A protocol for developing reporting guidelines for laboratory studies in Endodontology.

Laboratory-based research studies are the most common form of research endeavour and make up the majority of manuscripts that are submitted for publication in the field of Endodontology. The scientific information derived from laboratory studies can be used to design a wide range of subsequent studies and clinical trials and may have translational potential to benefit clinical practice. Unfortunately, the majority of laboratory-based articles submitted for publication fail the peer-review step, because unacceptable flaws or substantial limitations are identified. Even when apparently well-conducted laboratory-based articles are peer-reviewed, they can often require substantial corrections prior to the publication. It is apparent that some authors and reviewers may lack the training and experience to have developed a systematic approach to evaluate the quality of laboratory studies. Occasionally, even accepted manuscripts contain limitations that may compromise interpretation of data. To help authors avoid manuscript rejection and correction pitfalls, and to aid editors/reviewers to evaluate manuscripts systematically, the purpose of this project is to establish and publish quality guidelines for authors to report laboratory studies in the field of Endodontology so that the highest standards are achieved. The new guidelines will be named-'Preferred Reporting Items for Laboratory studies in Endodontology' (PRILE). A steering committee was assembled by the project leads to develop the guidelines through a five-phase consensus process. The committee will identify new items as well as review and adapt items from existing guidelines. The items forming the draft guidelines will be reviewed and refined by a PRILE Delphi Group (PDG). The items will be evaluated by the PDG on a nine-point Likert scale for relevance and inclusion. The agreed items will then be discussed by a PRILE face-to-face consensus meeting group (PFCMG) formed by 20 individuals to further refine the guidelines. This will be subject to final approval by the steering committee. The approved PRILE guidelines will be disseminated through publication in relevant journals, presented at congresses/meetings, and be freely available on a dedicated website. Feedback and comments will be solicited from researchers, editors and peer reviewers, who are invited to contact the steering committee with comments to help them update the guidelines periodically.

Association between missed canals and apical periodontitis.

AIM: To evaluate the frequency of post-treatment apical periodontitis associated with root filled teeth with at least one untreated root canal. METHODOLOGY: Eight hundred and seven cone beam computed tomography images containing at least one root filled tooth were selected from a collection of 1543 images from Brazilian individuals. Scans were taken using ICAT Classic devices (Imaging Sciences, Hatfield, PA, USA) in a private oral radiology clinic from January to April 2015. All root filled teeth were analysed for the presence of missed canals and apical periodontitis. The chi-square and odds ratio tests were used to verify if there were an association and risk relationship between the occurrence of untreated canals and apical periodontitis. RESULTS: A total of 2294 teeth with evidence of root fillings were identified. Two hundred and eighty-one teeth had at least one untreated missed canal (12%). The frequency of apical periodontitis in teeth with at least one untreated canal was significantly greater in comparison to teeth with all canals treated (274/281, 98% versus 1736/2013, 86%) (P < 0.01). The odds for apical periodontitis to be present was 6.25 times greater for teeth with an untreated canal. The mesiobuccal roots of maxillary first molars had the greatest frequency of untreated canals (114/154, 74%), with the second mesiobuccal canal being the most frequently missed (n = 106/114, 93%). CONCLUSION: Root filled teeth with at least one missed canal had a high prevalence of post-treatment apical periodontitis.

Influence of solvent and a supplementary step with a finishing instrument on filling material removal from canals connected by an isthmus.

