RESUMO
This study aimed to evaluate the expression of several differentiation markers in the apical papilla (AP) and dental pulp (DP) of human permanent teeth. Twenty young human teeth were extracted and classified according to three Moorrees tooth development stages: initial root formation (Ri), root length ½ (R1/2), and root length complete (Rc). Immunohistochemical assays were performed using STRO-1, VEGF Receptor-2, Neurofilament heavy (NFH), and Nestin antibodies and analyzed under light microscopy. Decalcified, formalin fixed paraffin embedded tooth sections stained with hematoxylin and eosin showed an apical cell rich zone between the DP and AP. The AP revealed fewer vascular and cellular components than the DP. STRO-1 was expressed on vascular and neuronal elements beneath the odontoblast (OB) and in the sub-odontoblastic (SOB) zone, and VEGFR-2 positive cells were observed in the endothelium, arterioles, and blood vessels. Neuroepithelial stem cell protein (Nestin) was highly expressed in differentiated odontoblasts in the predentin odontotoblast and odontoblast cell processes. Neurofilament heavy (NFH) was expressed in mature axons throughout the DP. STRO-1 and VEGFR-2 microvascular expression was higher at the stages Ri and R1/2 while STRO-1 and NFH expression showed strong spatial distribution of Rc neuronal elements as compared to Ri and R1/2. Differentiated OB and SOB cells showed Nestin expression, indicating a reservoir of newly differentiated odontoblast-like cells.
Assuntos
Polpa Dentária , Receptor 2 de Fatores de Crescimento do Endotélio Vascular , Humanos , Nestina , Antígenos de Diferenciação , Células-Tronco , BiomarcadoresRESUMO
OBJECTIVE: To evaluate the effect of several final irrigation protocols on radicular dentin microhardness, biochemical composition, and DMP1-CT expression. MATERIALS AND METHODS: A total of 140 single-rooted human teeth were prepared with WaveOne Gold files and randomly distributed into 7 groups (n = 20) according to the final irrigation protocol: distilled water (DW); sodium hypochlorite-EDTA (NaOCl-EDTA); EDTA (EDTA); EDTA-NaOCl (EDTA-NaOCl); EDTA-chlorhexidine (EDTA-CHX); passive ultrasonic irrigation (PUI:NaOCl-EDTA); and PUI:NaOCl-EDTA-NaOCl. Dentin microhardness (n = 10) was evaluated in the root canal lumen using Vickers hardness tester. Immunohistochemical analysis (n = 5) was used to evaluate DMP1-CT expression. Dentin ultrastructure and biochemical composition were evaluated by using Raman and energy dispersive X-ray analysis (EDAX) (n = 5) with a scanning electron microscope (SEM). Analysis of variance (ANOVA) and Tukey test were performed (pË0.05). RESULTS: Raman spectra of the organic content and DMP1-CT expression were lower at the lumen canal in EDTA-NaOCl, PUI:NaOCl-EDTA, and PUI:NaOCl-EDTA-NaOCl when compared to control (p < 0.05). EDAX showed reduced values for calcium and phosphorus in EDTA-NaOCl, PUI:NaOCl-EDTA, and PUI:NaOCl-EDTA-NaOCl. SEM microphotography's showed completely cleaned dentin, permeable tubules, and dentin erosion, mainly when PUI was used. NaOCl-EDTA presented significantly higher microhardness values than PUI:NaOCl-EDTA-NaOCl (p < 0.05). PUI:NaOCl-EDTA-NaOCl exhibited the lowest Vickers hardness values of all groups. CONCLUSION: The final irrigation protocols that used a final rinse with NaOCl and PUI showed a detrimental effect on radicular dentin DMP1-CT expression, biochemical composition, and microhardness. CLINICAL RELEVANCE: The adequate irrigation protocol could be advantageous to preserve the radicular dentin ultrastructure, promote adequate adhesion, and sustain favorable conditions for biomineralization and regeneration.
