without physiologic root resorption were treated successfully by the LSTR endodontic therapy.

Adolescent↗
PubMed1993

A comparison of endodontic treatment results at two dental schools.

Endodontic treatment results were assessed in a controlled clinical study in Arhus. Undergraduate students used either Kloroperka or AH26 as sealer in a randomized fashion. The findings were compared with the results from a previous study of identical design in Oslo, which involved Kloroperka, AH26 and ProcoSol. While Kloroperka performed slightly, but significantly less well than AH26 or ProcoSol in the Oslo study, no difference was found between Kloroperka and AH26 in the present study. The overall treatment results were poorer in Arhus than in Oslo, but this might be explained by a marked difference in the preoperative diagnoses and in the technical problems associated with the teeth selected for treatment. When results of endodontic treatment carried out at different locations are compared, variations in case selection and preoperative diagnoses must be corrected for in order to obtain an adequate basis for comparison.

Adult↗
PubMed1994

Retreatment or radiographic monitoring in endodontics.

UNLABELLED: The aim of this clinical study was to assess 1032 endodontically treated roots in relation to: (i) the success rate of retreatment (612 roots)--only cases that had recall examinations of 6 months or longer were evaluated; (ii) the influence of various factors on the technical and clinical results of the retreatment; and (iii) the consequences of radiographic monitoring of 420 asymptomatic roots when the root filling was radiographically deficient (short, overextended and/or permeable root fillings). Technical assessment of the retreatment showed that the root was adequately sealed in 52.3% of cases, the root filling was improved in 33.8%, was identical with the initial treatment in 11.1% and was worse than the first treatment in 2.8% of the canals. Clinical assessment of the retreatment of symptomatic roots showed that 71.8% of the retreatments were judged successful, 18.9% showed some healing and 9.3% had failed. The initial size of the periapical lesion, the use of rubber dam, the root filling technique and the apical level of the root filling had a statistically significant influence on the result of the retreatment. Monitoring radiographically (median time span 6 years) led to maintenance of the status quo in 94.8% of cases, healing in 2.4% and failure in 2.8% of the canals. Retreatment is clearly indicated when periapical radiolucency, clinical signs and/or symptoms are present with relative success of up to 91%. When no or little radiographic evidence of periapical pathology was present, when clinical signs and symptoms were absent or when the root filling was radiographically deficient, radiographic monitoring led to complications in only a limited number of cases. KEYWORDS: clinical decision, endodontic retreatment, radiographic evaluation.

Adult↗
PubMed1995

The guided tissue regeneration principle in endodontic surgery: one-year postoperative results of large periapical lesions.

Twenty patients with large endodontic lesions, which failed to respond to conventional endodontic therapy, were selected for this study. The lesions had a radiographic diameter of at least 10 mm, were removed by periradicular surgery, before retrofilling the apices with either super EBA or dessicated zinc oxide-eugenol. In 10 test sites large e-PTFE membranes (Gortex) were placed to cover the lesions, while at the control sites the lesions were not covered before resuturing. Radio-graphic analysis of the lesions at 3, 6, 9 and 12 months revealed that lesions covered with the membranes healed quicker than the control lesions, and that the quality and quantity of the regenerated bone was superior when membranes were used. Results of the study indicate that guided tissue regeneration (GTR) principles can be effectively applied to the healing of large periapical lesions, especially in through-and-through lesions.

Adult↗
PubMed1995

Factors associated with endodontic flare-ups: a prospective study.

The purpose of this prospective study was to assess the incidence of flare-ups (a severe problem requiring an unscheduled visit and treatment) among patients who received endodontic treatment by the two authors in their respective practices during a period of one year, and also to examine the correlation with pre-operative and operative variables. The results showed an incidence of 1.58% for flare-ups from 1012 endodontically treated teeth. Statistical analysis using the chi-square test (P<0.05) indicated that flare-ups were found to be positively correlated with multiple appointments, retreatment cases, periradicular pain prior to treatment, presence of radiolucent lesions, and patients taking analgesic or anti-inflammatory drugs. In contrast, there was no correlation between flare-up, and age, sex, different arch/tooth groups and the status of the pulp.

Acute Disease↗
PubMed1996

Variations in the susceptibilities of components of the endodontic microflora to biomechanical procedures.

