Efficacy of Endodontic Treatment for Endotoxin Reduction in Primarily Infected Root Canals and Evaluation of Cytotoxic Effects

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of the Effects of Endodontic Materials on Fibroblast Viability and Cytokine Production

Introduction: Recently, a new sealer composed of Portland cement named Endo-CPM-Sealer was developed. The aim of this study was to investigate the effects of Endo-CPM-Sealer (EGEO SRL, Buenos Aires, Argentina), Sealapex (Sybron Endo, Glendora, CA), and Angelus MTA (Angelus, Londrina, Brazil) on cell viability and cytokine (interleukin [IL]-1 beta and IL-6) production by mouse fibroblasts. Methods: Millipore culture plate inserts with polyethylene tubes filled with materials were placed into 24-well cell culture plates with mouse fibroblasts. Cells cultured with only empty polyethylene tubes were used as the control. After 24 hours, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was used to evaluate the cell viability. For cytokine assay, mouse fibroblasts were incubated in 24-well flat-bottom plates with set material disks at the bottom. Cells cultured without the material disks served as the negative control. After 24 hours of incubation, culture media were collected for cytokine evaluation by using an enzyme-linked immunosorbent assay. The data were statistically analyzed by analysis of variance and Bonferroni correction. Results: Endo-CPM-Sealer, Sealapex, and Angelus MTA did not inhibit the cell viability. All materials induced IL-6 releasing, but the amount was not statistically significant compared with the control group. Angelus MTA induced IL-1 beta releasing significantly more than the control. Conclusions: All materials were not considered cytotoxic in fibroblast culture. (J Endod 2009;35:1577-1579)

Methods of experimental induction of periapical inflammation. Microbiological and radiographic evaluation

Aim: To evaluate the influence of coronal filling and apical perforation on the induction of periapical inflammation. Methodology: Fifty-eight root canals in the teeth of dogs were divided into four groups. Groups I and II: root canals were exposed for 180 days; groups III and IV: root canals were exposed for 7 days and then the access cavity filled for 53 days. The root apices of groups I and III were perforated after the coronal opening, whilst those of groups II and IV remained intact. Standard radiographs were taken before and after the experimental periods. Digital images of the radiographs were created and then analysed by three examiners. After induction of periapical inflammation, the root canal contents were collected using paper points. Microbiologic evaluation of the type of microorganism was carried out by culture in different growth media. The radiographic and microbiologic data were statistically analysed using ANOVA at a 5% significance level. Results: There were a greater total number of microorganisms in groups I and II (P < 0.05). The number of anaerobes was greater than the number of aerobes (P < 0.05). The size of the periapical radiolucencies were not significantly different between the experimental groups. Conclusions: The different methods analysed induced similar areas of periapical radiolucency in dogs with predominantly anaerobic bacteria. However, the time required for induction was less when the method with coronal filling was used. © 2005 International Endodontic Journal.

Susceptibility of strict and facultative anaerobes isolated from endodontic infections to metronidazole and β-lactams

Endodontic infections are mixed aerobic-anaerobic infections and several microbial groups associated to these pathologies are also involved in orofacial infections. The goal of this study was to evaluate the susceptibility of microorganisms isolated from endodontic infections to β-lactams and metronidazole and verify the production of β-lactamases. Clinical specimens were collected from 58 endodontic infections of 52 patients. The microorganisms were isolated in selective and non-selective culture media, under anaerobiosis and aerobiosis, and identified using biochemical methods. In the susceptibility tests, it was used an agar dilution method, and Wilkins-Chalgren agar enriched with blood, hemin and menadione for the anaerobes, while Mueller-Hinton agar was employed for the facultative anaerobes. The production of β-lactamases was evaluated through the biological and chromogenic cephalosporin methods. All tested isolates were sensitive to imipenem and 99.3% to amoxicillin/clavulanate association, while 16.1% showed resistance to amoxicillin and penicillin G, and 4.89% to cefoxitin. Resistance to metronidazole was just found in facultative anaerobes. Production of β-lactamases was detected in 18.2% of the isolates and presented a correlation with resistance to β-lactams.

