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Introduction: This study characterized the bacteriome in primary endodontic infection (PEI) with apical periodontitis (AP), identified core and rare bacteriome species and community diversity metrics, and analyzed the relationship between the bacteriome composition, diversity and features, and patient variables., Methods: Twenty-seven patients with PEI and AP were sampled. The DNA was extracted and quantified using quantitative polymerase chain reaction. Raw V3-V4 amplicon sequencing data were processed with the DADA2 pipeline to generate amplicon sequence variants, and taxonomic assignment of the amplicon sequence variants up to the species level was done against the Human Oral Microbiome Database. Core bacteriome and differential abundance analyses were performed using ANCOM. Alpha diversity was determined using Chao1, Shannon, and Simpson indexes. LeFse analysis was used to identify abundant taxa. Sparse Estimation of Correlations among Microbiomes analysis estimated linear and nonlinear relationships among bacteria., Results: Of 27, 24 root canal samples were analyzed, and 3 root canal sampling were filtered out with a low read count. The bacterial phyla with top mean relative abundance were Bacteroidetes, Firmicutes, Synergistetes, Fusobacteria, and Actinobacteria. A total of 113 genera and 215 species were identified. The samples were gathered into 3 clusters. LefSe analysis identified differences in abundant taxa between distinct age, gender, symptomatology, and lesion size groups. Sparse Estimation of Correlations among Microbiomes distance analysis indicated Slackia exigua as the node with the highest degree., Conclusions: The bacteriome in PEI with AP among the patients in this study was complex and displayed high microbial heterogeneity. Moreover, age, gender, symptomatology, and lesion size were associated with differences in bacteriome features in PEI with AP., (Copyright © 2024 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: This study investigates the feasibility of a dynamic navigation system (DNS) small field of view workflow (DNS-SFOVw) for fiber-post removal and compares its accuracy and efficiency to the conventional large field of view workflow (DNS-LFOVw)., Methods: Fifty-six extracted human maxillary molars were divided into DNS-SFOVw (n = 28) and DNS-LFOVw (n = 28). The palatal canal was restored with an intraradicular RelyX fiber post and luted with RelyX Unicem; a core buildup was used. Teeth were mounted in a 3D-printed surgical jaw. A preoperative cone-beam computed tomography (CBCT) scan was taken with a 40 × 40 mm FOV for the DNS-SFOVw and a single arch CBCT scan for the DNS-LFOVw. The drilling entry point, trajectory, angle, and depth were planned in the X-guide software. The DNS registration method for the DNS-SFOVw was virtual-based registration on teeth, and the marker point-based method was used for the DNS-LFOVw. The fiber posts were drilled out under DNS guidance. A postoperative CBCT scan was taken. Three-dimensional deviations, angular deflection, number of mishaps, registration, and total operation time were calculated., Results: The DNS-SFOVw was as accurate as DNS-LFOVw (P > .05). The DNS-LFOVw registration time was less than DNS-SFOVw (P < .05). There was no difference in the number of mishaps (P > .05). Both DNS-SFOVw and DNS-LFOVw were time-efficient, with DNS-LFOVw taking less total operational time (P < .05)., Conclusion: Within the limitations of this in-vitro study, the DNS-SFOVw was as accurate as the DNS-LFOVw for fiberpost removal. Both DNS-LFOVw and DNS-SFOVw were time-efficient in removing fiber-posts., (Copyright © 2024 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: To investigate the bacteriome present in teeth with primary endodontic infection (PEI) and apical periodontitis (AP) and to determine quantitatively and qualitatively the impact of chemomechanical preparation (CMP) using 2.5% sodium hypochlorite NAOCl on the bacteriome found in PEI with AP using the Illumina MiSeq platform., Methodology: Thirty-six paired samples from 18 patients were successfully sequenced and analysed. Samples were collected at two sampling times: before (s1) and after (s2) CMP using 2.5% NaOCl. The DNA was extracted from s1 and s2 samples and quantified using quantitative PCR (qPCR). All 36 samples were sequenced using the Illumina MiSeq platform. Raw V3-V4 amplicon sequencing data were processed with the DADA2 pipeline to generate amplicon sequence variants (ASVs). Alpha diversity metrics representing abundance (Chao1) and diversity and evenness (Shannon, Simpson) were computed. The paired-sample Wilcoxon's test was used to compare alpha diversity metrics and qPCR counts between s1 and s2. The PERMANOVA method (with 999 permutations) was applied to compare community composition between sample types (s1 versus s2) and between patient IDs. ALDEx2 (ANOVA-like differential expression tool for high-throughput sequencing data) to investigate differentially abundant taxa between s1 and s2. A paired-sample Wilcoxon's test was used to compare alpha diversity metrics and qPCR counts between s1 and s2., Results: The qPCR counts were significantly higher in s1 compared to s2 (p = .0007). The Chao1 index indicated no difference in alpha diversity (p < .7019); whereas Shannon (p = .0056) and Simpson (p = .02685) indexes showed higher values in s2. The PERMANOVA test using Adonis2 showed a significant effect of sample time on community composition (R 2  = .0630, p = .012). Patient ID also showed a significant effect on community composition (R 2  = .6961, p = .001). At the genus level, Dialister, Mogibacterium, Prevotella, and Olsenella were differentially enriched at s1, while Actinomyces, Stenotrophomonas&#95;unclassified, Enterococcus&#95;unclassified, and Actinomyces&#95;unclassified were differentially enriched in s2., Conclusion: The bacteriome present in teeth with PEI with AP is complex and diverse. CMP using 2.5% NaOCl showed a high quantitatively and qualitatively disinfectant impact on the bacteriome present in PEI with AP., (© 2024 The Authors. International Endodontic Journal published by John Wiley & Sons Ltd on behalf of British Endodontic Society.)

