The effect of root canal preparation tapers on planktonic bacteria and biofilm reduction in the apical third: A correlative microtomography and microbiological laboratory study.

AIM: To evaluate the influence of different preparation tapers on the reduction in planktonic bacteria and biofilms of Enterococcus faecalis and Candida albicans in the apical third (4 mm) of the mesial roots of mandibular molars, correlating decontamination with canal shape. METHODOLOGY: After microtomography analysis for morphological standardization of the canals, 48 mandibular molar roots, each containing two canals (96 canals), were contaminated with E. faecalis and C. albicans and divided into four groups (n = 11) for canal instrumentation using ProDesign Logic 2 files with different tapers G (.03): # 25.03; G (.04): # 25.04; G (.05): # 25.05; and G (.06): # 25.06 and irrigation with 2.5% sodium hypochlorite. Four roots were examined under a scanning electron microscope (SEM) to qualitatively assess biofilm formation. Eight roots were used as the negative control group (samples were not contaminated). Bacteriological samples were taken exclusively from the apical third of the roots before and after chemical-mechanical preparation and bacterial counts were determined (CFU/mL). The final micro-CT scan was used to quantify the volume variation and unprepared canal area in the apical third. Statistical analysis was performed using the Kruskal-Wallis, Student-Newman-Keuls and Wilcoxon tests for analysis of microbiological data. anova and the Tukey or Games-Howell test were used for analysis of micro-CT data and Spearman's test for correlations (α = 5%). RESULTS: All groups showed a significant reduction in bacteria (p < .05), with no statistically significant difference between groups. There was no significant difference in per cent volume increase between groups. The unprepared area (Δ%) was affected by the file used (p = .026) and was significantly lower for G (.06) compared to G (.03). There was no statistically significant correlation among bacterial reduction, volume and unprepared area (p > .05). CONCLUSION: The different preparation tapers influenced root canal shaping in the apical third but did not improve decontamination in this region.

Microbiological aspects of traumatic injuries.

After traumatic injuries to teeth, microorganisms may invade the compromised pulp tissue and initiate pulp infection and periapical inflammation. In addition to bone resorption that typically accompanies pulp necrosis, root resorption frequently occurs. Root resorption has several variants that may occur shortly after the trauma or at a later stage. The pathological changes seen after traumatic injuries to teeth are invariably linked to the presence of microbial irritants. The presence of bacterial biofilms in the dental pulp space can be treated with regenerative or therapeutic endodontic procedures. However, necrosis of periodontal ligament is usually terminal for the tooth involved. In this review, the sources of bacteria after traumatic injuries are discussed. The types and role of microorganisms involved in the pathogenesis of endodontic pathosis after traumatic injuries are presented, and contemporary approaches for the management of these conditions are reviewed. Contemporary antimicrobial strategies are discussed. The rationale for the use of systemic and topical antimicrobials is presented. Finally, novel approaches to the use of antimicrobial therapies, particularly in regenerative procedures, are reviewed.

Microbiota in the apical root canal system of tooth with apical periodontitis.

BACKGROUND: Apical periodontitis (AP) is essentially an inflammatory disease of microbial etiology primarily caused by infection of the pulp and root canal system. Variation of the bacterial communities caused by AP, as well as their changes responding to dental therapy, are of utmost importance to understand the pathogensis of the apical periodontitis and establishing effective antimicrobial therapeutic strategies. This study aims to uncover the composition and diversity of microbiota associated to the root apex to identify the relevant bacteria highly involved in AP, with the consideration of root apex samples from the infected teeth (with/without root canal treatment), healthy teeth as well as the healthy oral. METHODS: Four groups of specimens are considered, the apical part of root from diseased teeth with and without root canal treatment, and wisdom teeth extracted to avoid being impacted (tooth healthy control), as well as an additional healthy oral control from biofilm of the buccal mucosa. DNA was extracted from these specimens and the microbiome was examined through focusing on the V3-V4 hypervariable region of the 16S rRNA gene using sequencing on Illumina MiSeq platform. Composition and diversity of the bacterial community were tested for individual samples, and between-group comparisons were done through differential analysis to identify the significant changes. RESULTS: We observed reduced community richness and diversity in microbiota samples from diseased teeth compared to healthy controls. Through differential analysis between AP teeth and healthy teeth, we identified 49 OTUs significantly down-regulated as well as 40 up-regulated OTUs for AP. CONCLUSION: This study provides a global view of the microbial community of the AP associated cohorts, and revealed that AP involved not only bacteria accumulated with a high abundance, but also those significantly reduced ones due to microbial infection.