The effect of endodontic materials on the optical density of dyes used in marginal leakage studies

The aim of this study was to determine the effect of the exposure of different endodontic materials to different dye solutions by evaluating the optical density of the dye solutions. Seventy-five plastic tubes were filled with one of the following materials: AH Plus, Sealapex, Portland cement, MTA (Angelus and Pro Root) and fifteen control plastic tubes were not. Each specimen of material and control was immersed in a container with 1 ml of each dye solution. A 0.1 ml-dye solution aliquote was removed before immersion and after 12, 24, 48 and 72 hours of each specimen immersion to record its optical density (OD) in a spectrophotometer. Statistical analysis was performed with ANOVA and Tukey tests (5%). No significant difference was found among any of the solution OD values for AH Plus cement. Portland cement promoted different OD values after 12 hours of immersion. MTA-Angelus cement presented different OD values only for 2% rhodamine B and the MTA-Pro Root cement presented different OD values in all 2% rhodamine B samples. Sealapex cement promoted a reduction in the India Ink OD values. Dye evaluation through OD seems to be an interesting method to select the best dye solution to use in a given marginal leakage study.

Histopathological Condition of the Remaining Tissues after Endodontic Infection of Rat Immature Teeth

Introduction: Recently, case reports have shown that immature teeth diagnosed with necrotic pulp and periapical periodontitis can be repaired through a regenerative endodontic procedure. True regeneration depends on the presence of stem cells in the remaining vital tissues. The aim of this study was to evaluate the histologic condition of the pulp tissue, root apical papilla, and periapical tissues after inducing endodontic infection in immature rat teeth for different periods. Methods: This study evaluated 18 first upper rat molars (36 roots). Periapical lesions were induced and were confirmed radiographically, and the animals were divided into 3 groups according to the days of pulp exposure for endodontic infection induction: 30, 60, and 90 days. Histologic analysis was performed in 5 different areas (ie, cervical, middle, and apical root canal thirds; the apical papilla; and the periapex surrounding the apical papilla). Results: At 30 days, one third of the specimens still showed vital but intensely inflamed pulp tissue in the apical third and vital apical papilla with varying degrees of inflammation. After 60 days, the results were similar with respect to the apical pulp tissue and apical papilla. Completely necrotic pulp tissue in the space canal and vital apical papilla were observed in about 67% of the cases after 90 days. Conclusions: Vital pulp tissue was observed in the apical third until 60 days and in the vital apical papilla until 90 days of infection in a rat model.

The impact of the addition of iodoform on the physicochemical properties of an epoxy-based endodontic sealer

Due to the low radiopacity of Sealer 26, iodoform is frequently empirically added to this sealer. Thus, the interference of this procedure with the physicochemical properties of Sealer 26 must be evaluated. Objective: This study evaluated the influence of the addition of iodoform on setting time, flow, solubility, pH, and calcium release of an epoxy-based sealer. Material and Methods: The control group was pure Sealer 26, and the experimental groups were Sealer 26 added with 1.1 g, 0.55 g or 0.275 g of iodoform. Setting time evaluation was performed in accordance with the ASTM C266-03 speciflcation. The analysis of flow and solubility was in accordance with the ISO 6876-2001 speciflcation. For the evaluation of pH and calcium ion release, polyethylene tubes were filled with the materials and immersed in flasks with 10 ml of deionized water. After 24 h, 7, 14, 21, 28, and 45 days pH was measured. In 45 days, the calcium released was evaluated with an atomic absorption spectrophotometer. Results: The addition of iodoform increased setting time in comparison with pure sealer (P < 0.05). As for flow, solubility, and calcium release, the mixtures presented results similar to pure sealer (p > 0.05). In the 24 h period, the mixture with 1.1 g and 0.55 g of iodoform showed lower pH than pure sealer and than sealer added with 0.275 g of iodoform (P < 0.05). Conclusions: The iodoform added to Sealer 26 interferes with its setting time and solubility properties. Further studies are needed to address the clinical signiflcance of this interference.