The role of the endodontic microflora in pulpal disease and in endodontic treatment failures is well established. Thus the need for effective microbial control is one of the important justifications for biomechanical procedures. However the efficacy of this stage of treatment is dependent upon the vulnerability of the involved species, which may not be uniform. The aim of this study was to investigate variations in the susceptibilities of members of the root canal microflora to routine biomechanical procedures. Forty-two root canals were investigated microbiologically. Samples were collected before and after instrumentation and the bacterial findings were compared. In 15 cases of 'primary' root canal therapy, despite changes in the population size, no significant change in the species composition of the microflora was observed. However in 27 cases 'secondary' treatment, a decrease in the number of isolations of Peptostreptococcus spp. was found (P = 0.008). When all 42 cases were considered together, significant decreases were found between first and second samples for anaerobes (P = 0.0117) and for Grampositive species (P = 0.008), especially Peptostreptococcus spp. (P = 0.02). It was therefore concluded that certain species are more resistant to the biomechanical procedures than others.

Bacteria↗
PubMed1996

The antimicrobial activity of endodontic sealers to anaerobic bacteria.

The effect of five root canal sealers and two root canal dressing materials on the growth of three anaerobic bacteria associated with endodontic infections was determined using the agar diffusion inhibitory test. Samples of the following endodontic sealers (Apexit, Ketac-Endo, Roth Sealer, Sealapex and Tubliseal) and root canal dressing materials (Pulpdent and Root-cal) were incubated for 48 h with the following anaerobic bacteria: Capnocytophaga ochracea, Porphyromonas gingivalis and Peptostreptococcus micros. Statistically significant zones of bacterial growth inhibition for all the bacteria tested were observed in descending order of antimicrobial activity: Roth Sealer, Ketac-Endo, Tubliseal, Apexit and Sealapex. Root-cal and Pulpdent also showed statistically significant antimicrobial activity, but only to Capnocytophaga ochracea, not to the other two bacteria tested, with Pulpdent being the least active.

Bacteria, Anaerobic↗
PubMed2004

Cleaning of rotary nickel-titanium endodontic instruments.

AIM: To develop and evaluate an effective cleaning procedure for rotary nickel-titanium (NiTi) endodontic instruments. METHODOLOGY: New rotary instruments (ProFile size 25/.04) were contaminated by preparing canals of extracted teeth. Three factors were evaluated to develop an effective cleaning sequence: dry or moist storage before cleaning; mechanical removal (brushing); and chemical dissolution in 1% NaOCl with ultrasonication. Debris on flutes was scored after staining in situ with Van Gieson's solution at x45 magnification. Debris was classified as stained or unstained particulate debris and organic film, and rated as none, slight, moderate or heavy. The effectiveness of a recommended cleaning sequence was tested on different instrument types and in private endodontic practices. RESULTS: All new instruments showed metallic spurs and fine particulate debris on the surfaces. After contamination, brushing alone removed most particulate debris, but did not remove organic film. NaOCl effectively removed organic film. Under laboratory conditions, the sequential cleaning procedures (moist storage, brushing followed by immersion in 1% NaOCl and ultrasonic cleaning) totally removed organic debris. Dry storage before cleaning or autoclaving with debris present reduced cleaning effectiveness (P<0.001, one-way ANOVA). In three private practices, the cleaning protocol substantially reduced biological contamination, but complete cleaning was not always achieved (87% clean). CONCLUSION: Complete removal of organic debris from instruments is feasible using a combination of mechanical removal and chemical dissolution, but requires meticulous attention to details.

Analysis of Variance↗
PubMed2004

Antimicrobial activity of varying concentrations of sodium hypochlorite on the endodontic microorganisms Actinomyces israelii, A. naeslundii, Candida albicans and Enterococcus faecalis.

AIM: To determine the resistance of microorganisms associated with refractory endodontic infections to sodium hypochlorite used as a root canal irrigant. METHODOLOGY: Two strains each of Actinomyces naeslundii, Candida albicans and Enterococcus faecalis were tested as late logarithmic phase inocula, against sodium hypochlorite adjusted to 0.5, 1.0, 2.5 and 5.25% w/v. Contact times used were 0, 10, 20, 30, 60 and 120 s. In the case of E. faecalis, additional experiments used contact times of 1.0, 2.0, 5.0, 10.0 and 30.0 min. Anti-microbial action was halted by sodium thiosulphate addition. Survivors were measured primarily using viable counts on drop plates. Additionally, pour plates were used to count low colony-forming units (cfu) and dilutions to 10(-6) were used to count high cfu. RESULTS: All concentrations of NaOCl lowered cfu below the limit of detection after 10 s in the case of A. naeslundii and C. albicans. However, E. faecalis proved to be more resistant to NaOCl. Using 0.5% NaOCl for 30 min reduced cfu to zero for both strains tested. This compares with 10 min for 1.0%, 5 min for 2.5% and 2 min for 5.25% (P < 0.001). Regression analysis for the dependent variable log(e)(count + 1) with log(e)(time + 1) and concentration as explanatory variables gave rise to a significant interaction between time and concentration (P < 0.001). CONCLUSION: The published association of E. faecalis with refractory endodontic infection may result, at least partially, from high resistance of this species to NaOCl. This does not appear to be the case with A. naeslundii or C. albicans.

