What Is the Microbial Profile in Persistent Endodontic Infections? A Scoping Review.

INTRODUCTION: This scoping review aimed to map the evidence about the microbiota found in persistent endodontic infections. METHODS: The study protocol was prospectively registered and is available at https://osf.io/3g2cp. The electronic search was performed in MEDLINE via PubMed, Lilacs, BBO, Scopus, Web of Science, Cochrane Library, and Embase. The eligibility criteria were based on the PCC acronym, where P (Population) represents patients with teeth presenting persistent endodontic infection, C (Concept) represents microbial profile, and C (Context) represents undergoing endodontic retreatment. Clinical studies that evaluated the microbial profile of samples collected from root canals of teeth undergoing retreatment, using classical or molecular methods, were included. Studies that did not show a minimum period of 1 year between primary endodontic treatment and retreatment or did not radiographically evaluate the quality of primary root canal filling were excluded. Two reviewers independently selected the articles and collected data. RESULTS: From a total of 957 articles, 161 were read in full, and 32 studies were included. The most prevalent species were Enterococcus faecalis, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Dialister invisus, Propionibacterium acnes, Tannerella forsythia, and Treponema denticola. Cases with symptomatology or inadequate root canal filling presented an increase in specific bacterial species compared to those with no symptomatology or adequate filling. A greater number of microorganisms was observed in teeth with inadequate coronal restoration compared to those with adequate restoration. CONCLUSIONS: Persistent endodontic infections have a polymicrobial profile identified by the commonly used methods for bacterial detection/identification and are subject to the limitations present in each of those methods.

Influence of variations in the environmental pH on the solubility and water sorption of a calcium silicate-based root canal sealer.

AIM: To evaluate the influence of pH variation on the solubility and water sorption of a premixed calcium silicate-based root canal sealer (EndoSequence BC Sealer) compared to the gold standard based on epoxy resin (AH Plus Jet) after immersion in distilled water and phosphate-buffered saline (PBS). METHODOLOGY: Solubility and water sorption were evaluated after immersion in distilled water or PBS at several pHs (5, 7 and 12) and the values were calculated as percentages of the original mass after 24 h, 7 and 30 days of immersion. The crystalline structures present in the sealers and surface precipitates were assessed by X-ray diffraction. The Shapiro-Wilk's test revealed that data were normally distributed; thus, statistical analysis was performed using one-way anova and Tukey's tests or independent t-test, assuming a 5% α-error. RESULTS: EndoSequence BC Sealer was associated with significantly greater water sorption and solubility compared to AH Plus Jet in all tested conditions (P < 0.05). The acid environment increased the 24 h solubility of EndoSequence BC Sealer immersed in PBS (P < 0.05) and did not induce significative changes in the water sorption (P > 0.05). Alkaline pH reduced the solubility of EndoSequence BC Sealer and increased that of AH Plus Jet at all experimental times and soaking media (P < 0.05). Alkaline environment also significantly increased the water sorption of AH Plus Jet immersed in PBS (P < 0.05). Immersion in PBS significantly reduced the solubility of EndoSequence BC Sealer and significantly increased that of AH Plus Jet (P < 0.05). Precipitates on the surface of EndoSequence BC Sealer corresponding to hydroxyapatite and calcium carbonate were detected after immersion in PBS at pH 5 and 7 for 30 days. CONCLUSIONS: EndoSequence BC Sealer had significantly greater solubility and water sorption than AH Plus Jet. Although the alkaline pH and soaking media directly influenced the solubility and water sorption of the sealers, the solubility of AH Plus Jet remained within the limits recommended by ISO 6876, whilst the solubility of the EndoSequence BC Sealer did not comply with ISO recommendations in all the conditions tested.

The impact of minimally invasive root canal preparation strategies on the ability to shape root canals of mandibular molars.

