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Laser ablation therapy (LA) uses Indocyanine Green dye (ICG) which efficiently absorbs laser energy and the increased temperature results in an instantaneous flame that chars tissue and microbes. Photodynamic therapy (PDT) uses different dyes that are activated by light to kill bacteria. This study evaluated the biocompatibility of the dye Curcumin (CUR), Methylene Blue (MB), and Indocyanine Green (ICG) before and after laser activation (ACT). Polyethylene tubes containing one of the dyes were implanted in the subcutaneous tissue of 32 rats (4 tubes per rat) which were divided into 8 groups: C - control (saline solution); C + ACT (Red Laser 660 nm); CUR; CUR + ACT (480 nm blue LED); MB; MB + ACT (Red Laser 660 nm); ICG; ICG + ACT (810 nm Infrared Laser). After 7 and 30 days (n = 8/time), the rats were euthanized and the tubes with the surrounding tissue were removed and processed for histological analysis of inflammation using H&E stain, and collagen fiber maturation using picrosirius red (PSR). A two-way analysis of variance statistical test was applied (p < 0.05). At 7 days, regardless of laser activation, the CUR group showed a greater inflammatory infiltrate compared to the ICG and control groups, and the MB group had a greater inflammation only in relation to the control (p < 0.05). At 30 days, CUR and MB groups showed a greater inflammatory infiltrate than the control (p < 0.05). ICG group was equal to the control in both periods, regardless of the laser activation (p > 0.05). Laser activation induced the proliferation of collagen immature fibers at 7 days, regardless of the dye (p < 0.05). The CUR group showed a lower percentage of immature and mature fibers at 7 days, compared to ICG and control (p < 0.05) and, at 30 days, compared to control (p < 0.05). Regardless of laser activation, the ICG showed the results of collagen maturation closest to the control (p > 0.05). It was concluded that all dyes are biocompatible and that laser activation did not interfere with biocompatibility. In addition, the maturity of collagen was adequate before and after the laser activation. These results demonstrate that the clinical use of dyes is safe even when activated with a laser., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Introduction: Laser ablation (LA) therapy is used as an adjunct to endodontic treatment to improve microbial reduction. However, studies evaluating the impact of LA with indocyanine green (ICG) are scarce. This study aimed to evaluate the antimicrobial efficacy of LA therapy with ICG in root canal treatment., Methods: Sixty patients with periapical lesions in teeth with a single canal and absence of pain, edema, and previous treatment were selected. Patients were randomly allocated into 3 groups according to the apical sizes used (n = 20); 25/04, 30/04, and 35/04 were the final sizes used. In half the patients of each group, 2.5% sodium hypochlorite was used as an irrigating solution, and in the other half, saline solution was used. After instrumentation, all patients received LA therapy with ICG. Root canal sampling was performed before (S1) and after (S2) root canal instrumentation and immediately after LA therapy with ICG (S3). Colony-forming units were counted, and statistical tests were applied (P < .05)., Results: There was a significant reduction in colony-forming units from S1 to S2 in all treatment protocols (P < .05); 2.5% sodium hypochlorite as an irrigating solution showed a greater microbial reduction compared with saline solution (P < .05). LA therapy with ICG further reduced the microbial counts significantly (S2 to S3 and S1 to S3) whether sodium hypochlorite or saline was used (P < .05)., Conclusions: LA therapy with ICG significantly increased microbial reduction in root canals regardless of instrumentation sizes or the irrigation solution used., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