Actinomyces↗
PubMed2004

Endodontic treatment of an invaginated maxillary lateral incisor with a periradicular lesion and a healthy pulp.

AIM: To report the healing of a periradicular lesion following nonsurgical root canal treatment of a dental invagination. SUMMARY: A case of dens invaginatus type 3 with a periradicular lesion and a healthy pulp is reported. Nonsurgical root canal treatment of the invagination was performed successfully and resolved an associated periradicular lesion. Despite the complex anatomy and a diagnosis of apical periodontitis, pulp health was retained after endodontic treatment of the invaginated canal. Clinical considerations and treatment are discussed and reported. KEY LEARNING POINTS: Dens invaginatus can present in a variety of forms, knowledge of which can usefully inform endodontic diagnosis and treatment. In certain cases, success can be achieved by treating the invagination alone. Pulp health may be preserved during and after treatment of an infected invagination.

Child↗
PubMed2004

Susceptibility to localized corrosion of stainless steel and NiTi endodontic instruments in irrigating solutions.

AIM: To evaluate the pitting and crevice corrosion characteristics of stainless steel (SS) and NiTi endodontic files in R-EDTA and NaOCl irrigating solutions. METHODOLOGY: The corrosion behaviour of two H-files produced from different SS alloys (Mani, AISI 303 SS, Dentsply Maillefer, AISI 304 SS) and one file produced from NiTi alloy (Maillefer) was determined in R-EDTA and NaOCl irrigating solutions by the cyclic potentiodynamic polarization method. The cutting flutes of 12 files of each material were embedded in an epoxy resin, polished, exposed to the irrigating solutions and used as an electrode. An Ag/AgCl electrode was used as a reference, a platinum plate was used as a counter electrode and polarization curves were obtained for all files in R-EDTA and NaOCl irrigating solutions in 37 degrees C with a potential scan rate of 5 mV min(-1). Corrosion potential (Ecorr), Corrosion current density (Icorr) and Pitting potential (Epit) were calculated from each curve. The results were statistically analysed with two-way anova and Student-Newman-Keuls (SNK) multiple comparison test with materials and irrigating solutions serving as discriminating variables (a = 0.05). RESULTS: Cyclic polarization curves presented negative hysteresis implying that pitting or crevice corrosion are not likely to occur for all the materials examined in both irrigating solutions. In NaOCl all materials showed significantly higher Ecorr (P = 0.011) as well as lower Icorr compared with R-EDTA reagent. Moreover, all materials demonstrated equal Epit in NaOCl, which was to be found significantly lower (P = 0.009) than the value of Epit in R-EDTA. CONCLUSIONS: None of the tested materials is susceptible to pitting or crevice corrosion in R-EDTA and NaOCl solutions and from this standpoint are appropriate for the production of endodontic files.

Analysis of Variance↗
PubMed2006

Longitudinal study of periapical and endodontic status in a Danish population.

AIM: To describe and discuss changes in periapical and endodontic status in a general Danish population. METHODOLOGY: In 1997, 616 randomly selected individuals had a full-mouth radiographic survey taken. In 2003, 77% of the participants attended for a new full-mouth radiographic examination. Information on endodontic treatment and periapical status was obtained. The periapical index (PAI) was used to assess apical periodontitis (AP). RESULTS: More participants had root filling(s) and AP in 2003 than in 1997. More teeth had AP and/or root fillings in 2003. Fewer of the root-filled teeth (RFT) had AP in 2003. Less than 3% of the teeth without root fillings (NRFT) that in 1997 had no AP, developed AP and/or received a root filling. Of the NRFT which in 1997 had AP, more than 35% still had AP and no root filling in 2003. Approximately, 30% of the NRFT with AP in 1997 received a root filling. Of the teeth that received a root filling, 40% had healed, whereas 60% had not. Approximately, 25% of the NRFT that in 1997 had AP had been extracted. In 1997 there were 618 RFT, 314 of the RFT had no AP in 1997. Almost 20% of the RFT that in 1997 were periapically sound, developed AP. Of the 304 RFT with AP in 1997, approximately 30% had healed, in 60% AP persisted, and 10% were extracted. CONCLUSIONS: The present study indicates that caution must be exercised when statements on the outcome of root canal treatment are made based on the cross-sectional studies.