Molecular Analysis of the Antibacterial Effects of Photodynamic Therapy in Endodontic Surgery: A Case Series.

In this case series, bacterial reduction promoted by antimicrobial photodynamic therapy (PDT) used during endodontic surgery was evaluated. Cases were also followed up, and the surgical outcome was reported. The study consisted of 19 teeth with posttreatment apical periodontitis that were consecutively treated by endodontic surgery. After apicoectomy, the root end was treated with PDT using methylene blue as the photosensitizer. Bacteriologic samples were taken from both the cut root surface and the root-end cavity before and after PDT. Samples were analyzed for the total bacterial and Streptococcus group counts using quantitative real-time polymerase chain reaction. EndoSequence BC-RRM Putty (Brasseler, Savannah, GA) was used as the root-end filling. Patients were followed up, and the surgical outcome was evaluated. The reduction in bacterial counts after the PDT approach was analyzed using the nonparametric Wilcoxon signed rank test. PDT significantly reduced the total bacterial and streptococcal counts in both root-end cavities and resected root surfaces (P < .05). The success rate for 15 teeth that were available for recall after 12 to 21 months was 93% under a loose evaluation criterion and 73% under a rigid one. Used during endodontic surgery, PDT significantly enhanced disinfection of the cut surface area and root-end cavity. Cases treated with PDT showed a high healing rate.

Management and Histobacteriological Findings of Mucosal Fenestration: A Report of 2 Cases.

This article describes 2 unusual cases of mucosal fenestration associated with necrotic infected teeth, resulting in exposure of the root apex to the oral cavity. Both cases consisted of maxillary incisors with pulp necrosis and radiographic/tomographic evidence of apical periodontitis. Clinically, the root apex was exposed to the oral cavity through a fenestration in both bone and mucosa and covered with bacterial plaque and calculus. These teeth were treated by a combination of nonsurgical and surgical endodontic treatment. During surgery, the root apices were resected to within the alveolus and the fenestrated area covered by the flap. Specimens consisting of the root apex and surrounding soft tissues were subjected to histopathological and histobacteriological analyses. Histobacteriological analysis revealed extensive resorptive defects on the root apices filled with thick bacterial biofilm, irregular detachment of the cementum layers with consequent infection of the underlying spaces, and heavy infection in the apical foramina. The soft tissue specimens exhibited no or minimal inflammation. The 2 cases showed satisfactory postsurgical healing of the hard and soft tissues. Both cases of mucosal fenestration showed root apices covered with dense bacterial biofilms and associated with a bone crypt with no significant inflammatory tissue therein. The 2 cases were successfully treated by conservative approaches involving a combination of nonsurgical and surgical endodontic treatment with root-end resection.

Bacteria and Hard Tissue Debris Extrusion and Intracanal Bacterial Reduction Promoted by XP-endo Shaper and Reciproc Instruments.

