Demineralized dentin matrix technique - a comparison of different demineralizing solutions.

This study aimed to evaluate the microstructure formed after the chemical treatment of teeth, for the development of autogenous grafts from the demineralized dentin matrix (DDM) technique, in order to identify the most efficient demineralizing solution. The specimens, originating from the root and coronal portion, were submitted to ultrasonic cleaning and drying in an oven for 1h at 100 ºC. Then, the density was determined by Archimedes' principle for each specimen, using distilled water as immersion liquid. The samples were separated into five groups: Control group: negative control, Distilled water;EDTA group: positive control, trisodium EDTA; NaOCl group: 2.5% sodium hypochlorite; HCl-0.6M group: 0.6M hydrochloric acid; and H2O2/H2SO4 group: hydrogen peroxide and sulfuric acid. Each specimen was immersed for 1h in the corresponding group descaling solution at 60 ºC. Subsequently, the mass loss and density of the treated specimens were determined by Archimedes' principle. Ultimately, the specimens of each group were characterized by microtomography, Scanning Electron Microscopy, and Energy Dispersive Spectrometry X-ray (SEM-EDS). The results demonstrated that the H2O2/H2SO4 solution allowed the formation of interconnected micropores, suggesting better pore structures for application in scaffolds, when compared to the other studied solutions.

Demineralized dentin matrix technique - a comparison of different demineralizing solutions

Abstract This study aimed to evaluate the microstructure formed after the chemical treatment of teeth, for the development of autogenous grafts from the demineralized dentin matrix (DDM) technique, in order to identify the most efficient demineralizing solution. The specimens, originating from the root and coronal portion, were submitted to ultrasonic cleaning and drying in an oven for 1h at 100 ºC. Then, the density was determined by Archimedes' principle for each specimen, using distilled water as immersion liquid. The samples were separated into five groups: Control group: negative control, Distilled water;EDTA group: positive control, trisodium EDTA; NaOCl group: 2.5% sodium hypochlorite; HCl-0.6M group: 0.6M hydrochloric acid; and H2O2/H2SO4 group: hydrogen peroxide and sulfuric acid. Each specimen was immersed for 1h in the corresponding group descaling solution at 60 ºC. Subsequently, the mass loss and density of the treated specimens were determined by Archimedes' principle. Ultimately, the specimens of each group were characterized by microtomography, Scanning Electron Microscopy, and Energy Dispersive Spectrometry X-ray (SEM-EDS). The results demonstrated that the H2O2/H2SO4 solution allowed the formation of interconnected micropores, suggesting better pore structures for application in scaffolds, when compared to the other studied solutions.

Resumo Este estudo teve como objetivo avaliar a microestrutura formada após o tratamento químico em dentes, para o desenvolvimento de enxertos autógenos a partir da técnica de matriz de dentina desmineralizada (DDM), a fim de identificar a solução desmineralizante mais eficiente. Os espécimes, provenientes da raiz e porção coronal, foram submetidos à limpeza ultrassônica e secagem em estufa por 1h a 100 ºC. Em seguida, a densidade foi determinada pelo princípio de Arquimedes para cada espécime, utilizando água destilada como líquido de imersão. As amostras foram separadas em cinco grupos: Controle: controle negativo, Água destilada; EDTA: controle positivo, EDTA trissódico; NaOCl: hipoclorito de sódio 2,5%; HCl-0.6M: ácido clorídrico 0,6M; e H2O2/H2SO4: peróxido de hidrogênio e ácido sulfúrico. Cada espécime foi imerso por 1h na solução descalcificante de grupo correspondente a 60 ºC. Posteriormente, a perda de massa e a densidade dos espécimes tratados foram determinadas pelo princípio de Arquimedes. Por fim, os espécimes de cada grupo foram caracterizados por microtomografia, microscopia eletrônica de varredura e espectrometria de energia dispersiva de raios-X (SEM-EDS). Os resultados demonstraram que a solução H2O2/H2SO4 permitiu a formação de microporos interligados, sugerindo melhores estruturas de poros para aplicação em scaffolds, quando comparada às demais soluções estudadas.

