EM evaluation of bacterial biofilm and microorganisms on the apical external root surface of human teeth

The aim of this study was to evaluate the presence of bacterial biofilm on the external surface of the root apex in teeth with pulp necrosis, with and without radiographically visible periapical lesions, and in teeth with a vital pulp. Twenty-one teeth were extracted, eight with pulp necrosis and periapical lesions, eight with pulp necrosis without radiographically visible periapical lesions, and five with a vital pulp. The roots were sectioned, and the root apexes (+/- 3 mm) were processed for scanning electron microscope evaluation. The surface of the apical root was evaluated for the presence of microorganisms, root resorption, and biofilm. There were no microorganisms on the apical root surface of either teeth with pulp vitality or with pulp necrosis with no radiographically visible periapical lesions. Microorganisms were always present in teeth with pulp necrosis and radiographically visible periapical lesions. These included cocci, bacilli, and filaments and the presence of an apical biofilm. Apical biofilm is clinically important because microbial biofilms are inherently resistant to antimicrobial agents and cannot be removed by biomechanical preparation alone. This may cause failure of endodontic treatment as a consequence of persistent infection.

In vitro penetration of bleaching agents into the pulp chamber

Aim: To investigate pulp chamber penetration of bleaching agents in teeth following restorative procedures. Methodology: Bovine lateral incisors were sectioned 3 mm apical to the cemento-enamel junction and the coronal pulpal tissue was removed. Teeth were divided into six groups (n = 10): G1, G2 and G3 were not submitted to any restorative procedure, while G4, G5 and G6 were submitted to Class V preparations and restored with composite resin. Acetate buffer was placed in the pulp chamber and treatment agents were applied for 60 min at 37°C as follows: G1 and G4, immersion into distilled water; G2 and G5, 10% carbamide peroxide (CP) exposure; G3 and G6, 35% CP bleaching. The buffer solution was removed and transferred to a glass tube where leuco crystal violet and horseradish peroxidase were added, producing a blue solution. The optical density of the blue solution was determined spectrophotometrically at 596 nm. A standard curve made with known amounts of hydrogen peroxide was used to convert the optical density values of the coloured samples into microgram equivalents of hydrogen peroxide. Data were submitted to ANOVA and Tukey's test (5%). Results: Amounts of hydrogen peroxide found in the pulp chamber of G2 and G5 specimens (0.1833 ± 0.2003 μg) were significantly lower (P = 0.001) when compared to G3 and G6 specimens (0.4604 ± 0.3981 μg). Restored teeth held significantly higher (P = 0.001) hydrogen peroxide concentrations in the pulp chamber than intact teeth. Conclusion: Higher concentrations of the bleaching agent produced higher levels of hydrogen peroxide in the pulp chamber, especially in restored teeth.

Secretion of tumor necrosis factor-alpha by mouse peritoneal macrophages in the presence of dental sealers, sealapex and endomethasone

After filling root canals, the healing process depends on the chemical composition or physical-chemical properties of the material used, among other factors. All root canal sealers, whether solid or plastic, are foreign matter for the body if they remain in permanent contact with apical and periapical tissues. As a result, the first organic reaction that occurs is an attempt to phagocytize the material. During phagocytosis, macrophages release a large number of cell mediators into the area, among which are cytokines that are essential in intercellular communication and in many physiological and pathophysiological processes. One of these cytokines is tumor necrosis factor-alfa (TNF-α), which acts through links to specific receptors on the cell membrane initiating a cascade of events leading to induction, activation, or inhibition of numerous cytokine-regulated genes in the cell nucleus. The release of TNF-α in a cell culture of mouse peritoneal macrophages incubated with three concentrations (25, 50, and 100 mg/ml) of two endodontic sealers was measured. The solutions containing the calcium hydroxide-based root canal sealer (Sealapex) released fewer units of TNF-α than solutions containing the zinc oxide and eugenol-based sealer (Endomethasone).

Physico-chemical properties of MTA and a novel experimental cement

Aim: To evaluate the release of calcium ions, pH and conductivity of a new experimental dental cement (EC) and to compare them with those of mineral trioxide aggregate (MTA-Angelus). Methodology: Five samples of each cement were prepared using plastic tubes 1 mm in diameter and 10 mm long. Each sample was sealed in a test tube containing 10 mL deionized water which was analysed after 24, 48, 72, 96, 192, 240 and 360 h for pH, electrical conductivity and calcium release. The concentration of calcium ions was obtained through atomic absorption spectroscopy technique. The data were analysed statistically using the analysis of variance (ANOVA) and the Student's test (t-test). Results: The pH of the storage solutions was not affected by the material and the interaction of material with time (P > 0.05). However, the time of immersion was significant (P < 0.01) for both materials. For the electric conductivity and calcium release, the interaction of material with time was statistically significant (P < 0.01), indicating that EC and MTA-Angelus did not behave in a similar manner. Conclusions: The experimental cement released calcium and increased the pH of the storage solutions in a similar manner to MTA-Angelus. However, EC showed significantly higher calcium release than commercial MTA-Angelus after 24 h. © 2005 International Endodontic Journal.