AIM: To evaluate the effectiveness of a solvent (eucalyptol) in improving filling material removal from canals connected by isthmuses, and the additional cleaning effect of a finishing instrument. METHODOLOGY: The mesial canals from 32 mandibular molars (Vertucci's type II morphology) were instrumented and filled with the single-cone technique using Reciproc R25 gutta-percha points (VDW, Munich, Germany) combined with Sealer 26 (Dentsply, Petrópolis, RJ, Brazil). Each root was then subjected to retreatment using the Mtwo instrument system (VDW), with or without a solvent (n = 16 per group). The volume of filling material in the canals was assessed by micro-computed tomographic (micro-CT) scans taken before and after retreatment. Canals with remnants of filling material received a supplementary procedure with the XP-endo Finisher R instrument (FKG Dentaire, La Chaux-de-Fonds, Switzerland), with or without eucalyptol, and another micro-CT scan was taken. All retreatment procedures were performed inside a cabinet under a controlled temperature (37 °C). Filling material removal was evaluated in the 5-mm apical canal system for the canal+isthmus space or the isthmus alone. Statistical analyses were performed to compare the removal of filling material with and without eucalyptol, and after a supplementary approach with XP-endo Finisher R. The level of significance was set at 5% for all statistical tests (P < 0.05). RESULTS: The amount of filling material removed from the canal+isthmus with Mtwo instruments was 83.2% when no solvent was used and 83.8% using the solvent (P > 0.05). When the isthmus area was evaluated separately, most specimens were associated with a reduction in the filling material, with no significant difference between the groups with or without using a solvent (P > 0.05). The supplementary step with XP-endo Finisher R significantly improved removal of filling material from both canal and isthmus area (P < 0.05), regardless of the use of a solvent (P > 0.05). CONCLUSION: The use of eucalyptol did not improve filling material removal from Vertucci's type II molar mesial canals and isthmuses. XP-endo Finisher R significantly enhanced removal of filling material from the canals and isthmuses.

Effects of preparation with the Self-Adjusting File, TRUShape and XP-endo Shaper systems, and a supplementary step with XP-endo Finisher R on filling material removal during retreatment of mandibular molar canals.

AIM: To compare the effects of three instrumentation systems, and a supplementary approach with a finishing instrument, on filling material removal during retreatment of mandibular molar canals. METHODS: Sixty mesial canals from mandibular molars (Vertucci's type IV anatomy) were instrumented, filled and subjected to retreatment. After initial removal of the root canal filling material using the D-RaCe system, the canals were randomly distributed into three groups (n = 20) according to the instrument system used for preparation: the Self-Adjusting File (SAF), TRUShape or XP-endo Shaper. The filling material volume in the apical 5 mm of the canals was assessed by means of micro-computed tomography (micro-CT) before and after retreatment. All specimens with residual filling material were subjected to a supplementary approach with the XP-endo Finisher R instrument and another micro-CT scan was taken. Data on the volumes of filling material and incidence of total removal were compared between groups by the general linear model for paired data and the Fisher's exact test. The effects of the refinement step were evaluated by the Wilcoxon Signed Ranks test. RESULTS: The amount of removed material was 92.4%, 96.9% and 96.9% for the SAF, TRUShape and XP-endo Shaper, respectively. There were no significant differences between them (P > 0.05). Canals were completely cleaned of filling material in 70% of the specimens for XP-endo Shaper, 55% for SAF and 30% for TRUShape; the difference between XP-endo Shaper and TRUShape was significant (P = 0.03). The supplementary step with the XP-endo Finisher R instrument was associated with additional filling material removal of 38% (P < 0.001). Six more canals were rendered free of filling material after using this finishing instrument. CONCLUSIONS: The tested systems were equally effective in removing the mass of filling material from the apical 5 mm of molar canals. The supplementary step with the XP-endo Finisher R instrument enhanced filling material removal.

Association between bacteria occurring in the apical canal system and expression of bone-resorbing mediators and matrix metalloproteinases in apical periodontitis.

AIM: To evaluate the association between the presence of selected bacterial species/groups in the apical root canal and expression of mediators of soft and bone tissue destruction in apical periodontitis lesions. Relationships between bacteria and some other features of apical periodontitis were also investigated. METHODOLOGY: Seventeen freshly extracted teeth with pulp necrosis and apical periodontitis were included. The apical root segment was sectioned and cryopulverized; DNA was extracted and evaluated for the presence of 9 bacterial species/groups using real-time polymerase chain reaction. Lesions were processed for histopathological and immunohistochemical analyses, which targeted matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9), receptor activator of NFκB (RANK), RANK ligand (RANKL) and osteoprotegerin (OPG). Associations of the target bacteria with expression of these mediators, presence of symptoms, lesion size and histopathological diagnosis were evaluated. Data were analysed using the chi-square, Fisher's exact, Mann-Whitney and Pearson tests. P values lower than 0.05 were considered significant. RESULTS: All pulverized apical root samples were positive for bacteria. The most prevalent taxa were Actinobacteria (53%), Streptococcus species (35%), Fusobacterium species and Parvimonas micra (18%). The target mediators exhibited a high mean expression in the lesions (MMP-2: 82%; MMP-9: 73%; RANK: 78%; RANKL; 81%; OPG; 83%). Mean RANKL:OPG ratio was significantly higher in granulomas than cysts (P < 0.05, Mann-Whitney test). Actinobacteria were associated with granulomas, higher MMP-2 expression, lower OPG expression, and higher RANKL:OPG ratio (P < 0.05 for all, Fisher's exact test or Mann-Whitney test). No other significant associations were found. CONCLUSION: Actinobacteria may play an important role in the active phase of soft and bone tissue destruction in apical periodontitis.

