α-IRAK-4 Suppresses the Activation of RANK/RANKL Pathway on Macrophages Exposed to Endodontic Microorganisms.

Periapical lesions are common pathologies affecting the alveolar bone, often initiated by intraradicular lesions resulting from microbial exposure to dental pulp. These microorganisms trigger inflammatory and immune responses. When endodontic treatment fails to eliminate the infection, periapical lesions persist, leading to bone loss. The RANK/RANKL/OPG pathway plays a crucial role in both the formation and the destruction of the bone. In this study, the objective was to inhibit the RANK/RANKL pathway in vitro within exposed Thp-1 macrophages to endodontic microorganisms, specifically Enterococcus faecalis, which was isolated from root canals of 20 patients with endodontic secondary/persistent infection, symptomatic and asymptomatic, and utilizing an α-IRAK-4 inhibitor, we introduced endodontic microorganisms and/or lipoteichoic acid from Streptococcus spp. to cellular cultures in a culture plate, containing thp-1 cells and/or PBMC from patients with apical periodontitis. Subsequently, we assessed the percentages of RANK+, RANKL+, and OPG+ cells through flow cytometry and measured the levels of several inflammatory cytokines (IL-1ß, TNF-α, IL-6, IL-8, IL-10, and IL-12p70) in the cellular culture supernatant through a CBA kit and performed analysis by flow cytometry. A significant difference was observed in the percentages of RANK+RANKL+, OPG+ RANKL+ cells in thp-1 cells and PBMCs from patients with apical periodontitis. The findings revealed significant differences in the percentages of the evaluated cells, highlighting the novel role of the IRAK-4 inhibitor in addressing this oral pathology, apical periodontitis, where bone destruction is observed.

4-META/MMA-TBB resin containing nano hydroxyapatite induces the healing of periodontal tissue repair in perforations at the pulp chamber floor.

We aimed to evaluate the materials based on 4-methacryloxyethyl trimellitate anhydride/methyl methacrylate tri-n-butylborane (Super-bond [SB]) and nano hydroxyapatite (naHAp) for the repair of perforation at pulp chamber floor (PPF) in vitro and in vivo models. SB and naHAp were mixed in the mass ratio of 10% or 30% to produce naHAp/SB. Human periodontal ligament stem cells (HPDLSCs) were cultured on resin discs of SB or naHAp/SB to analyze the effects of naHAp/SB on cell adhesion, proliferation, and cementoblastic differentiation. A rat PPF model was treated with SB or naHAp/SB to examine the effects of naHAp/SB on the healing of defected cementum and periodontal ligament (PDL) at the site of PPF. HPDLSCs were spindle-shaped and adhered to all resin discs. Changing the resin from SB to naHAp/SB did not significantly alter cell proliferation. Both 10% and 30% naHAp/SB were more effective than SB in promoting cementoblastic differentiation of HPDLSCs. In the rat PPF model, 30% naHAp/SB was more effective than SB in promoting the formation Sharpey's fiber-like structures with expression of the PDL-related marker and cementum-like structures with expression of cementum-related markers. In conclusion, 30% naHAp/SB can be the new restorative material for PPF because it exhibited the abilities of adhering to dentin and healing of defected periodontal tissue.

Effect of whitening toothpastes with different hydrogen peroxide concentrations: Penetration into the pulp chamber and color change.