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.

Ex vivo biocompatibility tests of regular and white forms of mineral trioxide aggregate

Aim: To examine the genotoxicity and cytotoxicity of regular and white mineral trioxide aggregate (MTA) ex vivo by the single-cell gel (comet) assay and trypan blue exclusion test, respectively. Methodology: Aliquots of 1 × 10 4 Chinese hamster ovary cells were incubated at 37°C for 3 h with grey and white forms of MTA at final concentrations ranging from 1 to 1000 μg mL -1. The negative control group was treated with vehicle control phosphate buffer solution for 3 h at 37°C and the positive control group was treated with methyl metasulfonate (at 1 μg mL -1) for 1 h at 37°C. After incubation, the cells were centrifuged at 180 g for 5 min and washed twice with fresh medium and resuspended with fresh medium. Each individual treatment was repeated three times consecutively to ensure reproducibility. Parameters from single-cell gel (comet) and cytotoxicity assays were assessed by the Kruskal-Wallis nonparametric test. Results: Neither compounds produced genotoxic effects with respect to the single-cell gel (comet) assay in all concentrations evaluated. In the same way, the dose-response relationships of all compounds tested at concentrations ranging from 1 to 1000 μg mL -1 on cell viability assessed by the trypan blue assay displayed no statistically significant differences (P > 0.05) for either endodontic material. Conclusions: Regular (grey) and white MTA are not genotoxins and do not induce cellular death. © 2006 International Endodontic Journal.

In vivo microbiological evaluation of the effect of biomechanical preparation of root canals using different irrigating solutions

The aim of this study was to evaluate the antimicrobial effect of biomechanical preparation using different irrigating solutions. Seventy-eight root canals from premolars of four dogs were used. After experimental induction of periapical lesions, the root canals were prepared using the following solutions for irrigation: Group 1) 2.5% sodium hypochlorite (NaOCl); Group 2) 2% chlorhexidine (CHX); Group 3) saline solution and Group 4) control group with no biomechanical preparation. The microbiological evaluation of the root canals was performed by counting the colony forming units (CFUs) using different culture mediums. Two absorbent paper cones were used in each root canal in order to collect the microbiological samples before, and thirty days after the biomechanical preparation. The culture plates were incubated in aerobic, anaerobic and microaerophilic environment. Statistical evaluation was carried out using analysis of variance, Tukey and Student tests. The results demonstrated that there was reduction in the number of microorganisms in the NaOCl and CHX groups (p<0.05). There was greater effectiveness in the chlorhexidine group. The group that used saline solution and the control group presented an increased number of microorganisms. It can be concluded that the use of antimicrobial irrigating solutions during biomechanical preparation promotes the reduction of endodontic microbiota. However, a considerable number of microorganisms were still observed.

Associação do hidróxido de cálcio e metronidazol no tratamento de dentes de cães com lesão periapical crônica

One of the primary objectives of endodontic treatment of teeth with pulp necrosis is the elimination of microorganisms from the root canal system, as effectively as possible, especially in cases with chronic periapical lesions. AIM: The purpose of this study was to analyze the response of the periapical tissue of dogs' teeth with chronic periapical lesions to endodontic treatment performed with utilization of metronidazole, calcium hydroxide, and an association of both as root canal dressings. METHODOLOGY: Forty root canals were submitted to pulpectomy and the root canals were kept exposed to the oral environment for 6 months. Then, they were submitted to biomechanical preparation and divided into 4 study groups with 10 specimens: group I - no root canal dressing; group II - calcium hydroxide; group III - metronidazole; group IV - calcium hydroxide associated to metronidazole. After 15 days, the root canals were filled with Fill Canal sealer. After 90 days, the animals were killed and the especimens processed for histological analysis. RESULTS: Calcium hydroxide dressing provided a significantly better outcome compared to other experimental groups (α = 0.01). Also, the results of the association of metronidazole and calcium hydroxide were similar to those observed for the metronidazole group. The worst results were obtained by the no root canal dressing group. CONCLUSION: The use of metronidazole alone or associated with Calcium hydroxide, did not improve periapical healing when compared to Calcium hydroxide dressing.