Introduction: The aim of this study was to investigate the feasibility of a real-time three-dimensional dynamic navigation system (3D-DNS) for post space preparation (PSP) in root canal-treated teeth and to compare the accuracy and efficiency of 3D-DNS to freehand (FH) for PSP., Methods: Fifty-four maxillary molars were divided into two groups: 3D-DNS (n = 27) and FH group (n = 27). Cone beam computed tomography (CBCT) scans were taken preoperatively and postoperatively. The drilling path for the PSP was virtually planned in the preoperative CBCT scan in the X-guide software (X-Nav Technologies, Lansdale, PA). For the 3D-DNS group, the PSP drilling was conducted under dynamic navigation. The 3D deviations and angular deflections were calculated. The residual dentin thickness (RDT) was determined after PSP. The operation time and the total number of mishaps were recorded. Shapiro-Wilk, t-test or Mann-Whitney rank sum, weighted Cohen's kappa, and Fisher exact tests were used (P < .05)., Results: The PSP was completed in all samples (54/54). The 3D-DNS was more accurate than the FH, with significantly fewer 3D deviations and angular deflections (all, P < .05). The 3D-DNS required less operating time than the FH (P < .05). For the 3D-DNS, no teeth had RDT < 1 mm, whereas 6/27 in the FH showed RDT < 1 mm after the PSP. There was no difference in the total number of mishaps (P > .05)., Conclusion: Within the limitations of this in vitro study, the 3D-DNS is feasible for PSP. The 3D-DNS improved the accuracy and efficiency of PSP. The dynamic navigation system can potentially become a safe and reliable technology for PSP., (Copyright © 2024 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: This study evaluates the feasibility of an augmented reality (AR) head-mounted device (HMD) displaying a dynamic navigation system (DNS) in the surgical site for fiber postremoval in maxillary molars and compares it to the DNS technique., Methods: Fifty maxillary first molars were divided into 2 groups: AR HMD + DNS (n = 25) and DNS (n = 25). The palatal canal was restored with RelyX fiber post (3M ESPE) luted with RelyX Unicem (3M ESPE). A core buildup was performed using Paracore (Coltene/Whaledent). Cone beam computed tomography (CBCT) scans were taken before and after postremoval. The drilling trajectory and depth were planned under X-guide software (X-Nav Technologies, Lansdale, PA). For the AR HMD + DNS group, the AR HMD (Microsoft HoloLens 2) displayed the DNS in the surgical site. The three dimensional (3D) deviations (Global coronal deviation [GCD] and global apical deviation [GAD]) and angular deflection (AD) were calculated. The number of mishaps and operating time were recorded., Results: Fiber post was removed from all samples (50/50). The AR HMD + DNS was more accurate than DNS, showing significantly lower GCD and GAD deviations and AD (P < .05). No mishap was detected. The AR HMD + DNS was as efficient in time as DNS (P > .05)., Conclusions: Within the limitations of this in vitro study, the AR HMD can safely display DNS in the surgical site for fiber post-removal in maxillary molars. AR HMD improved the DNS accuracy. Both AR HMD + DNS and DNS were time-efficient for fiber postremoval in maxillary molars., (Copyright © 2024 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: This study investigates the feasibility of integrating a piezoelectric device (PIEZO) into a dynamic navigation system (DNS) for bone-window guided surgery. It compares the accuracy and efficiency of PIEZO + DNS to PIEZO + Freehand (FH) procedure for bone-window cutting and root-end resection (RER)., Methods: Forty-eight mandibular molars of 3D-printed surgical jaw models were divided into two groups: PIEZO + DNS (n = 24) and PIEZO + FH (n = 24). Cone-beam computed tomography scans were taken before and after the procedure. The procedure was virtually planned on X-guide software. The bone-window cutting and RER were conducted with a PIEZO under dynamic navigation in the PIEZO + DNS group and using the dental operating microscope in the PIEZO + FH group. The 2D- and 3D-accuracy deviations and angular deflection were measured for the bone window cut. The root length resected and resection angle were calculated. The bone window cut, RER, total operating time, and number of mishaps were recorded., Results: PIEZO + DNS was more accurate than PIEZO + FH for bone-window cutting, showing fewer 2D and 3D deviations and less angular deflection (P < .05). The resection angle was lower in the PIEZO + DNS (P < .05). The bone-window cut and total operating time were significantly reduced using a DNS (P < .05). There was no difference in the number of mishaps (P > .05)., Conclusions: Within the limitations of this in vitro study, the integration of a PIEZO into a DNS is feasible for bone-window guided surgery. The DNS improved the accuracy and efficiency of bone-window cutting., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: Augmented reality (AR) superimposes high-definition computer-generated virtual content onto the existing environment, providing users with an enhanced perception of reality. This study investigates the feasibility of integrating an AR head-mounted device into a 3-dimensional dynamic navigation system (3D-DNS) for osteotomy and root-end resection (RER). It compares the accuracy and efficiency of AR + 3D-DNS to 3D-DNS for osteotomy and RER., Methods: Seventy-two tooth roots of 3D-printed surgical jaw models were divided into two groups: AR + 3D-DNS (n = 36) and 3D-DNS (n = 36). Cone-beam computed tomography scans were taken pre and postoperatively. The osteotomy and RER were virtually planned on X-guide software and delivered under 3D-DNS guidance. For the AR + 3D-DNS group, an AR head-mounted device (Microsoft HoloLens 2) was integrated into the 3D-DNS. The 2D- and 3D-deviations were calculated. The osteotomy and RER time and the number of procedural mishaps were recorded., Results: Osteotomy and RER were completed in all samples (72/72). AR + 3D-DNS was more accurate than 3D-DNS, showing lower 2D- and 3D-deviation values (P < .05). The AR + 3D-DNS was more efficient in time than 3D-DNS (P < .05). There was no significant difference in the number of mishaps (P > .05)., Conclusions: Within the limitations of this in vitro study, the integration of an AR head-mounted device to 3D-DNS is feasible for osteotomy and RER. AR improved the accuracy and time efficiency of 3D-DNS in osteotomy and RER. Head-mounted AR has the potential to be safely and reliably integrated into 3D-DNS for endodontic microsurgery., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: This study compared the accuracy and efficiency of fully guided static and dynamic computer-assisted surgical navigation techniques for osteotomy and root-end resection (RER)., Methods: Fifty roots from cadaver heads were divided into two groups: fully guided static computer-assisted endodontic microsurgery (FG sCAEMS) and dynamic computer-assisted endodontic microsurgery (dCAEMS) (all, n = 25). Cone-beam computed tomography scans were taken pre and postoperatively. The osteotomy and RER were planned virtually in the preoperative cone-beam computed tomography scan and guided using 3D-printed surgical guides in the FG sCAEMS and 3D-dynamic navigation system in the dCAEMS. The 2D and 3D deviations and angular deflection were calculated. The osteotomy volume, resected root length, and resection angle were measured. The osteotomy and RER time and the number of procedural mishaps were recorded., Results: FG sCAEMS was as accurate as dCAEMS, with no difference in the 2D and 3D deviation values or angular deflection (P > .05). The osteotomy and RER time were shortened using FG sCAEMS (P < .05). The FG sCAEMS showed a greater number of incomplete RERs than dCAEMS. Osteotomy volume, RER angle, and root length resected were similar in both groups (P > .05). FG sCAEMS and dCAEMS were feasible for osteotomy and RER., Conclusions: Within the limitations of this cadaver-based study, FG sCAEMS was as accurate as dCAEMS. Both FG sCAEMS and dCAEMS were time-efficient for osteotomy and RER., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This study aimed to evaluate Nd:YAG laser, calcium phosphate, and adhesive system effect as different pretreatments in different protocols on dentin permeability (DP) and bond strength (BS). Fifty human dentin discs were used (4 mm in diameter and 1,5 mm in height). Specimens were divided into five groups ( n  = 10): (A): adhesive system (control); (AL): adhesive system + Nd:YAG laser; (LAL): Nd:YAG laser + adhesive system + Nd:YAG laser; (PAL): calcium phosphate-based dentin desensitizer TeethMate + adhesive system + Nd:YAG laser; and group (PLAL): Nd:YAG laser + calcium phosphate-based dentin desensitizer + adhesive system + Nd:YAG laser. All materials were used according to the manufacturers' instructions. The specimens were submitted to artificial aging (5,000 thermal cycles and 12 × 10 4 mechanical cycles) then a bond test was performed. DP was measured using the split chamber model. Data were submitted to one-way analysis of variance (ANOVA), paired t -test, RM ANOVA, and Tukey test ( p < 0.05). All treatments were effective in DP reduction. For BS, the groups PAL and PLAL had improved BS with a statistically significant difference of the control group (A). Nd:Yag laser irradiation and calcium phosphate-based desensitizing agents significantly reduced DP, and the association between them could improve the BS on resin-human dentin interface., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2023 Amjad Abu Hasna et al.)

This scoping review (SCR) was conducted to map the existing literature on dynamic navigation system (DNS), to examine the extent, range, and nature of research activity. Additionally, this SCR disseminates research findings, determines the value of conducting a full systematic review with meta-analysis, and identifies gaps in the existing literature and future directions. This SCR followed Arksey and O'Malley's five stages framework. The electronic search was performed in PubMed (Medline), Scopus (Elsevier), and Web of Science (Clarivate Analytics) databases using a search strategy. Five themes emerged during the descriptive analysis that captured the DNS application in endodontics. The DNS has been explored for creating access cavities (8/18, 44.44%), locating calcified canals (4/18, 22.2%), microsurgery (3/18, 16.6%), post removal (2/18, 11.1%), and intraosseous anesthesia (1/18, 5.5%). Out of the 18 studies included, 12 are in vitro (66.6%), five are in vivo (case report) (27.7%), and one is ex vivo (5.5%). The DNS demonstrated accuracy and efficiency in performing minimally invasive access cavities, locating calcified canals, and performing endodontic microsurgery, and it helped target the site for intraosseous anesthesia., Competing Interests: None declared., (The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/).)