The aim of this study was to evaluate the biocompatibility and immunoinflammatory response of the Sealepox and Sealepox-RP, based on interleukin (IL)-6, tumor necrosis factor (TNF)-α, and CD5 immunolabelling. The ProRoot MTA (PRMTA) was used for comparison. Polyethylene tubes (1.0-mm internal, 1.6-mm external diameter, and 10.0-mm length; ISO 10993) with or without (control) materials were randomly implanted in the dorsum of 35 rats (4 per rat). After 7, 15, 30, 60, and 90 days (n = 7), the tubes were removed for histological and immunohistochemical analysis. The Kruskal-Wallis and Dunn's test for non-parametric data and, ANOVA and Tukey test for parametric data were used (P < 0.05). Hematoxylin and eosin staining revealed that the concentration of inflammatory cells decreased over time with no differences between groups in all periods (P > 0.05). Regarding IL-6 immunostaining, there was no difference at 7 days (P > 0.05); all groups decreased over time, being faster for the PRMTA group and also, with no differences between groups in the last period (P > 0.05). For TNF-α, at 7 days there was no difference between groups (P > 0.05); there was an increase at 15 days for PRMTA and, at 30 and 60 days, for PRMTA and Sealepox compared to the control (P < 0.05). At 90 days, Sealepox RP showed the lowest immunostaining being similar to the control (P > 0.05). Regarding CD5 cells, at 7 days, there was high immunostaining for PRMTA compared to the control (P < 0.05); and significant reduction over time with difference for all groups at 30 and 60 days. (P < 0.05); Sealepox was similar to the control in all periods (P > 0.05). Sealepox RP showed the highest immunostaining at 15 days, being different from the control and PRMTA (P < 0.05); in the other periods it was similar to the control (P > 0.05). It can be concluded that Sealepox and Sealepox-RP were biocompatible and demonstrated similar immunoinflammatory response regarding IL-6, TNF-α, and CD5 compared to PRMTA., (© 2022. The Author(s), under exclusive licence to The Society of The Nippon Dental University.)

Background: Antimicrobial photodynamic therapy (aPDT) is used as an adjunct to endodontic treatment to enhance microbial reduction in the root canal system. However, studies evaluating the impact of laser ablation with Indocyanine Green (ICG) are scarce. Thus, this in vitro study aimed to evaluate the efficiency of laser ablation with ICG using different laser parameters compared with aPDT using photosensitizer methylene blue (MB) and curcumin (CUR) on the reduction of E. faecalis biofilms on root canals., Methods: Forty-nine human premolars were used after biomechanical instrumentation for standardization. The root canals were contaminated with E. faecalis for 10 days to form biofilms, and divided into 7 groups (n = 7): 0.01% MB activated by red laser - MB+RL; 0.05% CUR activated by blue LED - CUR+BL; 0.05% ICG activated by infrared diode laser (2.5 W power, 30 ms interval, and 30 ms duration) - ICG+DL 2.5/30/30; 0.05% ICG activated by infrared diode laser (2.5 W power, 300 ms interval, and 100 ms duration) - ICG+DL 2.5/300/100; 0.05% ICG activated by infrared diode laser (3 W power, 300 ms interval, and 100 ms duration) - ICG+DL 3/300/100; sterile saline solution (negative control) - NC; and 2.5% NaOCl (positive control) - PC. Root canal sampling was performed prior to and immediately after the different treatment protocols. Data were submitted to One- and Two-Way ANOVA, followed by Student-Newman-Keuls test or Fisher LSD's test (α = 0.05)., Results: All aPDT protocols promoted significant CFU reductions compared with the NC; the highest CFU reduction was observed for PC (p < 0.05). Among the protocols the highest CFU reduction was promoted by laser ablation with ICG+DL 3/300/100 (p < 0.05) except compared with aPDT using CUR+BL (p > 0.05)., Conclusions: Laser ablation protocol using ICG+DL proved to be efficient in reducing E. faecalis biofilms, especially when activated at 3/300/100 configuration., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Objectives: The aim of this study was to assess the physicochemical properties, cytotoxicity and penetration into dentinal tubules of ChlorCid™ Surf (3% sodium hypochlorite [NaOCl] with surfactant) in comparison to ChlorCid™ (3% NaOCl without surfactant)., Materials and Methods: The physicochemical properties evaluated were pH, surface tension, free available chlorine (FAC) and contact angle. Cytotoxicity was evaluated in L929 fibroblasts exposed to the solutions by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red assays. Assessment of penetration into dentinal tubules was performed by staining single-rooted permanent human teeth with crystal violet ( n = 9), which were irrigated with the solutions and analyzed in cervical, middle and apical segments. Data were analyzed by one-way analysis of variance (ANOVA) and Tukey's post -test, 2-way ANOVA and Bonferroni's post -test or t -test (α = 0.05)., Results: ChlorCid™ Surf and ChlorCid™ FAC values were close to those indicated by the manufacturer. ChlorCid™ Surf showed lower surface tension and contact angle on dentin, and higher pH than ChlorCid™ ( p < 0.05). The penetration of ChlorCid™ Surf was higher in cervical and middle segments, compared with ChlorCid™ ( p < 0.05). There was no difference in irrigant cytotoxicity ( p > 0.05)., Conclusions: ChlorCid™ Surf showed lower surface tension, lower contact angle on root canal dentin, higher penetration into dentinal tubules and more alkaline pH, compared with ChlorCid™. However, both solutions showed similar cytotoxicity and FAC content., Competing Interests: Conflict of Interest: No potential conflict of interest relevant to this article was reported., (Copyright © 2020. The Korean Academy of Conservative Dentistry.)