INTRODUCTION: This study used a multipurpose analytic approach to compare the levels of apically extruded bacterial and hard tissue debris as well as intracanal bacterial reduction after root canal preparation with either XP-endo Shaper (FKG Dentaire, La Chaux-de-Fonds, Switzerland) or Reciproc (VDW, Munich, Germany) instruments. METHODS: Distobuccal canals from extracted maxillary molars were contaminated with Enterococcus faecalis and randomly distributed into 2 groups according to the instrumentation system: the XP-endo Shaper or Reciproc. Teeth were mounted in an apparatus that simulates the apical resistance offered by the periapical tissues and permitted to collect debris extruded during preparation. Saline was used as the irrigant during preparation, and all treatment procedures were performed inside a cabinet under a controlled temperature of 37°C. DNA extracts from samples taken from the canal before and after preparation were subjected to quantitative real-time polymerase chain reaction for E. faecalis counting. The volume of extruded debris was evaluated by micro-computed tomographic imaging. DNA was extracted from the extruded hard tissue debris and analyzed by quantitative real-time polymerase chain reaction. RESULTS: Mechanical intracanal bacterial reduction was significantly more pronounced when using the XP-endo Shaper (P < .001). Although both instruments produced a similar volume of extruded debris (P > .05), extruded bacteria counts were significantly lower with Reciproc than the XP-endo Shaper (P < .001). No correlation was observed between the extruded bacterial counts and debris volume. CONCLUSIONS: Although bacterial extrusion was lower with Reciproc, the intracanal bacterial reduction was higher with the XP-endo Shaper. Both techniques produced a similar volume of hard tissue debris extrusion.

Detection of the unknown components of the oral microflora of teeth with periapical radiolucencies in a Turkish population using next-generation sequencing techniques.

AIM: To detect the unknown components of the oral microbiome and the effects of root canal treatment in a Turkish population and to evaluate the changes in microbial diversity in the root canals before and after treatment. METHODOLOGY: Single-rooted central and lateral maxillary incisors with one canal were chosen from 20 patients. Baseline samples of intact intracanal microbiota were collected from 20 root canals of permanent teeth with necrotic pulps using sterile paper points. After root canal preparation, the root canals were filled with a calcium hydroxide paste for 7 days. Calcium hydroxide was removed from root canal with 2.5% sodium hypochlorite and 17% EDTA using passive ultrasonic irrigation (PUI). A second bacteriologic samples were taken with sterile paper points prior to root filling. The samples were processes with DNase-I treatment using next-generation sequencing (NGS). Reduction in bacterial numbers during root canal treatment was evaluated using real-time quantitative PCR (qPCR). All statistical analyses were conducted using the MINITAB 17 software (Minitab Ltd. Co., Coventry, UK). A one-sample t-test was used to analyse the data. Statistical significance was accepted at P < 0.05. RESULTS: Relative abundances of Mycoplasma sp., Paludibacter sp., Tannerella sp., Prevotella spp. and an uncultured species from the order Bacteroidales decreased with root canal preparation and medication (98.7%, 99.8%, 98.8%, 97.7% and 99.3%, respectively), whilst the relative abundances of Methylobacterium sp., Corynebacterium sp. and Streptococcus infantis increased (93.1%, 94.8% and 99.4%, respectively). Considerable numbers of Streptophyta species were detected before and after treatment. The ratio of Agrobacterium sp. in the treated teeth community and the ratio of order Streptophyta in the infected canals had negative correlations with the success of bacterial elimination. CONCLUSIONS: The combination of NGS and qPCR techniques resulted in detection of previously unknown components of the oral microbiome and the effects of root canal treatment on their relative abundance in a Turkish population.

Molecular Typing of Enterococcus faecalis from Persistent Endodontic Infections / Tipificación Molecular de Enterococcus faecalis Provenientes de Infecciones Endodónticas Persistentes

ABSTRACT: Molecular techniques that provide valuable information about the epidemiology of oral strains. The purpose of this study was to determine the genetic relatedness of 83 Enterococcus faecalis strains isolated from treated root canals. These strains were obtained from patients who were treated for persistent endodontic infections. E. faecalis isolates were molecular typed by Pulsed Field Gel Electrophoresis using Smal. Ten clonal groups and 13 pulse types with 38.7 % similarity for the less related strains were identified. Genetic heterogeneity among strains from different patients and a high level of genetic homogeneity among intrapatient strains were observed. Therefore, restriction endonuclease fingerprinting of genomic DNA from E. faecalis strains confirmed the polyclonality of the isolates obtained from the root canals of patients diagnosed with persistent endodontic infections, compared with other reports. These results provide additional data for a better understanding of the epidemiological aspects of root canal infections by E. faecalis.