AIM: To evaluate the use of a reduced taper endodontic instrument system (Bassi Logic™ .03 taper) and expandable heat-treated system (XP-endo Shaper) on the ability to shape canals in mandibular molars, by means of microcomputed tomography (micro-CT) analysis. The Reciproc system was used as the reference for comparison. METHODOLOGY: Twenty-four mandibular molars were scanned in a micro-CT, matched based on similar anatomical features and sorted into three groups (n = 8 per group), according to root canal preparation system: Bassi Logic™ .03, XP-endo Shaper and Reciproc. The teeth were mounted onto a mannequin and the pulp chambers were accessed with traditional access cavities. In Bassi Logic™ .03 and Reciproc groups, mesial canals were prepared with size 25, .03 taper or R25 (size 25, .08v taper) instruments and the distal root canal with size 25, .03 taper and size 40, .03 taper or R25 and R40 (size 40, .06v taper), respectively. In XP-endo Shaper groups, all root canals were prepared with XP-endo Shaper (size 30, .04v taper). After root canal preparation, the teeth were rescanned. The percentage of untouched canal areas and the percentage of removed dentine were evaluated separately for mesial and distal root canals. The data were analysed using one-way anova and Tukey tests (P < 0.05). RESULTS: The Bassi Logic™ .03 group was associated with a greater percentage of untouched canal areas when compared to XP-endo Shaper and Reciproc groups for mesial and distal root canals (P < 0.05), but no differences were found between XP-endo Shaper and Reciproc groups (P > 0.05). No significant difference was observed in the percentage of dentine removed amongst the groups for mesial and distal root canals (P > 0.05). CONCLUSION: The use of a reduced taper system (Bassi Logic™ .03) during root canal preparation resulted in a greater percentage of untouched canal areas when compared to XP-endo Shaper and Reciproc instruments. No differences were observed amongst the systems regarding the percentage of dentine removed.

The influence of endodontic access cavity design on the efficacy of canal instrumentation, microbial reduction, root canal filling and fracture resistance in mandibular molars.

AIM: To assess the impact of conservative endodontic access cavities (CEC) and truss access cavities (TAC) during root canal treatment performed on mandibular molars in terms of: ability to shape and fill root canals, microbial reduction in canals, and cleaning of the pulp chamber. In addition, the fracture resistance of the teeth after coronal restoration was assessed. Traditional endodontic cavities (TEC) were used as a reference technique for comparison. METHODOLOGY: Thirty extracted intact mandibular molars were scanned in a microcomputed tomography device (micro-CT), matched based on similar anatomical features and assigned to TEC, CEC or TAC groups (n = 10). The specimens were accessed accordingly, and root canals were contaminated with bacterial suspensions of Enterococcus faecalis (21 days). Subsequently, the first microbial sample was collected from root canals (S1). The canals were initially prepared with Reciproc Blue R25 instrument followed by a second instrumentation using Reciproc Blue R40. Eight mL of 0.5% NaOCl were used as an irrigant for each instrument. A final irrigation protocol was performed with 2 mL of 0.5% NaOCl, 2 mL of 17% EDTA and another 2 mL of 0.5% NaOCl. Microbial samples were collected from root canals after R25 (S2), R40 (S3) and final irrigation (S4). The teeth were rescanned after S4. Then, root canals were filled, rescanned, restored and the teeth subjected to fracture resistance tests. The statistical analysis was performed with type I negative binomial and beta 0-1 inflation regression models for microbiological analysis. Instrumentation, filling and resistance to fracture results were subjected to anova and Tukey tests (P < 0.05). RESULTS: S4 revealed no significant variations in microbial reduction amongst the groups (P > 0.05). TEC had a significantly lower percentage of unprepared surface area than CEC (P < 0.05). No differences were found regarding the percentage of dentine removed, transportation, centring ability and filling voids amongst the groups (P > 0.05). The TEC group had a significantly lower volume of remaining root filling material within the pulp chamber than CEC and TAC groups (P < 0.05). There was no difference regarding fracture resistance amongst the groups (P > 0.05). CONCLUSION: Conservative access cavities did not offer any advantage in comparison with the traditional endodontic cavities in any of the parameters considered. Furthermore, conservative methods were associated with larger percentages of unprepared canal surface area and larger volumes of remaining root filling material within the pulp chamber.

A laboratory study of the impact of ultraconservative access cavities and minimal root canal tapers on the ability to shape canals in extracted mandibular molars and their fracture resistance.