OBJECTIVES: This study evaluated the efficacy of simulated brushing with toothpastes containing different concentrations of hydrogen peroxide (HP) in pulp chamber penetration and color change. Also, physical-chemical properties (concentration, pH and viscosity) were evaluated. METHODS: Forty-nine premolars were divided into seven groups (n = 7): untreated (control); whitening gel (White Class 6 %, 6 %BG) with one 90  min application (6 %BG 90  min) and 14 applications of 90  min (6 %BG 14×90 min); toothpastes (Colgate Luminous White Glow 3 %, 3 %TP; Crest 3D White Brilliance 4 %, 4 %TP; Colgate Optic White Pro-Series 5 %, 5 %TP) and 6 %BG toothbrushing for 14 applications of 90 s. HP penetration into the pulp chamber was measured through UV-Vis spectrophotometry and color change with a spectrophotometer (ΔEab, ΔE00, and ΔWID). Initial concentration, pH, and viscosity were measured through Titration, Digital pH-meter, and Rheometer, respectively. Statistical analysis used one-way ANOVA and Tukey's test (α = 0.05). RESULTS: 6 %BG (14×90 min) and 4 %TP groups showed acidic pH and higher concentrations of HP in the pulp chamber compared to the other groups (p < 0.05). On the other side, 3 %TP and 5 %TP groups showed alkaline pH, higher viscosity between the toothpastes and lower HP penetration (p < 0.05). The 6 %BG AH (14×90 min) group exhibited the most significant color change (ΔEab, ΔE00, and ΔWID) (p < 0.05). CONCLUSIONS: Brushing with whitening toothpaste with an acidic pH leads to greater HP penetration into pulp chamber; but, even when a high concentrated HP whitening toothpaste was used, a lower whitening effect was observed when compared to a two-week at-home bleaching. CLINICAL SIGNIFICANCE: Whitening toothpastes containing up to 5 % HP produced lower whitening effect than two-week at-home bleaching. Additionally, HP was detected within the pulp chamber which can potentially impact in tooth sensitivity.

Reduced C-reactive protein levels after root canal treatment in clinically healthy young apical periodontitis individuals at cardiovascular risk. A prospective study.

AIM: To determine the systemic inflammatory burden, including hsCRP and its monomeric forms, in patients with apical lesions of endodontic origin treated with root canal treatment (RCT). METHODOLOGY: Prospective pre-/post-study. Apical periodontitis (AP) individuals aged 16-40 were included (N = 29). Individuals received RCT and were followed at 1 and 6 months. Fasting blood samples were obtained. Apical lesions of endodontic origin (ALEO) diameter (mm), and periapical index (PAI), were recorded. The serum concentrations of total hsCRP were determined by turbidimetry. Tumour necrosis factor (TNF)-α, interleukin (IL)-6, IL-10, IL-1ß, and soluble (s) E-selectin were assessed by Multiplex assay. Additionally, mCRP forms were determined in the serum of AP patients with a baseline moderate to high cardiovascular risk based on hsCRP stratification (hsCRP ≥1 mg/L) by immunowestern blot (n = 15). Also, CRP isoforms were explored in ALEOs from AP individuals (n = 4). Data were analysed with StataV16. RESULTS: Periapical index and ALEO sizes were reduced at both follow-up visits after RCT (p < .05). Serum levels of TNF-α, IL-6, IL-10, IL-1ß, and sE-selectin did not show significant differences. CRP was borderline reduced at 1 month (p = .04); however, in AP individuals at cardiovascular risk (hsCRP ≥ 1 mg/L), hsCRP and its monomeric isoform significantly decreased at 1 and 6 months (p < .05). CONCLUSIONS: High-sensitivity CRP and mCRP are reduced after RCT in AP individuals at cardiovascular risk.

Analysis of bacterial communities of infected primary teeth in a Mexican population

BACKGROUND: The objective of this study was to describe the bacterial communities associated with pediatric patients with endodontic infections of temporal teeth by targeting the 16S rRNA gene using pyrosequencing. MATERIAL AND METHODS: Microbiological samples were obtained from the lower primary molars of thirteen 13 pediatric patients with dental infections. An aspiration method for microbiological sampling was used. The identification of microbiota employing the pyrosequencing method by targeting the 16S gene was performed. RESULTS: Ribosomal 16S RNA gene sequences were amplified, obtaining a total of 16,182 sequences from 13 primary infected molars (13 different individuals) by pyrosequencing. Bacteroidetes phyla (35.15%) were the most abundant followed by Firmicutes (33.3%) and Fusobacteria (10.05%); the presence of specific pathogenic bacteria was determined as well. CONCLUSIONS: The infected root canal of primary teeth contains a high diversity of anaerobic bacteria, and Bacteroidetes, Firmicutes, and Fusobacteria phyla were the most abundant; Prevotella and Streptococcus genera were the most prevalent

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Root canal contamination or exposure to lipopolysaccharide differentially modulate prostaglandin E 2 and leukotriene B 4 signaling in apical periodontitis.