Effect of root canal preparation, type of endodontic post and mechanical cycling on root fracture strength

Objective: To evaluate the impact of the type of root canal preparation, intraradicular post and mechanical cycling on the fracture strength of roots. Material and Methods: Eighty human single rooted teeth were divided into 8 groups according to the instruments used for root canal preparation (manual or rotary instruments), the type of intraradicular post (fiber posts-FRC and cast post and core-CPC) and the use of mechanical cycling (MC) as follows: Manual and FRC; Manual, FRC and MC; Manual and CPC; Manual, CPC and MC; Rotary and FRC; Rotary, FRC and MC; Rotary and CPC; Rotary, CPC and MC. The filling was performed by lateral compactation. All root canals were prepared for a post with a 10 mm length, using the custom # 2 bur of the glass fiber post system. For mechanical cycling, the protocol was applied as follows: an angle of incidence of 45 degrees, 37 degrees C, 88 N, 4 Hz, 2 million pulses. All groups were submitted to fracture strength test in a 45 degrees device with 1 mm/min cross-head speed until failure occurred. Results: The 3-way ANOVA showed that the root canal preparation strategy (p<0.03) and post type (p<0.0001) affected the fracture strength results, while mechanical cycling (p=0.29) did not. Conclusion: The root canal preparation strategy only influenced the root fracture strength when restoring with a fiber post and mechanical cycling, so it does not seem to be an important factor in this scenario.

Signaling Pathways Activation by Primary Endodontic Infectious Contents and Production of Inflammatory Mediators

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

18-Year Follow-up of Dens Invaginatus: Retrograde Endodontic Treatment

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Tooth Discoloration Induced by Endodontic Phenothiazine Dyes in Photodynamic Therapy

Objective: This study sought to assess if discoloration of tooth structures occurs after photodynamic therapy (PDT) and to determine the efficacy of a protocol to remove the photosensitizers. Background data: PDT has been used in root canal treatment to enhance cleaning and disinfection of the root canal system. PDT uses a low power laser in association with a dye as a photosensitizer. Photosensitizers can induce staining of the dental structures, resulting in an unaesthetic appearance. Methods: Forty teeth were randomly divided into four groups according to the photosensitizer used and pre-irradiation time: 0.01% methylene blue for 5 min (MB5); 0.01% methylene blue for 10 min (MB 10); 0.01% toluidine blue for 5 min (TB5); and 0.01% toluidine blue for 10 min (TB 10). Specimens were irradiated with a 660 nm diode laser with a 300 mu m diameter optical fiber, at 40 mW power setting for 3 min. Immediately after, the photosensitizers were removed with Endo-PTC cream +2.5% sodium hypochlorite (NaOCl). The shade was measured by a Vita Easyshade spectrophotometer based on the CIELAB color system (L*a*b* values) at three different experimental times: before PDT (T0), immediately after PDT (T1), and after removal of the photosensitizer (T2). Results: The results showed a decrease in the averages of the L*a*b* coordinate values after PDT (T1) in all the groups, when compared with the number at T0, with a significant statistical difference in group MB10. After photosensitizer removal (T2), all the values of the coordinates increased with significant statistical differences (p < 0.05) between T1 and T2 in L* and a*. Conclusions: It can be concluded that both methylene blue and toluidine blue dyes cause tooth discoloration, and that Endo-PTC cream associated with 2.5% NaOCl effectively remove these dyes, regardless of the pre-irradiation time used for PDT.

Two-and tridimensional analysis of periapical repair after endodontic surgery

Objective The aim of this study was to evaluate repair after endodontic surgery using two- and tridimensional imaging methods. Materials and methods Periapical radiographs and cone beam computed tomography (CBCT) were performed before the surgeries and after 48 h (baseline), 4 months, and 8 months. The area (square millimeters) of periapical lesions in CBCT and in radiographs was compared regarding the percentage of repair. In the CBCT, multiple areas were converted to volume. Repeated-measures analyses and paired t tests (α=0.05) were used to compare the methods. Correlation coefficients were calculated between the periods of evaluation within the CBCT volumetric analysis. Bland-Altman plots were used to compare the methods, based on the 95 % limits of agreement for the difference of the means. Results Baseline showed a larger lesion volume (192.54 mm3 ) than 4-month (79.79 mm3 ) and 8-month (47.51 mm3 ) periods. No differences were found in the percentage of repair in the first 4 months and after 8 months. The volumetric analysis showed a higher percentage of repair when the first and last 4 months were compared. No differences were found in the percentage of repair by area in the CBCTs. Repair of 73 % was obtained after 8 months. Similar results were observed by the Bland-Altman agreement analyses. Conclusions The percentage of repair varied after 8 months, when lower values were obtained by volumetric evaluation. Clinical relevance Considering the outcome at follow-up periods over 4 months, tridimensional evaluation by CBCT is more capable of determining the absence of periapical bone repair than conventional two-dimensional radiographs. Therefore, the use of CBCT would be suggested only for more complex cases with slower evolution of repair or for the association of factors that make prognosis difficult after surgery.