Assuntos
Cavidade Pulpar , Irrigantes do Canal Radicular , Dentina , Ácido Edético/farmacologia , Humanos , Microscopia Eletrônica de Varredura , Irrigantes do Canal Radicular/farmacologia , Preparo de Canal Radicular/métodos , Hipoclorito de Sódio/farmacologia , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVE: Matrix metalloproteinases (MMPs) are present in radicular dentin and can convert structural matrix proteins into signaling molecules; thus, these enzymes play an essential role in dentin biomineralization and tissue regeneration therapies. Their expression on radicular dentin may be affected by the irrigation solutions used during root canal treatments. This study aimed to evaluate the effects of the most common irrigants on radicular dentin MMP expression. MATERIALS AND METHODS: The experimental solutions were distilled water (control), 5% sodium hypochlorite (NaOCl), 18% ethylenediaminetetraacetic acid (EDTA), and 2% chlorhexidine (CHX). Samples were prepared from extracted human teeth. For zymography analysis, root sections were powderized, and dentin proteins were extracted to observe gelatinolytic activity. Root dentin slices were treated with the experimental solutions for immunohistochemical analysis using anti-MMP-2 and anti-MMP-9 antibodies. ANOVA and the Tukey test were performed. RESULTS: Zymograms revealed the presence of MMP-2, MMP-8, and MMP-20 in the control group and the EDTA-treated group. Immunohistochemistry confirmed the presence of MMP-2 and MMP-9 mainly associated with the dentinal tubule lumens and occasionally with intertubular dentin. NaOCl- and CHX-treated groups showed lower expression of MMPs than the control group. Immuno-staining for both proteinases in the EDTA-treated group showed higher expression compared to the other experimental groups. CONCLUSION: Our results showed that most common irrigants affect MMP expression on radicular dentin. Treatment with NaOCl and chlorhexidine resulted in lower expression of MMPs, while EDTA increased their expression in root canal dentin.
Assuntos
Clorexidina/farmacologia , Dentina , Ácido Edético/farmacologia , Metaloproteinase 2 da Matriz , Hipoclorito de Sódio/farmacologia , Dentina/efeitos dos fármacos , Humanos , Metaloproteinase 2 da Matriz/efeitos dos fármacos , Metaloproteinase 2 da Matriz/metabolismoRESUMO
OBJECTIVE: To evaluate the push-out bond strength of premixed and powder-liquid bioceramic sealers with or without gutta-percha (GP) cone. MATERIALS AND METHODS: Radicular dentin samples were prepared from 80 single-rooted human teeth. After root canal preparation using ProTaper® and irrigation with NaOCl and EDTA, teeth were divided according to the root canal sealer (n = 20): AH Plus®, EndoSequence® BC Sealer™, ProRoot® Endo Sealer, and BioRoot™ RCS. Samples were randomly divided into two subgroups (n = 10): GP-S: root canal filling using the single-cone technique, or S: filling with only sealer. Specimens were kept at 37°C and 100% humidity in calcium-free PBS for 30 days. The push-out bond strength was measured in MPa. Fractured specimens were observed at 25x to evaluate the type of failure. pH and calcium ion release were measured at different experimental periods. Raman and SEM-EDAX analyses were performed for root canal sealers. Data were analysed using three-way analysis of variance (ANOVA) and post hoc Tukey test at a significance of P < 0.05. RESULTS: Push-out bond strength was greater for samples obturated with only sealers (S) than samples obturated with the single-cone technique (GP-S) (P < 0.05). BioRoot™ RCS had greater bond strength than EndoSequence® BC Sealer™. Adhesive failures between cement and gutta-percha cone (87.5%) were predominant in the GP-S. Cohesive failures were predominant for S (80%). BioRoot™ RCS and ProRoot® ES presented higher alkalinization potential than the premixed sealer (EndoSequence® BC Sealer™). Powder-liquid bioceramic sealers (BioRoot™ RCS and ProRoot® ES) released the highest cumulative amount of calcium (28.46 mg/L and 20.05 mg/L). CONCLUSION: Push-out test without gutta-percha cone presents higher bond strength for bioceramic sealers. Powder-liquid calcium silicate-based sealers present greater bioactivity related to alkalinization potential and calcium ion release.
Assuntos
Guta-Percha , Materiais Restauradores do Canal Radicular , Resinas Epóxi , Humanos , Pós , Obturação do Canal RadicularRESUMO
Abstract In December 2019, a novel coronavirus disease (COVID-19) presenting with pneumonia emerged in Wuhan, China. The ongoing spread of COVID-19 worldwide is creating enormous uncertainty. In particular, the dental profession is facing a major challenge. Due to the characteristics of dental settings, the risk of cross-infection is elevated between patients and dental staff. The present review aimed to briefly discuss the overview of the disease, including the basic science, epidemiology, symptoms, routes of transmission and several topics that the authors believe are binding and relevant to the dental practice. The guidelines adopted by governments and international organizations also address all dental associations to protect the health of the community and to contain the spread of COVID-19 infection until a vaccine or an effective/validated treatment becomes available.
Resumen En diciembre del 2019, surgió una nueva enfermedad por coronavirus (COVID-19) que se presentó como una neumonía con etiología desconocida en Wuhan, China. La propagación continua de COVID-19 en todo el mundo está creando una enorme incertidumbre. En particular, la profesión dental se enfrenta a un gran desafío. Debido a las características de los consultorios dentales, el riesgo de infección cruzada es elevado entre los pacientes y el personal. La presente revisión tuvo como objetivo discutir brevemente la visión general de la enfermedad, incluyendo conceptos de ciencia básica, la epidemiología, vías de transmisión, síntomas y varios temas que los autores consideran vinculantes y relevantes para la práctica dental. Las directrices adoptadas por los gobiernos y las organizaciones internacionales también involucran a todas las asociaciones dentales para proteger la salud de la comunidad y contener la propagación de la infección por COVID-19 hasta que una vacuna o un tratamiento eficaz/validado esté disponible.