RESUMEN: Las técnicas moleculares proporcionan información valiosa sobre la epidemiología de aislados orales. El propósito de este estudio fue determinar la relación genética de 83 cepas de Enterococcus faecalis aisladas de conductos radiculares tratados. Estas cepas se obtuvieron de pacientes que fueron tratados por infecciones endodónticas persistentes. Los aislados de E. faecalis se tipificaron molecularmente por electroforesis en gel de campo pulsado usando Smal. Se identificaron diez grupos clonales y 13 pulsotipos con un 38,7 % de similitud para las cepas menos relacionadas. Se observó heterogeneidad genética entre las cepas de diferentes pacientes y un alto nivel de homogeneidad genética entre las cepas intrapacientes. Por lo tanto, la toma de huellas dactilares a traves de restricción de ADN genómico de cepas de E. faecalis confirmó la policlonalidad de los aislados obtenidos de los conductos radiculares de pacientes diagnosticados con infecciones endodónticas persistentes, en comparación con otros informes. Estos resultados proporcionan datos adicionales para una mejor comprensión de los aspectos epidemiológicos de las infecciones del conducto radicular por E. faecalis.

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.

Bacterial colonization in the apical part of extracted human teeth following root-end resection and filling: a confocal laser scanning microscopy study.

OBJECTIVES: The purpose of this study was to evaluate Enterococcus faecalis colonization at the apical part of root canals following root-end resection and filling using confocal laser scanning microscopy (CLSM). MATERIALS AND METHODS: The apical 3-mm root-ends of 55 extracted single rooted human teeth were resected, and 3-mm retrograde cavities were prepared and filled using either mineral trioxide aggregate (MTA), intermediate restorative material (IRM), or Biodentine (n = 10 each); 25 teeth served as controls. The roots were placed in an experimental model, sterilized, and coronally filled with E. faecalis bacterial suspension for 21 days. Then, the apical 3-mm segments were cut to get two slabs (coronal and apical). The slabs were stained using LIVE/DEAD BacLight Bacterial Viability Kit and evaluated using CLSM. RESULTS: The fluorescence-stained areas were larger in the bucco-lingual directions compared with the mesio-distal directions (p < 0.05). The mean and maximal depths of bacterial colonization into the dentinal tubules were 755 and 1643 µm, respectively, with no differences between the root-end filling materials (p > 0.05). However, more live bacteria were found in the MTA group in comparison to IRM and Biodentine groups (p < 0.05). CONCLUSIONS: CLSM can be used to histologically demonstrate bacterial root-end colonization following root-end filling. This colonization at the filling-dentine interfaces and deeper into the dentinal tubules may be inhomogeneous, favoring the bucco-lingual aspects of the root. CLINICAL RELEVANCE: Following root-end resection and filling bacterial colonization may lead to inflammatory reactions at the periapical tissues; the viability of the colonized bacteria may be affected by the type of root-end filling material.

Quantitative Assessment of apically Extruded Bacteria using different Instrumentation Techniques and Preparation Taper.

