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.

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.

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.

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.

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.

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.

ß-lactam substituted polycyclic fused pyrrolidine/pyrrolizidine derivatives eradicate C. albicans in an ex vivo human dentinal tubule model by inhibiting sterol 14-α demethylase and cAMP pathway.

BACKGROUND: Further quest for new anti-fungal compounds with proven mechanisms of action arises due to resistance and dose limiting toxicity of existing agents. Among the human fungal pathogens C. albicans predominate by infecting several sites in the body and in particular oral cavity and root canals of human tooth. METHODS: In the present study, we screened a library of ß-lactam substituted polycyclic fused pyrrolidine/pyrrolizidine compounds against Candida sp. Detailed molecular studies were carried out with the active compound 3 on C. albicans. Morphological damage and antibiofilm activity of compound 3 on C. albicans was studied using scanning electron microscopy (SEM). Biochemical evidence for membrane damage was studied using flow cytometry. In silico docking studies were carried out to elucidate the mechanism of action of compound 3. Further, the antifungal activity of compound 3 was evaluated in an ex vivo dentinal tubule infection model. RESULTS: Screening data showed that several new compounds were active against Candida sp. Among them, Compound 3 was most potent and exerted time kill effect at 4h, post antifungal effect up to 6h. When used in combination with fluconazole or nystatin, compound 3 revealed an minimum inhibitory concentration (MIC) decrease by 4 fold for both drugs used. In-depth molecular studies with compound 3 on C. albicans showed that this compound inhibited yeast to hyphae (Y-H) conversion and this involved the cAMP pathway. Further, SEM images of C. albicans showed that compound 3 caused membrane damage and inhibited biofilm formation. Biochemical evidence for membrane damage was confirmed by increased propidium iodide (PI) uptake in flow cytometry. Further, in silico studies revealed that compound 3 docks with the active site of the key enzyme 14-α-demethylase and this might inhibit ergosterol synthesis. In support of this, ergosterol levels were found to be decreased by 32 fold in compound 3 treated samples as analyzed by high performance liquid chromatography (HPLC). Further, the antifungal activity of compound 3 was evaluated in an ex vivo dentinal tubule infection model, which mimics human tooth root canal infection. Confocal laser scanning microscopy studies showed 83% eradication of C. albicans and a 6 log reduction in colony forming unit (CFU) after 24h treatment in the infected tooth samples in this model. CONCLUSION: Compound 3 was found to be very effective in eradicating C. albicans by inhibiting cAMP pathway and ergosterol biosynthesis. GENERAL SIGNIFICANCE: The results of this study can pave the way for developing new antifungal agents with well deciphered mechanisms of action and can be a promising antifungal agent or medicament against root canal infection.

Electrochemical dissolution of fractured nickel-titanium instruments in human extracted teeth.

AIM: To assess the effect of sodium chloride concentration in fluoridated solutions during the electrochemical dissolution of fractured rotary endodontic instruments. METHODOLOGY: Two solutions were assessed (solution 1: NaF 12 g L-1  + NaCl 1 g L-1 , pH = 5.0; and solution 2: NaF 12 g L-1  + NaCl 180 g L-1 , pH = 5.0) using two tests: the ProTaper Universal F1 (PTU F1) instrument polarization test and the polarization test for intracanal PTU F1 fragments fractured in mandibular incisors. In the first test, two sets of five instruments were separately and partially immersed in each solution, and the electrical current was evaluated over 30 min. In the second test, 45 PTU F1 instruments were fractured within the root canals of mandibular incisors and subjected to potentiodynamic polarization for 30 min. The electrical current and the variations in the length of PTU F1 fragments were measured. The data were analysed statistically (anova and Wilcoxon and Mann-Whitney tests, respectively). RESULTS: Solution 2 was associated with more corrosive effects in both tests. In the first test, the PTU F1 instruments immersed in solution 2 had a higher electrical current (P < 0.001) and had a total dissolution time of approximately 540 s. In the second test, a larger difference between the baseline and final lengths of the fragments was noted in solution 2 (P = 0.011). CONCLUSION: Saturation of fluoridated solution with sodium chloride led to an increase in electrical current and microscopic reductions in the length of fractured instrument fragments subjected to electrochemical dissolution.