Pathological Evaluation by Microtomography of Ranula. Clinical Case Report

ABSTRACT: This study aims to demonstrate, through a clinical case report, the applicability of the use of microtomography (µCT) in the histopathological evaluation of a ranula lesion on the oral floor and to evaluate the use of 2 % elemental iodine solution as a contrast agent, in order to obtain a better contrast effect in a tissue sample, thus facilitating the identificat ion of anatomical structures, the histomorphological evaluation and the potential use of µCT in the imaging diagnosis of lesions. Different parameters were evaluated for obtaining images in SkyScan 1172, in a biopsied piece, when impregnated in a 10 % formalin solution and in a 2 % elemental iodine solution, when impregnated for 24h and 48h. Contrast agent impregnation was evaluated using the Hounsfield unit. The use of µCT allowed the identification of sialoliths dispersed inside the biopsy specimen, while the impregnation with Elemental iodine 2 % for 24h resulted in a better contrast when compared to the other conditions. The use of the Hounsfield unit allowed an adequate evaluation of the contrast obtained when the different parameters of impregnation and image acquisition were applied. The comparison between the 3D images with and without a specific marker highlighted a better evidencing of the soft tissues, with an improvement in the contrast of the images, also allowing the ident ification of the glandular duct obstructed by the sialoliths, allowing a conclusive histopathological evaluation of the biopsied lesion.

RESUMEN: Este estudio tiene como objetivo demostrar, a través de un reporte de caso clínico, la aplicabilidad del uso de la microtomografía (µCT) en la evaluación histopatológica de una lesión de ránula en el piso de la cavidad oral y evaluar el uso de solución de yodo elemental al 2 % como agente de contraste, con el fin de obtener un mejor efecto de contraste en una muestra de tejido, facilitando así la identificación de estructuras anatómicas, la evaluación histomorfológica y el potencial uso de µCT en el diagnóstico por imágenes de lesiones. Se evaluaron diferentes parámetros para la obtención de imágenes en SkyScan 1172, en una pieza biopsiada, cuando se impregna en una solución de formalina al 10 % y en una solución de yodo elemental al 2 %, durante 24 h y 48 h. La impregnación del agente de contraste se evaluó utilizando la unidad Hounsfield. El uso de µCT permitió la identificación de sialolitos dispersos dentro de la muestra de la biopsia, mientras que la impregnación con Yodo Elemental al 2 % durante 24 h resultó en un mejor contraste en comparación con las otras condiciones. El uso de la unidad Hounsfield permitió una adecuada evaluación del contraste obtenido cuando se aplicaron los diferentes parámetros de impregnación y adquisición de imágenes. La comparación entre las imágenes 3D con y sin marcador específico destacó una mejor evidenciación de los tejidos blandos, con una mejora en el contraste de las imágenes, permitiendo además identificar el conducto glandular obstruido por los sialolitos, permitiendo una evaluación histopatológica concluyente de la lesión sometida a biopsia.

Complications in the guided bone regeneration technique associated with the d-PTFE membrane: case report / Complicações na técnica de regeneração óssea guiada associada à membrana de PTFE-d: relato de caso

ABSTRACT Guided bone regeneration aims to gain vertical and horizontal bone volume in atrophic ridges, using different regenerative techniques associated with biomaterials, with occasional post-surgical complications. The objective of the case report was to describe the successive approaches to minimize and eliminate the complications presented in the postoperative period of a patient submitted to the guided bone regeneration technique. In the first surgery, the dense expanded polytetrafluoroethylene membrane (d-PTFE), supported by the tent technique and autogenous platelet graft, was used to perform the guided bone regeneration technique. After three months, the membrane was exposed, with the membrane and the retaining screws removed in the seventh month, with the installation of three Internal Hexagon implants in the areas of teeth 13, 14 and 15. The exposed threads were covered with hydroxyapatites resorbable and covered with d-PTFE membrane. A four-month postoperative panoramic radiograph suggested implant osseointegration. The guided bone regeneration technique associated with the d-PTFE membrane enabled bone neoformation, enabling the installation of osseointegrated implants in an aesthetic and functional position. The exposure of the edges of the membrane allowed the penetration of fluids and contamination, suggesting the worsening of the signs of infection and purulent secretion. On the contrary, the exposure of central areas did not cause inflammatory and infectious signs.