Effects of increased apical enlargement on the amount of unprepared areas and coronal dentine removal: a micro-computed tomography study.

AIM: To evaluate the effects of progressive apical enlargement on the amount of unprepared root canal surface area and remaining dentine thickness. METHODOLOGY: The root canals of 30 extracted mandibular incisors with Vertucci's type I configuration were instrumented with rotary HyFlex CM instruments (Coltene-Whaledent, Altstätten, Switzerland) up to 4 instruments larger than the first one that bound at the working length (WL). Teeth were scanned in a micro-computed tomography (micro-CT) device before canal preparation and after instrumentation with the 2nd, 3rd and 4th larger instruments. The amount of unprepared surface area in the full canal or in the apical 4 mm as well as the remaining dentine thickness at 10 mm from the WL were calculated and compared. The general linear model for repeated measures adjusted by Bonferroni's post hoc test was used for statistic analysis. RESULTS: There was a significant reduction in the amount of unprepared areas after each increase in preparation size (P < 0.01). This was observed for both the full canal length and the 4-mm apical segment. The amount of remaining dentine was also significantly reduced after each file size (P < 0.01). However, dentine thickness always remained greater than 1 mm, even after using the largest instrument. CONCLUSION: Apical preparations up to 4 instruments larger than the first one to bind at the WL caused a significant progressive reduction in the unprepared canal area.

Bacteremia after supragingival scaling and dental extraction: Culture and molecular analyses.

OBJECTIVE: To study the incidence and magnitude of bacteremia after dental extraction and supragingival scaling. SUBJECTS AND METHODS: Blood samples were taken before and 5 and 30 min after dental extraction and supragingival scaling from individuals at high (n = 44) or negligible risk (n = 51) for infective endocarditis. The former received prophylactic antibiotic therapy. Samples were subjected to aerobic and anaerobic culture and quantitative real-time polymerase chain reaction to determine the incidence of bacteremia and total bacterial levels. RESULTS: Patients who did not receive prophylactic antibiotic therapy had a higher incidence of positive blood cultures (30% 5 min after extraction) than patients who received prophylactic antibiotic therapy (0% 5 min after extraction; p < .01). Molecular analysis did not reveal significant differences in the incidence or magnitude of bacteremia between the two patient groups either 5 or 30 min after each of the procedures evaluated. Extraction was associated with higher incidence of bacteremia than supragingival scaling by blood culture (p = .03) and molecular analysis (p = .05). CONCLUSIONS: Molecular methods revealed that dental extraction and supragingival scaling were associated with similar incidence of bacteremia in groups receiving or not prophylactic antibiotic therapy. However, blood culture revealed that antibiotic therapy reduced viable cultivable bacteria in the bloodstream in the extraction group.

What happens to unprepared root canal walls: a correlative analysis using micro-computed tomography and histology/scanning electron microscopy.