Purpose To evaluate the kinetics of apical periodontitis development in vivo , induced either by contamination of the root canals by microorganisms from the oral cavity or by inoculation of bacterial lipopolysaccharide (LPS) and the regulation of major enzymes and receptors involved in the arachidonic acid metabolism. Methodology Apical periodontitis was induced in C57BL6 mice (n=96), by root canal exposure to oral cavity (n=48 teeth) or inoculation of LPS (10 µL of a suspension of 0.1 µg/µL) from E. coli into the root canals (n= 48 teeth). Healthy teeth were used as control (n=48 teeth). After 7, 14, 21 and 28 days the animals were euthanized and tissues removed for histopathological and qRT-PCR analyses. Histological analysis data were analyzed using two-way ANOVA followed by Sidak's test, and qRT-PCR data using two-way ANOVA followed by Tukey's test (α=0.05). Results Contamination by microorganisms led to the development of apical periodontitis, characterized by the recruitment of inflammatory cells and bone tissue resorption, whereas inoculation of LPS induced inflammatory cells recruitment without bone resorption. Both stimuli induced mRNA expression for cyclooxygenase-2 and 5-lipoxygenase enzymes. Expression of prostaglandin E 2 and leukotriene B 4 cell surface receptors were more stimulated by LPS. Regarding nuclear peroxisome proliferator-activated receptors (PPAR), oral contamination induced the synthesis of mRNA for PPARδ, differently from inoculation of LPS, that induced PPARα and PPARγ expression. Conclusions Contamination of the root canals by microorganisms from oral cavity induced the development of apical periodontitis differently than by inoculation with LPS, characterized by less bone loss than the first model. Regardless of the model used, it was found a local increase in the synthesis of mRNA for the enzymes 5-lipoxygenase and cyclooxygenase-2 of the arachidonic acid metabolism, as well as in the surface and nuclear receptors for the lipid mediators prostaglandin E2 and leukotriene B4.

Pretreatment of Root Canal with Photodynamic Therapy Facilitates Adhesion, Viability and Differentiation of Stem Cells of the Apical Papilla.

This study was to test the hypothesis that root canal pretreated with photodynamic therapy (PDT) would promote stem cells from the apical papilla (SCAP) adhesion, proliferation and differentiation without affecting smear layer removal and microhardness of root canal. Standardized root canals were randomized into four groups (n = 30/group): (1) sodium hypochlorite(NaOCl) group, (2) NaOCl + ethylene diaminetetraacetic acid (EDTA) group, (3) NaOCl + PDT group, (4) NaOCl + EDTA + PDT group. After treatments, smear layer removal and microhardness of root canal were evaluated. SCAP with hydroxyapatite-based scaffolds were seeded into root canals for 7 days. SCAP adhesion was observed by scanning electron microscope (SEM), and viable cells were calculated by CellTiter-Glo Luminescent kit. Platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) expression of SCAP were evaluated by Quantitative Reverse Transcriptase-Polymerase Chain Reaction. There was no significant difference in the smear layer removal and microhardness of root dentin between the groups with and without PDT treatment (P > 0.05). SCAP with elongated cytoplasmic processes and cell-cell contact were observed on the dentin surfaces treated with PDT. Elevated cell viability, PDGF and VEGF expression were found in root canal treated with PDT (P < 0.05). Under the experimental conditions, PDT could provide positive microenvironment for SCAP growth.

Comparison of apical irrigant solution extrusion among conventional and laser-activated endodontic irrigation.

The aim of this study was to determine the amount of extruded endodontic irrigant among needle-syringe irrigation (NSI) and laser-activated irrigation (LAI) regimens. Twenty extracted maxillary central incisors were prepared utilizing GT professional rotary files (size 40, taper 0.06). Irrigation was performed with two 27 G irrigation needles (notched open ended (ON) and single side vented (SV)) each at two different irrigant volumetric flow rates (VFR)-0.05 ml/s (3 ml/min) and 0.10 ml/s (6 ml/min). LAI was performed with Er:YAG (erbium-doped yttrium aluminum garnet) using different fiber types (X-Pulse-14/400 cylindrical tip, Preciso- 14/300 flat cylindrical tip, PIPS- 14/400 quartz tapered tip). The Er:YAG laser with a wavelength of 2940 nm (Lightwalker AT, Fotona, Ljubljana, Slovenia) was used according to the following protocol: 10 mJ per pulse, 15 Hz, pulse duration 50 µs. Irrigation time was 60 s for all protocols. Precision syringe pump (PSP) maintained constant irrigant volumetric flow rate. Apically extruded irrigant was collected and net weighed for each protocol (N = 10). Data were analyzed by t tests and Kruskal-Wallis. All LAI regimens had statistically significant lower irrigant extrusion compared with NSI except for the SV 27 G needle used with 0.05 ml/s VFR when compared with the Preciso fiber tip (p = 0,230). The largest amount of extruded irrigant was with the ON 27 G needle at the 0.10 ml/s VFR, while the smallest was after LAI with PIPS fiber tip. The lower quantity of apically extruded irrigant during LAI (X-Pulse and PIPS) points out a safer endodontic irrigation method compared with conventional irrigations.