BACKGROUND: Cleaning and shaping of the pulp canal is one of the most important steps of endodontic therapy. Serious complications occur by the apical extrusion of bacteria during the instrumentation procedures. Both crown-down (CD) and full-length linear motion (FM) techniques are routinely used as a component of taper rotary instrument procedures for achievement of thorough cleaning and shaping of the pulp canal space. Hence, we aimed for this study to assess the change in the amount of apically extruded bacteria using CD and FM instrumentation techniques produced by differences in taper between the instruments used during biomechanical preparation of root canals. MATERIALS AND METHODS: The present study included assessments of 132 extracted maxillary central incisor teeth. To achieve a uniform teeth length of 21 mm, the height of the tooth crown was reduced for preserving the coronal portion of teeth. A modified glass vial model was constructed for the estimation of amount of bacterial extrusion through the apical region. For filling of each pulp canal specimen, 20 mL of Enterococcus faecalis suspension was used followed by the use of a number 10 K-file for carrying the bacteria down the lengths of pulp canals. All the contaminated teeth specimens were divided into six study groups with groups I to III containing specimens prepared in the CD manner, while groups IV to VI contained specimens prepared in the FM manner. Six teeth were taken as negative control with three specimens with each technique, and another six specimens were taken as positive controls. Cultivable bacterial counts were determined by evaluating 100 mL saline solution from each vial followed by its inoculation on blood agar. All the colony-forming unit (CFU) values were log-transformed (base 10), and the results were analyzed by Statistical Package for the Social Sciences software. RESULTS: A significantly lower quantity of CFU values was observed during CD instrumentation procedures with 0.02 files in comparison with all other study groups. However, while comparing both the instrumentation procedures when different taper files, other than 0.02 taper, were used for biomechanical preparation of root canal, nonsignificant results were obtained. CONCLUSION: With 0.02 taper preparations, significantly less amount of extrusion of bacteria is associated when done with CD technique. CLINICAL SIGNIFICANCE: No change in the amount of apical extrusion of bacteria will be seen by changing the type of instrumentation procedures. Amount of bacteria extruded can be minimized using 0.02 taper. Key words: Bacteria, Instrumentation, Taper.

Effect of Instrumentation Techniques and Preparation Taper on Apical Extrusion of Bacteria.

INTRODUCTION: The aim of this in vitro study was to evaluate the effects of different root canal instrumentation techniques and preparation tapers on the amount of apically extruded bacteria. METHODS: The root canals of 98 extracted human mandibular incisors were contaminated with Enterococcus faecalis suspension. After incubation at 37°C for 24 hours, the root canals were instrumented with K3 rotary files in a crown-down (CD) or full-length linear instrumentation technique (FL) by using 3 different root canal tapers (0.02, 0.04, and 0.06). During instrumentation, apically extruded bacteria were collected into vials containing saline solution. The microbiological samples were taken from the vials and incubated in brain-heart agar medium for 24 hours, and the numbers of colony-forming units (CFUs) were determined. The obtained results were analyzed with t test and one-way analysis of variance for the comparisons between the instrumentation techniques (CD and FL) and the preparation tapers (0.02, 0.04, and 0.06), respectively. Tukey honestly significant difference test was used for pairwise comparisons. RESULTS: The preparation taper had no effect on the number of CFUs when a FL instrumentation technique was used (P > .05). There was a statistically significant difference in the CFUs between FL and CD techniques when the preparation taper was 0.02 (P < .05). There was no statistically significant difference between the 0.04 and 0.06 preparation tapers in any of the instrumentation techniques (P > .05). CONCLUSIONS: Using a 0.02 taper in a CD manner results in the least amount of bacterial extrusion. The instrumentation technique did not seem to affect the amount of bacterial extrusion when 0.04 and 0.06 taper instruments were used for cleaning and shaping the root canal space.

Evaluation of Bacteriological Profile in the Apical Root Segment of the Patients with Primary Apical Periodontitis.