AIM: To evaluate the influence of ultraconservative endodontic access cavities (UEC) and the use of instruments with various tapers (.03 or .05) and tip diameters (25 or 40) on the ability to shape canals in mandibular molars and their fracture resistance in comparison with traditional endodontic access cavities (TEC). METHODOLOGY: Thirty-two intact extracted mandibular molars were scanned in a micro-computed tomography device, matched based on similar anatomical features and assigned to UEC or TEC groups and to subgroups according to mechanical canal preparation with Bassi Logic™ rotary NiTi instruments with .03 or .05 taper (n = 8 per group). The teeth were mounted onto a mannequin head, and the pulp chambers were accessed and the root canals subjected to instrumentation. In the first stage, size 25 endodontic instruments were used, with .03 or .05 tapers. The teeth were re-scanned. Subsequently, a second canal instrumentation was performed with the size 40 instruments with .03 or .05 tapers and scanned again. The root canals were filled and then the teeth restored before being subjected to fracture resistance tests. The data on untouched canal areas, canal transportation, centring ability, the percentage of removed dentine and maximum load at fracture were analysed. One-way anova and Tukey's tests were performed for inter-group analysis. The paired-samples t-test was performed on the results between size 25 and size 40 apical preparations for each individual specimen (P < 0.05). RESULTS: No significant difference was observed for the percentage of untouched canal area when comparing UEC and TEC groups or between the .03 and .05 tapers with the same instrument tip size (P > 0.05). There was a significantly lower percentage of untouched canal area after instrumentation with size 40 instruments when compared to size 25 (P < 0.05). No significant difference in the percentage of dentine removed was observed between the access cavities or the different tapers (P > 0.05). Size 40 apical preparations removed significantly more dentine than size 25 preparations (P < 0.05). No significant differences were observed in canal transportation, centring ability and fracture resistance in any of the tested groups (P > 0.05). CONCLUSIONS: Ultraconservative endodontic access cavities did not offer any advantages in comparison with traditional endodontic access cavities on the ability to shape canals, nor the fracture resistance of mandibular molars. No differences were observed in shaping ability and fracture resistance between .03 and .05 taper canal preparations. Apical preparation with larger instruments resulted in significantly less untouched canal area in all groups.

Effects of alcohol and nicotine consumption on the development of apical periodontitis in rats: a correlative micro-computed tomographic, histological and immunohistochemical study.

AIM: To evaluate the effects of alcohol and nicotine, when used alone or simultaneously, in the development of apical periodontitis induced in rats, using a correlative analytic approach with micro-CT, histological and immunohistochemical analysis. METHODOLOGY: Twenty-eight male Wistar rats were arranged into four groups: Control, Nicotine, Alcohol and Alcohol + Nicotine. The alcohol groups were exposed to self-administration of a 25% alcohol solution, whilst the other groups drunk only filtered water. The nicotine groups received daily intraperitoneal injections of a solution with 0.19 µL of nicotine per mL, whilst the other groups received saline solution. The pulps of the left mandibular first molars were exposed for 28 days to induce periapical lesions. Throughout the experiment, drug administration was maintained, and the animals had their weight and solid and liquid consumption measured. After euthanasia, the mandibles were removed and the area, volume and major diameter of the periapical lesions were measured using micro-computed tomography images. The samples were submitted to histopathological evaluation and immunohistochemistry for RANKL and PTHrP. Statistical analysis was undertaken with a significance level of 5%. Nonparametric data were analysed using the Kruskal-Wallis test followed by Dunn's test, whilst one-way anova followed by Tukey's test was performed for parametric data. RESULTS: The alcohol groups had lower solid and liquid consumption and gained less weight when compared to the nonalcohol groups (P < 0.05). The Alcohol + Nicotine group had lesions with significantly larger volume and area when compared to the other groups (P < 0.05), whilst the Alcohol or Nicotine groups had significantly larger lesions than the control group (P < 0.05). There was no significant difference in the largest diameter of the lesions amongst groups (P > 0.05). The experimental groups had greater inflammatory response scores than the control group (P < 0.05), and the representative samples had more pronounced immunoreaction against RANKL and PTHrP antibodies. CONCLUSIONS: Alcohol and nicotine consumption exacerbated the inflammatory response and the development of periradicular lesions in rats. The association of both substances enhanced their harmful effects.