Root canal contamination or exposure to lipopolysaccharide differentially modulate prostaglandin E 2 and leukotriene B 4 signaling in apical periodontitis

Abstract Purpose To evaluate the kinetics of apical periodontitis development in vivo , induced either by contamination of the root canals by microorganisms from the oral cavity or by inoculation of bacterial lipopolysaccharide (LPS) and the regulation of major enzymes and receptors involved in the arachidonic acid metabolism. Methodology Apical periodontitis was induced in C57BL6 mice (n=96), by root canal exposure to oral cavity (n=48 teeth) or inoculation of LPS (10 µL of a suspension of 0.1 µg/µL) from E. coli into the root canals (n= 48 teeth). Healthy teeth were used as control (n=48 teeth). After 7, 14, 21 and 28 days the animals were euthanized and tissues removed for histopathological and qRT-PCR analyses. Histological analysis data were analyzed using two-way ANOVA followed by Sidak's test, and qRT-PCR data using two-way ANOVA followed by Tukey's test (α=0.05). Results Contamination by microorganisms led to the development of apical periodontitis, characterized by the recruitment of inflammatory cells and bone tissue resorption, whereas inoculation of LPS induced inflammatory cells recruitment without bone resorption. Both stimuli induced mRNA expression for cyclooxygenase-2 and 5-lipoxygenase enzymes. Expression of prostaglandin E 2 and leukotriene B 4 cell surface receptors were more stimulated by LPS. Regarding nuclear peroxisome proliferator-activated receptors (PPAR), oral contamination induced the synthesis of mRNA for PPARδ, differently from inoculation of LPS, that induced PPARα and PPARγ expression. Conclusions Contamination of the root canals by microorganisms from oral cavity induced the development of apical periodontitis differently than by inoculation with LPS, characterized by less bone loss than the first model. Regardless of the model used, it was found a local increase in the synthesis of mRNA for the enzymes 5-lipoxygenase and cyclooxygenase-2 of the arachidonic acid metabolism, as well as in the surface and nuclear receptors for the lipid mediators prostaglandin E2 and leukotriene B4.

Periapical bone response to bacterial lipopolysaccharide is shifted upon cyclooxygenase blockage.

OBJECTIVES: Infection, inflammation and bone resorption are closely related events in apical periodontitis development. Therefore, we sought to investigate the role of cyclooxygenase (COX) in osteoclastogenesis and bone metabolism signaling in periapical bone tissue after bacterial lipopolysaccharide (LPS) inoculation into root canals. METHODOLOGY: Seventy two C57BL/6 mice had the root canals of the first molars inoculated with a solution containing LPS from E. coli (1.0 mg/mL) and received selective (celecoxib) or non-selective (indomethacin) COX-2 inhibitor. After 7, 14, 21 and 28 days the animals were euthanized and the tissues removed for total RNA extraction. Evaluation of gene expression was performed by qRT-PCR. Statistical analysis was performed using analysis of variance (ANOVA) followed by post-tests (α=0.05). RESULTS: LPS induced expression of mRNA for COX-2 (Ptgs2) and PGE2 receptors (Ptger1, Ptger3 and Ptger4), indicating that cyclooxygenase is involved in periapical response to LPS. A signaling that favours bone resorption was observed because Tnfsf11 (RANKL), Vegfa, Ctsk, Mmp9, Cd36, Icam, Vcam1, Nfkb1 and Sox9 were upregulated in response to LPS. Indomethacin and celecoxib differentially modulated expression of osteoclastogenic and other bone metabolism genes: celecoxib downregulated Igf1r, Ctsk, Mmp9, Cd36, Icam1, Nfkb1, Smad3, Sox9, Csf3, Vcam1 and Itga3 whereas indomethacin inhibited Tgfbr1, Igf1r, Ctsk, Mmp9, Sox9, Cd36 and Icam1. CONCLUSIONS: We demonstrated that gene expression for COX-2 and PGE2 receptors was upregulated after LPS inoculation into the root canals. Additionally, early administration of indomethacin and celecoxib (NSAIDs) inhibited osteoclastogenic signaling. The relevance of the cyclooxygenase pathway in apical periodontitis was shown by a wide modulation in the expression of genes involved in both bone catabolism and anabolism.