AIM: To microscopically examine the cleanliness of root canal walls that remained unprepared as revealed by micro-CT. METHODOLOGY: The root canals of 10 freshly extracted mandibular premolars with necrotic pulps and apical periodontitis along with the mesiobuccal canals of 11 mandibular molars with vital pulps were prepared using Reciproc instruments R40 and R25, respectively, and 2.5% sodium hypochlorite irrigation. Specimens were scanned in micro-CT before and after preparation, and the unprepared areas were identified. The outer root surface corresponding to the untouched areas was marked on each root third to guide further analysis using histological (for teeth with vital pulps) and scanning electron microscopic (SEM; for necrotic teeth) examination. In the teeth with vital pulps, the root canal area occupied by tissue remnants was calculated. In SEM analysis of teeth with necrotic pulps, scores were attributed for the amount of debris on the untouched areas. RESULTS: The proportion of unprepared areas in the mesiobuccal molar canals was 18.1% and 9.6% over the full canal length and apical canal, respectively. In premolars, corresponding figures were 34.6% and 17.6%, respectively. Histological analysis of canals with vital pulps revealed tissue remnants over the untouched walls almost exclusively in the apical canal. SEM analysis of the canals with necrotic pulps revealed debris along the untouched walls in all root canal thirds. CONCLUSION: The areas that remain untouched by Reciproc instruments used with 2.5% NaOCl irrigation as revealed by micro-CT analysis were usually covered with debris, in the form of pulp tissue remnants, bacteria and dentine chips, especially in the apical root canal.

Influence of oestrogen deficiency on the development of apical periodontitis.

AIM: To evaluate the effects of a long period of oestrogen deficiency on the development of apical periodontitis in rats. METHODOLOGY: Wistar rats (n = 24), 3 months old, evaluated by vaginal cytology, were included in the study. Twelve animals were ovariectomized (OVX group) and the other 12 were sham operated (control group). One hundred and twenty days after castration, the pulps of the left mandibular first molars were exposed to induce the development of apical periodontitis. Body mass was verified on a weekly basis. Following 21 and 40 days of lesion induction, the animals were sacrificed. Blood was collected for biochemical analysis, and mandibles were removed for radiographic analysis. Comparative analysis of the data was performed by the nonparametric Kruskal-Wallis and Dunn's multiple-comparisons tests. The t-test was applied to compare the oestrogen levels between control and OVX groups. RESULTS: Radiographs revealed that apical periodontitis lesions were significantly larger in the 40-day OVX group when compared with both 40-day (P < 0.05) and 21-day (P < 0.001) control groups. Serum oestrogen levels were significantly lower in the OVX group (P < 0.01), confirming the efficacy of castration. Oestrogen deficiency resulted in significantly greater body mass gain (P < 0.01) in 40-day OVX group when compared with 40-day control group. Serum concentrations of calcium were similar between groups (P > 0.05). Alkaline phosphatase levels, although higher in the OVX groups (21 and 40 days), were not significantly different. CONCLUSIONS: Ovariectomized rats had significantly larger apical periodontitis lesions after 40 days of pulp exposure when compared with controls. These findings suggest that bone alterations as a result of long periods of oestrogen deficiency can influence the progression of apical periodontitis.

Micro-CT evaluation of the efficacy of hard-tissue removal from the root canal and isthmus area by positive and negative pressure irrigation systems.

AIM: To evaluate the removal of accumulated hard-tissue debris (AHTD) from the root canal system of mandibular molars by positive and negative pressure irrigation systems, using micro-CT imaging analysis. METHODOLOGY: Mandibular molars with a single canal in the distal root and 2 canals connected by an isthmus in the mesial root were matched based on similar morphological dimensions using micro-CT evaluation and assigned to 2 experimental groups (n = 20 mesial and 10 distal canals), according to the irrigation protocol: apical positive (conventional irrigation) or negative (EndoVac system) pressure. Changes in root canal volume and surface area as well as percentage of uninstrumented canal wall surface and accumulated hard-tissue debris (AHTD) after canal preparation were compared statistically using the independent sample t-test and Mann-Whitney U-test, with the significance level set at 5%. RESULTS: Volume, surface area and percentage of static voxels in either mesial or distal root canal systems were not significantly different between groups before or after root canal preparation (P > 0.05). After preparation, AHTD was not observed in the distal canal of both groups. However, in the mesial root canal system, the conventional irrigation group was associated with a significantly higher median percentage of AHTD (11.48%; IQR: 5.9-22.6; range: 1.86-41.98) than the EndoVac group (3.40%; IQR: 1.5-7.3; range: 0.82-12.84) (P < 0.05). CONCLUSIONS: Neither irrigation protocol succeeded in rendering the mesial canal system free of AHTD; however, apical negative pressure irrigation resulted in lower levels of AHTD than conventional irrigation.