Influence of calcium hydroxide dressing and acid etching on the push-out bond strengths of three luting resins to root canal dentin.

OBJECTIVES: This study aims to investigate the effects of calcium hydroxide (Ca(OH)2) dressing in root canals and the effects of subsequent acid etching on the adhesion of luting resins to root canals. MATERIALS AND METHODS: Root specimens were prepared from extracted human permanent molars. Specimen canals were (1) filled with etch-and-rinse (Nexus® third generation (NX3)) and two self-adhesive (RelyX Unicem, Maxcem Elite) luting resins, respectively; (2) dressed with Ca(OH)2 before Ca(OH)2 removal and luting resin filling; (3) dressed with Ca(OH)2 before Ca(OH)2 removal and post-cementation; or (4) treated as described in item (2) except that the canals were further etched with phosphoric acid before luting resin filling. Push-out bond strengths were measured and analyzed using one-way analysis of variance, and Fisher's multiple comparison tests provided a follow-up comparison among these four canal treatments. Attenuated total reflectance-Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM) were used to analyze the specimen surfaces. RESULTS: Ca(OH)2 dressing adversely affected the bond strengths to canal dentin of the three luting resins tested. Acid etching did not increase the bond strengths. Infrared analysis revealed that Ca(OH)2 dressing caused no structural changes on the dentin surface. XPS and SEM analyses revealed Ca(OH)2 remnants as the ultimate chemical cause leading to the decrease in bond strength. CONCLUSIONS: The bond strength of luting resin to dentin was affected by Ca(OH)2 dressing. Acid etching treatment could not increase the bond strength. CLINICAL RELEVANCE: Adhesion of the fiber post to the root canal wall may be compromised after Ca(OH)2 dressing. An effective method for complete removal of Ca(OH)2 dressing or increase of bond strength for luting resin needs to be developed.

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.

Comparison of irrigant penetration up to working length and into simulated lateral canals using various irrigating techniques.

AIM: To evaluate the effect of an apical negative pressure system, a passive ultrasonic irrigation system and a combination of both apical negative pressure and passive ultrasonic irrigation on the penetration of the irrigating contrast solution (ICS) up to working length and into simulated lateral canals. METHODOLOGY: The root canals of 64 single-rooted teeth were instrumented using the ProTaper rotary system. In each sample, three simulated lateral canals were created at 2, 4 and 6 mm levels from the root apex using a 06-size C+ file (Dentsply Maillefer, Ballaigues, Switzerland). Samples were randomly assigned into 4 experimental groups (n = 16): group I - conventional needle irrigation, group II - passive ultrasonic irrigation, group III - apical negative irrigation system and group IV - combination of passive ultrasonic irrigation and apical negative pressure irrigation system. To examine irrigating solution penetration, Indian ink was mixed with 5.25% NaOCl and delivered into the root canals. Samples were then assessed by direct observation of the images taken using Canon EOS rebel T3. The depth of penetration of ICS up to the working length and into the simulated lateral canals was analysed using chi-squared tests. RESULTS: The combination (ANP and PUI) and ANP group had significantly deeper ICS penetration up to the working length (P < 0.001). The combination (ANP and PUI) and the PUI group exhibited significantly greater ICS penetration into lateral canals at the 6 mm level (P < 0.001). At the 4 and 2 mm levels, the combination of ANP and PUI had significantly greater ICS penetration into the lateral canals than the other groups (P < 0.001). CONCLUSIONS: The combination of ANP and PUI was the only group able to achieve irrigating contrast solution penetration both up to the working length and into lateral canals.

The effect of surface tension reduction on the clinical performance of sodium hypochlorite in endodontics.