A novel patient-specific three-dimensional drug delivery construct for regenerative endodontics.

Evoked bleeding (EB) clinical procedure, comprising a disinfection step followed by periapical tissue laceration to induce the ingrowth of undifferentiated stem cells from the periodontal ligament and alveolar bone, is currently the only regenerative-based therapeutic approach to treating pulp tissue necrosis in undeveloped (immature) permanent teeth approved in the United States. Yet, the disinfection step using antibiotic-based pastes leads to cytotoxic, warranting a biocompatible strategy to promote root canal disinfection with no or minimal side-effects to maximize the regenerative outcomes. The purpose of this investigation was to develop a tubular three-dimensional (3D) triple antibiotic-eluting construct for intracanal drug delivery. Morphological (scanning electron microscopy), chemical (Fourier transform infrared spectroscopy), and mechanical (tensile testing) characteristics of the polydioxanone-based triple antibiotic-eluting fibers were assessed. The antimicrobial properties of the tubular 3D constructs were determined in vitro and in vivo using an infected (Actinomyces naeslundii) dentin tooth slice model and a canine method of periapical disease, respectively. The in vitro data indicated significant antimicrobial activity and the ability to eliminate bacterial biofilm inside dentinal tubules. In vivo histological findings demonstrated that, using the EB procedure, the tubular 3D triple antibiotic-eluting construct allowed the formation of an appropriate environment that led to apex closure and the ingrowth of a thin layer of osteodentin-like tissue into the root canal. Taken together, these findings indicate that our novel drug delivery construct is a promising biocompatible disinfection strategy for immature permanent teeth with necrotic pulps. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1576-1586, 2019.

Periapical bone response to bacterial lipopolysaccharide is shifted upon cyclooxygenase blockage

Abstract Objectives: Infection, inflammation and bone resorption are closely related events in apical periodontitis development. Therefore, we sought to investigate the role of cyclooxygenase (COX) in osteoclastogenesis and bone metabolism signaling in periapical bone tissue after bacterial lipopolysaccharide (LPS) inoculation into root canals. Methodology: Seventy two C57BL/6 mice had the root canals of the first molars inoculated with a solution containing LPS from E. coli (1.0 mg/mL) and received selective (celecoxib) or non-selective (indomethacin) COX-2 inhibitor. After 7, 14, 21 and 28 days the animals were euthanized and the tissues removed for total RNA extraction. Evaluation of gene expression was performed by qRT-PCR. Statistical analysis was performed using analysis of variance (ANOVA) followed by post-tests (α=0.05). Results: LPS induced expression of mRNA for COX-2 (Ptgs2) and PGE2 receptors (Ptger1, Ptger3 and Ptger4), indicating that cyclooxygenase is involved in periapical response to LPS. A signaling that favours bone resorption was observed because Tnfsf11 (RANKL), Vegfa, Ctsk, Mmp9, Cd36, Icam, Vcam1, Nfkb1 and Sox9 were upregulated in response to LPS. Indomethacin and celecoxib differentially modulated expression of osteoclastogenic and other bone metabolism genes: celecoxib downregulated Igf1r, Ctsk, Mmp9, Cd36, Icam1, Nfkb1, Smad3, Sox9, Csf3, Vcam1 and Itga3 whereas indomethacin inhibited Tgfbr1, Igf1r, Ctsk, Mmp9, Sox9, Cd36 and Icam1. Conclusions: We demonstrated that gene expression for COX-2 and PGE2 receptors was upregulated after LPS inoculation into the root canals. Additionally, early administration of indomethacin and celecoxib (NSAIDs) inhibited osteoclastogenic signaling. The relevance of the cyclooxygenase pathway in apical periodontitis was shown by a wide modulation in the expression of genes involved in both bone catabolism and anabolism.

Butyrate Stimulates Histone H3 Acetylation, 8-Isoprostane Production, RANKL Expression, and Regulated Osteoprotegerin Expression/Secretion in MG-63 Osteoblastic Cells.