RESUMO A regeneração óssea guiada visa o ganho de volume ósseo vertical e horizontal em rebordos atróficos, sendo utilizadas diferentes técnicas regenerativas associadas a biomateriais, podendo apresentar ocasionais complicações pós-cirúrgicas. O objetivo do relato de caso foi descrever as sucessivas abordagens para minimizar e eliminar as complicações apresentadas no pós-operatório de paciente submetida à técnica de regeneração óssea guiada. Na primeira cirurgia foi utilizada a membrana de politetrafluoretileno expandido denso (PTFE-d), suportada pela técnica da tenda e enxerto plaquetário autógeno, para realizar a técnica de regeneração óssea guiada. Após três meses, constatou-se a exposição da membrana, sendo removida a membrana e os parafusos de contenção no sétimo mês, com instalação de três implantes Hexágono Interno nas áreas dos dentes 13, 14 e 15. As roscas expostas foram recobertas com as hidroxiapatitas reabsorvíeis e recobertas com membrana PTFE-d. A radiografia panorâmica pós-operatória de quatro meses sugeriu osseointegração dos implantes. A técnica de regeneração óssea guiada associada à membrana de PTFE-d possibilitou a neoformação óssea propiciando a instalação de implantes osseointegrados em posição estética e funcional. A exposição das bordas da membrana permitiu a penetração de fluídos e contaminação, sugerindo o agravamento dos sinais de infecção e secreção purulenta. Ao contrário, a exposição de áreas centrais, não acarretou sinais inflamatórios e infecciosos.

Leukocyte-platelet rich fibrin on the treatment of a large paradental cyst: a novel regenerative approach.

Leukocyte-platelet rich fibrin (PRF) is an autologous biomaterial formed by platelets, cytokines, growth factors and cells imprisoned on a fibrin mesh, produced according to Choukroun's protocol. The aim of the present article was to report the use of PRF, associated with a bone substitute, on the regenerative treatment of a large bone defect resulting from the enucleation of a paradental cyst involving the posterior mandible. The treatment resulted in the maintenance of the bone volume, and radiographic evaluation showed new bone formation after 40 days, suggesting an osteogenic and osteoinductive effect. Also, the current literature was reviewed.

Biocorrosive behavior of sulphate-reducing bacteria in kerr endodontic files: Determination of the corrosion.

AIMS: This study determined the corrosion rate by mass loss caused by oral strains of sulphate-reducing bacteria (SRB) in Kerr endodontic files (KF), aiming the development of a biopharmaceutical that facilitates the removal of endodontic limb fragments from root canals. MATERIALS AND METHODS: Nine new KF were analyzed after immersion in the modified Postgate E culture medium inoculated with Desulfovibrio desulfuricans oral (84 days), Desulfovibrio fairfieldensis in the consortium (84 days) and environmental D. desulfuricans (119 days). RESULTS: Optical microscopy revealed corrosion suggestive areas in all files submitted to immersion in SRB cultures, presenting a statistical difference (P < 0.05) between the samples environmental D. desulfuricans and KF control and between oral D. desulfuricans and KF control. Epifluorescence microscopy revealed an active SRB biofilm over the entire metal surface of the KF, as evidenced by the SYTO® 9 fluorophore. CONCLUSION: SRB were capable of promoting biocorrosion in Kerr type endodontic files, but with low rate.

Análise de limas endodônticas submetidas a biocorrosão por bactérias redutoras de sulfato in vitro / Analysis of endodontic files submitted to biocorrosion by sulfate reducing bacteria in vitro

Objetivo: Avaliar as alterações químicas presentes na superfície metálica de limas endodônticas fraturadas em canais radiculares, in vitro, após a inoculação intrarradicular de culturas de BRS de três cepas microbianas, Desulfovibrio desulfuricans (uma cepa oral e outra ambiental) e Desulfovibrio fairfieldensis. Métodos: foram analisadas 5 limas kerr #90, sendo uma Lima Kerr nova, sem tratamento, e as outras 4 limas fraturadas dentro de canais radiculares in vitro, com posterior inoculação de Desulfovibrio desulfuricans, cepa oral e ambiental, e Desulfovibrio fairfieldensis e um grupo controle sem inoculação bacteriana, por 477 dias. Os grupos foram analisados no modo EDS (Espectrometria de Energia Dispersiva de Raios-x) do microscópio eletrônico de varredura (FEI-Inspect-S50). Resultados:A presença do S, Cl e O foram relacionados ao processo biocorrosivo, assim como a redução dos elementos de liga nesta área. Conclusão:As análises no modo EDS demonstraram biocorrosão ao longo da superfície metálica das limas quando empregado o biofármaco BACCOR, nas três diferentes cepas empregadas, indicada pela redução dos elementos formadores da liga metálica, Fe, Ni e Cr, com a associação da presença de elementos indicadores de biocorrosão como O, Cl e S. (AU)

