Antimicrobial activity and enterococcus faecalis biofilm formation on chlorhexidine varnishes

Objective: To evaluate, in vitro, the antimicrobial activity and biofilm formation of three chlorhexidine varnishes in four Enterococcus faecalis strains: E. faecalis ATCC 29212, E. faecalis EF-D1 (from failed endodontic treatment), E. faecalis 072 (cheese) and E. faecalis U-1765 (nosocomial infection), and one Enterococcus durans strain (failed endodontic treatment). Study Design: The direct contact test was used to study the antimicrobial activity. Bacterial suspensions were exposed for one hour to EC40, Cervitec (CE) and Cervitec Plus (CEP) varnishes. "Eradication" was defined as 100% bacterial kill. The formation of enterococci biofilms was tested on the surface of the varnishes after 24 hours of incubation and expressed as percentage of biofilm reduction. Results: EC40 eradicated all strains except E. faecalis ATCC 29212, where 98.78% kill was achieved. CE and CEP showed antimicrobial activity against all the strains, but most clearly against E. durans and E. faecalis 072. EC40 completely inhibited the formation of biofilm of E. faecalis ATCC 29212, E. faecalis 072 and E. durans. CE and CEP led to over 92% of biofilm reduction, except in the case of E. faecalis U-1765 on CEP (76.42%). Conclusion: The three varnishes studied were seen to be effective in killing the tested strains of enterococci and in inhibiting the formation of biofilm, the best results being observed with EC40.

Short implants (6 mm) installed immediately into extraction sockets: An experimental study in dogs

Aim: To evaluate the effect of implant length (6 mm vs. 11 mm) on osseointegration (bone-toimplant contact) of implants installed into sockets immediately after tooth extraction. Material and methods: In six Labrador dogs, the pulp tissue of the mesial roots of P-3(3) was removed and the root canals were filled. Flaps were elevated bilaterally, the premolars hemisectioned and the distal roots removed. Recipient sites were prepared in the distal alveolus and a 6 mm or an 11 mm long implant was installed at the test and control sites, respectively. Non-submerged healing was allowed. After 4 months of healing, block sections of the implant sites were obtained for histological processing and peri-implant tissue assessment. Results: No statistically significant differences were found between test and control sites both for hard and soft tissue parameters. The bone-to-implant contact evaluated at the apical region of the implants was similar as well. Although not statistically significant, the location of the top of the bony crest at the buccal aspect was more apical in relation to the implant shoulder at the test compared with the control sites (2.0 +/- 1.4 and 1.2 +/- 1.1 mm, respectively). Conclusions: Shorter implants (6 mm) present with equal osseointegration than do longer implants (11 mm).

Effect of pre-flaring and file size on the accuracy of two electronic apex locators

Objective: This ex vivo study evaluated the effect of pre-flaring and file size on the accuracy of the Root ZX and Novapex electronic apex locators (EALs). Material and methods: The actual working length (WL) was set 1 mm short of the apical foramen in the palatal root canals of 24 extracted maxillary molars. The teeth were embedded in an alginate mold, and two examiners performed the electronic measurements using #10, #15, and #20 K-files. The files were inserted into the root canals until the "0.0" or "APEX" signals were observed on the LED or display screens for the Novapex and Root ZX, respectively, retracting to the 1.0 mark. The measurements were repeated after the pre-flaring using the S1 and SX Pro-Taper instruments. Two measurements were performed for each condition and the means were used. Intra-class correlation coefficients (ICCs) were calculated to verify the intra-and inter-examiner agreement. The mean differences between the WL and electronic length values were analyzed by the three-way ANOVA test (p<0.05). Results: ICCs were high (>0.8) and the results demonstrated a similar accuracy for both EALs (p>0.05). Statistically significant accurate measurements were verified in the pre-flared canals, except for the Novapex using a #20 K-file. Conclusions: The tested EALs showed acceptable accuracy, whereas the pre-flaring procedure revealed a more significant effect than the used file size.