Histomicrobiologic aspects of the root canal system and periapical lesions in dogs' teeth after rotary instrumentation and intracanal dressing with Ca(OH)2 pastes

OBJECTIVE: The purpose of this study was to evaluate the distribution of microorganisms in the root canal system (RCS) and periapical lesions of dogs' teeth after rotary instrumentation and placement of different calcium hydroxide [Ca(OH)2]-based intracanal dressings. MATERIALS AND METHODS: Chronic periapical lesions were experimentally induced in 80 premolar roots of four dogs. Instrumentation was undertaken using the ProFile rotary system and irrigation with 5.25% sodium hypochlorite. The following Ca(OH) 2-based pastes were applied for 21 days: group 1 - Calen (n=18); group 2 - Calen+CPMC (n=20); group 3 - Ca(OH)2 p.a. + anaesthetic solution (n=16) and group 4 - Ca(OH)2 p.a.+ 2% chlorhexidine digluconate (n=18). Eight root canals without endodontic treatment constituted the control group. Histological sections were obtained and stained with Brown & Brenn staining technique to evaluate the presence of microorganisms in the main root canal, ramifications of the apical delta and secondary canals, apical cementoplasts, dentinal tubules, areas of cemental resorption and periapical lesions. The results were analyzed statistically by the Mann-Whitney U test (p<0.05). RESULTS: The control group showed the highest prevalence of microorganisms in all sites evaluated. Gram-positive cocci, bacilli and filaments were the most frequent morphotypes. Similar microbial distribution patterns in the RCS and areas of cementum resorption were observed in all groups (p>0.05). The percentage of RCS sites containing microorganisms in groups 1, 2, 3, 4 and control were: 67.6%, 62.5%, 78.2%, 62.0% and 87.6%, respectively. CONCLUSION: In conclusion, the histomicrobiological analysis showed that the rotary instrumentation and the different calcium hydroxide pastes employed did not effectively eliminate the infection from the RCS and periapical lesions. However, several bacteria seen in the histological sections were probably dead or were inactivated by the biomechanical preparation and calcium hydroxide-based intracanal dressing.

Avaliação radiográfica, densitométrica e histológica do uso de perfurações ósseas para o estímulo de consolidação de fraturas do terço distal do rádio de cães

To stimulate the bone callus development process from distal third of radius, 24 adult mongrel dogs used were from both sexes. These dogs were separated in two experimental groups of 12 animals each, named control and treated, divided in 4 moments (M1=15 days; M2=30 days; M3=45 days; M4=60 days), who underwent were performed surgical fractures. In treated group, it was performed bone perforations on proximal and distal edges, craniolateral and mediolateral to the fracture site. At the end of each moment, control and treated animals were evaluated by radiography, histology, and bone mineral densitometry (BMD) was determined on fracture site. According to the radiographic data of treated dogs, it was verified on days 15 and 30 more intense bone regeneration than control group. During M3 and M4, it wasn't detected any difference in bone reparation process betweencontrol and treated groups. In densitometric study, BMD values were greater in treated animals than in control dogs. Histological studies revealed at 15 and 30 days chondrocyte hyperplasia and initial endochondral ossification on drilled limbs; control group showed sustainment connective tissue and initial chondrocyte hiperplasia. At M3 and M4 of the treated group, were verified development and remodeling of periosteal callus in more advanced phases when comparing with limbs from control group. It can be concluded that using perforations enhances blood flow supply and activation of osteogenic cells on fracture site, stimulating the beginning of fracture consolidation process.

Intraosseous schwannoma mimicking a periapical lesion on the adjacent tooth: Case report

Aim: To present an additional case of intraosseous schawannoma involving the apical area in the mandibular alveolar bone mimicking an inflammatory periapical lesion. Summary: This article describes a case of schwannoma periapically located mimicking an inflammatory periapical lesion in the mandible of a 34-year-old female. Diagnostic and therapeutic problems can occur when this lesion is misinterpreted as being endodontic in origin. The diagnosis, radiograph, immunohistochemical aspects and treatment are also discussed. Key learning points: • Intraosseous schwannoma is a rare unilocular radiolucency that when located periapically could be misdiagnosed as an endodontic lesion and result in unnecessary root canal treatment. • The vitality of the pulp is an important test to exclude lesions of inflammatory origin. • Histological examination is important to establish the diagnosis of lesions in the periradicular region. © 2007 International Endodontic Journal.

In vitro antimicrobial activity of different gutta-percha points and calcium hydroxide pastes

The aim of this study was to evaluate the antimicrobial activity of different trademarks and compositions of gutta-percha points and calcium hydroxide pastes used in endodontic therapy. The evaluated material consisted of gutta-percha points containing calcium hydroxide (Roeko™), gutta-percha points containing chlorhexidine (Roeko™), two convencional gutta-percha points (Endo Points™ and Roeko™) and two calcium hydroxide pastes (Calen™ and Calen/PMCC™). Antimicrobial tests included five species of microorganisms: Escherichia coli (ATCC10538), Staphylococcus epidermidis (ATCC12228), Staphylococcus aureus (ATCC6538), Pseudomonas aeruginosa (ATCC27853), and Micrococcus luteus (ATCC9341). The Agar difusion method was employed. The plates were kept at room temperature for 2 h for prediffusion and then incubated at 37°C for 24 h. The triphenyltetrazolium chloride gel was added for optimization and the zones of inhibition were measured. Statistical evaluation was carried out using analysis of variance and Tukey Test. The obtained results showed that all microbial species used in the study were inhibited by the gutta-percha points containing chlorhexidine and by the calcium hydroxide pastes (Calen™ and Calen/ PMCC™), with similar results (p > 0.05). No antimicrobial activity was observed for the other groups. It was concluded that the gutta-percha points containing chlorhexidine presented antimicrobial activity, whereas the gutta-percha points containing calcium hydroxide did not.