Sodium hypochlorite (NaOCl) is recommended as an endodontic irrigant in view of its broad antimicrobial and tissue dissolution capacities. To enhance its penetration into inaccessible areas of root canals and to improve its overall effect, the addition of surface-active agents has been suggested. The aim of this investigation was to review the effect of the reduction of the surface tension on the performance of NaOCl in endodontics. A search was performed in the Medline electronic database (articles published up to 28 July 2012, in English) with the search terms and combinations as follows: 'sodium hypochlorite AND surface tension or interfacial force or interfacial tension or surface-active agent or amphiphilic agent or surface active agent or surfactant or tenside or detergent'. The purpose of this search was to identify publications that compared NaOCl alone and NaOCl modified with the addition of a surface-active agent in endodontics. A hand search of articles published online ('in-press' and 'early view'), and appearing in the reference list of the articles included, was further performed, using the same search criteria as the electronic search. The search identified 302 publications, of which 11 fulfilled the inclusion/exclusion criteria of the review. The evidence available suggests that surface-active agents improve the penetration of NaOCl in the main canal and have no effect on its pulp tissue dissolution ability. There are, however, insufficient data to enable a sound conclusion to be drawn regarding the effect of modifying NaOCl's surface tension on lubrication, antimicrobial and smear layer or debris removal abilities.

The relationship of hydrogen peroxide exposure protocol to bleaching efficacy.

The purpose of this study was to compare two in-office bleaching methods with respect to tooth color change and level of hydrogen peroxide penetration into the pulp cavity and to evaluate relationships between penetration level and color change. Eighty extracted canines were exposed to two different bleaching regimens (conventional vs sealed bleaching technique). After exposure to 38% hydrogen peroxide gel for one hour, hydrogen peroxide amount was estimated spectrophotometrically. Color change was measured per Commission Internationale de l'Eclairage methodology. Linear regression was used to evaluate factors affecting color change, including bleaching technique. The conventional and sealed bleaching groups showed no difference for any color change parameters (ΔL, Δa, Δb, ΔE); however, there was significantly greater hydrogen peroxide penetration in the conventional bleaching group (p<0.05). Linear modeling of the change in lightness (ΔL) showed that the increase in lightness tended to be greater for teeth with lower initial L* values (r=-0.32, p<0.05). After adjustment for initial L*, there was no evidence that ΔL differed with hydrogen peroxide penetration levels (p>0.05) or bleaching technique (mean group difference in ΔL=0.36; p>0.05).

Revascularization and tissue regeneration of an empty root canal space is enhanced by a direct blood supply and stem cells.

BACKGROUND: Regenerative endodontics is an innovative treatment concept aiming to regenerate pulp, dentin and root structures. In the diseased or necrotic tooth, the limitation in vascular supply renders successful tissue regeneration/generation in a whole tooth challenging. The aim of this study is to evaluate the ability of vascularized tissue to develop within a pulpless tooth using tissue engineering techniques. MATERIALS AND METHODS: A pulpless tooth chamber, filled with collagen I gel containing isolated rat dental pulp cells (DPC) and angiogenic growth factors, was placed into a hole created in the femoral cortex or into its own tooth socket, respectively. The gross, histological and biochemical characteristics of the de novo tissue were evaluated at 4 and 8 weeks post-transplantation. RESULTS: Tooth revascularization and tissue generation was observed only in the femur group, confirming the important role of vascular supply in tissue regeneration. The addition of cells and growth factors significantly promoted connective tissue production in the tooth chamber. CONCLUSION: Successful revascularization and tissue regeneration in this model demonstrate the importance of a direct vascular supply and the advantages of a stem cell approach.

Hydrogen peroxide penetration into the pulp chamber and dental permeability after bleaching.

This study sought to quantify the concentration of hydrogen peroxide (HP) in the pulp chamber and evaluate changes on dental permeability after bleaching with 3 HP concentrations (10%, 35%, and 50%). This study was divided into 2 experiments and the bleaching treatments consisted of 3 applications of HP for 30 minutes during a single session. The first experiment tested HP penetration into the pulp chamber of 4 experimental groups (n = 10) of bovine crowns, which were divided by HP concentration: an unbleached control group (0% HP), 10% HP, 35% HP, and 50% HP. Acetate buffer solution was placed into the pulp chamber and after each application of HP. This solution was collected to determine spectrophotometrically the concentration of HP that reached the pulp chamber. The second experiment evaluated dental permeability. Bovine crowns were divided into 3 groups (n = 10). The crowns were connected to a permeability device and the initial permeability was measured at 10 psi. Three different concentrations of HP gels (10%, 35% and 50%) were applied to the buccal enamel surfaces and the dental permeability was measured after the first, second, and third applications of HP. The data were analyzed by 2-way ANOVA and Tukey test (P ≤ 0.05). All concentrations of HP reached the pulp chamber, although no significant differences were noted between the 3 concentrations tested (P > 0.05). However, the increase of dental permeability in the group that received 50% HP was significantly higher than the 10% HP group (P < 0.05). The results indicate that the HP bleaching treatments increased dental permeability.