Introduction: This study compared the accuracy and efficiency of a novel static computer-aided surgical technique using a 3-dimensional (3D)-printed surgical guide (3D-SG) with a fully guided drill protocol (3D-SG FG) to the freehand (FH) osteotomy and root-end resection (RER)., Methods: Forty-six roots from 2 cadaver heads were divided into 2 groups: 3D-SG FG (n = 23) and FH (n = 23). Cone-beam computed tomographic scans were taken preoperatively and postoperatively. The endodontic microsurgery was planned in Blue Sky Bio software, and the 3D-SG was designed and 3D printed. The osteotomy and RER were conducted using a guided twist drill diameter of 2 mm and an ascending tapered drill with diameters of 2.8/3.2, 3.2/3.6, 3.8/4.2, and 4.2 mm with respective guided drill guides. Two-dimensional and three-dimensional virtual deviations and angular deflection were calculated. Linear osteotomy measures and root resection angle were obtained. The osteotomy and RER time and the number of mishaps were recorded., Results: Two-dimensional and three-dimensional accuracy deviations and angular deflection were lower in the 3D-SG FG protocol than in the FH technique (P < .05). The height, length, and depth of the osteotomy and root resection angle were less in the 3D-SG FG protocol than in the FH technique (P < .05). The osteotomy and RER time with the 3D-SG FG protocol were less than the FH method (P < .05)., Conclusions: Within the limitations of this cadaver-based study using denuded maxillary and mandibular jaws, 3D-SG FG protocol showed higher accuracy than FH osteotomy and RER. Moreover, the 3D-SG FG drill protocol significantly reduced the surgical time., (Copyright © 2023 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Objective: To compare 2 different resin composites and 2 adhesive systems used in a new restorative protocol (partial restoration) to treat non-carious cervical lesions associated with gingival recession type 1 (RT1)., Material and Methods: Eighty combined defects (CDs) were treated with a partial restoration and periodontal plastic surgery for root coverage. The CDs were randomly assigned to one of the following groups: NP + TE (n = 20), nanofilled composite and 2-step total-etch adhesive system; NP + UA (n = 20), nanofilled composite and universal adhesive system; MH + TE (n = 20), microhybrid composite and 2-step total-etch adhesive; MH + UA (n = 20), microhybrid composite and universal adhesive. Restorations were assessed using the United States Public Health Service (USPHS) criteria at 1 week (baseline) and 6, 12, and 24 months. Survival rate, periodontal parameters, dentin hypersensitivity (DH), and aesthetics were also evaluated., Results: After 24 months, only the MH + TE group did not lose any restoration, with no significant differences between groups. For surface roughness parameter, MH presented 83.3% of the restorations scoring Bravo, whereas NP presented 48.5% of the restorations scoring Bravo. All groups presented restorations with marginal discoloration. All periodontal parameters behaved similarly, regardless of the restorative material. All groups presented significant reductions of dentin hypersensitivity and improved aesthetic perceptions (p < 0.05)., Conclusion: Both resin composites and adhesives tested can be combined for partial restorations to treat CDs., Clinical Relevance: This new restorative-surgical protocol to treat CDs presents satisfactory outcomes. The partial restorations can be successfully executed with both combinations of adhesives and resin composites evaluated in this investigation. TRN : ClinicalTrial.gov: NCT03215615; registration date July 12, 2017., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: This RCT investigated the impact of N-acetylcysteine (NAC) and calcium hydroxide [Ca(OH) 2 ] intracanal medications (ICMs) in primary endodontic infection with apical periodontitis (PEIAP)., Materials and Methods: Thirty-six teeth with PEIAP were randomly divided into groups according to the ICM: NAC, Ca(OH) 2  + saline solution (SSL), and Ca(OH) 2  + 2% chlorhexidine-gel (2% CHX-gel) (all, n = 12). Root canal samples (RCSs) were collected before (s1) and after instrumentation (s2) and after 14 days of ICM (s3). Chemomechanical preparation (CMP) was performed with a Reciproc file and 2.5% NaOCl. Checkerboard DNA-DNA hybridization was used to assess 40 target bacteria species., Results: At s1, bacterial DNA was detected in 100% of RCSs (36/36). All 40 bacterial species were found in PEIAP. The mean number of species per RCS was 17.92 ± 13.18. The most frequent bacteria were S. mitis (65%), E. nodatum (63%), E. faecalis (63%), F. nucl sp vicentii (58%), T. forsythia (58%), and F. periodonticum (56%). CMP reduced the mean number of species per RCS to 6.8 ± 2.36 (p < 0.05). At s3, the intragroup analysis revealed a broader antimicrobial activity for Ca (OH) 2  + 2% CHX-gel and NAC than Ca(OH) 2  + SSL (p < 0.05). NAC eliminated 8/12 bacteria species resistant to both Ca (OH) 2 ICMs, including P. micra, P. nigrescens, T. denticola, A. israelii, P. endodontalis, P. acnes, C. ochracea, and E. corrodens., Conclusions: Ca (OH)2 + 2% chlorhexidine gel (2% CHX gel) showed a greater bacterial elimination over the number of bacterial species; however, NAC eliminated 8/12 bacteria species resistant to both Ca (OH)2 ICMs (RBR-3xbnnn)., Clinical Relevance: The use of intracanal medication with a broad antimicrobial activity can optimize root canal disinfection. Ca(OH) 2  + 2% CHX gel and NAC showed a broader antimicrobial activity than Ca(OH) 2  + SSL against endodontic pathogens in primary root canal infection., Trial Registration: Brazilian Registry of Clinical Trials (REBEC), No. RBR-3xbnnn., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study devised a dual-species biofilm model to investigate bacteria, lipoteichoic acid (LTA), and lipopolysaccharide (LPS) simultaneously, and compared the efficacy of conventional and contemporary disinfection protocols. Seventy single-rooted mandibular premolars were included. Fourteen teeth were negative control, and 56 teeth were infected with 3-week-old E. faecalis and E. coli GFP biofilm. Fourteen/56 teeth were positive control, with seven teeth processed for CLSM analysis and seven teeth sampled with paper points (PPs) and cryogenically ground for bacterial, LTA, and LPS analyses. Forty-two teeth were randomly divided into three groups: GWS (GentleWave system) + MIT (minimally invasive technique), PUI (passive ultrasonic irrigation) + CIT (conventional instrumentation technique), and XP-EF (XP-endo Finisher) + CIT (All, n = 14). Samples were collected before (s1) and after disinfection (s2) with PPs and after cryogenically ground (s3). CFUs were counted, and LTA and LPS were quantified with LTA-ELISA and LAL assay, respectively. XP-EF was as effective as PUI (p > 0.05). GWS + MIT was the most effective disinfection protocol against bacteria, LTA, and LPS (p < 0.05). In conclusion, PUI, XP-EF, and GWS were highly effective against bacteria, LTA, and LPS, with GWS being the most effective., (© 2023. The Author(s).)

Introduction: The aim of the study is to investigate whether the 3-dimensional dynamic navigation system (3D-DNS) can improve experienced endodontists' (EEs') and novice endodontists' (NEs') accuracy and efficiency in osteotomy and root-end resection (RER) and to verify that the 3D-DNS enables NEs to perform osteotomy and RER as accurately and efficiently as EEs., Methods: Seventy-six roots in cadaver heads were randomly divided into 4 groups: 3D-DNS-NE, 3D-DNS-EE, freehanded (FH)-NE, and FH-EE (all, n = 19). Cone-beam computed tomography scans were taken preoperatively and postoperatively. Osteotomy and RER were planned virtually in the X-guided software (X-Nav Technologies, Lansdale). Accuracy was calculated by measuring the 2-dimensional and 3D virtual deviations and angular deflection using superimposing software (X-Nav technologies). Efficiency was determined by the time of operation and the number of mishaps., Results: Accuracy deviations were significantly fewer in the 3D-DNS-EE group than those in the FH-EE group (P < .05). We found less 2-dimensional and 3D accuracy deviations comparing the 3D-DNS-NE group to the FH-NE group (P < .05). The time required for osteotomy and RER with the 3D-DNS was ∼ ½ of that required for the FH method for both EEs and NEs (P < .05). We found no difference in the number of mishaps between the 3D-DNS and FH groups for EEs and NEs (P > .05)., Conclusions: The 3D-DNS improved EEs' and NEs' accuracy and efficiency in osteotomy and RER. The NEs were as efficient as the EEs using the 3D-DNS. Notably, the 3D-DNS improved the NEs' accuracy compared to the FH method, but the 3D-DNS did not enable the NEs to perform osteotomy and RER as accurately as the EEs., (Copyright © 2022 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: The lipopolysaccharides-dentine-infection (LPS-dentine-infection) models and sampling techniques frequently used to evaluate LPS disinfection have limitations. In this study, a LPS-dentine-infection model was devised using fluorescent conjugate LPS. Secondly, a sampling technique using cryogenic grinding for intraradicular LPS analysis was evaluated. Thirdly, the effectiveness of the XP-endo Finisher (XP-EF) was compared with passive ultrasonic irrigation (PUI) in removing LPS from root canal system., Methodology: Sixty-nine mandibular premolars were submitted to dentine pretreatment and inoculated with fluorescent LPS conjugate (Alexa Fluor® 594). Twenty-three teeth were analysed under confocal laser scanning microscopy (CLSM) to validate this modified LPS-dentine-infection model. Forty-six teeth were randomly divided into two experimental groups: XP-EF (n = 23) and PUI (n = 23). All teeth were instrumented with XP-endo shaper (XPS; FKG Dentaire) and 2.5% NaOCl. The root canals were sampled with paper points before (s1) and after (s2) instrumentation and after supplemental treatment (s3) with XP-EF and PUI. After s3, all roots were cryogenically ground for intraradicular LPS analysis (s4). Limulus amebocyte lysate assay was used for LPS quantification. The Friedman test was used for differences in LPS among four time-points (s1, s2, s3, and s4). Dunn's test was used for pairwise testing of time-points. The significance level was set at 5% (p < .05)., Results: Fluorescent LPS conjugate was detected in 100% of the samples under CLSM with a penetration depth of approximately 400 μm into dentine. Chemo-mechanical preparation using XPS files significantly reduced LPS levels (p < .05). Both the XPS and PUI improved the LPS disinfection (p < .05), with no difference between them (p > .05). LPS was recovered from all samples after cryogenic grinding. The residual amount of LPS detected using the cryogenically sampling technique at s4 was approximately three times greater than with the paper-point sampling technique at s3., Conclusion: This study established a modified LPS-dentine-infection model using fluorescent conjugate LPS, and validated a LPS sampling technique for using cryopulverization intraradicular LPS analysis. Moreover, both the XP-EF and PUI further improved LPS disinfection from the root canals, and the innovative XP-EF was as effective as PUI., (© 2022 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Introduction: This Consolidated Standards of Reporting Trials randomized clinical trial investigated T helper (Th1, Th2, Th9, Th17, and Tfh) and regulatory T (Treg) cell-type cytokines and their networks in apical periodontitis (AP). We also assessed the effects of calcium hydroxide [Ca(OH)2] intracanal medications (ICMs) on helper T and Treg cell-type cytokines., Methods: Twenty teeth with primary endodontic infection and apical periodontitis were randomly divided into two groups: Ca(OH)2 + saline solution (n = 10) and Ca (OH)2 + 2% chlorhexidine-gel (n = 10). Samples were collected from the periradicular tissue fluid (PTF) before (PTFs1) and after 14 days of ICMs (PTFs2). The Human High Sensitivity T Cell Panel was used to quantify target T-helper (Th)1: interelukin (IL)-2, IL-12, and interferon-gamma (IFN-γ); Th2: IL-4, IL-5, and IL-13; Th9: IL-9; Th17: IL-17; T follicular helper cells (Tfh): IL-21; and Treg-cell-type cytokine: IL-10., Results: Th1-type cytokines were higher than Th2-type ones, at PTFs1. Positive (+) associations were found among all Th1-type cytokines and all Th2-type cytokines. There were negative (-) correlations between all Th1- and Th2-type cytokines. Size of radiolucent lesions and symptoms (tenderness to percussion and/or pain on palpation) were positively correlated with Th1-type cytokines, IL-17, and IL-21 but negatively correlated with Th2-type cytokines and IL-10 (all, P < .001). Both ICMs increased Th2-type cytokines and IL-10 (P < .05) and decreased Th1-type cytokines, IL-17, and IL-21 (P < .05), with no differences among them (P > .05)., Conclusions: Complex T-cell cytokine networks are involved in AP. Both Ca(OH)2 ICMs effectively increased IL-4, IL-5, IL-10, and IL-13 and lowered IL-2, IL-12, IL-17, IL-21, and IFN-γ., (Copyright © 2022 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: This study evaluated the accuracy and efficiency of the 3-dimensional dynamic navigation system (3D-DNS) to perform minimally invasive osteotomy (MIO) and root end resection (RER) in endodontic microsurgery (EMS) and investigated the viability of root end cavity preparation (RECP) and root end fill (REF) in MIO., Methods: Forty-eight tooth roots were divided in cadaver heads into 2 groups: 3D-DNS (n = 24) and freehand (n = 24). Cone-beam computed tomographic scans were taken before and after surgery. First, virtual 3D-DNS accuracy was verified using 3 outcome measures: 2-dimensional and 3-dimensional virtual deviations and angular deflection. Second, the accuracy of 3D-DNS for performing MIO was investigated in 2 outcome measures: osteotomy size and volume. Third, the 3D-DNS accuracy was determined for RER in 3 outcomes: resected root length, root length after resection, and resection angle. The viability of RECP and REF was investigated and REF depth and volume measured as well, and procedural time and the number of mishaps were recorded., Results: Two- and 3-dimensional virtual deviations and the angular deflection were lower in the 3D-DNS group than the freehand group (P < .05). Osteotomy height, length, and volume were all reduced when using 3D-DNS (P < .05). The resection angle was lower for 3D-DNS (P < .05). RECP and REF were completed in 100% of the roots. The REF depth achieved was ∼3 mm. Osteotomy time, RER time, and the total procedure time were all significantly shortened using 3D-DNS (P < .05)., Conclusions: 3D-DNS enabled our surgeon to perform accurate and efficient EMS with minimally invasive osteotomy and RER. The surgeon was also able to conduct RECP with adequate REF in minimally invasive osteotomy performed using 3D-DNS guidance., (Copyright © 2022 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: This study evaluated the effectiveness of the GentleWave System (GWS; Sonendo, Laguna Hills, CA) and passive ultrasonic irrigation (PUI) in removing Enterococcus faecalis lipoteichoic acid (LTA) from infected root canals with a minimally invasive technique (MIT) and the conventional instrumentation technique (CIT)., Methods: Sixty extracted human maxillary first premolars were included. All teeth were dentin pretreated and inoculated with LTA prepared from E. faecalis. First, 12 teeth were cryogenically ground to investigate the viability of recovering intraradicular E. faecalis LTA. Afterward, 48 teeth were randomly divided into the following groups: GWS + MIT, GWS + CIT, PUI + MIT, and GWS + CIT (all n = 12). Teeth were instrumented with a Vortex Blue (Dentsply Sirona, Ballaigues, Switzerland) rotary file size 15/.04 for MIT and 35/.04 for CIT. Samples were collected before and after a root canal procedure with sterile/apyrogenic paper points and after cryogenically ground for intraradicular LTA analysis. LTA was quantified with an LTA enzyme-linked immunosorbent assay kit., Results: E. faecalis LTA was recovered from 100% of the samples (48/48) before the root canal procedure. GWS + MIT and GWS + CIT were the most effective protocols against E. faecalis LTA, with no difference between them (P > .05). PUI + CIT was more effective than PUI + MIT (P < .05) but less effective than GWS + MIT and GWS + CIT (P < .05). The GWS groups showed more root canals with undetected E. faecalis LTA after treatment than all groups tested., Conclusions: GWS + MIT and GWS + CIT were the most effective protocols against E. faecalis LTA in infected root canals., (Copyright © 2022 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This study compared the effectiveness of GentleWave system (GWS) and passive ultrasonic irrigation (PUI) in removing lipopolysaccharides (LPS) from infected root canals after minimally invasive (MIT) and conventional instrumentation (CIT) techniques. Sixty first premolars with two roots were inoculated with fluorescent LPS conjugate (Alexa Fluor 594). Of those, twelve were dentin pretreated, inoculated with fluorescent LPS conjugate, and submitted to confocal laser scanning microscopy (CLSM) to validate the LPS-infection model. Forty-eight teeth were randomly divided into treatment groups: GWS + MIT, GWS + CIT, PUI + MIT, and PUI + CIT (all, n = 12). Teeth were instrumented with Vortex Blue rotary file size 15/0.04 for MIT and 35/0.04 for CIT. Samples were collected before (s1) and after a root canal procedure (s2) and after cryogenically ground the teeth (s3) for intraradicular LPS analysis. LPS were quantified with LAL assay (KQCL test). GWS + MIT and GWS + CIT were the most effective protocols against LPS, with no difference between them (p > 0.05). PUI + CIT was more effective than PUI + MIT (p < 0.05) but less effective than GWS + MIT and GWS + CIT. GWS was the most effective protocol against LPS in infected root canals using MIT and CIT techniques., (© 2022. The Author(s).)