Adult↗
PubMed2006

Nonsurgical treatment of extensive cyst-like periapical lesion of endodontic origin.

AIM: To report the repair of an extensive periapical lesion of endodontic origin, following nonsurgical treatment. SUMMARY: Clinical and radiographic examination revealed an extensive periapical lesion related to tooth 22, extending from the distal surface of tooth 21 to the mesial surface of 26. The patient reported a previous history of dental trauma involving this quadrant and had been under orthodontic treatment for a year. Intraoral examination revealed an asymptomatic bony hard swelling, mainly confined to the palate. During root canal exploration irregular walls associated with 3 mm of apical calcification were noted. After apical patency was obtained 1 mL of bloody serous exudate was drained. Intracanal aspiration provided a further 2 mL of yellow serous exudate. Following biomechanical preparation, a dressing of calcium hydroxide with anaesthetic solution was applied and replaced four times over a period of 12 months. The clinical-pathological picture demonstrated resolution of the lesion during this period of time. The 14-month clinical and radiographic examinations revealed normal bony contour and a significant resolution of the maxillary radiolucency. KEY LEARNING POINTS: Periapical lesions of endodontic origin may develop asymptomatically and become large. Proper biomechanical preparation followed by calcium hydroxide medication renewed periodically represents a nonsurgical approach to resolve extensive inflammatory periapical lesions.

Adult↗
PubMed2006

Bactericidal effect of Nd:YAG laser irradiation on some endodontic pathogens ex vivo.

AIM: To define the role of neodymium:yttrium-aluminum-garnet (Nd:YAG) lasers in root canal disinfection along with a minimally invasive treatment concept. METHODOLOGY: The hypothesis was tested ex vivo that Nd:YAG laser irradiation has a bactericidal effect on endodontic pathogens inoculated in root canals. Resultant colony-forming unit counts were associated with observations of bacterial cell structural changes using conventional scanning electron microscopy (CSEM) and environmental scanning electron microscopy (ESEM) on inoculated dentine surfaces, following indirect and direct Nd:YAG laser irradiation, respectively. RESULTS: The Nd:YAG laser irradiation (1.5 W, 15 Hz, four times for 5 s) of Enterococcus faecalis inoculated canals resulted in a significant reduction (P < 0.05, Wilcoxon signed rank test) of the bacterial load, meaning a 99.7% kill, but no sterilization. The CSEM procedure verified that the extent of radiation damage was in line with the total amount of laser energy applied. After 2 h of incubation and three cycles of indirect laser treatment (i.e. through a 1-mm-thick dentine disc), no morphologically intact bacteria of Actinomyces naeslundii or Streptococcus anginosus were discernible. However, when micro-colonies of S. anginosus and specially biofilms of E. faecalis were present after 2 days, the in situ experiment using ESEM and direct laser treatment showed that bacterial eradication was reduced in deep layers. CONCLUSIONS: The Nd:YAG laser irradiation is not an alternative but a possible supplement to existing protocols for canal disinfection as the properties of laser light may allow a bactericidal effect beyond 1 mm of dentine. Endodontic pathogens that grow as biofilms, however, are difficult to eradicate even upon direct laser exposure.

Actinomyces↗
PubMed2006

Delayed type allergic reaction following the use of nonlatex rubber dam during endodontic treatment.

AIM: To describe the development of a delayed type allergic reaction 24 h following the use of nonlatex rubber dam during endodontic treatment. The symptoms were alleviated with adequate medication; however, the patient rejected any further analysis. SUMMARY: In recent years, allergic reactions because of latex rubber dam and gloves have received more attention. Nonlatex products are advocated in clinical use in cases where the patient presents an allergic background. Although rare, adverse reactions may be encountered even with these products. In this article, a very rare case is presented in which a patient developed signs of allergy 24 h after the utilization of nonlatex rubber dam and gloves during endodontic treatment. Signs including visible redness on the mucosa, submucosal oedema and desquamation in the vestibular mucosa were observed. With proper medication, the signs and symptoms were alleviated. Although the patient was referred to an allergy specialist for patch testing, she did not accept any further analysis. KEY LEARNING POINTS: This case emphasizes the necessity of precautions during the management of latex-allergic patients in clinical practice. The unusual response encountered in this patient does not undermine the usefulness of nonlatex rubber dam and gloves in sensitive individuals.

Dermatitis, Allergic Contact↗