Aim: To assess the penetration of sodium hypochlorite (NaOCl) gel or NaOCl solutions with surfactants, and the effect of passive ultrasonic irrigation (PUI) on penetration into dentinal tubules., Methodology: Bovine incisor root canals were instrumented, the roots sectioned and the dentine blocks obtained were stained with crystal violet. Dentine blocks (n = 10 per group) were exposed to 3% NaOCl gel or 3% NaOCl solution for 10 and 20 min. Other dentine blocks (n = 10 per group) were exposed to Chlor-Extra (6% NaOCl + surfactant), 6% NaOCl, 2.5% NaOCl with 0.2% cetrimide and 2.5% NaOCl for 10 and 20 min. The penetration depth of irrigants into dentinal tubules was measured in micrometres by viewing the bleached crystal violet under a stereomicroscope. Additionally, bovine incisor root canals, instrumented and stained with crystal violet, were distributed into two groups (n = 10) and irrigated with 2.5% NaOCl with PUI or conventional syringe irrigation (CSI). The penetration depth of irrigants into dentinal tubules was assessed 3 and 7 mm from the apex. Statistical analysis was performed by ANOVA and Tukey tests (α = 0.05)., Results: There was significantly greater penetration of 3% NaOCl solution into dentinal tubules compared with the gel form (P < 0.05). There was no difference (P > 0.05) between 6% NaOCl and Chlor-Extra, and between 2.5% NaOCl and 2.5% NaOCl + cetrimide. PUI significantly increased the penetration depth of NaOCl into dentinal tubules when compared with CSI (P < 0.05)., Conclusions: In extracted bovine incisors, NaOCl gel penetrated less into dentinal tubules than NaOCl solution. The addition of surfactants did not increase the penetration depth. The use of PUI significantly increased NaOCl penetration into dentinal tubules., (© 2018 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Aim: To evaluate the influence of powder-to-gel ratio (0.19 g powder to 50 μL of gel, thick MTA Flow, and 0.06 g powder to 50 μL of gel, fluid MTA Flow) on biocompatibility of MTA Flow (Ultradent Products Inc., South Jordan, UT, USA, lot: 2015122901) and compare it with Biodentine (Septodont Inc., Saint-Maur-des-Fossés, France, lot: B18542A)., Methodology: The materials were manipulated and inserted into polyethylene tubes for implantation in twenty rats. After 7, 15, 30 and 60 days, the specimens were removed and embedded in paraffin. Haematoxylin and eosin sections were used to count the number of inflammatory cells (IC) and fibroblasts mm -2 (Fb). In the Masson's trichrome-stained sections, the fibrous capsule thickness was measured; picrosirius red-stained sections were used for birefringent collagen quantification. The data were submitted to two-way ANOVA and Tukey test (P ≤ 0.05)., Results: A significantly lower number of IC and consequently higher number of Fb were observed in the capsules adjacent to thick MTA Flow at all periods, in comparison with other materials (P ≤ 0.05). At 60 days, the quantity of birefringent collagen was significantly greater in the tissue in contact with thick MTA Flow, when compared with fluid MTA Flow and Biodentine., Conclusions: Although thick MTA Flow induced a less intense inflammatory response, all evaluated materials are biocompatible because they allowed regression of this process after 60 days., (© 2018 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Aim: To evaluate pH and antibacterial activity of pastes with calcium hydroxide [Ca(OH)2] and zinc oxide (ZnO) microparticles (micro) or nanoparticles (nano) and association with 0.4% chlorhexidine against Enterococcus faecalis., Materials and Methods: The following pastes were analyzed: Ca(OH)2/ZnO micro, (2) Ca(OH)2/ZnO nano, (3) Ca(OH)2/ ZnO micro + 0.4% chlorhexidine, (4) Ca(OH)2/ZnO nano + 0.4% chlorhexidine. Antibacterial activity against E. faecalis was evaluated by agar diffusion test. The direct contact test on planktonic cells of E. faecalis was performed for 30 and 60 seconds. Root canals from bovine teeth were filled with the pastes and pH was evaluated after 1, 7, 14, 21, 30 and 60 days. The data obtained were submitted to the statistical tests analysis of variance (ANOVA) and Tukey or Kruskal-Wallis and Dunn test, with a 5% significance level., Results: Calcium hydroxide and zinc oxide nano, and the pastes with 0.4% chlorhexidine were more effective in agar diffusion test. In the direct contact test, the pastes with chlorhexidine showed the highest effect after 30 seconds. All pastes eliminated E. faecalis after 60 seconds. All pastes promoted an increase in pH. The highest increase in pH was observed with nanoparticle medications after 1 and 7 days (p < 0.05). After this period, the pastes presented similar pH increase., Conclusion: It was concluded that calcium hydroxide and zinc oxide nanoparticles promoted greater initial alkalinization. The antimicrobial activity of the pastes against E. faecalis is favored by the association with chlorhexidine., Clinical Significance: Although nanoparticles of calcium hydroxide and zinc oxide promoted antibacterial effect, the activity against E. faecalis is favored by association with chlorhexidine.