Ethanol is inefficient to remove endodontic sealer residues of dentinal surface

Endodontic sealer residues on dentinal surface have negative effects on adhesion of adhesives system and/or can cause discoloration of the dental crown. Objective: To evaluate the efficacy of 95% ethanol in removal of residues of epoxy-based (AH Plus), methacrylate-based (Epiphany SE) or calcium-based (Sealapex) sealers on dentinal surface. Material and methods: Thirty-two bovine incisor dental crown fragments (0.5 mm x 0.5 mm) were treated with 17% EDTA and 2.5% NaOCl. The specimens were divided into three experimental groups (n = 10): G1 (AH Plus), G2 (Epiphany SE) and G3 (Sealapex). In each group was applied a coating of one endodontic sealer type and were left undisturbed for 5 minutes. After this period, the specimens were cleaned with 95% ethanol. The control group was composed by two specimens that did not receive any sealer or cleaning treatment. The sealer residues persistence after cleaning with 95% ethanol was evaluated by scanning electron microscopy (x500) and a score system was applied. Data obtained were analyzed by Kruskal-Wallis and Dunn tests (α = 5%). Results: Moderate amount of endodontic sealer residues were observed in all groups, regardless of the endodontic sealer compositions. G1, G2 and G3 presented similar amount of sealer residues on dentinal surface after cleaning with 95% ethanol (p > 0.05). Conclusion: 95% ethanol was inefficiency to remove completely AH Plus, Epiphany SE and Sealapex residues of sealer-contaminated dentin.

Ethanol is inneficient to remove endodontic sealer residues of dentinal surface

Introduction: Endodontic sealer residues on dentinal surface have negative effects on adhesion of adhesives system and/or can cause discoloration of the dental crown. Objective: To evaluate the efficacy of 95% ethanol in removal of residues of epoxy-based (AH Plus), methacrylate-based (Epiphany SE) or calcium-based (Sealapex) sealers on dentinal surface. Material and methods: Thirty-two bovine incisor dental crown fragments (0.5 mm x 0.5 mm) were treated with 17% EDTA and 2.5% NaOCl. The specimens were divided into three experimental groups (n = 10): G1 (AH Plus), G2 (Epiphany SE) and G3 (Sealapex). In each group was applied a coating of one endodontic sealer type and were left undisturbed for 5 minutes. After this period, the specimens were cleaned with 95% ethanol. The control group was composed by two specimens that did not receive any sealer or cleaning treatment. The sealer residues persistence after cleaning with 95% ethanol was evaluated by scanning electron microscopy (x500) and a score system was applied. Data obtained were analyzed by Kruskal-Wallis and Dunn tests (α = 5%). Results: Moderate amount of endodontic sealer residues were observed in all groups, regardless of the endodontic sealer compositions. G1, G2 and G3 presented similar amount of sealer residues on dentinal surface after cleaning with 95% ethanol (p > 0.05). Conclusion: 95% ethanol was inefficiency to remove completely AH Plus, Epiphany SE and Sealapex residues of sealercontaminated dentin.

A method for retrieving endodontic or atypical non-endodontic separated instruments from the root canal: a report of two cases

Aim: This clinical report presents a new method for retrieving separated instruments from the root canal with minimally invasive procedures. Background: The presence of separated instrument in root canal may interfere in the endodontic treatment prognosis. There are several recommended methods to retrieve separated instruments, but some are difficult in clinically practice. Case report: This study describes two cases of separated instrument removal from the root canal using a stainless-steel prepared needle associated with a K-file. Case 1 presented a fractured gutta-percha condenser within the mandibular second premolar, it was separated during incorrect intracanal medication calcium hydroxide placement. Case 2 had a fractured sewing needle within the upper central incisor that the patient used to remove food debris from the root canal. After cervical preparation, the fractured instruments were fitted inside a prepared needle and then an endodontic instrument (#25 K-file) was adapted with clockwise turning motion between the needle inner wall and the fragment. Conclusion: The endodontic or atypical nonendodontic separated instrument may be easily pull on of the root canal using a single and low cost device. Clinical significance: The methods for retrieving separated instruments from root canal are difficult and destructive procedures. The present case describes a simple method to solve this problem.