INTRODUCTION: Apical periodontitis usually results from bacterial accumulation and contamination occurring in the root-canal system, and extending beyond the apical foramen to involve the periapical tissues. Literature has a paucity of the studies that stress on the division and analysis of the pulp canal segments. The reason for this disparity might be the technique used for collecting the samples from the pulp canals. Hence, we carried out the present study to evaluate the microbial flora in the apical part of the roots with necrotic pulp canals. MATERIALS AND METHODS: The present study included the assessment of 40 freshly extracted teeth that had necrotized pulpal tissue along with the presence of periapical periodontal lesions. Removal of the soft tissue lesions attached to the root portion of the teeth along with apical periodontal lesions was done with the help of scalpel blade, after rinsing them with a sterile solution of saline. Thorough cleaning of the root surfaces was done with hydrogen peroxide followed by rapid disinfection with the help of sodium hypochlorite at varying concentrations. Sectioning of the root portion of all the specimens with the help of a disk was done perpendicular to the long axis of the teeth at a distance of roughly 5 to 6 mm from the teeth's apicalmost point. Cryotubes were used for transferring the specimens of apical portions containing 1 mL of buffer and were subjected to immediate frozen processing at a temperature of -20°C. A 10 K-type file was used for the initial collection of the samples followed by subsequent incubation of the files and paper pints in the incubation cabinet. Subsequent deoxyribonucleic acid (DNA) extraction from the samples was done following the procedure described by Siqueira et al. Paster et al's modification of the reverse-capture checkerboard assay was used in the present study. Semiquantitative data were used for overcoming the difficulties arising due to obtaining the counts of the polymerase chain reaction (PCR)-based analysis of specimens. RESULTS: A positive result for the 16S ribosomal ribonucleic acid (rRNA) gene primer was observed only in two examined specimens of all the samples of the apical portion of the root canals in the present study. Negative result was shown by all the control group specimens, which were sterile samples. Presence of bacteria was confirmed by PCR in 38 out of 40 examined specimens. Amount of bacterial taxa, out of these 24 samples, ranged up to 6. Pseudoramibacter alactolyticus, Porphyromonas endodontalis, Dialister oral species, Bacteroidetes species, Streptococcus species, Olsenella uli, Synergistes species, Fusobacterium nucleatum, Parvimonas micra, Treponema denticola, and Filifactor alocis were the specific species detected. Bacteroidetes species was the only species that were detected at levels at or above 105. Heavy bacterial infections were noticed in more than 45% of the cases at the periradicular part of the root canals. CONCLUSION: Microbial flora of the apical segment of the root with necrotized pulp tissue comprises a vast variety of pathogenic bacteria. CLINICAL SIGNIFICANCE: For better prognosis of the treatment of such cases, adequate knowledge of the microbial flora of the root, especially the apical portion is necessary.

Pulp and apical tissue response to deep caries in immature teeth: A histologic and histobacteriologic study.

Descriptions of the pathologic changes in the pulp and associated apical structures of human immature teeth in response to deep caries are lacking in the literature. OBJECTIVES: This article describes the histologic events associated with the radicular pulp and the apical tissues of human immature teeth following pulp inflammation and necrosis. METHODS: Twelve immature teeth with destructive caries lesions were obtained from 8 patients. Two intact immature teeth served as controls. Teeth were extracted for reasons not related to this study and immediately processed for histopathologic and histobacteriologic analyses. Serial sections were examined for the pulp conditions and classified as reversible or irreversible pulp inflammation, or pulp necrosis. Other histologic parameters were also evaluated. RESULTS: In the 3 cases with reversible pulp inflammation, tissue in the pulp chamber showed mild to moderate inflammation and tertiary dentin formation related to tubules involved in the caries process. Overall, the radicular pulp tissue, apical papilla and Hertwig's epithelial root sheath (HERS) exhibited characteristics of normality. In the 3 cases with irreversible pulp inflammation, the pulps were exposed and severe inflammation occurred in the pulp chamber, with minor areas of necrosis and infection. Large areas of the canal walls were free from odontoblasts and lined by an atubular mineralized tissue. The apical papilla showed extremely reduced cellularity or lack of cells and HERS was discontinuous or absent. In the 6 cases with pulp necrosis, the coronal and radicular pulp tissue was necrotic and colonized by bacterial biofilms. The apical papilla could not be discerned, except for one case. HERS was absent in the necrotic cases. CONCLUSION: While immature teeth with reversible pulpitis showed histologic features almost similar to normal teeth in the canal and in the apical region, those with irreversible pulpitis and necrosis exhibited significant alterations not only in the radicular pulp but also in the apical tissues, including the apical papilla and HERS. CLINICAL SIGNIFICANCE: Alterations in the radicular pulp and apical tissues help explain the outcome of current regenerative/reparative therapies and should be taken into account when devising more predictable therapeutic protocols for teeth with incomplete root formation.

Apical Extrusion of Intracanal Bacteria following use of Two Engine-driven Instrumentation Techniques: An in vitro Study.