Objective: The goal of this study was to evaluate the efficacy of the Self-Adjusting File (SAF) and ProTaper for removing calcium hydroxide [Ca(OH)2] from root canals., Material and Methods: Thirty-six human mandibular incisors were instrumented with the ProTaper system up to instrument F2 and filled with a Ca(OH)2-based dressing. After 7 days, specimens were distributed in two groups (n=15) according to the method of Ca(OH)2 removal. Group I (SAF) was irrigated with 5 mL of NaOCl and SAF was used for 30 seconds under constant irrigation with 5 mL of NaOCl using the Vatea irrigation device, followed by irrigation with 3 mL of EDTA and 5 mL of NaOCl. Group II (ProTaper) was irrigated with 5 mL of NaOCl, the F2 instrument was used for 30 seconds, followed by irrigation with 5 mL of NaOCl, 3 mL of EDTA, and 5 mL of NaOCl. In 3 teeth Ca(OH)2 was not removed (positive control) and in 3 teeth canals were not filled with Ca(OH)2 (negative control). Teeth were sectioned and prepared for the scanning electron microscopy. The amounts of residual Ca(OH)2 were evaluated in the middle and apical thirds using a 5-score system., Results: None of the techniques completely removed the Ca(OH)2 dressing. No difference was observed between SAF and ProTaper in removing Ca(OH)2 in the middle (P=0.11) and the apical (P=0.23) thirds., Conclusion: The SAF system showed similar efficacy to rotary instrument for removal of Ca(OH)2 from mandibular incisor root canals.

Aim: To compare the ex vivo antibacterial effectiveness of the Endox Plus system and sodium hypochlorite (NaOCl) in combination with BioPure MTAD (Tulsa Dental, Tulsa, OK, USA) or with EDTA in Enterococcus faecalis-contaminated root canals., Methodology: After initial preparation, the root canals of 70 single-rooted human teeth were inoculated with E. faecalis (ATCC 29212) and incubated for 21 days. Specimens were divided into five groups: Endox Plus/saline; 2.5% NaOCl/MTAD; 2.5% NaOCl/EDTA; saline (positive control); negative control (root canals not prepared, nor irrigated). Samples were collected using paper points. Microbiological analysis evaluated the number of CFUs. Data were analysed by anova and Tukey tests at 0.05 significance., Results: All specimens had bacterial growth after the incubation period, with similar CFU per mL counts (P > 0.05). After chemo-mechanical preparation, the number of bacteria in all groups reduced, except for the negative control. No significant differences were observed between 2.5% NaOCl/MTAD and 2.5% NaOCl/EDTA, but these groups had lower CFU counts than the other groups (P < 0.05). In the final samples, an increase in the bacterial counts was observed for Endox Plus/saline, 2.5% NaOCl/MTAD, 2.5% NaOCl/EDTA and saline (P < 0.05) with no significant differences between these groups., Conclusions: This ex vivo study revealed that the Endox Plus system was associated with a reduced antibacterial effectiveness compared with conventional irrigation using 2.5% NaOCl/MTAD and 2.5% NaOCl/EDTA. All irrigation procedures allowed recovery of bacteria 7 days after treatment, demonstrating persistence of contamination within the root canal system., (© 2012 International Endodontic Journal.)