Aim:To evaluate the chemical alterations present on the metallic surface of root canal fractured endodontic files in vitro after the intraradicular inoculation of BRS cultures of three microbial strains, Desulfovibrio desulfuricans (one oral and one environmental strain), and Desulfovibrio fairfieldensis. Methods: Five kerr #90 files were analyzed, one new untreated Kerr file and the other 4 files fractured within root canals in vitro, with a subsequent inoculation of Desulfovibrio desulfuricans (oral and environmental strains), and Desulfovibrio fairfieldensis, as well as a control group without bacterial inoculation for 477 days. The groups were analyzed using the scanning electron microscope (FEI-Inspect-S50) EDS (X-ray Dispersive Energy Spectrometry) mode. Results:The presence of S, Cl, and O were related to the biocorrosive process, as well as the reduction of alloying elements in this area. Conclusion: The EDS mode analysis showed biocorrosion along the metallic surface of the files when the BACCOR biopharmaceutical was used in the three different strains employed in this study, indicated by the reduction of the alloying elements ­ Fe, Ni, and Cr ­ with the association of the presence of indicator elements of biocorrosion, such as O, Cl, and S. (AU)

Sulfate-Reducing Bacteria: Biofilm Formation and Corrosive Activity in Endodontic Files.

AIM: This study describes the biofilm formation and the corrosive capacity of sulfate-reducing bacteria (SRB) on the metallic structure of used endodontic files. METHODS: Sulfate-reducing bacteria (SRB) (Desulfovibrio desulfuricans oral and Desulfovibrio fairfieldensis or D. desulfuricans environmental) were inoculated into the culture media (Postgate C culture medium or modified Postgate E culture medium). The biocorrosive potential of these bacteria will be an important component of a biopharmaceutical under development called BACCOR. Afterwards, four used endodontic files (UEFs) were separately inoculated into a specific culture media for 445 days at 30°C in an incubator. The four UEFs were placed in a scanning electron microscope (SEM) and analyzed by the energy-dispersive X-ray spectrometry (EDS). RESULTS: The confocal laser scanning microscopic images indicate the presence of biofilm in the four samples. The SEM and SEM-EDS revealed the presence of rough, irregular structures adhering along the metallic surface of the used endodontic files, suggesting a mature calcified biofilm with a high concentration of Ca, P, C, and S. CONCLUSION: The formation of SRB biofilms on used endodontic files shows characteristics that may contribute to the biocorrosion of these files, and the results may also provide complementary data for a biopharmaceutical, which is still under development to assist in the removal of fractured endodontic files inside root channels.

Initial cytotoxicity assays of media for sulfate-reducing bacteria: An endodontic biopharmaceutical product under development.

This study assessed the cell viability of the inoculation vehicle of BACCOR (a combination of sulfate-reducing bacteria plus a culture media for bacteria), a biopharmaceutical product under development for dental use as aid in fractured endodontic file removal from the root canal. Different culture media for bacteria were evaluated: modified Postgate E (MCP-E mod), Modified Postgate E without Agar-agar (MCP-E w/Ag), Postgate C with Agar-agar (MCP-C Ag) and Postgate C without Agar-agar (MCP-C w/Ag). Cytotoxicity was quantified by the MTT test, exposing L929 and Vero cell lines to the vehicles over 24 h. The exposure of L929 cell line to MCP-E w/Ag resulted in biocompatibility (52% cell viability), while the exposure of the Vero kidney line revealed only MCP-E mod as cytotoxic. When diluted, all the vehicles showed biocompatibility with both cell lines. MCP-E w/Ag was the vehicle chosen for BACCOR, because of its biocompatibility with the cells used.