Butyric acid as a histone deacetylase (HDAC) inhibitor is produced by a number of periodontal and root canal microorganisms (such as Porphyromonas, Fusobacterium, etc.). Butyric acid may affect the biological activities of periodontal/periapical cells such as osteoblasts, periodontal ligament cells, etc., and thus affect periodontal/periapical tissue destruction and healing. The purposes of this study were to study the toxic effects of butyrate on the matrix and mineralization marker expression in MG-63 osteoblasts. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Cellular apoptosis and necrosis were analyzed by propidium iodide/annexin V flow cytometry. The protein and mRNA expression of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) were analyzed by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). OPG, soluble RANKL (sRANKL), 8-isoprostane, pro-collagen I, matrix metalloproteinase-2 (MMP-2), osteonectin (SPARC), osteocalcin and osteopontin (OPN) secretion into culture medium were measured by enzyme-linked immunosorbant assay. Alkaline phosphatase (ALP) activity was checked by ALP staining. Histone H3 acetylation levels were evaluated by immunofluorescent staining (IF) and Western blot. We found that butyrate activated the histone H3 acetylation of MG-63 cells. Exposure of MG-63 cells to butyrate partly decreased cell viability with no marked increase in apoptosis and necrosis. Twenty-four hours of exposure to butyrate stimulated RANKL protein expression, whereas it inhibited OPG protein expression. Butyrate also inhibited the secretion of OPG in MG-63 cells, whereas the sRANKL level was below the detection limit. However, 3 days of exposure to butyrate (1 to 8 mM) or other HDAC inhibitors such as phenylbutyrate, valproic acid and trichostatin stimulated OPG secretion. Butyrate stimulated 8-isoprostane, MMP-2 and OPN secretion, but not procollagen I, or osteocalcin in MG-63 cells. Exposure to butyrate (2⁻4 mM) for 3 days markedly stimulated osteonectin secretion and ALP activity. In conclusion, higher concentrations of butyric acid generated by periodontal and root canal microorganisms may potentially induce bone destruction and impair bone repair by the alteration of OPG/RANKL expression/secretion, 8-isoprostane, MMP-2 and OPN secretion, and affect cell viability. However, lower concentrations of butyrate (1⁻4 mM) may stimulate ALP, osteonectin and OPG. These effects are possibly related to increased histone acetylation. These events are important in the pathogenesis and repair of periodontal and periapical destruction.

Regenerating a monoblock to obturate root canalsvia a mineralising strategy.

To develop a novel strategy for sealing and obturating dental root canals by tooth-like tissue regeneration, premolars with mature root apices were freshly collected, and root canals were prepared by following the clinical protocols in vitro. The teeth were immersed in supersaturated calcium and phosphate solution containing gallic acid and fluoride. At certain intervals, the dental roots were taken out, and their mineral precipitates were characterised by scanning electron microscopy, energy-dispersive spectroscopy mapping, X-ray diffraction and transmission electron microscopy. The cytocompatibility of the mineralizing products were evaluated with rabbit bone-marrow-derived mesenchymal stem cells in vitro. Results showed that the precipitates were mainly composed of fluoridated hydroxyapatite with ahexagonal prism morphology. Fluoridated hydroxyapatite initially nucleated and grew from the root canal dentine surface to the root canal centre. The fluoridated hydroxyapatite precipitate and root canal dentine intergraded together such that the interface became hardly distinguishable. The fluoridated hydroxyapatite precipitate grew into and obturated the dentinal tubules. In the root canal, the regenerated fluoridated hydroxyapatite densely packed and bundled together with a c-axis extension. After 7 days of mineralisation, the root canal was completely obturated, and the apical foramen was sealed. The mineralizing products had good biocompatibility with the cells, and the cells grew well on the mineralized surface. Biomimetic mineralisation strategy provides a novel means to regenerate tooth-like tissue to seal the root canal system permanently other than by passive synthetic material filling.

Effect of Bleaching Gel Viscosity on Tooth Whitening Efficacy and Pulp Chamber Penetration: An In Vitro Study.