Introduction: This study compared the absorbency capacity of paper points (PPs) with positively charged and noncharged polyvinylidene fluoride membranes (PVDFMs) and investigated the ability of PPs and PVDFMs to bind and remove endotoxin., Methods: Three commercially available PPs were compared with PVDFM (Millipore Sigma, Burlington, MA) prototype points. We recorded the initial dry weight for each PP and PVDFM using a digital balance to ±0.0001 precision for absorbency. PPs and PVDFMs were then immersed in deionized water and weighed to obtain the wet weight. The absorbency was calculated with the following formula: percent increase = ([wet weight - dry weight]/dry weight) × 100. For endotoxin removal, we first quantified endotoxin remaining in wells after immersing PPs and PVDFMs in a 24-well plate containing 10 endotoxin units/mL Escherichia coli O55:B5 (Lonza, Walkersville, MD). We then extracted and quantified endotoxin from PPs and PVDFMs. Endotoxin was quantified using the Kinetic-QCL test (Lonza)., Results: The absorbencies for the positively charged and noncharged PVDFMs were higher than the PPs (P < .05), with no difference between them (P > .05). The positively charged PVDFMs removed more endotoxin than the noncharged PVDFMs and the PPs (P < .05). Moreover, the noncharged PVDFMs bound and removed more endotoxin than any PPs (P < .05)., Conclusions: PVDFM prototype points are more absorbent than PPs. Moreover, the positively charged PVDFM points are more effective in binding and removing endotoxin than noncharged PVDFMs and PPs. This study suggests that positively charged PVDFMs with a 0.22-μm pore size could potentially replace PPs used in endodontics., (Copyright © 2021 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Objectives: This study investigated the influence of calcium hydroxide intracanal medications on the levels of metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in apical periodontitis (AP)., Materials and Methods: Twenty primarily infected root canals with AP were randomly divided into two groups: Ca(OH) 2  + sterile saline solution (SSL) group and Ca(OH) 2  + 2% chlorhexidine gel (CHX gel) group. We collected samples from the periradicular tissue fluid (PTF) before (s1) and after 14 days of intracanal medication (s2). MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 were measured by ELISA assay., Results: MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 were detected in all PTF samples at s1 and s2 (20/20). At s1, MMP-2 and MMP-9 were detected at higher levels than MMP-1 (p < .05). Higher levels of TIMP-1 than TIMP-2 were found in AP (p < .05). Additionally, we detected higher MMP-1, MMP-2, and MMP-9 over TIMP-1 and TIMP-2 levels in AP (p < .05). At s2, Ca(OH) 2  + SSL was as effective as Ca(OH) 2  + 2% CHX gel in lowering the levels of MMP-1, MMP-2, and MMP-9 after 14 days of intracanal medication, with no significant difference between them (p > .05). Both Ca(OH) 2 intracanal medications had no significant impact on the levels of TIMP-1 and TIMP-2 (both p > .05). At s2, TIMP-1 levels were higher than TIMP-2 (p < .05). Moreover, there were positive correlations between the levels of MMP-1 and TIMP-1 and MMP-1 and TIMP-2 (p < .05)., Conclusions: Calcium hydroxide medications effectively lowered the levels of MMP-1, MMP-2, and MMP-9 in periapical tissues after 14 days of treatment, with no difference between them. Moreover, the calcium hydroxide intracanal medications tested here had no impact in TIMP-1 and TIMP-2 in periapical tissues., Clinical Relevance: MMPs and TIMPs play an essential role in the degradation of the extracellular matrix. The imbalance MMPs and TIMPs can cause periapical tissue destruction. Therefore, the reestablishment of the balance between activated MMPs and TIMPs with root canal therapy is essential to restore tissue homeostasis., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Introduction: The purpose of this study was to investigate the accuracy and efficiency of the 3-dimensional dynamic navigation system (DNS) compared with the freehand technique (FH) when removing fiber posts from root canal-treated teeth., Methods: Twenty-six maxillary teeth were included. Teeth were root canal treated and restored with Parapost Taper Lux (Coltene/Whaledent, Altstätten, Switzerland) luted with RelyX Unicem (3M ESPE, St Paul, MN). A core buildup was then performed using Paracore (Coltene/Whaledent). Teeth were mounted in tissue-denuded cadaver maxillae. Teeth were divided into 2 groups: the DNS group (n = 13) and the FH group (n = 13). Cone-beam computed tomographic scans were taken pre- and postoperatively. The drilling path and depth were planned virtually using X-guide software (X-Nav Technologies, Lansdale, PA) in both groups. For the DNS group, drilling was guided with X-Nav software and the FH group under a dental operating microscope. Global coronal and apical deviations, angular deflection, operation time, and the number of mishaps were compared between the groups to determine the accuracy and efficiency. The 3-dimensional volume (mm 3 ) of all teeth was calculated before and after post removal using the Mimics Innovation Suite (Materialise NV, Leuven, Belgium). The Shapiro-Wilk, 1-way analysis of variance, and Fisher exact tests were used (P < .05)., Results: The DNS group showed significantly less global coronal and apical deviations and angular deflection than the FH group (P < .05). DNS required less operation time than FH. Moreover, the DNS technique had significantly less volumetric loss of tooth structure than the FH technique (P < .05)., Conclusions: The DNS was more accurate and efficient in removing fiber posts from root canal-treated teeth than the FH technique., (Copyright © 2021 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: We verified the association between selected bacterial profiles and levels of cytokines, chemokines, and the expression of signs and symptoms of primary endodontic infection with apical periodontitis., Methods: Samples were collected from 21 root canals, and macrophages were stimulated for 24 hours. Tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-10, IL-12p70, interferon gamma, and chemokine (C-C motif) ligand 2 (CCL2) were measured using cytometric bead array. We investigated the overlapping networks between cytokines and chemokines with regression analysis. Checkerboard DNA-DNA hybridization was used to assess 40 target bacteria species. Using factor analysis, bacterial species aggregated in 2 factors. The association of bacteria species-based factors on cytokine and chemokine levels and clinical features was estimated with regression analysis., Results: A negative relationship between IL-10 (anti-inflammatory cytokine) and CCL2, TNF-α, and IFN-γ (proinflammatory cytokines) (all P < .05) was observed. CCL2 was positively correlated with TNF-α (P < .01). Thirty-eight bacteria species were detected in primary endodontic infection with apical periodontitis. The first bacteria species-based factor was associated with the size of the radiolucent area (coefficient = 15.42) and tenderness to percussion/pain on palpation (coefficient = 20.79). The second factor was associated with CCL2 levels (coefficient = 1.28)., Conclusions: Different bacterial profiles can be differentially related to the expression of inflammatory proteins and the experience of clinical features., (Copyright © 2021 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: To determine and compare the accuracy and efficiency of a dynamic navigation system (DNS) with a freehand (FH) technique when conducting root-end resection in a human cadaver model., Methodology: Forty roots in cadaver heads were included and divided into two groups: DNS (n = 20) and FH (n = 20). Cone beam computed tomography (CBCT) scans of all teeth were taken. The drilling path and depth were planned virtually to using the X-guide software (X-Nav Technologies, Lansdale, PA, USA). Osteotomy and root-end resection were done under navigation in the DNS group, and freehand under the dental operating microscope in the FH group. Post-operative CBCTs were taken. Linear deviations, angular deflection, time of operation and number of mishaps were compared with determine the accuracy and efficiency. Shapiro-Wilk, one-way ANOVA and Fisher exact tests were used (P < 0.05)., Results: Linear deviations, angular deflection and operation time were significantly less in the DNS group (P < 0.05). The number of mishaps was not different between the two groups (P > 0.05). Subgroup analyses revealed that the distance of >5 mm from buccal cortical plate was significantly associated with lower accuracy, increased operation time and greater incidence of mishaps in the FH group (P < 0.05), but not in the DNS group., Conclusions: The dynamic navigation system was more accurate and more efficient in root-end resection in a cadaver model than the freehand technique. The distance of the roots from the buccal cortical plate had a significant negative impact on the accuracy and efficacy of the root-end resection procedure when using the freehand technique. The dynamic navigation system has the potential to be a safe and reliable technological addition to endodontic microsurgery., (© 2020 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Introduction: The aims of this study were to investigate aerosolized microorganisms generated during endodontic emergencies and nonsurgical root canal therapy (NSRCT), to assess the spread of airborne microbes, and to verify the spatial distribution of airborne microbial spread., Methods: A total of 45 endodontic procedures were sampled, including full pulpotomy (n = 15), pulpectomy (n = 15), and NSRCT (n = 15). Samples were collected during room resting and after treatment. The passive air sampling technique using settle plates was applied. Agar plates were set at different locations in the operatory. The colony-forming unit (CFU) was counted in brain-heart infusion blood agar plates. A set of agar plates containing selective chromogenic culture media was used for the isolation and presumptive identification of target microorganisms. Fungi were investigated using Sabouraud dextrose agar., Results: Pulpotomy generated the lowest mean CFU count (P < .05). There was no difference between the mean CFU counts found in pulpectomy and NSRCT (P > .05). A higher mean CFU count was found close to the patient's mouth (0.5 m) than at a 2-m distance in pulpectomy and NSRCT (P < .05). There was no difference between the mean CFU count found in front of the patient's mouth versus diagonal in pulpectomy and NSRCT (P > .05). Staphylococcus aureus (22/45, 48.8%) was the most frequent bacteria species. Longer treatment times were associated with higher CFU counts., Conclusions: Our findings indicated that pulpotomy generates less aerosolized microorganisms than pulpectomy and NSRCT. The proximity to the patient's mouth and the treatment duration were implicated in the level of contamination., (Copyright © 2021 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Sepsis is a leading cause of death in the United States, with a mortality rate in excess of 215,000 deaths per year. It may lead to septic shock, a complex pathophysiological process with microbial and host response events that progress to multisystem derangement. There is poor documentation of the relationship between dental infection and septic shock, with only a few case reports of septic shock secondary to dentoalveolar abscess. Presented is a case of sepsis/septic shock in a 23-year-old man with signs and symptoms of pulpal necrosis, acute apical abscess, and canine space infection that rapidly progressed to an altered mental state, hyperthermia, tachycardia, hypotension, acute respiratory failure, diarrhea, renal insufficiency, lactic acidosis, leukocytosis, and hyperglycemia. Once septic shock develops, the mortality rate is nearly 50%. Early antimicrobial intervention is associated with surviving severe sepsis, making it critical for dentists to understand local factors leading to the crisis and the signs and symptoms of the sepsis-septic shock continuum., (Copyright © 2021 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This study aimed to determine the mean distances between apexes of the maxillary posterior teeth and the maxillary sinus, between apexes of the mandibular posterior teeth and the mandibular canal, and between the root apexes of all teeth and the adjacent cortical plates. A total of 800 cone-beam computed tomography (CBCT) scans (400 maxillary and 400 mandibular) were obtained from patients indicated for several treatments. The proximity between apexes and anatomical structures, and the relationship between apexes and adjacent cortical plates were assessed together with the risk of over-instrumentation. Paired-sample comparisons were performed by using the paired t-test. The means were compared by ANOVA, Kruskal-Wallis and Dwass-Steel-Critchlow-Fligner tests. a) Most of the apexes classified as A (high-risk proximity) were observed in maxillary first and second molars, in mandibular first and second molars, and in second premolars in relation to near anatomical structures. b) A predominance of class A (86.42%) was noticed in the first premolars, between apexes of maxillary teeth and adjacent cortical plates. c) The distance between apexes of mandibular teeth and buccal cortical plates showed a predominance of medium-risk proximity (B) in all the groups, except the first premolars, with the highest risk (82.22%), and the second molars, with low-risk proximity (C) to distal and mesiobuccal apexes (91.77% and 89.62%). CBCT images are important for endodontic diagnosis and treatment, since many teeth have high risk proximity to near anatomical structures and adjacent cortical plates.