Sealing ability of two root-end filling materials in a bacterial nutrient leakage model.

AIM: To compare in vitro the sealing ability of root-end fillings with mineral trioxide aggregate (MTA) and EndoSequence BioCeramic Root Repair Material-Fast Set (BC-RRM) Putty using a novel bacterial nutrient leakage model, which provides information on whether or not intracanal bacteria are receiving nutrients from serum via leakage channels. METHODOLOGY: Sixty single-rooted decoronated mandibular incisors with instrumented root canals were subjected to root-end resection and ultrasonic preparation. The root specimens were mounted in the experimental apparatus, and the root-end cavities filled with the test materials. The positive control group used warm Gutta-percha and no sealer. In the negative controls, the entire resected surface was covered with varnish. After sterilization in ethylene oxide, the root canal was inoculated with 1.5 × 10(5) washed cells of Enterococcus faecalis. The apparatus was filled with foetal bovine serum, leaving only the apical root immersed. After 30-day incubation, samples were taken from the canal, cultured and the colony-forming units (CFUs) counted. Statistical analysis was performed using the Mann-Whitney test for quantitative and the Fisher exact test for qualitative data. RESULTS: In the MTA group, 10 of 20 (50%) specimens still had detectable viable bacteria in the canals (mean, 8.97 × 10(3)  CFUs). In the BC-RRM Putty group, 5 of 18 (28%) specimens were positive for bacterial growth (mean, 2.88 × 10(4)  CFUs). There was no significant difference when comparing the quantitative or presence/absence data from the MTA and BC-RRM Putty groups. Positive and negative controls yielded the expected results. CONCLUSIONS: MTA and BC-RRM Putty had similar sealing ability. The experimental model was effective in determining whether or not residual intracanal bacteria could survive by receiving nutrients from outside.

Subgingival bacterial community profiles in HIV-infected Brazilian adults with chronic periodontitis.

BACKGROUND AND OBJECTIVE: To compare the subgingival microbial diversity between non-HIV-infected and HIV-infected individuals with chronic periodontitis using denaturing gradient gel electrophoresis (DGGE). MATERIAL AND METHODS: Thirty-two patients were selected: 11 were HIV-infected and 21 were non-HIV-infected, and all had chronic periodontitis. Periodontal measurements included probing depth, clinical attachment level, visible supragingival biofilm and bleeding on probing. Subgingival biofilm samples were collected from periodontal sites (50% with probing depth ≤ 4 mm and 50% with probing depth ≥ 5 mm) and whole-genomic-amplified DNA was obtained. The DNA samples were subjected to amplification of a 16S rRNA gene fragment using universal bacterial primers, followed by DGGE analysis of the amplified gene sequences. RESULTS: The non-HIV-infected group presented higher mean full-mouth visible supragingival biofilm (p = 0.004), bleeding on probing (p = 0.006), probing depth (p < 0.001) and clinical attachment level (p = 0.001) in comparison with the HIV-infected group. DGGE analysis revealed 81 distinct bands from all 33 individuals. Banding profiles revealed a higher diversity of the bacterial communities in the subgingival biofilm of HIV-infected patients with chronic periodontitis. Moreover, cluster and principal component analyses demonstrated that the bacterial community profiles differed between these two conditions. High interindividual and intra-individual variability in banding profiles were observed for both groups. CONCLUSION: HIV-infected patients with chronic periodontitis present greater subgingival microbial diversity. In addition, the bacterial communities associated with HIV-infected and non-HIV-infected individuals are different in structure.

Causes and management of post-treatment apical periodontitis.

Endodontic treatment failure is usually characterised by the presence of post-treatment apical periodontitis, which may be persistent, emergent or recurrent. The major aetiology of post-treatment disease is persistent intraradicular infection, but in some cases a secondary intraradicular infection due to coronal leakage or an extraradicular infection may be the cause of failure. Understanding the causes of endodontic treatment failure is of paramount importance for the proper management of this condition. Teeth with post-treatment apical periodontitis can be managed by either nonsurgical endodontic retreatment or periradicular surgery, both of which have very high chances of restoring the health of the periradicular tissues and maintaining the tooth function in the oral cavity. This review article focuses on the aetiological factors of post-treatment apical periodontitis and discusses the indications and basics of the procedures for optimal clinical management of this condition.