OBJECTIVES: Whitening efficacy has been related to hydrogen peroxide (HP) diffusion into tooth structure. However, little information is available relating rheological properties to whitening efficacy. The purpose was to evaluate the whitening efficacy and HP penetration level of a 10% HP gel at three different viscosities and to compare them to a strip delivery system. METHODS AND MATERIALS: Extracted molars (n=120) were randomly assigned into five groups (n=24/ group): NC_MED (negative control; median): medium viscosity gel without HP; LOW: 10% HP gel (low viscosity experimental gel, Ultradent Products Inc); MED: 10% HP gel (medium viscosity experimental gel, Ultradent); HIGH: 10% HP gel (high viscosity gel, Ultradent); and CWS: Crest 3D Whitestrips 1-Hour Express (Procter & Gamble). All teeth were subjected to five 60-minute whitening sessions. Instrumental color measurements were performed at baseline (T0), and 1-day after each application (T1-T5), and 1-month after whitening (T6). HP penetration was estimated with leucocrystal violet and horseradish peroxidase. A Kruskal-Wallis test and post hoc Bonferroni test were performed to assess the difference in tooth color change and HP penetration among the groups (α=0.05). RESULTS: Hydrogen peroxide penetration levels and overall color changes at T6 were 0.24 µg/mL / 2.80; 0.48 µg/mL / 8.48; 0.44 µg/mL / 7.72; 0.35 µg/mL / 8.49; 0.36 µg/mL / 7.30 for groups NC, LOW, MED, HIGH, and CWS, respectively. There was a significant difference for HP penetration, while there was no significant difference among the four experimental groups for tooth color change. CONCLUSION: Rheological properties should be considered when developing new whitening formulations.

Association between bacteria occurring in the apical canal system and expression of bone-resorbing mediators and matrix metalloproteinases in apical periodontitis.

AIM: To evaluate the association between the presence of selected bacterial species/groups in the apical root canal and expression of mediators of soft and bone tissue destruction in apical periodontitis lesions. Relationships between bacteria and some other features of apical periodontitis were also investigated. METHODOLOGY: Seventeen freshly extracted teeth with pulp necrosis and apical periodontitis were included. The apical root segment was sectioned and cryopulverized; DNA was extracted and evaluated for the presence of 9 bacterial species/groups using real-time polymerase chain reaction. Lesions were processed for histopathological and immunohistochemical analyses, which targeted matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9), receptor activator of NFκB (RANK), RANK ligand (RANKL) and osteoprotegerin (OPG). Associations of the target bacteria with expression of these mediators, presence of symptoms, lesion size and histopathological diagnosis were evaluated. Data were analysed using the chi-square, Fisher's exact, Mann-Whitney and Pearson tests. P values lower than 0.05 were considered significant. RESULTS: All pulverized apical root samples were positive for bacteria. The most prevalent taxa were Actinobacteria (53%), Streptococcus species (35%), Fusobacterium species and Parvimonas micra (18%). The target mediators exhibited a high mean expression in the lesions (MMP-2: 82%; MMP-9: 73%; RANK: 78%; RANKL; 81%; OPG; 83%). Mean RANKL:OPG ratio was significantly higher in granulomas than cysts (P < 0.05, Mann-Whitney test). Actinobacteria were associated with granulomas, higher MMP-2 expression, lower OPG expression, and higher RANKL:OPG ratio (P < 0.05 for all, Fisher's exact test or Mann-Whitney test). No other significant associations were found. CONCLUSION: Actinobacteria may play an important role in the active phase of soft and bone tissue destruction in apical periodontitis.

Increased Root Canal Endotoxin Levels are Associated with Chronic Apical Periodontitis, Increased Oxidative and Nitrosative Stress, Major Depression, Severity of Depression, and a Lowered Quality of Life.

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.

Influence of Cervical Preflaring on the Incidence of Root Dentin Defects.