This case report describes the resolution of a 20-year misdiagnosed nasal sinus tract after root canal therapy with multiple sessions of calcium hydroxide (Ca[OH] 2 ) intracanal medication. Clinical evaluation, including diagnostic testing and sinus tract tracing, was performed followed by a cone-beam computed tomographic scan and 3-dimensional reconstruction of the apical lesion. Bacteria and endotoxin analyses were performed from the nasal sinus tract and paired root canal infection before (s1) and after instrumentation (s2) and after 7 (s3), 14 (s4), and 21 (s5) days of Ca(OH) 2 medication. The bacteria analysis was performed using the checkerboard DNA-DNA hybridization method and endotoxin quantified by the limulus amebocyte lysate method. A similar microbiota profile was found in the sinus tract and paired root canal infection. No target bacterial species were detected in the root canal at s2, s3, and s5. In contrast, Actinomyces israellii and Eubacterium nodatum were detected at s4. Differences in bacterial detection were found between s1 × s2, s3 × s4, and s4 × s5 (all P < .05). Endotoxin was detected in the root canal at all sampling times. Differences in the levels of endotoxin were found between s1 × s2, s2 × s3, and s3 × s4 (all P < .05).The bacterial analysis revealed similar microbiota profiles present in the nasal sinus tract and paired root canal infection with the participation of a wide variety of gram-positive and -negative species. Additionally, root canal therapy with multiple sessions of Ca(OH) 2 intracanal medication for 21 days was effective in disinfecting the root canal system and resolving the nasal sinus tract., (Copyright © 2020 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This systematic review (SR) addressed the following common clinical question: What is more effective in reducing or eliminating endotoxin in endodontic infections-single or multiple-session treatments using calcium hydroxide medications? Literature searches of Medline/PubMed, Embase, Cochrane Library, Scielo, Science Direct, Web of Knowledge, Scopus, and Google Scholar databases. Two reviewers independently assessed the eligibility for inclusion, extracted data, and evaluated the quality of the studies using the risk of bias tools. Electronic searches resulted in 358 articles, of which 32 studies were included for full-text assessment, and nine were included in this review. Meta-analysis pooling all the nine studies revealed lower levels of endotoxin for multiple-session treatment (P < 0.001). The sub-group analysis indicated no difference between single-session and 7 days of Ca(OH) 2 medication (SMD - 0.32; P = 0.22). However, 14-days (I 2  = 80.5%, P < 0.001) and 30-days (I 2  = 78.9%, P < 0.01) of Ca(OH) 2 medication was more effective than single-session treatment (both, p < 0.001). Overall, Overall, this SR provides evidence to support that multiple-session disinfection protocols with the placement of Ca(OH) 2 medications are more effective in reducing the levels of endotoxin from root canal infections compared to single-session when applied for 14 and 30 days.