Does tobacco smoking predispose to apical periodontitis and endodontic treatment need? A systematic review and meta-analysis.

BACKGROUND: Controversial findings exist in the literature regarding the association between tobacco smoking and development of apical periodontitis or need for root canal treatment, with some studies reporting an increase in the prevalence of these outcomes in smokers, whilst others reporting no association. AIM: To evaluate if there is scientific evidence to support an association between tobacco smoking and a greater prevalence of apical periodontitis and/or root canal treatments. DATA SOURCES: A systematic search was performed using MeSH terms and free terms in the PubMed, Scopus, Virtual Health Library (VHL), Cochrane library and Open Gray databases. STUDY ELIGIBILITY CRITERIA, PARTICIPANTS AND INTERVENTIONS: Observational studies that evaluated the association between smoking and the development or healing of periapical lesion and/or the prevalence of root canal treatment in humans were included. STUDY APPRAISAL AND SYNTHESIS METHODS: A quality assessment of included studies was performed, and the relevant information and findings were evaluated. A meta-analysis using nine included studies was performed through RevMan software, and the certainty of evidence was evaluated through GRADE. RESULTS: Fifteen studies were included, of which 10 were classified as low risk of bias, 4 were considered as moderate risk of bias and 1 as high risk of bias. Ten studies reported that smoking was associated with a greater prevalence of periapical periodontitis and/or root canal treatment. Five studies reported no association. Nine studies were included in the pooled meta-analysis, comprising two subgroups: apical periodontitis and root canal treatment. From these nine studies, eight studies were included in the apical periodontitis subgroup meta-analysis and demonstrated that this condition was significantly more prevalent in smokers when compared to the nonsmokers (odds ratio = 2.78[CI: 1.60, 4.85], P < 0.001; I2  = 79%; no adjusted odds ratio). Three studies were included in the root canal treatment subgroup meta-analysis and demonstrated an increased prevalence in smokers when compared to the nonsmokers (odds ratio = 2.73 [CI: 1.06, 2.83], P < 0.001; I2  = 72%; no adjusted odds ratio). The pooled meta-analysis demonstrated that smokers had twice the chance of having apical periodontitis and/or root canal treatment when compared to nonsmokers (odds ratio of 2.42 [CI: 1.59, 3.68], P < 0.01; I2  = 85%). The GRADE analysis demonstrated moderate certainty of evidence. LIMITATIONS: Several confounding factors were identified such as misinformation regarding the period of time subjects had smoked, as well as the frequency of smoking and the number of cigarettes consumed. No adjustment in odds ratio data for confounding was performed. CONCLUSIONS AND IMPLICATIONS OF KEY FINDINGS: This systematic review and meta-analysis indicate that tobacco smokers have an increased prevalence of periapical periodontitis and root canal treatments with moderate certainty of evidence.

Do smear-layer removal agents affect the push-out bond strength of calcium silicate-based endodontic sealers?

AIM: To evaluate the effect of 17% ethylenediaminetetraacetic acid (EDTA), 2.25% peracetic acid (PA) or 10% citric acid (CA) on the push-out bond strength of calcium silicate-based endodontic sealers [MTA Fillapex (Angelus, Londrina, Brazil) and Total Fill BC Sealer (FKG, La Chaux-de-Fonds, Switzerland)] to root dentine after different periods of immersion in phosphate-buffered saline (PBS). AH Plus (Dentsply DeTrey, Konstaz, Germany) was used as a reference material for comparison. METHODOLOGY: Thirty human maxillary incisors were selected and three 1-mm discs were obtained from the middle third of each root. On the axial surface of each disc, three 0.8-mm-diameter holes were drilled through the dentine. After standardized irrigation, each dentine disc from the same root was treated for smear-layer removal with one of the three tested chelating solutions: EDTA, PA or CA. After that, the holes were dried with paper points and filled with one of the three test root canal sealers. The filled dental slices were further immersed in PBS solution (pH 7.2) for 7 days (T1) or 30 days (T2) at 37 °C, before the push-out assay. Friedman, Kruskal-Wallis and Mann-Whitney tests were used respectively to estimate the impact of the independent variables (solutions, sealers and period of PBS immersion) on the push-out bond strength. The level for significance was set at α = 5% and data were further subjected to a Bonferroni correction. RESULTS: Solutions and periods of immersion did not significantly influence the displacement resistance (P = 0.285, and P = 0.104, respectively) of sealers to root dentine. On the other hand, the type of sealer significantly affected the push-out bond strength (P = 0.000). AH Plus was associated with greater resistance to dislodgment compared with Total Fill BC Sealer and MTA Fillapex (P = 0.000). MTA Fillapex resulted in the lowest push-out bond strength (P = 0.000). CONCLUSION: The use of different chelating agents did not influence the push-out bond strength of endodontic sealers. Calcium silicate-based sealers had lower push-out bond strength values compared with a conventional epoxy resin-based sealer (AH Plus).