Aim: To determine the influence of filling technique and root canal area on the percentage of gutta-percha (PGP) in laterally compacted root fillings., Methodology: Sixty extracted canine teeth were accessed and the root canals instrumented to the same size. They were then divided in three groups and filled with laterally compacted gutta-percha cones and AH Plus using different techniques. A variation of cold lateral compaction using a sequence of spreaders prior to accessory cone placement was compared to two commonly-used techniques. Twenty additional canines with prepared root canals were used as negative controls in which gutta-percha was introduced into the canals but no compaction applied. The roots were sectioned horizontally at 3 and 6 mm from the apex and micro-photographs taken. Using software, the area of the canals and gutta-percha at each level were measured and PGP calculated. A Multivariate analysis was used to determine the variables influencing PGP. A linear regression test was used to verify the variation in PGP explained by canal area., Results: At each level the largest canal was two to three times wider than the smallest. Canal area significantly influenced the PGP at both levels (P < 0.05), however, the variation in PGP was only partially explained by canal area (r(2) = 0.154, 6 mm; r(2) = 0.119, 3 mm). The PGP at the 3 mm level was lower than at 6 mm (P = 0.003). The spreader-sequence technique achieved a higher PGP than the other two techniques (P = 0.00002). The control group had the lowest area of GP., Conclusions: Variations in root canal filling technique and canal area influenced the percentage of gutta-percha of laterally compacted root fillings. The percentage of gutta-percha was lower at the 3 mm level compared to the 6 mm level.

This study evaluated in vitro the cytotoxicity of four root canal sealers (Topseal, EndoRez, TubliSeal and Kerr Pulp Canal Sealer E.W.T.) and their effects on reactive oxygen/nitrogen intermediate induction by mouse peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 mL 10 mM phosphate-buffered saline, washed twice and resuspended (10(6) cells/mL) in appropriate medium for each test. Cytotoxicity was determined by the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. Sealer suspensions were obtained in two different concentrations from each material: 18 mg/mL and 9 mg/mL, established according to compatibility parameters following MTT assay. Comparing the sealers, H2O2 release at concentrations of 9 mg/mL and 18 mg/mL was similar: Topseal > positive control (medium + cells + 5 mg/mL zimozan solution) > EndoRez > TubliSeal > Kerr Pulp E.W.T. > negative control (medium + cells). NO release at concentration of 9 mg/mL was: positive control (medium + cells + 10 microg/mL LPS solution) > Topseal > Kerr Pulp E.W.T. > TubliSeal = EndoRez > negative control (medium + cells); at concentration of 18 mg/mL was: positive control > Topseal > Kerr Pulp E.W.T > TubliSeal > EndoRez > negative control. Based on the results, it may be concluded that Topseal presented the highest cytotoxicity among the tested sealers, releasing higher concentrations of NO and H2O2 in macrophage culture.

Aim: To report the diagnosis and successful clinical management of a patient with anatomical variation in maxillary first and second premolars., Summary: Maxillary premolars have a highly variable root canal morphology, but it is rare, especially in the second premolar, to find three canals. This article describes the diagnosis and clinical management of first and second premolars with three canals and three separate roots, drawing particular attention to radiographic interpretation and access refinements., Key Learning Points: Clinicians should be aware of anatomical variations in maxillary premolars and be able to apply this knowledge in radiographic and clinical interpretation. Access cavity refinements may be required for stress-free entry to complex anatomy. Complex premolar anatomy may be predictably managed following its identification and negotiation.

The cytotoxicity of four calcium hydroxide-based root canal sealers (Sealapex, CRCS, Apexit, and Sealer 26) and one zinc oxide-eugenol-based sealer (Fill Canal) was evaluated microscopically for morphological changes in rat peritoneal macrophages. The least cytotoxic sealer was Fill Canal, followed in increasing order of cytotoxicity by CRCS, Sealer 26, Apexit, and Sealapex.

A study of the subcutaneous connective tissue response of 24 white rats to three different formulations of gutta-percha was undertaken. The prepared specimens were examined under the light microscope after intervals of 7, 21, 60 and 120 days. The results showed identical tissue responses after the initial period of 7 days. However, after 120 days the gutta-percha supplied with the Ultrafil system presented mature granulation tissue with neither oedema nor vascular congestion, in contrast to the responses observed with the McSpadden and Obtura formulations.