This survey investigated the effect of the coronavirus disease 2019 (COVID-19) pandemic on the clinical practice of endodontics among the American Association of Endodontists (AAE) members by evaluating the impact on clinical activities, patient screening, infection control measurements, potential transmission, clinical protocols, as well as psychological concerns. A descriptive, cross-sectional survey was developed to query AAE members from all 7 districts. The survey consisted of 24 questions, 8 demographic questions and 16 questions related to the COVID-19 pandemic impact on the clinical practice. A total of 454 AAE members participated in the survey. As of July 2020, most endodontists were active in front-line treatment of dental patients (82%). N95 respirator face mask was described by 83.1% of the participants as special measures beyond the regular personal protective equipment. Rubber dam isolation was recognized by the majority of the participants at some level to reduce the chance of COVID-19 cross infection. Most of the endodontist participants acknowledged trauma followed by swelling, pain, and postoperative complication to be emergencies. The majority of respondents reported being concerned about the effect of COVID-19 on their practice. No differences in worries about COVID-19 infection were related to demographics (P > .05). The majority of the endodontists are aware of the COVID-19 pandemic, are taking special precautions, and are concerned about contracting and spreading the virus. Despite the conflict between their roles as health care providers and family members with the potential risk of exposing their families, most of them remain on duty providing front-line care for dental treatment., (Copyright © 2020 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: To investigate the presence of resolvins E1 (RvE1) and D2 (RvD2) in teeth with primary endodontic infections and apical periodontitis, and to assess the influence of calcium hydroxide medication [Ca(OH) 2 ], in association with 2% chlorhexidine gel (2% CHX gel), and N-acetylcysteine (NAC) on the levels of RvE1 and RvD2 in periapical tissues., Methodology: Thirty-six single-rooted teeth with primary endodontic infections and apical periodontitis were selected and randomly divided into three groups according to the medication: [Ca(OH) 2 ] + saline solution (SSL) [Ca(OH) 2  + SSL group] (n = 12), Ca(OH) 2  + 2% chlorhexidine gel [Ca(OH) 2  + 2% CHX gel group] (n = 12) and NAC [NAC group] (n = 12). Samples were collected from the periapical interstitial fluid at two different sampling times: before (S1) and after 14 days of intracanal medications (S2). Resolvins were measured using the enzyme-linked immunosorbent assay. Data were analysed using paired t-test, Wilcoxon test and Kruskal-Wallis test, followed by Dunn's post hoc test; all statistical tests were performed at a significance level of 5%., Results: RvE1 and RvD2 were detected in 100% of the samples (36/36) at S1 and S2. Ca(OH) 2 medication did not increase the levels of RvE1 or RvD2 (both P > 0.05); however, NAC significantly increased the levels of RvE1 and RvD2 after 14 days of treatment (P < 0.05)., Conclusions: RvE1 and RvD2 were detected in periapical tissues from teeth with root canal infections. Moreover, calcium hydroxide medication did not increase the levels of resolvins in apical periodontitis. In contrast, the use of NAC intracanal medication significantly increased the levels of RvE1 and RvD2 after 14 days of treatment., (© 2020 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Herein, we investigated a biointeractive tooth sealing material consisted of dimethylaminohexadecyl methacrylate (DMAHDM) and amorphous calcium phosphate nanoparticles (NACPs) to address the above issues simultaneously. Of note, 5% DMAHDM was incorporated into the resin blend, and 20% NACP was added to inorganic filler content of dental formulations intended as dental sealants. The sealing materials were used to seal human extracted teeth. The sealed teeth were subjected to an early childhood caries (ECC) key pathogen (Candida albicans and Streptococcus mutans) biofilm model using a dynamic caries tooth model (CDC reactor). The biofilm growth over the sealed teeth was assessed via colony-forming unit counting metabolic activity assays. The enamel surface hardness loss, degree of conversion, shear bond strength (SBS), and cytotoxicity were also investigated. Formulations having DMAHDM displayed antibacterial efficiency of 2.8-3.5 and 1.4-4.0 log inhibition for Streptococcus mutans and Candida albicans, respectively. Furthermore, the metabolic activity was reduced on the top of the sealed tooth with the biointeractive sealing materials (p < .05). The degree of conversion values was acceptable. The enamel surface hardness loss decreases (36 ± 9.8%) when in contact with the biointeractive tooth sealing material. The SBS of the combined formulation (5% DMAHDM + 20% NACP) was lower than commercial sealant but similar to experimental control. The investigated sealing material holds valuable dual antibacterial and antifungal activities associated with a reduced mineral loss against the cariogenic challenge promoted by ECC key pathogens., (© 2020 Wiley Periodicals LLC.)

Caries-related biofilms and associated complications are significant threats in dentistry, especially when biofilms grow over dental restorations. The inhibition of cariogenic biofilm associated with the onset of carious lesions is crucial for preventing disease recurrence after treatment. This in vitro study defined optimized parameters for using a photosensitizer, toluidine blue O (TBO), activated via a red light-emitting diode (LED)-based wireless device to control the growth of cariogenic biofilms. The effect of TBO concentrations (50, 100, 150, and 200 μg/mL) exposed to light or incubated in the dark was investigated in successive cytotoxicity assays. Then, a mature Streptococcus mutans biofilm model under sucrose challenge was treated with different TBO concentrations (50, 100, and 150 μg/mL), different light energy doses (36, 108, and 180 J/cm 2 ), and different incubation times before irradiation (1, 3, and 5 min). The untreated biofilm, irradiation with no TBO, and TBO incubation with no activation represented the controls. After treatments, biofilms were analyzed via S. mutans colony-forming units (CFUs) and live/dead assay. The percentage of cell viability was within the normal range compared to the control when 50 and 100 μg/mL of TBO were used. Increasing the TBO concentration and energy dose was associated with biofilm inhibition ( p < 0.001), while increasing incubation time did not contribute to bacterial elimination ( p > 0.05). Irradiating the S. mutans biofilm via 100 μg/mL of TBO and ≈180 J/cm 2 energy dose resulted in ≈3-log reduction and a higher amount of dead/compromised S. mutans colonies in live/dead assay compared to the control ( p < 0.001). The light energy dose and TBO concentration optimized the bacterial elimination of S. mutans biofilms. These results provide a perspective on the determining parameters for highly effective photo-killing of caries-related biofilms and display the limitations imposed by the toxicity of the antibacterial photodynamic therapy's chemical components. Future studies should support investigations on new approaches to improve or overcome the constraints of opportunities offered by photodynamic inactivation of caries-related biofilms.

Objective: This review clusters the growing field of nano-based platforms for antimicrobial photodynamic therapy (aPDT) targeting pathogenic oral biofilms and increase interactions between dental researchers and investigators in many related fields. Background data: Clinically relevant disinfection of dental tissues is difficult to achieve with aPDT alone. It has been found that limited penetrability into soft and hard dental tissues, diffusion of the photosensitizers, and the small light absorption coefficient are contributing factors. As a result, the effectiveness of aPDT is reduced in vivo applications. To overcome limitations, nanotechnology has been implied to enhance the penetration and delivery of photosensitizers to target microorganisms and increase the bactericidal effect. Materials and methods: The current literature was screened for the various platforms composed of photosensitizers functionalized with nanoparticles and their enhanced performance against oral pathogenic biofilms. Results: The evidence-based findings from the up-to-date literature were promising to control the onset and the progression of dental biofilm-triggered diseases such as dental caries, endodontic infections, and periodontal diseases. The antimicrobial effects of aPDT with nano-based platforms on oral bacterial disinfection will help to advance the design of combination strategies that increase the rate of complete and durable clinical response in oral infections. Conclusions: There is enthusiasm about the potential of nano-based platforms to treat currently out of the reach pathogenic oral biofilms. Much of the potential exists because these nano-based platforms use unique mechanisms of action that allow us to overcome the challenging of intra-oral and hard-tissue disinfection.