Effects of curing protocol and storage time on the micro-hardness of resin cements used to lute fiber-reinforced resin posts

Objectives: To determine the micro-hardness profile of two dual cure resin cements (RelyX - U100 (R), 3M-ESPE and Panavia F 2.0 (R), Kuraray) used for cementing fiber-reinforced resin posts (Fibrekor (R) - Jeneric Pentron) under three different curing protocols and two water storage times. Material and methods: Sixty 16mm long bovine incisor roots were endodontically treated and prepared for cementation of the Fibrekor posts. The cements were mixed as instructed, dispensed in the canal, the posts were seated and the curing performed as follows: a) no light activation; b) light-activation immediately after seating the post, and; c) light-activation delayed 5 minutes after seating the post. The teeth were stored in water and retrieved for analysis after 7 days and 3 months. The roots were longitudinally sectioned and the microhardness was determined at the cervical, middle and apical regions along the cement line. The data was analyzed by the three-way ANOVA test (curing mode, storage time and thirds) for each cement. The Tukey test was used for the post-hoc analysis. Results: Light-activation resulted in a significant increase in the microhardness. This was more evident for the cervical region and for the Panavia cement. Storage in water for 3 months caused a reduction of the micro-hardness for both cements. The U100 cement showed less variation in the micro-hardness regardless of the curing protocol and storage time. Conclusions: The micro-hardness of the cements was affected by the curing and storage variables and were material-dependent.

Topography of four different endodontic rotary systems, before and after being used for the 12th time

Root canal preparation may damage NiTi instruments resulting in wear and deformation. The aim of this study was to make a comparative evaluation of the surface topography of the cervical third of four different rotary systems, before and after being used twelve times, in 1.440 resin blocks with simulated root canals with standardized 45 degrees curvatures, and analyzed by atomic force microscopy AFM. The blocks were divided into four groups and prepared according to the manufacturers recommendations: Group 1 - K3 (R); Group 2 - Protaper Universal (R); Group 3 - Twisted Files (R) and Group 4 - Biorace (R). After each preparation, the instruments were washed and autoclaved. A total of 240 instruments were selected, being 30 new instruments and 30 after having been used for the 12th time, from each group. These instruments were analyzed by AFM and for quantitative evaluation, the mean RMS (Root mean square) values of the cervical third of the specimens from the four groups were used. The result showed that all the rotary files used for the 12th time suffered wear with change in the topography of the cervical region of the active portion of the file (ANOVA p < 0.01). Classifying the specimens in increasing order, from the least to the greatest wear suffered, Group 3 (2.8993 nm) presented the least wear, followed by Group 4 (12.2520 nm), Group 1 (36.0043 nm) and lastly, Group 2 (59.8750 nm) with the largest amount of cervical surface wear. Microsc. Res. Tech. 75:97-102, 2012. (c) 2011 Wiley Periodicals, Inc.

Back-scattered electron imaging for leakage analysis of four retrofilling materials

This study aimed to evaluate, ex vivo, the nanoleakage in dentinal tubules, the linear infiltration of silver nitrate in the dentin wall/root-end filling material interface, and the presence of gaps in this interface in root-end cavities filled with 4 filling materials. Forty-eight disto-buccal root canals of maxillary molars were instrumented and filled. Retrograde cavities were prepared with ultrasonic points (apical 2 mm). The samples were divided into 2 control groups (n = 4) and 4 experimental groups (n = 10): Group I white mineral trioxide aggregate (MTA); Group II Super EBA; Group III Portland cement; and Group IV Sealer 26. After 1 week, the specimens were subjected to silver nitrate and prepared for SEM (backscattered electrons). In the apical-apical segment, an area with significantly higher leakage was observed for Super EBA, followed by Portland cement, MTA, and Sealer 26 (P = 0.0054). In the medium and cervical segments, all materials showed the same leakage behavior (P = 0.1815 and P = 0.1723, respectively). The linear infiltration at the dentin wall/root-end filling material interface was higher with Super EBA than the other groups. No differences in the percentage of gaps along the 3 mm of dentin wall/root-end filling material interface between the 4 materials were evident (P > 0.05). Nanoleakage occurred mainly in the apical segment of the samples, and Super EBA showed the highest values. The area and linear leakage were lower in the middle and coronal segments, regardless of the root-end filling material. No material perfectly sealed the root-end cavities, which allowed for the leakage occurrence. Microsc. Res. Tech. 75:796800, 2012. (C) 2011 Wiley Periodicals, Inc.