AIM: The aim of the present study was to compare in vitro the amount of debris extruded apically from extracted teeth, using Twisted files and ProTaper rotary during two different instrumentation systems. MATERIALS AND METHODS: Forty-five human single-rooted mandibular premolar teeth were randomly divided into three groups and contaminated with Enterococcus faecalis. The teeth in experimental groups were instrumented until reaching the working length with ProTaper rotary instruments and Twisted files with XSmart and XSmart Dual groups. Debris extruded from the apical foramen was collected into glass vials and the amount of bacteria was calculated. The data obtained were analyzed using Kruskal-Wallis one-way analysis of variance and Mann-Whitney U tests, with p = 0.05 as the level for statistical significance. RESULTS: The XSmart Dual group extruded comparatively lesser bacteria compared to the XSmart group. Lesser amount of bacterial extrusion was seen when Twisted files were used compared to the ProTaper files (p < 0.05). CONCLUSION: Under the circumstances of this in vitro study, it can be concluded that all instrumentation techniques produced measurable apical extrusion of debris. So, it is upon the practitioner to decide which system best fits their individual needs and their level of skill and experience that will provide the best possible endodontic care for our patients. CLINICAL SIGNIFICANCE: The newer instrument designs, including noncutting tips, different cross sections, radial lands, and variable tapers, are better for the clinicians to improve working safety, to reduce the working time, and to create a greater flare within the preparations.

Adjunctive Steps for Disinfection of the Mandibular Molar Root Canal System: A Correlative Bacteriologic, Micro-Computed Tomography, and Cryopulverization Approach.

INTRODUCTION: This study evaluated the disinfecting ability of chemomechanical preparation with rotary nickel-titanium instruments, followed by 2 distinct adjunctive procedures in the root canals of extracted mandibular molars by means of a correlative analytical approach. METHODS: Twenty-two extracted mandibular molars were selected and anatomically matched between groups on the basis of micro-computed tomographic analysis. In the first phase of the experiment, root canals were contaminated with Enterococcus faecalis and subjected to chemomechanical preparation with BT RaCe instruments and 2.5% NaOCl irrigation. Then either XP-Endo Finisher instrument or passive ultrasonic irrigation was used to supplement disinfection. Micro-computed tomography was used to show whether the percentage of unprepared areas correlated to bacterial counts. In the second phase, the same teeth were contaminated once again, and the adjunctive procedures were used. Samples from the isthmus area of mesial roots and the apical 5-mm fragment of distal roots were obtained by cryopulverization. Samples taken before and after treatment steps in both phases were evaluated by quantitative polymerase chain reaction and statistically analyzed. RESULTS: In phase 1, preparation in both groups resulted in substantial decrease of bacterial counts (P < .001). The adjunctive approaches led to a further small bacterial reduction, which was significant for XP-Endo Finisher (P < .05). No significant differences were observed between groups for persisting bacterial counts. Correlative analysis revealed no statistically significant relationship between bacterial reduction and the percentage of unprepared areas (P > .05). In phase 2, both methods had significant antibacterial effects in the main canal, but none of them could predictably disinfect the isthmus/recess areas. CONCLUSIONS: Both XP-Endo Finisher and passive ultrasonic irrigation exhibited antibacterial effectiveness, but only the former caused a significant reduction in the bacterial counts after chemomechanical preparation. None of them were effective in predictably disinfecting the isthmus/recess areas.

Efficacy of 4 Irrigation Protocols in Killing Bacteria Colonized in Dentinal Tubules Examined by a Novel Confocal Laser Scanning Microscope Analysis.