Assuntos
Aerossóis , Consultórios Odontológicos , COVID-19/prevenção & controleRESUMO
Abstract: The use of phosphate buffered saline (PBS) as an intracanal medication triggers a biomineralization process within mineral trioxide aggregate (MTA) apical plugs during the apexification process in teeth with incomplete rhizogenesis. However, no consensus is available in the literature regarding a restorative protocol for this type of treatment. Thus, the objective of this study was to use scanning electron microscopy (SEM) to evaluate the processes of biomineralization and adhesion in a restorative protocol for teeth with simulated incomplete rhizogenesis. Methodology: Root sections with a thickness of 2mm and cavities with a diameter of 2mm were used. The sections were randomly prepared and filled with the following materials: Group 1 (n=12), ProRoot MTA; and Group 2 (n=12): MTA Exp. Subsequently, the samples were immersed in PBS for 35 days. Every 5 days, the PBS was replaced, and the precipitates were collected, dried, and weighed. Two samples from each group were analyzed by SEM. Moreover, 24 single-rooted teeth were standardized, incomplete rhizogenesis was simulated, and 5-mm-long apical plugs were created with Pro Root MTA. As an intracanal medication, PBS was used for different periods of time: Group 1:48 h; Group 2:7 days; and Group 3:15 days. Then, fiberglass posts were cemented with the REBILDA® Post System. The samples were prepared and analyzed by SEM. Results: ProRoot MTA and MTA Exp effectively promoted the formation of carbonated apatite precipitates and biomineralization with dentin. ProRoot MTA yielded more carbonated apatite precipitates compared to MTA Exp (p=0.0536). The use of PBS as an intracanal medication for 7 and 15 days promoted intratubular mineralization (MIT), and treatment for 15 days was more effective (p < 0.05). The REBILDA® Post System effectively promoted the microimbrication of the adhesive system and the formation of resinous tags with lateral adhesive branches. Conclusion: Apexification with MTA associated with the use of PBS as an intracanal medication for 15 days, in addition to the use of the REBILDA® Post System, seems to be a feasible restorative protocol.
Resumen: El uso de solución salina tamponada con fosfato (PBS) como medicamento intracanal desencadena un proceso de biomineralización en los plugs apicales con agregado de trióxido mineral (MTA) durante el proceso de apexificación en dientes con rizogénesis incompleta. Sin embargo, no hay consenso disponible en la literatura sobre un protocolo restaurador para este tipo de tratamiento. Por lo tanto, el objetivo de este estudio fue utilizar microscopía electrónica de barrido (MEB) para evaluar los procesos de biomineralización y adhesión en un protocolo restaurador para dientes con rizogénesis incompleta simulada. Metodología: Se utilizaron secciones de raíz con un espesor de 2mm y se realizaron cavidades con un diámetro de 2 mm. Las cavidades en las secciones se obturaron con: Grupo 1 (n=12), ProRoot MTA; y Grupo 2 (n=12): MTA Exp. Posteriormente, las muestras se sumergieron en PBS durante 35 días. Cada 5 días, se reemplazó el PBS y se recogieron los precipitados, se secaron y pesaron. Dos muestras de cada grupo fueron analizadas por MEB. Además, se estandarizaron 24 dientes de raíz única, se simuló la rizogénesis incompleta y se crearon tapones apicales de 5mm de longitud con Pro Root MTA. Como medicamento intracanal, se utilizó PBS durante diferentes períodos de tiempo: Grupo 1:48 h; Grupo 2:7 días; y Grupo 3:15 días. Posteriormente, los postes de fibra de vidrio se cementaron con el sistema de postes REBILDA®. Las muestras fueron preparadas y analizadas por MEB. Resultados: ProRoot MTA y MTA Exp promovieron efectivamente la formación de precipitados de apatita carbonatada y la biomineralización con dentina. ProRoot MTA produjo más precipitados de apatita carbonatada en comparación con MTA Exp (p=0.0536). El uso de PBS como medicamento intracanal durante 7 y 15 días promovió la mineralización intratubular (MIT), siendo el tratamiento durante 15 días más efectivo (p <0.05). El sistema de postes REBILDA® promovió efectivamente la microimbricación del sistema adhesivo y la formación de tags resinosos. Conclusión: La apexificación con MTA asociada con el uso de PBS como medicación intracanal durante 15 días, además del uso del sistema de postes REBILDA®, parece ser un protocolo factible y eficaz en este tipo de tratamientos.