Clinical antibacterial effectiveness of the self-adjusting file system.

AIM: To evaluate in vivo the antibacterial effectiveness of the self-adjusting file (SAF) using molecular methods. METHODOLOGY: Root canals from single-rooted teeth with apical periodontitis were instrumented using the SAF system under continuous irrigation with 2.5% NaOCl. DNA extracts from samples taken before and after instrumentation were subjected to quantitative analysis of total bacteria counts and levels of streptococci by quantitative real-time polymerase chain reaction (qPCR). The reverse-capture checkerboard assay was also used to identify 28 bacterial taxa before (S1) and after (S2) SAF instrumentation. SAF was also compared with a conventional hand nickel-titanium instrumentation technique for total bacterial reduction. Data from qPCR were analysed statistically within groups using the Wilcoxon matched pairs test and between groups using the Mann-Whitney U-test and the Fisher's exact test, with significance level set at P < 0.05. RESULTS: Self-adjusting file significantly reduced the total bacterial counts from a mean number of 1.96 × 10(7) cells to 1.34 × 10(4) cells (P < 0.001). Quantitatively, the 99.9% reduction in total bacterial counts associated with the SAF system was significantly superior to the 95.1% reduction obtained by hand instrumentation (P < 0.001). Qualitatively, SAF resulted in significantly more cases with negative PCR results for bacteria (54.5%) than hand instrumentation (4.5%) (P < 0.001). The SAF system succeeded in significantly reducing the streptococcal levels, but four cases still harboured these bacteria in S2. Checkerboard analysis revealed that not only streptococci but also some anaerobic and even as-yet-uncultivated bacteria may resist the effects of chemomechanical procedures. CONCLUSION: The SAF instrumentation system was highly effective in reducing bacterial populations from infected root canals and performed significantly better than hand instrumentation. However, because half of the samples still had detectable bacteria after preparation with SAF, supplementary disinfection is still required to maximize bacterial elimination.

Antibacterial, physicochemical and mechanical properties of endodontic sealers containing quaternary ammonium polyethylenimine nanoparticles.

AIM: To evaluate the antibacterial, physicochemical and mechanical properties of two endodontic sealers incorporating quaternary ammonium polyethylenimine (QPEI) nanoparticles at concentrations of 1% and 2% (w/w). METHODOLOGY: The sealers tested were AH Plus and Pulp Canal Sealer EWT in the commercial unmodified form or containing 1% or 2% QPEI. Antibacterial activity was evaluated using a direct contact test (DCT) against Enterococcus faecalis ATCC 29212 and two endodontic isolates (RW35 and RN44). Sealers freshly mixed or set for 7 days were exposed to bacterial suspensions for 10, 30 and 60 min. Setting time, flow test, solubility, apparent porosity, dimensional change following setting, wettability, zeta potential and compressive strength were assessed according to the International Standard Organization 6876:2001 (ISO 6876). RESULTS: DCT results revealed that both freshly prepared sealers had antibacterial effects unaffected by QPEI incorporation. Both unmodified sealers had lost much of their antibacterial effects after 7 days. However, for Pulp Canal Sealer EWT incorporated with 1% and 2% QPEI nanoparticles, the antibacterial effects against all test E. faecalis strains within 30 and 60 min of contact were significantly greater than the unmodified formula. Addition of QPEI resulted in no significant increase in the antibacterial effects of AH Plus after ageing. As for physicochemical and mechanical tests, setting time, wettability and zeta potential were influenced by the presence of QPEI nanoparticles. CONCLUSION: Incorporation of QPEI nanoparticles can improve the long-term antibacterial activity of Pulp Canal Sealer EWT without relevant changes in physicochemical and mechanical properties.

Reduction in bacterial counts in infected root canals after rotary or hand nickel-titanium instrumentation--a clinical study.