Influence of working length and apical preparation size on apical bacterial extrusion during reciprocating instrumentation.

AIM: To evaluate the influence of reciprocating single-file instrumentation with different working lengths (WL) and apical preparation sizes on apical bacterial extrusion. METHODOLOGY: Sixty-eight human single-rooted pre-molars were used. Conventional access cavities were prepared, and the root canals were contaminated with an Enterococcus faecalis suspension and incubated at 37°C for 30 days. Teeth were then divided into four groups of 15 specimens each (Reciproc size 25, .08 taper and Reciproc size 40, .06 taper instruments were used at the foramen; Reciproc size 25, .08 taper and Reciproc size 40, .06 taper instruments were used 1 mm short of the foramen). Positive and negative controls consisted of four infected and four uninfected pre-molars that were instrumented according to each experimental group. Bacteria extruded from the apical foramen during instrumentation were collected into vials containing 0.9% NaCl. The microbiological samples were then incubated in a brain-heart agar medium for 24 h. The resulting bacterial titre, in colony-forming units (CFU) per mL, was determined, and these data were analysed using a Wilcoxon matched-pairs signed rank test and a Kruskal-Wallis H-test. The level of significance was set at α = 0.05. RESULTS: No growth was observed in the negative control group. All positive controls demonstrated bacterial growth after the experimental time interval. No significant difference was found in the number of CFU amongst all experimental groups (P = 0.95). CONCLUSIONS: This study showed that the WL and the apical preparation size did not have a significant effect on bacterial extrusion when performing reciprocating instrumentation.

Microbial diversity in persistent root canal infections investigated by checkerboard DNA-DNA hybridization.

INTRODUCTION: The aim of the present study was to investigate the composition of the root canal microbiota in endodontic failures in order to identify and quantify these microorganisms. METHODS: Microbiological samples were taken from 36 root canals with persistent endodontic infection. The presence, levels, and proportions of 79 bacterial species were determined by checkerboard DNA-DNA hybridization. The Pearson correlation coefficient was used to investigate the relations between bacterial counts and clinical conditions (P ≤ .05). RESULTS: Enterococcus faecium (36%), Streptococcus epidermidis (36%), Eubacterium saburreum (28%), Parvimonas micra (28%), Streptococcus sanguis (28%), Capnocytophaga sputigena (28%), Leptotrichia buccalis (28%), Enterococcus faecalis (28%), and Staphylococcus warneri (28%) were the most prevalent species; and there was a low prevalence of Treponema socranskii (3%), Fusobacterium periodonticum (3%), Capnocytophaga gingivalis (3%), and Spiroplasma ixodetis (3%). The highest mean levels were found for the following species: E. faecium, Dialister pneumosintes, Staphylococcus epidermidis and Helicobacter pylori. There was a statistically significant difference between the levels of gram-negative species and gram-positive species (13.5 × 10(5) vs 6.5 × 10(5), respectively). A positive correlation was found between the area of the periapical lesion and the levels of gram-negative and rod species (P < .05). CONCLUSIONS: The microbiota from teeth with persistent apical periodontitis presents a mixed and complex profile, hosting E. faecium and S. epidermidis as the most highly prevalent species. No correlation was found between any of the species tested and clinical findings; however, periapical lesions with the largest areas presented higher counts of gram-negative and rod species.

Apical extrusion of bacteria when using reciprocating single-file and rotary multifile instrumentation systems.