INTRODUCTION: This study evaluated the influence of cervical preflaring on the incidence of root dentin defects after root canal preparation. METHODS: Extracted human maxillary central incisors were selected and allocated to 1 control group and 12 experimental groups (n = 15). Teeth in the control group were left unprepared, whereas the others were prepared using 2 reciprocating single-file systems (Reciproc and WaveOne [WO]), 3 full-sequence rotary systems (ProTaper Universal, ProTaper Next [PTN], and ProFile), and K-files driven by an oscillatory system, with and without cervical preflaring. Roots were then horizontally sectioned at 4, 8, and 12 mm from the apex, stained with 1% methylene blue, and viewed through a stereomicroscope at ×25 magnification. Slices were inspected and the absence/presence of defects (fractures, partial cracks, and craze lines) recorded. Data were analyzed using Kolmogorov-Smirnov and Levene tests followed by the Tukey post hoc test at a significance level of P < .05. RESULTS: No root dentin defects were observed in the control group. WO was associated with a significantly higher number of defects than K-files, ProFile, and PTN (P < .05), but was not significantly different from Reciproc or ProTaper Universal (P > .05). Cervical preflaring significantly reduced the incidence of fractures and other defects in the WO and PTN groups (P < .05). CONCLUSIONS: All instruments caused root dentin defects, regardless of the enlargement or not of the cervical portion. Cervical preflaring was associated with a lower incidence of defects, mainly in root canals prepared with WO and PTN.

Does the Orifice-directed Dentin Conservation Access Design Debride Pulp Chamber and Mesial Root Canal Systems of Mandibular Molars Similar to a Traditional Access Design?

INTRODUCTION: Despite the increasing reports on mechanical aspects of contracted endodontic access cavities (CECs), we believe that the biological aspects (debridement) have not been adequately investigated. This study examined if 1 type of CEC (orifice-directed dentin conservation [DDC] access) was able to debride the pulp chamber, root canals, and isthmus of mesial roots of mandibular molars similar to a traditional endodontic access cavity (TEC). METHODS: Mandibular molars (N = 32) were selected and divided randomly into 2 experimental groups (n = 12) after micro-computed tomographic scanning (group 1: TEC and group 2: DDC) and histologic controls (n = 8). After instrumentation to a size 30/0.06 taper using 3% sodium hypochlorite as irrigant, specimens were processed for histologic evaluation, and the remaining pulp tissue (RPT) was measured from the pulp chamber, root canal, and isthmus at all root thirds. Data were analyzed using 1-way analysis of variance, Kruskal-Wallis, and appropriate post hoc tests (P = .05). RESULTS: The RPT in the pulp chamber was significantly higher in DDC compared with TEC (P < .05). Comparing the root thirds in each group, there was no significant difference in the RPT within the root canals or the isthmus (P > .05). The RPT within the root canals and isthmus was not significantly different between the 2 access cavity designs at any root third (P > .05). CONCLUSIONS: Debridement of the pulp chamber was significantly compromised in DDC. The type of access cavity did not influence the amount of RPT in the root canals and isthmus.

Evaluation of Various Pretreatments to Fiber Post on the Push-out Bond Strength of Root Canal Dentin.

INTRODUCTION: The aim of this in vitro study was to compare the effect of different pretreatments (fiber post) with the laser-activated irrigation (LAI) technique (for removal of the smear layer) on root canal dentin in terms of push-out bond strength (PBS) in a fiber post. METHODS: Fifty freshly extracted mandibular single-rooted premolars were prepared, and LAI was applied to all roots (17% EDTA was 5 mL for 120 seconds with an erbium, chromium:yttrium-scandium-gallium-garnet laser [0.50 W, 20 Hz, 25 mJ]). In addition, 50 quartz fiber posts were randomly assigned to 5 groups (n = 10) according to the surface treatments as follows: group S (sandblasting), group N1 and group N2 (neodymium:yttrium-aluminum-garnet laser irradiation [2 W, 200 mJ, 10 Hz, with pulse durations of 180 or 320 microseconds), group HF (9.7% hydrofluoric acid etched), and group C (control with no treatment). Dual-cure resin cement was adhered onto the fiber posts after they were covered with a silane agent, and then the posts were placed into the canal space using a Lentulo spiral. The PBS test was performed after all specimens were transversally sectioned (root slices of 1-mm thickness). Data were analyzed with 1-way analysis of variance/Tukey post hoc test (α = 0.05). RESULTS: The highest PBS value was observed in group S (middle part), and the lowest value was observed in group C (apical part). There were no statistical differences among the groups regardless of the part. Furthermore, when the PBS values of the different parts of dentin were compared within treated groups, significant differences were observed in all groups except group N2 (P < .05). CONCLUSIONS: Within the limitations of this study, it can be concluded that the LAI technique when used with 17% EDTA had a significant effect on the amount of smear layer removed from the root canal dentin, which was also detected in the fracture pattern (adhesive failure [resin-post interface]). However, the various treatments of the fiber post did not improve the PBS of the root dentin.