Dens invaginatus is a malformation affecting mainly the superior lateral incisors. It is defined as an infolding of the crown hard tissues, including the enamel and dentin, and can extend up to the root apex. Root canal treatment of this abnormality is considered difficult due to the complex anatomy presented by these teeth. This case series presents nonsurgical endodontic treatment in two cases of dens invaginatus (type II and III) in maxillary lateral incisors. This nonsurgical or conventional endodontic treatment results in healing of the periapical lesions associated with both cases, with no need for extra intervention e.g. surgical or invasive management. The manual instrumentation associated with sodium hypochlorite and calcium hydroxide were able to completely heal the lesions.  Radiographic exams were carried out to control and asses the healing. Nonsurgical treatment was successful in both cases with adequate repair after a 6-year follow-up with radiographic and tomographic assessments., Competing Interests: No competing interests were disclosed., (Copyright: © 2019 Abu Hasna A et al.)

This study was carried out to investigate the microbial profile and endotoxin levels of endodontic-periodontal lesions of periodontal origin. Periodontal and endodontic samples were taken from periodontal pockets and necrotic root canals of 10 teeth with endodontic-periodontal lesions. Evidencing of 40 different bacterial species were determined in each endodontic and periodontal sample using the checkerboard DNA-DNA hybridization method and Kinetic chromogenic LAL assay was used for quantification of endotoxins. Fisher's exact test correlated the bacterial species with the endodontic or periodontal microbiota. The endotoxin levels (EU/mL) found in samples of the root canal and periodontal pocket were compared by the Wilcoxon test (p<0.05). Bacteria and LPS units were found in 100% of the endodontic and periodontal samples. The species E. faecium, P. acnes, G. morbillorum, C. sputigena and L. buccalis were strongly correlated with the endodontic microbiota and P. nigrescens with the periodontal microbiota. P. intermedia, P. endodontalis and V. parvula were more prevalent in both endodontic and periodontal microbiots. The endotoxin levels in the periodontal pocket (89600 EU/mL) were significantly higher than in the root canal (2310 EU/mL). It was concluded that the microbiota present in the periodontal and endodontic tissues is similar, with a higher prevalence of species of the orange complex and a higher level of endotoxin in the periodontal pockets.

The present study aimed to compare the effectiveness of QMiX and 17% EDTA associated to passive ultrasonic irrigation (PUI) or manual agitation (MA) on the reduction of E. faecalis, E. coli and LPS from root canals. Forty single rooted human teeth were randomly divided into four groups (n=10), according to the final irrigation protocol: EDTA+MA, QMiX+MA, EDTA+PUI, QMiX+PUI. Sample collections were obtained from the root canal content immediately before preparation (baseline-S1), after instrumentation (S2), after final irrigation protocol (S3) and 7 days after instrumentation and final irrigation (S4). The antimicrobial effectivity and on endotoxin content were analyzed by culture procedure (CFU/mL) and LAL assay (EU/mL), respectively. The results were statistically analyzed by Kruskal-Wallis and Friedman test (α=5%). QMiX+MA and QMiX+PUI reduced 100% of E. coli and E. faecalis bacteria and also prevented E. faecalisregrowth at S4. EDTA significantly reduced E. coli, but it was not effective in reducing E. faecalis. All protocols reduced EU/mL when compared to S1, however at S4 there was a significant reduction of EU/mL only in the QMiX+MA and QMiX+PUI groups in relation to S3 and S2, respectively. Final irrigation with QMiX associated with MA or PUI had superior antibacterial efficacy compared to EDTA, eliminating 100% of E. coli and E. faecalis strains. In addition, QMiX+PUI reduced 97.61% of the initial content of LPS.

This clinical study was conducted to correlate the levels of endotoxins and culturable bacteria found in primary endodontic infection (PEI) with the volume of root canal determined by using Cone Beam Computed Tomography (CBCT); and to evaluate the bacterial diversity correlating with clinical features. Twenty patients with PEI were selected and clinical features were recorded. The volume (mm3) of root canal was determined by CBCT analysis. Root canal samples were analyzed by using kinetic LAL-assay test to determine the levels of endotoxins and anaerobic technique to determine the bacterial count (CFU/mL). DNA was extracted from all samples to determine bacterial diversity and quantified by using Checkerboard-DNA-DNA- Hybridization. Culturable bacteria and endotoxins were detected in 100% of the root canal samples. Linear regression analysis revealed a correlation between root canal volume and presence of anaerobic bacteria (p<0.05). Positive correlations were found between bacteria species and presence of different clinical features (p<0.05). After grouping the bacteria species into bacterial complexes, positive associations were found between green, orange and red complexes with presence of sinus tract (p<0.05). This clinical study revealed that larger root canals hold higher levels of culturable bacteria in PEI. Thus, the interaction of different virulent bacteria species in complexes seems to play an important role in the development of clinical features.

This clinical study compared the effectiveness of two rotary systems: HyFlex CM (Coltene-Whaledent, Altstetten, Switzerland) and ProTaper Next (Dentsply Sirona, Ballaigues, Switzerland) on the removal of cultivable bacteria and endotoxins from primarily infected root canals. This study was designed as a randomized single-blinded, 2-arm, clinical trial. Twenty-four primarily infected root canals were selected and randomly divided into 2 groups: HyFlex CM (n = 12); and ProTaper Next (n = 12). Samples were collected before and after the biomechanical preparation and inoculated in specific flasks. Irrigation was performed using 2.5% sodium hypochlorite. A kinetic turbidimetric lysate assay of limulus amoebocytes was used to quantify endotoxins. Microbiological culture technique was used to determine the count of bacterial colony forming units (CFU/mL). Data collected were statistically analyzed using SigmaPlot 12.0 for Windows. The Two-Way ANOVA statistical test was performed and the level of significance was 5%. In the samples before the biomechanical preparation, cultivable bacteria and endotoxins were evidenced in 100% of the cases. The culture analysis revealed that there was no statistically significant difference in the bacterial reduction between the two instrumentation systems. Endotoxins were present in 100% of the canals after instrumentation and there was no statistical difference between the two systems in endotoxin reduction. Thus, it was concluded that both instrumentation systems were effective in reducing root canal bacteria and endotoxins with primary endodontic infection and that there was no statistical difference between them. However, no system was able to eliminate 100% of the bacteria and their by-products.

Evidence indicates that major depression is accompanied by increased translocation of gut commensal Gram-negative bacteria (leaky gut) and consequent activation of oxidative and nitrosative (O&NS) pathways. This present study examined the associations among chronic apical periodontitis (CAP), root canal endotoxin levels (lipopolysaccharides, LPS), O&NS pathways, depressive symptoms, and quality of life. Measurements included advanced oxidation protein products (AOPP), nitric oxide metabolites (NOx), lipid peroxides (LOOH), -sulfhydryl (SH) groups, total radical trapping antioxidant parameter (TRAP), and paraoxonase (PON)1 activity in participants with CAP, with and without depression, as well as healthy controls (no depression, no CAP). Root canal LPS levels were positively associated with CAP, clinical depression, severity of depression (as measured with the Hamilton Depression Rating Scale (HDRS) and the Beck Depression Inventory) and O&NS biomarkers, especially NOx and TRAP. CAP-related depression was accompanied by increased levels of NOx, LOOH, AOPP, and TRAP. In CAP participants, there was a strong correlation (r = 0.734, p < 0.001) between root canal LPS and the HDRS score. There were significant and positive associations between CAP or root canal endotoxin with the vegetative and physio-somatic symptoms of the HDRS as well as a significant inverse association between root canal endotoxin and quality of life with strong effects on psychological, environmental, and social domains. It is concluded that increased root canal LPS accompanying CAP may cause depression and a lowered quality of life, which may be partly explained by activated O&NS pathways, especially NOx thereby enhancing hypernitrosylation and thus neuroprogressive processes. Dental health and "leaky teeth" may be intimately linked to the etiology and course of depression, while significantly impacting quality of life.

Objectives: The aim of this study is to improve the understanding of interleukin mechanisms during osseointegration to enhance the monitoring of implant failure and success. Clinical parameters, implant stability, and cytokine levels in peri-implant crevicular fluid (PICF) during early bone healing after implant placement were investigated., Material and Methods: Sixty narrow implants were placed in mandible anterior region of 30 edentulous patients (67.23 ± 7.66 years). Bone type, insertion torque, and primary stability were registered during surgery. Clinical measurements of peri-implant health and the secondary implant stability quotient (ISQ) were recorded. Samples from the PICF were collected 1, 2, 4, 8, and 12 weeks after surgery and analyzed for IL-1β, IL-6, IL-10, and TNF-α levels using ELISAs., Results: The gingival index increased significantly during the first week (p = 0.05), while the plaque index increased significantly between 4 to 8 and 8 to 12 weeks (p < 0.05). The probing depth and the ISQ also reduced significantly (p < 0.05) over time. The TNF-α release increased significantly after the 2nd week for non-atrophic patients and 4th week for atrophic patients (p < 0.05). The IL-1β concentrations showed a short-lived peak after 1st week (p = 0.003), specially in atrophic patients and sites with bone type I (p = 0.034; p = 0.007). The IL-6 concentrations peaked during the 1st and 2nd weeks (p < 0.05; p = 0.005) in atrophic patients and in bone type II (p = 0.023; p = 0.003). The IL-10 concentrations increased gradually over time, showing the highest concentrations at the 12th week (p < 0.005). A total of 12 implants failed at different periods., Conclusion: While the clinical measurements presented differences between the evaluation periods, these were not indicative of early dental implant failure or peri-implant diseases. Smoking, bone atrophy, and bone type can greatly influence the cytokines concentrations during the healing time.