Quantification of cultivable bacteria and endotoxin in post-treatment apical periodontitis before and after chemo-mechanical preparation.

This clinical study was conducted to quantify cultivable bacteria and endotoxin in root canals with post-treatment apical periodontitis by correlating their levels with clinical features and to evaluate the effect of chemo-mechanical preparation (CMP) with 2 % chlorhexidine gel + 17 % EDTA on bacterial and endotoxin removal/elimination. Moreover, target strict Gram-negative anaerobic bacteria were investigated by polymerase chain reaction (PCR). Fifteen teeth with post-treatment apical periodontitis were sampled before (s1) and after (s2) CMP. Culture techniques determined the number of colony-forming units (CFU). PCR (16S rDNA) and limulus amebocyte lysate (LAL) assay were used for bacterial and endotoxin detection, respectively. Prevotella nigrescens (4/15), Prevotella intermedia (2/15), and Tannerella forsythia (2/15) were the most frequently detected species. Endotoxin was recovered in 100 % of the samples. At s1, bacteria and endotoxin were detected at a median value of 5.14 × 10(3) CFU/mL and 3.96 EU/mL, respectively. Higher levels of endotoxin were related to a larger size of radiolucent area (>5 mm) (p < 0.05). CMP was more effective in reducing bacteria (99.61 %) than endotoxin (60.6 %) (both p < 0.05). Our findings indicated that the levels of endotoxin found in infected root canals were related to a larger size of radiolucent area in the periapical region. Moreover, CMP was effective in reducing both bacterial and endotoxin contents in post-treatment apical periodontitis.

The effect of using an alternative irrigant between sodium hypochlorite and chlorhexidine to prevent the formation of para-chloroaniline within the root canal system.

AIM: To determine if the formation of para-chloroaniline (PCA) can be avoided by using an alternative irrigant following sodium hypochlorite but before chlorhexidine. METHODOLOGY: Fifty-five single-rooted teeth were decoronated, instrumented to size 40, .06 taper whilst being irrigated with 14% ethylene-diamine-tetra-acetic acid (EDTA) and 6% NaOCl. Samples were then randomly divided into three experimental and two control groups. Group 1 was irrigated with saline followed by 2% chlorhexidine gluconate (CHX). Group 2 was irrigated with 50% citric acid (CA) followed by 2% CHX. Group 3 was irrigated with 14% EDTA followed by 2% CHX. The chemical identity and quantification of the PCA in the formed precipitate was determined using gas chromatography/mass spectrometry (GC/MS). RESULTS: All experimental groups contained PCA. The mean level of PCA for group 1 (sterile saline) was 229 ng mL(-1), group 2 (citric acid) 72 ng mL(-1) and group 3 (EDTA) 400 ng mL(-1), respectively. A significant difference was found between the saline and EDTA groups and the negative control (P < 0.05). Although no statistical significance was found between the negative control and citric acid group, PCA was still present in this experimental group. CONCLUSIONS: Citric acid used as the intermittent irrigant had the least amount of PCA formation in the canal system. Until the threshold required to cause biological damage in humans is determined, the combination of NaOCl and CHX in root canal treatment should be avoided.

Dynamic model of hydrogen peroxide diffusion kinetics into the pulp cavity.