Influence of pulp condition on the accuracy of an electronic foramen locator in posterior teeth: an in vivo study

The aim of this study was to assess, in vivo, the accuracy of the NovApex (R) electronic foramen locator in determining working length (WL) in vital and necrotic posterior teeth. The NovApex (R) was used in 144 canals: 35 teeth with vital pulps (68 canals) and 42 teeth with necrotic pulps (76 canals). WL was measured with the NovApex (R) locator and confirmed using the radiographic method. Differences between electronic and radiographic measurements ranging between 0.0 and 0.4 millimeters were classified as acceptable; differences equal to or greater than 0.5 millimeter were considered unacceptable. Pearson's chi-square test was used to assess the influence of pulp condition on the accuracy of NovApex (R) (alpha = 0.05). Regardless of pulp condition, differences between electronic and radiographic WL measurements were acceptable in 73.61% of the canals. No statistically significant differences in accuracy were observed when comparing vital and necrotic canals (p > 0.05). There were 38 unacceptable measurements. In none of these cases was the file tip located beyond the radiographic apex; in 32, it was located short of the NovApex (R) measurement. Pulp condition had no significant effect on the accuracy of NovApex (R).

Bactericidal effects of two parameters of Er:YAG laser intracanal irradiation: ex-vivo study

The success of endodontic treatment depends on the complete elimination of microorganisms from the root canal system, thus the search for new procedures to eliminate them is justified. The aim of this study was to assess bacterial reduction after intracanal irradiation with the Er:YAG laser. The canals of 70 extracted human maxillary canines were prepared up to file #40 using 1% NaOCl, irrigated with 17% EDTA, and then washed with physiological solution activated by ultrasound. The roots were sterilized by autoclaving, inoculated with 10 mu l of a suspension containing 1.5 x 10(8) CFU/ml of Enterococcus faecalis ATCC 29212 and incubated at 37A degrees C for 72 h. The canals were irradiated with the Er:YAG laser using two energy settings: 60 mJ and 15 Hz, and 100 mJ and 10 Hz. The remaining bacteria were counted immediately and 48 h after laser irradiation. The results showed a high bacterial reduction at both time points. With 60 mJ and 15 Hz there was an immediate reduction of 99.73% and the reduction was 77.02% after 48 h, and with 100 mJ and 10 Hz there was an immediate reduction of 99.95% and the reduction was 84.52% after 48 h. Although the best results were observed with 100 mJ of energy, the difference between the two settings was not statistically significant. The count performed 48 h after irradiation showed that E. faecalis were able to survive, and can grow even from small numbers.

Antimicrobial Photodynamic Therapy for the Treatment of Teeth with Apical Periodontitis: A Histopathological Evaluation