AIM: To compare the antibacterial efficacy of two instrumentation techniques, one using hand nickel-titanium (NiTi) instruments and the other using rotary NiTi instruments, in root canals of teeth with apical periodontitis. METHODOLOGY: Root canals from single-rooted teeth were instrumented using either hand NiTi instruments in the alternated rotation motion technique or rotary BioRaCe instruments. The irrigant used in both groups was 2.5% NaOCl. DNA extracts from samples taken before and after instrumentation were subjected to quantitative analysis by real-time polymerase chain reaction (qPCR). Qualitative analysis was also performed using presence/absence data from culture and qPCR assays. RESULTS: Bacteria were detected in all S1 samples by both methods. In culture analysis, 45% and 35% of the canals were still positive for bacterial presence after hand and rotary NiTi instrumentation, respectively (P > 0.05). Rotary NiTi instrumentation resulted in significantly fewer qPCR-positive cases (60%) than hand NiTi instrumentation (95%) (P = 0.01). Intergroup comparison of quantitative data showed no significant difference between the two techniques. CONCLUSION: There was no significant difference in bacterial reduction in infected canals after instrumentation using hand or rotary NiTi instruments. In terms of incidence of positive results for bacteria, culture also showed no significant differences between the groups, but the rotary NiTi instrumentation resulted in more negative results in the more sensitive qPCR analysis.

Clinical antimicrobial efficacy of NiTi rotary instrumentation with NaOCl irrigation, final rinse with chlorhexidine and interappointment medication: a molecular study.

AIM: To evaluate clinically the antibacterial effects of root canal treatment procedures using molecular microbiology analyses. METHODOLOGY: Samples were taken from 14 necrotic root canals of teeth with apical periodontitis before (S1) and after instrumentation with NaOCl irrigation (S2), a final rinse with chlorhexidine (CHX) (S3) and then one-week interappointment medication with calcium hydroxide/CHX paste (S4). The parameters examined included the following: incidence of positive broad-range PCR results for bacterial presence; impact on bacterial community structures evaluated by PCR-Denaturing Gradient Gel Electrophoresis (DGGE); quantitative bacterial reduction determined by real-time PCR; and identification of bacterial persisters by cloning and sequencing. Data from the different tests were subjected to statistical analyses and diversity indicator calculations. RESULTS: All S1 samples were positive for bacteria in all tests. Treatment procedures promoted a decrease in microbial diversity and significantly reduced the incidence of positive results and the bacterial counts (P < 0.05). In general, each subsequent treatment step improved disinfection. No specific taxon or community pattern was associated with post-treatment samples. CONCLUSION: Supplementary steps consisting of a final rinse with CHX followed by calcium hydroxide interappointment medication promoted further decrease in the bacterial bioburden to levels significantly below those achieved by the chemomechanical procedures alone. Because the long-term outcome of root canal treatment is dependent upon maximal bacterial reduction, the present results are of clinical relevance.

Quantitative molecular and culture analyses of bacterial elimination in oval-shaped root canals by a single-file instrumentation technique.

AIM: Bacterial reduction in oval-shaped root canals by a single-instrument technique was compared ex vivo with a conventional nickel-titanium rotary technique. Data obtained from two quantification methods, quantitative real-time polymerase chain reaction (qPCR) and culture, were also compared. METHODOLOGY: Oval-shaped canals of extracted teeth contaminated with Enterococcus faecalis were instrumented using either a single Reciproc instrument or the BioRaCe instrument series. Bacteriological samples were taken before (S1) and after instrumentation (S2). Bacterial quantification was performed using qPCR and culture. RESULTS: Intragroup analysis showed that both protocols promoted a highly significant bacterial reduction (P < 0.001). Intergroup analysis (S2 samples) showed no significant differences between the two instrumentation systems (P > 0.05). As for the quantification methods, qPCR revealed significantly higher counts of E. faecalis in S1 than culture (P < 0.05), but no significant differences occurred for S2 (P > 0.05). CONCLUSION: The single-file technique was comparable with the conventional technique in oval-shaped canals provided the width of apical preparation, volume of irrigants and duration of irrigation are kept similar. No significant difference was observed for qPCR and culture in post-instrumentation samples, indicating that both methods can be reliably used for studies of antibacterial effectiveness.