AIM: To evaluate ex vivo, apical bacterial extrusion associated with two reciprocating single-file systems (WaveOne and Reciproc) compared with a conventional multifile rotary system (BioRace). METHODOLOGY: Forty-five human single-rooted mandibular incisors were used. Endodontic access cavities were prepared, and root canals were contaminated with an Enterococcus faecalis suspension. Following incubation at 37 °C for thirty days, the contaminated teeth were divided into three groups of 15 specimens each (G1 - Reciproc, G2 - WaveOne and G3 - BioRace). Positive and negative controls consisted of 5 infected teeth and 3 uninfected incisors that were instrumented with one of the tested NiTi systems, respectively. Bacteria extruded from the apical foramen during instrumentation were collected into vials containing 0.9% NaCl. The microbiological samples were taken from the vials and incubated in brain heart agar medium for 24 h. The resulting bacterial titre, in colony-forming units (CFU) per mL, was determined, and these data were analysed by Wilcoxon matched-pairs signed rank test and Kruskal-Wallis H-test. The level of significance was set at α = 0.05. RESULTS: No significant difference was found in the number of CFU between the two reciprocating systems (P = 0.41). The conventional multifile rotary system group was associated with significantly higher CFU than both of the two reciprocating groups (P = 0.01). CONCLUSIONS: All instrumentation systems extruded bacteria beyond the foramen. However, both reciprocating single-file systems extruded fewer bacteria apically than the conventional multifile rotary system.

Microbiological evaluation of primary endodontic infections in teeth with and without sinus tract.

AIM: To examine the microbiological status of primary endodontic infections in teeth with and without a sinus tract. METHODOLOGY: Samples were collected by means of a size 15 H-type file and two sterile paper points from 30 cases of primary endodontic infections with (n = 15) or without (n = 15) a sinus tract. The presence of 40 bacterial species was determined by the checkerboard DNA-DNA hybridization method. RESULTS: The species found at the highest levels and prevalence were Fusobacterium nucleatum sp. vincentii, Porphyromonas gingivalis, Veillonella parvula, Enterococcus faecalis, Campylobacter gracilis and Neisseria mucosa. Total bacterial counts were similar between teeth with (44 x 10(5)) and without (50 x 10(5)) a sinus tract (t-test: P > 0.05). E. faecalis, Streptococcus anginosus, Capnocytophaga sputigena and Capnocytophaga gingivalis had significantly higher counts in the absence of sinus tract (Mann-Whitney test, P < 0.05). Higher levels of P. gingivalis and Fusobacterium nucleatum sp. nucleatum were observed in cases with a sinus tract. Leptotrichia buccalis (OR = 1.83; CI 95%) and Porphyromonas endodontalis (OR = 2.15; CI 95%) were associated with an increased chance of subjects having a sinus tract. CONCLUSIONS: Primary endodontic infections were associated with a large variety of bacterial species. Specific differences between the composition of the microbiota of primary root canal infections were observed in cases with or without a sinus tract.

The influence of organic load on the antimicrobial activity of different concentrations of NaOCl and chlorhexidine in vitro.

AIM: To evaluate bacterial growth after contact with sodium hypochlorite (NaOCl; 1 and 5%) and chlorhexidine (CHX; 0.12, 0.5 and 1%) in vitro with or without the addition of organic material (bovine serum albumin (BSA) 0.5%). METHODOLOGY: Bacterial samples (American Type Culture Collection (ATCC)) of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Porphyromonas gingivalis and Fusobacterium nucleatum were kept in contact with each irrigating solution for varying intervals of time: immediately (t0), 5 min (t5), 15 min (t15) and 30 min (t30). Each test was repeated 10 times. In half of the specimens, 0.5% BSA was added as organic material in an attempt to simulate the organic tissue present in the root canal system. Bacterial growth under appropriate condition of incubation was evaluated and compared for each microorganism at all time intervals. RESULTS: A 0.12% CHX solution did not eliminate E. faecalis at any time interval. One percent CHX eliminated all strains, as did both NaOCl concentrations. BSA did not interfere substantially with the antimicrobial activity of any of the irrigating solutions. CONCLUSIONS: Under the condition of this study, a 0.12% CHX solution was ineffective at killing E. faecalis.