Introduction: This study evaluated the in vivo response of apical and periapical tissues of dogs' teeth with apical periodontitis after one-session endodontic treatment with and without antimicrobial photodynamic therapy (aPDT). Methods: Sixty root canals with experimentally induced apical periodontitis were instrumented and assigned to 4 groups receiving aPDT and root canal filling (RCF) or not: group aPDT+/RCF- (n = 20): aPDT (photosensitizer phenothiazine chloride at 10 mg/mL for 3 minutes and diode laser [2 = 660 nm, 60 mW/cm(2)] for 1 minute) and RCF in the same session; group aPDT+/RCF (n = 10); group aPDT /RCF+ (n = 20), and group aPDT /RCF (n = 10). Teeth were restored, and the animals were killed after 90 days. Sections from the maxillas and mandibles were stained with hematoxylin-eosin and Mallory trichrome and examined under light microscopy. Descriptive (ie, newly formed apical mineralized tissue, periapical inflammatory infiltrate, apical periodontal ligament thickness, and mineralized tissue resorption) and quantitative (ie, periapical lesion size and number of inflammatory cells) microscopic analysis was performed. Quantitative data were analyzed by the Kruskal-Wallis and Dunn tests (alpha =.05). Results: In the aPDT-treated groups, the periapical region was moderately/severely enlarged with no inflammatory cells, moderate neoangiogenesis and fibrogenesis, and the smallest periapical lesions. Conclusions: Although apical closure by mineralized tissue deposition was not achieved, the absence of inflammatory cells, moderate neoangiogenesis, and fibrogenesis in the periapical region in the groups treated with aPDT indicate that this can be a promising adjunct therapy to cleaning and shaping procedures in teeth with apical periodontitis undergoing one-session endodontic treatment. (J Endod 2012;38:360-366)

Clinical, radiographic and histologic analysis of the effects of pulp capping materials used in pulpotomies of human primary teeth

Aim: To compare the clinical, radiographic and histological responses of the pulp to mineral trioxide aggregate (MTA), calcium hydroxide (CH) and Portland cement (PC) when used as a pulpotomy agent in human primary teeth. Study design: Forty-five mandibular primary molar teeth were randomly assigned to CH, MTA or PC groups and treated by pulpotomy technique. Methods: The teeth were treated by conventional pulpotomy technique, differing only in the capping material for each group. Clinical and radiographic evaluations were recorded at 6-, 12- and 24-month follow-up. Teeth in the regular exfoliation period were further processed for histologic analysis. Statistics: The teeth were treated by conventional pulpotomy technique, differing only in the capping material for each group. Clinical and radiographic evaluations were recorded at 6-, 12- and 24-month follow-up. Teeth in the regular exfoliation period were further processed for histologic analysis. Statistics: Clinically and radiographically, the MTA and PC groups showed 100 % success rates at 6, 12 and 24 months. In CH group, several teeth presented clinical and radiographic failures detected throughout the follow-up period, and internal resorption was a frequent radiographic finding. Histologic analysis revealed the presence of dentine-like mineralised material deposition obliterating the root canal in the PC and MTA groups. CH group presented, in most of the sections, necrotic areas in the root canals. Conclusions: MTA and PC may serve as effective materials for pulpotomies of primary teeth as compared to CH. Although our results are very encouraging, further studies and longer follow-up assessments are needed in order to determine the safe clinical indication of Portland cement.

An in vivo radiographic evaluation of the accuracy of Apex and iPex electronic Apex locators

The aim of this study was to evaluate in vivo the clinical applicability of two electronic apex locators (EALs) - Apex (Septodont) and iPex (NSK) - in different groups of human teeth by using radiography. The working lengths (WLs) of 100 root canals were determined electronically. The EAL to be used first was chosen randomly and a K-file was inserted into the root canal until the EAL display indicated the location of the apical constriction (0 mm). The K-file was fixed to the tooth and a periapical radiograph was taken using a radiographic film holder. The K-file was removed and the WL was measured. The same procedure was repeated using the other EAL. Radiographs were examined with the aid of a light-box with lens of ×4 magnification by two blinded experienced endodontists. The distance between the file tip and the root apex was recorded as follows: (A) +1 to 0 mm, (B) -0.1 to 0.5 mm, (C) -0.6 to 1 mm, (D) -1.1 to 1.5 mm, and (E) -1.6 mm or greater. For statistical purposes, these scores were divided into 2 subgroups according to the radiographic apex: acceptable (B, C, and D) and non-acceptable (A and E). Statistically significant differences were not found between the results of Apex and iPex in terms of acceptable and non-acceptable measurements (p>0.05) or in terms of the distance recorded from file tip and the radiographic apex (p>0.05). Apex and iPex EALs provided reliable measurements for WL determination for endodontic therapy.