Bisphosphonate-related osteonecrosis of the jaws: Report of a case using conservative protocol.

Bisphosphonates have been the first-line treatment option for osteometabolic diseases, such as osteoporosis, hypercalcaemia in malignant bone diseases, and in bone metastasis. It is possible to observe a growing number of cases of osteonecrosis of the jaws in patients using this medication, called bisphosphonate-related osteonecrosis of the jaws. The purpose of this study was to report a conservative treatment for bisphosphonate-related osteonecrosis of the jaws--Stage 2, using antibacterial solution and low-level laser therapy. At the end of the treatment, the patient presented improvement of the lesion with the healing of the mucosa. The literature still lacks successful definite protocols, thus the present case may contribute with another option for conservative management for bisphosphonate-related osteonecrosis of the jaws. More research is necessary in order to develop a good protocol management for bisphosphonate-related osteonecrosis of the jaws.

Biocorrosion of Endodontic Files through the Action of Two Species of Sulfate-reducing Bacteria: Desulfovibrio desulfuricans and Desulfovibrio fairfieldensis.

AIM: This study assessed the biocorrosive capacity of two bacteria: Desulfovibrio desulfuricans and Desulfovibrio fairfieldensis on endodontic files, as a preliminary step in the development of a biopharmaceutical, to facilitate the removal of endodontic file fragments from root canals. MATERIALS AND METHODS: In the first stage, the corrosive potential of the artificial saliva medium (ASM), modified Postgate E medium (MPEM), 2.5 % sodium hypochlorite (NaOCl) solution and white medium (WM), without the inoculation of bacteria was assessed by immersion assays. In the second stage, test samples were inoculated with the two species of sulphur-reducing bacteria (SRB) on ASM and modified artificial saliva medium (MASM). In the third stage, test samples were inoculated with the same species on MPEM, ASM and MASM. All test samples were viewed under an infinite focus Alicona microscope. RESULTS: No test sample became corroded when immersed only in media, without bacteria. With the exception of one test sample between those inoculated with bacteria in ASM and MASM, there was no evidence of corrosion. Fifty percent of the test samples demonstrated a greater intensity of biocorrosion when compared with the initial assays. CONCLUSION: Desulfovibrio desulfuricans and D. fairfieldensis are capable of promoting biocorrosion of the steel constituent of endodontic files. CLINICAL SIGNIFICANCE: This study describes the initial development of a biopharmaceutical to facilitate the removal of endodontic file fragments from root canals, which can be successfully implicated in endodontic therapy in order to avoiding parendodontic surgery or even tooth loss in such events.

Detection of Sulphate-Reducing Bacteria and Others Cultivable Facultative Bacteria in Dental Tissues.

AIM: To detect for the presence of sulphate-reducing bacteria (SRB) and evaluate the possible association between SRB and cultivable facultative bacterial of oral sites with different periodontal conditions. METHODS: The study was carried out on 9 samples from different oral sites in 8 patients (two samples were collected from the same patient). Material was collected using modified Postgate E culture medium, indicated for the growth and isolation of SRB. In addition, a reducing solution for anaerobic bacteria was used as a transport solution for facultative bacteria and identified by polymerase chain reaction amplification (PCR) and sequencing of the 16S rRNA gene. RESULTS: SRB was found in 3 patient samples: the first in a root fragment, the second in a root fragment and a healthy tooth with vertical bone loss and a mobility degree of 3; and the third in a healthy tooth extracted for orthodontic treatment. In the final patient, the cultivable facultative species Lactobacillus casei was identified. Other facultative bacterial species were identified in patient 5 (Kurthia Gibsonii) and patient 7 (Pseudomonas aeruginosa). CONCLUSIONS: The detection of SRB in different dental tissues with distinct periodontal features demonstrated that new studies need to be developed in order to determine the true role of SRB in the oral microbiota. In addition, it was possible to verify the presence of Lactobacillus casei together with SRB in one sample.

Detection of sulphate-reducing bacteria in human saliva.