INTRODUCTION: The aim of this study was to determine the efficiency of 4 irrigation systems in eliminating bacteria in root canals, particularly in dentinal tubules. METHODS: Roots of human teeth were prepared to 25/04, autoclaved, and inoculated with Enterococcus faecalis for 3 weeks. Canals were then disinfected by (1) standard needle irrigation, (2) sonically agitating with EndoActivator, (3) XP Endo finisher, or (4) erbium:yttrium aluminum garnet laser (PIPS) (15 roots/group). The bacterial reduction in the canal was determined by MTT assays. For measuring live versus dead bacteria in the dentinal tubules (4 teeth/group), teeth were split open and stained with LIVE/DEAD BackLight. Coronal, middle, and apical thirds of the canal dentin were scanned by using a confocal laser scanning microscope (CLSM) to determine the ratio of dead/total bacteria in the dentinal tubules at various depths. RESULTS: All 4 irrigation protocols significantly eliminated bacteria in the canal, ranging from 89.6% to 98.2% reduction (P < .001). XP Endo had the greatest bacterial reduction compared with other 3 techniques (P < .05). CLSM analysis showed that XP Endo had the highest level of dead bacteria in the coronal, middle, and apical segments at 50-µm depth. On the other hand, PIPS had the greatest bacterial killing efficiency at the 150-µm depth in all 3 root segments. CONCLUSIONS: XP Endo appears to be more efficient than other 3 techniques in disinfecting the main canal space and up to 50 µm deep into the dentinal tubules. PIPS appears to be most effective in killing the bacteria deep in the dentinal tubules.

Host-Bacterial Interactions in Post-treatment Apical Periodontitis: A Metaproteome Analysis.

INTRODUCTION: This study evaluated the bacterial and human metaproteome of root apexes and the matched inflammatory lesions from teeth with post-treatment apical periodontitis. METHODS: Root apexes and matched inflammatory lesions from root canal-treated teeth with apical periodontitis were obtained during periradicular surgery. All root canal fillings were rated as adequate on the basis of radiographs and cone-beam computed tomography. The specimens were cryopulverized and subjected to metaproteomic analysis for human and bacterial proteins by using a mass spectrometry platform that is based on nanoflow liquid chromatography coupled with linear ion trap quadrupole Velos Orbitrap. RESULTS: The metaproteome analyses revealed the presence of viable and metabolically active human and bacterial cells in both apexes and lesions. Several bacterial proteins of interest for pathogenicity and therapeutics were identified in both apexes and lesions, including proteins related to antibiotic resistance, proteolytic function, stress response, adhesion, and virulence. Many human proteins related to immune defense mechanisms were also detected in both root apex and lesion specimens, including immunoglobulins, complement system, and proteins linked to T-cell and B-cell activation, neutrophil microbicidal processes, antigen recognition/presentation, bone resorption, and protection against tissue damage. CONCLUSIONS: Occurrence of host defense factors from the innate and adaptive immune responses and bacterial virulence, survival, and resistance proteins in matched root apexes/periradicular inflammatory lesions indicates a complex and dynamic host-pathogen interaction in teeth with post-treatment apical periodontitis.

A Comparison of Apical Bacterial Extrusion in Manual, ProTaper Rotary, and One Shape Rotary Instrumentation Techniques.

INTRODUCTION: Apical extrusion of irrigants and debris is an inherent limitation associated with cleaning and shaping of root canals and has been studied extensively because of its clinical relevance as a cause of flare-ups. Many factors affect the amount of extruded intracanal materials. The purpose of this study was to assess the bacterial extrusion by using manual, multiple-file continuous rotary system (ProTaper) and single-file continuous rotary system (One Shape). METHODS: Forty-two human mandibular premolars were inoculated with Enterococcus faecalis by using a bacterial extrusion model. The teeth were divided into 3 experimental groups (n = 12) and 1 control group (n = 6). The root canals of experimental groups were instrumented according to the manufacturers' instructions by using manual technique, ProTaper rotary system, or One Shape rotary system. Sterilized saline was used as an irrigant, and bacterial extrusion was quantified as colony-forming units/milliliter. The results obtained were statistically analyzed by using one-way analysis of variance for intergroup comparison and post hoc Tukey test for pair-wise comparison. The level for accepting statistical significance was set at P < .05. RESULTS: All the instrumentation techniques resulted in bacterial extrusion, with manual step-back technique exhibiting significantly more bacterial extrusion than the engine-driven systems. Of the 2 engine-driven systems, ProTaper rotary extruded significantly more bacteria than One Shape rotary system (P < .05). CONCLUSIONS: The engine-driven nickel-titanium systems were associated with less apical extrusion. The instrument design may play a role in amount of extrusion.