Effect of photodynamic therapy with different formulations of methylene blue in teeth contaminated by Enterococcus faecalis

The aim of this study was to compare the disinfection of dentine using photodynamic therapy with methylene blue in different formulations. Thirty bovine teeth roots were autoclaved and incubated with a suspension of Enterococcus faecalis. The specimen were randomly divided into three groups: G1, the roots were filled with 10 mM methylene blue dissolved in water; G2, the roots were filled with 10 mM methylene blue dissolved in a mixture of glycerol: ethanol: water; G3, roots filled with 100 mM methylene blue dissolved in water. The groups were irradiated with a 660 nm diode laser with an output power of 100 mW for 4 min, energy density of 850 J/cm2 and after this procedure, the sensitizer was removed and microbial samples were collected from within the root canals. The samples were plated on mEnterococcus to count the colony-forming units (CFU/mL). The means were: Group 1=513×103, Group 2=1431×103 and Group 3=2.96×103. The statistical analysis detected higher disinfection achieved by G3 when compared with groups G1 and G2, and no significant difference between the groups G1 and G2 (P>0.05). The increase of the concentration of methylene blue dye achieved higher disinfection in photodynamic therapy.

Cone beam computed tomography in mandibular molars referred for apical surgery

The purpose of the present study was to compare conventional intraoral periapical radiographs (PA) with limited cone beam computed tomography (CBCT) for evaluation of mandibular molars prior to apical surgery. The apical extent and homogeneity of the root canal fillings (RCF) as well as the number of root canals were examined.

A 5-year review of teeth filled with the noninstrumentation technology

AIM: The aim of this Case Series was to evaluate the radiographic quality of root fillings performed 5 years previously using the noninstrumentation technology (NIT)-obturation method and to assess radiographically the outcome of these root canal treatments. METHODOLOGY: Seventeen patients requiring root canal treatment participated in this study and were re-evaluated after 5 years. After instrumentation with K-Flexofiles, Calcium-Hydroxide inter-appointment dressing, re-entry and copious irrigation with NaOCl, the teeth were root filled using the NIT. RESULTS: Immediately after obturation the root fillings were (-0.78 +/- 0.11 mm) short when taking the radiographic apex as a reference point. After 60 months these values were -0.85 +/- 0.11 mm. No statistical difference was found (P > 0.05). In the periapical region, PAI rating 1 and 2 increased from 20.1% to 75.6% after 60 months. CONCLUSIONS: * This prospective Case Series demonstrated the performance of the NIT-obturation method in vivo. * Root canals filled by the reduced-pressure method using sealer combined with gutta-percha cones showed good radiographic quality. * Periapical healing after 5 years was comparable with conventional filling techniques.

Comparative antibacterial efficacies of hydrodynamic and ultrasonic irrigation systems in vitro

INTRODUCTION To ensure root canal treatment success, endodontic microbiota should be efficiently reduced. The in vitro bactericidal effects of a hydrodynamic system and a passive ultrasonic irrigation system were compared. METHODS Single-rooted extracted teeth (n = 250) were contaminated with suspensions of Enterococcus faecalis ATCC 29212, mixed aerobic cultures, or mixed anaerobic cultures. First, the antibacterial effects of the hydrodynamic system (RinsEndo), a passive ultrasonic irrigation system (Piezo smart), and manual rinsing with 0.9% NaCl (the control) were compared. Colony-forming units were counted. Second, the 2 systems were used with 1.5% sodium hypochlorite (NaOCl) alone or NaOCl + 0.2% chlorhexidine (CHX). The colony-forming units in the treated and untreated roots were determined during a period of 5 days. RESULTS Both irrigation systems reduced bacterial numbers more effectively than manual rinsing (P < .001). With NaCl, ultrasonic activated irrigation reduced bacterial counts significantly better than hydrodynamic irrigation (P = .042). The NaOCl + CHX combination was more effective than NaOCl alone for both systems (P < .001), but hydrodynamic irrigation was more effective with NaOCl + CHX than the passive ultrasonic irrigation system. CONCLUSIONS Both irrigation systems, when combined with NaOCl + CHX, removed bacteria from root canals.