OBJECTIVE: The aim of the current study was to investigate the presence of sulphate-reducing bacteria (SRB) in human saliva and correlate with oral and systemic conditions. METHODS: Saliva samples were collected from 118 patients and inoculated in 2 ml of modified Postgate's E medium culture. After 28 days of incubation at 30°C the presence of SRB was identified by the production of sulphide. RESULTS: Of 118 saliva samples collected, 35 were positive for the presence of SRB. Three positive samples were randomly chosen to identify the species of SRB by PCR and sequenced. The three selected samples were identified as Desulfovibrio fairfieldensis, Desulfovibrio desulfuricans and Raoultella ornithinolytica. Gastritis (14.4%) was the most prevalent systemic disease, followed by diabetes (3.4%), while periodontitis (11%) and traumatic fibroma (4.2%) were the oral manifestations most frequently found. A bivariate analysis was performed to examine for the presence of SRB and the most prevalent systemic and oral manifestations. Only periodontitis showed a statistically significant association (p = 0.0003). CONCLUSIONS: The results showed SRB can be found in oral microbiota of healthy patients. Regarding the several conditions studied, there was a higher prevalence of SRB in patients with gastritis and patients with periodontal disease, with a possible correlation between the presence of SRB in the oral microbiota and periodontal disease.

Avaliação estereomicroscópica de limas endodônticas tipo K / Stereomicroscope evaluation of endodontics files K-type

As superfícies metálicas de dezesseis limas endodônticas tipo Kerr, novas, foram avaliadas através do uso de um microscópio esterioscópico. Todas as limas estudadas revelaram a existência de defeitos superficiais oriundos do processo de fabricação: partículas ao longo de todo o instrumento e/ou ranhuras de usinagem. A importância de caracterizar estes defeitos se reflete como um risco estrutural resultando em uma fratura inesperada do instrumento, obstrução do conduto radicular e o transporte destes fragmentos para a região perirradicular devido ao deslocamento das partículas presentes sobre a superfície metálica da lima.

The metallic surfaces of sixteen new Kerr-type endodontics files were evaluated through use of a stereomicroscope. All studied files showed surface defects from the manufacturing process: particles throughout the instrument and / or machining grooves. The importance of characterizing these defects is reflected as a structural risk resulting in an unexpected fracture of the instrument, obstruction of the root canal and transport of these fragments into the periradicular region due to displacement of the particles on the surface of the metal file.

Processos fisiológicos e patológicos das bactérias redutoras de sulfato do gênero desulfovibrio sp / Physiologic and pathological processes of the sulphate-reducing bacteria of the genus desulfovobrio sp

As bactérias redutoras de sulfato do gênero Desulfovibrio sp. podem ser encontradas normalmente formando parte da biota intestinal e oral de seres humanos saudáveis, participando, direta ou indiretamente, com seus produtos metabólicos, de diversas afecções como: periodontites, câncer colorretal, infecções e sepsemias. Propõe-se com esta revisão avaliar os aspectos normais e as possíveis alterações patológicas correlacionadas com as bactérias redutoras de sulfato no organismo humano. As conclusões levam a crer que o desequilíbrio na biota oral e intestinal pode levar a aumento no número de bactérias redutoras de sulfato e na produção de sulfeto, como produto metabólico final, podendo representar um fator adicional no desenvolvimento daquelas afecções. Além disso, por haver forte propensão para formar associações bacterianas, aumentando seu potencial patológico, pode ser difícil a identificação do seu verdadeiro papel nas morbidades em que estão envolvidas assim como o seu potencial virulento.

Sulfate-reducing bacteria of the genus Desulfovibrio spp. can be routinely detected as member of the normal intestinal and oral microbiota in health individuals. This bacterial group produces metabolic byproducts, which participate, direct or indirect, in several diseases, such as periodontitis, colorectal cancer, infections and sepsis. The purpose of the present study was to assess the association between sulfate-reducing bacteria and normal conditions and pathology alterations in human. In conclusion, it is possible that alteration of the oral and intestinal flora can result in increase of sulfate-reducing bacteria levels and products of sulfide as final metabolic. Therefore, these conditions can represent an important fact associated with those diseases. In addition, this bacterial group presents a great tendency in to associate with other microorganisms. Like that, it can increase the pathologic potential and can difficult the identification of the true involvement with several diseases as well as its virulent potential.