AIM: To measure the time course hydrogen peroxide penetration into the pulp cavity and evaluate short-term tooth color changes after bleaching. MATERIALS AND METHODS: Twenty extracted human canines were sectioned, pulp tissue removed and the cavity enlarged. Teeth were painted with nail varnish to leave a 6-mm diameter circle on the buccal surface. Baseline color was measured spectrophotometrically. Teeth were randomized into a control group (n = 10) treated with 30 µl of glycerin base and a bleaching group (n = 10) exposed to 30 µl of 40% hydrogen peroxide for 1 hour. A linear low density polyethylene wrap was placed to prevent evaporation of the material. Acetate buffer was placed into the cavity and replenished every 10 minutes and placed into plastic tubes. Hydrogen peroxide amount was estimated spectrophotometrically using leukocrystal violet and horseradish peroxidase. Specimen color was remeasured immediately after bleaching, 1 hour, 1 day 1, 2 and 6 weeks postbleaching. Color change was measured per Commission Internationale de l'Eclairage methodology. Mann-Whitney procedure was used to assess baseline color measurements and total hydrogen peroxide penetration amount. Friedman's test was used to assess within group differences for color change and hydrogen peroxide penetration. RESULTS: There was significantly greater hydrogen peroxide penetration in the bleaching group (p < 0.05). Hydrogen peroxide penetration levels were constant throughout the 1-hour evaluation period in the bleaching group. The groups showed no difference at baseline with respect to any of L*a*b color measurements (p > 0.05). The postbleaching color measurement showed an increase of change in overall color (ΔE) and lightness (ΔL) up to 1 week followed by a gradual stabilization up to 6 weeks. CONCLUSION: This dynamic model provided information about the time course diffusion kinetics into the pulp cavity, demonstrating constant penetration of hydrogen peroxide into the pulp cavity during a 1-hour bleaching session. CLINICAL SIGNIFICANCE: A prolonged application of 40% hydrogen peroxide bleaching material for 1 hour produces constant penetration of hydrogen peroxide into the pulp cavity in vitro.

Uptake of calcium and silicon released from calcium silicate-based endodontic materials into root canal dentine.

AIM: To compare Biodentine and White ProRoot mineral trioxide aggregate (MTA) with regard to Ca and Si uptake by adjacent root canal dentine in the presence of phosphate-buffered saline (PBS). METHODOLOGY: Root canals of bovine incisor root segments were instrumented, filled with either Biodentine or MTA (n = 20 each) and then immersed in Ca-and Mg-free PBS for 1, 7, 30 or 90 days (n = 5 each). Unfilled, unimmersed dentine specimens (n = 5) served as controls. The specimens were sectioned longitudinally, and the ultrastructure of the dentine-material interface and the elemental composition/distribution in the material-adjacent dentine were analysed using a wavelength-dispersive X-ray spectroscopy electron probe microanalyser with image observation function. Data were statistically analyzed using one-way anova and Tukey's honestly significant difference test or the Mann-Whitney U-test. RESULTS: Along the material-dentine interface, both materials formed a tag-like structure that was composed of either Ca- and P-rich crystalline deposits or the material itself. The width of a Ca- and Si-rich layer detected along the dentine layer of the material-dentine interface showed increases over time. The Ca- and Si-rich layer width was significantly larger (P < 0.05) in Biodentine than MTA at 30 and 90 days. CONCLUSIONS: Both Biodentine and MTA caused the uptake of Ca and Si in the adjacent root canal dentine in the presence of PBS. The dentine element uptake was more prominent for Biodentine than MTA.

Reaction rate of NaOCl in contact with bovine dentine: effect of activation, exposure time, concentration and pH.

AIM: To determine the influence of activation method (ultrasound or laser), concentration, pH and exposure time on the reaction rate (RR) of NaOCl when in contact with dentinal walls. METHODOLOGY: The walls from standardized root canals in bovine incisors were exposed to a standardized volume of sodium hypochlorite (NaOCl) with different concentrations (2% and 10%), pH (5 and 12) and exposure times (1 and 4min). Two irrigation protocols were tested: passive ultrasonic irrigation or laser activated irrigation with no activation as the control. The activation interval lasted 1min followed by a rest interval of 3 min with no activation. The RR was determined by measuring the iodine concentration using an iodine/thiosulfate titration method. RESULTS: Exposure time, concentration and activation method influenced the reaction rate of NaOCl whereas pH did not. CONCLUSIONS: Activation is a strong modulator of the reaction rate of NaOCl. During the rest interval of 3min, the consumption of available chlorine increased significantly. This effect seems to be more pronounced after irrigant activation by laser. pH did not affect the reaction rate of 2% NaOCl.