Objective: To investigate the microbial diversity existing in oral cavity and respiratory tract samples (from mini-bronchoalveolar lavage (BAL), endotracheal aspirate, and orotracheal tube) of patients on mechanical ventilation by using the checkerboard DNA-DNA hybridisation. Also, the study aimed to evaluate whether the microbial profile in the oral cavity is found in respiratory tract samples, at different periods of mechanical ventilation time (12h, 48h, 96h) in attempt to identification of relationship between VAP (ventilator-associated pneumonia) and bacterial species studied. The last objective was to analyses correlation between blood cultures and VAP., Design: The samples were collected from ten patients in intensive care unit with medical prescription of orotracheal intubation and mechanical ventilation. Clinical data were tabulated and blood cultures were performed according medical indication. For checkerboard samples collection, chosen sites were the dorsal side of the tongue and gingival sulcus at 12h, 48h, 96h, BAL, at 12h, endotracheal aspirate at 48 and 96h, and orotracheal tube at extubation time, when feasible., Results: It was possible to identify the presence of bacterial species in mouth and in the BAL/endotracheal aspirate. The data demonstrated an increase in the quantity of bacterial associated with prolonged use of mechanical ventilation (48 and 96h)., Conclusions: Bacterial species may migrate rapidly from mouth and upper airways during orotracheal intubation which contributes to the pathogenesis of VAP. There were associations between VAP and Enterococcus faecalis, Fusobacterium periodonticum, Gemella morbillorum, Neisseria mucosa, Propionibacterium acnes, Prevotella melaninogenica, Streptococcus oralis, Streptococcus sanguinis, Treponema denticola, Treponema socransckii, and Veillonella parvula., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objective: This clinical study compared the effectiveness of 7- and 14-day intracanal medications in the reduction of bacteria/endotoxins from primarily infected root canals and determined their antigenicity against macrophages through the levels of cytokines., Methods: Seventy-two primarily infected teeth were randomly divided into six groups according to medication and time of application: 7-day groups = G1, Ca(OH) 2 + saline solution (SSL); G2, Ca(OH) 2 + 2% chlorhexidine (CHX) gel; and G3, 2% CHX gel and 14-day groups = G4, Ca(OH) 2 + SSL; G5, Ca(OH) 2 + 2% CHX gel; and G6, 2% CHX gel (all groups, n = 12). Bacterial and endotoxin samples were collected from root canals and inflammatory cytokines of macrophages supernatants. Culture techniques were used to determine bacterial counts and limulus amebocyte lysate (LAL) assay to quantify endotoxins. IL-1β, TNF-α, and PGE 2 were measured by ELISA-assay., Results: With regard to the bacterial reduction, no differences were found between all protocols tested (p > 0.05). The CHX protocols (G3 and G6) exhibited the lowest effectiveness against endotoxins (p < 0.05). All protocols were effective in lowering the levels of IL-1β, TNF-α, and PGE 2 (p < 0.05), with no difference between the medications tested on days 7 or 14 (p > 0.05). Particularly, the 7-day CHX-protocol (G3) exhibited the lowest effectiveness in lowering the levels of most cytokines compared to the 14-day protocols (G6) (p < 0.05)., Conclusions: All the 7- and 14-day intracanal medications were effective in reducing bacteria and endotoxins as well as in lowering the levels of inflammatory cytokines, with CHX showing limited effectiveness against endotoxins. Moreover, 7-day CHX-protocol exhibited the lowest effectiveness in lowering the levels of most cytokines compared to the 14-day protocols., Clinical Significance: Seven-day CHX protocol is the less effective protocol and should be carefully applied by the clinician.

Aim: To establish an SV40 T-Ag-transfected cell line of human pulp-derived cells in order to compare the cytotoxicity, genotoxicity and to investigate the activities of immunological biomarkers of several endodontic sealers., Methodology: Primary human pulp cells and transfected cells were cultured. Cell morphology and proliferation were analysed, and the expression of cell-specific gene transcripts and proteins was detected by RT-PCR and immunohistochemistry. Transfection of human pulp-derived cells resulted in an immortalized cell line retaining phenotypic characteristics from the primarily cells tested. The SV40 T-Ag-transfected cells were cultured and stimulated by sealers (Apexit Plus, Real Seal, AH Plus, and EndoREZ) to evaluate the cytotoxicity and genotoxicity by MTT and MTN assays, respectively. Immunological inflammatory biomarkers (IL6, IL8 and TNF-α) were determined by ELISA assay. The differences between median values were statistically analysed using Kruskal-Wallis and Dunn's tests at 5% significance level., Results: The cytotoxicity assay revealed that multimethacrylate (Real Seal) was the most cytotoxic sealer (P < 0.05) and exhibited the highest inflammatory potential against the SV40 T-Ag-transfected cells (P < 0.05). All root canal sealers tested were able to stimulate the immortalized pulp cells to produce IL-6, IL-8 and TNF-α, with differences in relation to the control group (P < 0.05). Higher levels of IL-6, IL-8 and TNF-α were found in cell supernatant after stimulation with multimethacrylate (Real Seal) compared to all other sealers tested (P < 0.05). No differences were found comparing epoxy resin-based sealer (AHPlus), single-methacrylate sealer (EndoREZ) and calcium hydroxide-based sealer (Apexit Plus), regardless of the cytokine investigated (all P > 0.05)., Conclusions: A SV40 T-Ag-transfected cell line of human pulp-derived cells was established. The methacrylate resin-based sealer (Real Seal) exhibited the greatest cytoxicity and inflammatory potential against immortalized pulp cells compared to an epoxy resin-based sealer (AH Plus), a methacrylate-based sealer (EndoRez) and a calcium hydroxide-based sealer (Apexit)., (© 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Aim: This clinical study was conducted to investigate the influence of 17% ethylenediaminetetraacetic acid (EDTA) ultrasonic activation after chemomechanical preparation (CMP) on eliminating/reducing oral bacterial lipopolysaccharides (known as endotoxins) and cultivable bacteria in teeth with pulp necrosis and apical periodontitis., Methodology: Samples were taken from 24 root canals at several clinical periods: S1 - before CMP; S2 - after CMP; S3 - after EDTA: G1 - with ultrasonic activation (n = 12) and G2 - without ultrasonic activation (n = 12). Root canals were instrumented using Mtwo rotary files. Culture techniques were used to determine the number of colony-forming units (CFU). Limulus amebocyte lysate (LAL) was used to measure endotoxin levels. Friedman's and Wilcoxon signed-rank tests were used to compare the amount of bacteria and endotoxin levels in each period (P < 0.05)., Results: Endotoxins and cultivable bacteria were recovered in 100% of the initial samples (S1). CMP was effective in reducing endotoxins and bacterial load (all with P < 0.05). Higher values of endotoxin reduction were achieved with EDTA ultrasonic activation [G1, 0.02 EU mL -1 (range 0.01-0.75)] compared with the no activation group [G2, 1.13 EU mL -1 (range 0.01-8.34)] (P < 0.05). Regarding bacterial reduction, no statistically significant difference was found in S3, regardless of the group (G1, G2, P > 0.05)., Conclusions: Chemomechanical preparation was effective in reducing bacteria and endotoxins, but could not completely eliminate them. The ultrasonic activation of EDTA was effective in further reducing endotoxin levels in the root canals of teeth with pulp necrosis and apical periodontitis., (© 2016 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Aim: To evaluate the effectiveness of supplemental photodynamic therapy (PDT) in optimizing the removal of bacteria and endotoxins from primarily infected root canals after one-visit and two-visit treatments., Methodology: Twenty-four primarily infected root canals with apical periodontitis were selected and randomly divided into one-visit (n=12) and two-visit treatment groups (n=12). Chemo-mechanical preparation (CMP) was performed by using the single-file reciprocating technique+2.5% NaOCL and a final rinse with 17% EDTA. The photosensitizer agent (methylene blue 0.1mg/mL) was applied to root canals for 60s before application of laser with a potency of 60mW and energy density of 129J/cm 2 for 120s after CMP in the one-visit treatment and after 14-day inter-appointment medication with Ca(OH) 2 +Saline solution (SSL) in the two-visit treatment. Samples were collected before and after root canal procedures. Endotoxins were quantified by chromogenic limulus amebocyte lysate assay. Culture techniques were used to determine bacterial colony-forming unit counts., Results: Bacteria and endotoxins were detected in 100% of the initial samples, with median values of 1.97×10 5 CFU/mL and 24.983EU/mL, respectively. The CMP using single-file reciprocating technique was effective in the reduction of bacteria and endotoxins (All, p<0.05). The supplemental PDT was effective in reducing bacterial load in the one-visit (p<0.05) but not in the two-visit treatment after use of Ca(OH) 2 medication for 14days (p>0.05). In the two-visit group, after 14days of inter-appointment medication with Ca(OH) 2 , a significant reduction in the median levels of endotoxins was found in comparison to CMP alone (from 1.041 to 0.094EU/mL) (p<0.05). Despite the type of treatment, the supplemental PDT was not effective against endotoxins (p>0.05)., Conclusions: The photodynamic therapy optimized the disinfection of bacteria from root canals in one-visit but not for two visit treatment modality with the accomplishment of calcium hydroxide medication. Despite the type of treatment, the supplemental PDT was not effective against endotoxins., (Copyright © 2017 Elsevier B.V. All rights reserved.)