Evaluation of the thermoplasticity of different gutta-percha cones and Resilon®

The goal of this study was to evaluate the thermoplasticity of conventional and thermoplastic gutta-percha and Resilon®, a polyester polymer-based material. Specimens with standardised dimensions were made from the following materials: conventional and thermoplastic gutta-percha (Dentsply), conventional and thermoplastic gutta-percha (Endopoints) and Resilon®. After 24 h, the specimens were placed in water at 70°C for 60 s, and thereafter positioned between two glass slabs. Each set was compressed by a 5-kg weight. Digital images of the specimens before and after compression were obtained and analysed. The thermoplasticity of each material was confirmed by the difference between final and initial areas of each sample. The data were analysed statistically by ANOVA and Tukey's test at a 5% significance level. Resilon® had the highest thermoplasticity means (P < 0.05). Among the gutta-percha cones, Endopoints TP (thermoplastic) presented the highest thermoplasticity means and differed significantly from the other commercial brands (P < 0.05). Resilon® had good thermoplasticity, endorsing its use as a thermoplastic root canal filling material. © 2007 The Authors. Journal compilation © 2007 Australian Society of Endodontology.

Estudo morfológico e histológico dos ovos de seis espécies do gênero Triatoma (Hemiptera: Reduviidae)

The objective of this study was to analyze the morphological and histological characteristics of eggs from six triatomine species in order to obtain a generic and specific characterization of this group of Trypanosoma cruzi vectors and better understand their phylogenetic and taxonomic aspects. The eggs of the respective species came from the collection of the Laboratório de Triatomíneos e Culicídeos da Faculdade de Saúde Pública/USP, from the Insetário do Serviço Especial de Saúde de Araraquara - SESA and from the Triatomine insectary of the Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos (FIOCRUZ), in Rio de Janeiro. Morphological studies were done with optical microscopy and scanning electron microscopy. Histological analyses used only optical microscopy. All of the species analyzed showed a predominance of hexagonal cells. The exchorion of Triatoma breyeri Del Ponte, T. costalimai Verano & Galvão, and T. tibiamaculata (Pinto) consists of unornamented cells, overlapping or slightly separated, with a smooth, padded appearance. Meanwhile, in T. matogrossensis Leite & Barbosa, T. sherlocki Papa, Juberg, Carcavallo, Cerqueira & Barata, and T. williami Galvão, Souza & Lima, the exchorion cells are ornamented on their entire surface with perforations and fissures. Egg histology showed undulations, pores, and orifices. The study helps to expand the generic and specific knowledge of the Triatominae subfamily. The characteristics identified in the exchorion of the eggshells can help separate close species. These new parameters will back the elaboration of future dichotomous keys, help to determine each vector species' role and expand knowledge of the various species.

Comparative Analysis of 2 Techniques of Double-Gloving Protection During Arch Bar Placement for Intermaxillary Fixation

Purpose: This study was conducted to comparatively evaluate, in a prospective and randomized manner, 2 techniques for providing double-gloving protection during arch bar placement for intermaxillary fixation. Materials and Methods: A total of 42 consecutive patients in whom application of an Erich bar was indicated for intermaxillary fixation were equally divided into 2 groups. In group 1, 2 sterile surgical gloves were used; in group 2, a nonsterile disposable inner glove was used under a sterile surgical glove. Wilcoxon, Mann-Whitney, Kruskal-Wallis, and binomial statistical tests were used to analyze the findings. Results: A total of 103 perforations were found in the outer gloves (47 in group 1 and 56 in group 2), along with 5 perforations in inner gloves in both groups (α = .01). No significant statistical difference was found between groups in terms of inner glove perforations (α = .05). The nondominant hand presented with 70.9% of the perforations, statistically significant to 1%. Conclusions: Both double-gloving techniques were found to provide effective clinician protection. The use of a nonsterile disposable glove under the surgical glove is possible for less-invasive procedures, offering the same safety as using 2 sterile surgical gloves while decreasing operational costs. This method does not eliminate the need to change gloves when a perforation is suspected or noted during the surgery, however. © 2007 American Association of Oral and Maxillofacial Surgeons.