Impact of root canal disinfection on the bacteriome present in primary endodontic infection: A next generation sequencing study.

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 (R2 = .0630, p = .012). Patient ID also showed a significant effect on community composition (R2 = .6961, p = .001). At the genus level, Dialister, Mogibacterium, Prevotella, and Olsenella were differentially enriched at s1, while Actinomyces, Stenotrophomonas_unclassified, Enterococcus_unclassified, and Actinomyces_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.

Real-Time Three-dimensional Dynamic Navigation for Post Space Preparation in Root Canal-Treated Teeth: An In vitro Study.

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.

Augmented Reality Head-Mounted Device and Dynamic Navigation System for Postremoval in Maxillary Molars.

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.

Piezoelectric Device and Dynamic Navigation System Integration for Bone Window-Guided Surgery.

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.

Augmented Reality and 3-Dimensional Dynamic Navigation System Integration for Osteotomy and Root-end Resection.

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.

A Cadaver-based Comparison of Sleeve-guided Implant-drill and Dynamic Navigation Osteotomy and Root-end Resections.

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.

Comparison of a Novel Static Computer-aided Surgical and Freehand Techniques for Osteotomy and Root-end Resection.

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.

Comparison of conventional and contemporary root canal disinfection protocols against bacteria, lipoteichoic acid (LTA), and lipopolysaccharide (LPS).

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.

Impact of N-acetylcysteine (NAC) and calcium hydroxide intracanal medications in primary endodontic infection: a randomized clinical trial.

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.

A modified dentine infection model with fluorescent lipopolysaccharide and lipopolysaccharides sampling technique to compare XP-Endo finisher and passive ultrasonic irrigation.

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.

Comparison of the Accuracy and Efficiency of a 3-Dimensional Dynamic Navigation System for Osteotomy and Root-end Resection Performed by Novice and Experienced Endodontists.

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.

Real-time 3-dimensional Dynamic Navigation System in Endodontic Microsurgery: A Cadaver Study.

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.

Effects of Calcium Hydroxide Intracanal Medications on T Helper (Th1, Th2, Th9, Th17, and Tfh) and Regulatory T (Treg) Cell Cytokines in Apical Periodontitis: A CONSORT RCT.

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.

Efficacy of GentleWave System and Passive Ultrasonic Irrigation with Minimally Invasive and Conventional Instrumentation Technique against Enterococcus faecalis Lipoteichoic Acid in Infected Root Canals.

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.

Comparison of GentleWave system and passive ultrasonic irrigation with minimally invasive and conventional instrumentation against LPS in infected root canals.

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.

Clinical influence of calcium hydroxide intracanal medications on matrix metalloproteinases and tissue inhibitors of metalloproteinases in apical periodontitis.

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.

Positively Charged Polyvinylidene Fluoride Membrane: A Potential Alternative for Absorbent Paper Points in Endodontics.

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.

Accuracy and Efficiency of 3-dimensional Dynamic Navigation System for Removal of Fiber Post from Root Canal-Treated Teeth.

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 (mm3) 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.

Influence of Bacterial Profiles in Cytokine and Clinical Features of Endodontic Disease.

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.

Aerosols Generated during Endodontic Treatment: A Special Concern during the Coronavirus Disease 2019 Pandemic.

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.