Assuntos
Apexificação , Biomineralização , Abscesso PeriapicalRESUMO
The biomineralisation of radicular dentin involves complex molecular signalling. Providing evidence of protein binding sites for calcium ions and mineral precipitation is essential for a better understanding of the remineralisation process. This study aimed to evaluate the functional relationship of metalloproteinases (MMPs) and non-collagenous proteins (NCPs) with mineral initiation and maturation during the biomineralisation of radicular dentin. A standardized demineralisation procedure was performed to radicular dentin slices. Samples were remineralised in a PBS-bioactive material system for different periods of time. Assessments of ion exchange, Raman analysis, and energy dispersive X-ray analysis (EDAX) with a scanning electron microscope (SEM) were used to evaluate the remineralisation process. Immunohistochemistry and zymography were performed to analyse NCPs and MMPs expression. SEM evaluation showed that the mineral nucleation and growth occurs, exclusively, on the demineralised radicular dentin surface. Raman analysis of remineralised dentin showed intense peaks at 955 and 1063 cm-1, which can be attributed to carbonate apatite formation. Immunohistochemistry of demineralised samples revealed the presence of DMP1-CT, mainly in intratubular dentin, whereas DSPP in intratubular and intertubular dentin. DMP1-CT and DSPP binding sites control carbonate apatite nucleation and maturation guiding the remineralisation of radicular dentin.
RESUMO
Apical periodontitis is frequently associated with the presence of bacteria biofilm, which has an indisputable impact on the prognosis of endodontic therapy due to the high resistance to adverse environmental conditions, chemicals, and antibiotic therapy that characterize bacteria within biofilm. The biofilm matrix acts as a protective shield over the encased microorganisms. The aim of this investigation was to identify the main biochemical components of biofilm matrix from endodontic mono- and dual-species biofilms. Enterococcus faecalis and Actinomyces naeslundii were cultured as mono- and dual-species biofilms for 14 days. Crude extracellular polymeric substances (EPSs) from biofilm matrices were extracted using chemical and physical methods. High-performance liquid chromatography, gas chromatography, and mass spectrometry were used to determine the carbohydrate, protein, and fatty acid components. Chemical analysis of the biofilm matrices revealed that they were mainly composed of stachyose, maltose, and mannose carbohydrates. The protein profile in all biofilm samples showed abundant oxidoreductases and chaperone proteins and some virulence- associated proteins mainly located in the membrane surface. High percentages of saturated and monounsaturated fatty acids were identified in all biofilm matrices, with a major prevalence of palmitic, stearic, and oleic acids. Based on the results, it was possible to obtain for the first time a general overview of the biochemical profile of endodontic biofilm matrices.
Assuntos
Actinomyces/fisiologia , Biofilmes/crescimento & desenvolvimento , Enterococcus faecalis/fisiologia , Periodontite Periapical/microbiologia , Proteínas de Bactérias/análise , Proteínas de Bactérias/metabolismo , Metabolismo dos Carboidratos , Carboidratos/análise , Ácidos Graxos Monoinsaturados/análise , Ácidos Graxos Monoinsaturados/metabolismo , Humanos , Chaperonas Moleculares/análise , Chaperonas Moleculares/metabolismo , Oxirredutases/análise , Oxirredutases/metabolismoRESUMO
Abstract One of the major approaches on dental research in this century is the development of biological strategies (tissue engineering) to regenerate/biomineralize lost dental tissues. During dentin- pulp regeneration, the interaction between stem cells, signaling molecules, biomaterials and the microenvironment in the periapical area drives the process for pulp tissue engineering. Understanding the signaling mechanisms and interactions involved with the biological process for the formation of a new tissue is essential. The knowledge of the micro-environment is the key for the application of tissue engineering. The present article is a short review of the current state of this topic, with the purpose of showing insights of pulp regeneration.
Resumen Actualmente la investigación en odontología se orienta al desarrollo de estrategias basadas en principios biológicos (ingeniería de tejidos) para la regeneración/biomineralización de estructuras dentales perdidas. El proceso de regeneración del complejo dentino-pulpar está guiado por la compleja interacción entre las células indiferenciadas de origen dental (DTSC), moléculas de señalización y biomateriales con el microambiente donde se va a restablecer. Es esencial comprender detalladamente, los mecanismos de señalización e interacciones involucradas en los procesos biológicos para la formación de un nuevo tejido, además de la identificación de los componentes presentes en los tejidos dentales implicados en este proceso (características del microambiente), ya que representan la base sobre la cual se debe emplear la ingeniería de tejidos. El presente articulo es una breve revisión del estado actual del tema, con el fin de entender el proceso de regeneración pulpar, basado en la comprensión de los fundamentos biológicos.