Cytotoxicity, and antimicrobial and physicochemical properties of sealers incorporated with Uncaria tomentosa

Abstract This study evaluated the cytotoxicity, the antimicrobial and physicochemical properties of root canal sealers incorporated with phytotherapic Uncaria tomentosa (UT). Unmodified AH Plus (Dentsply, DeTrey, Germany) and MTA Fillapex (Angelus, Londrina, Brazil) were used as controls. UT was incorporated into AH Plus and MTA Fillapex, at concentrations of 2% and 5% of the total weight of these sealers (w/w). Flowability, setting time, and solubility were evaluated following ISO requirements. The pH values were measured at periods of 12, 24, 48 hours, and 7 days. The antimicrobial activity of the sealers against Enterococcus faecalis was analyzed by both direct contact tests in freshly prepared sealers, and after 7 days. The cytotoxicity of the samples was evaluated by the MTT assay, to check Balb/c 3T3 cell viability. The statistical analysis was performed by one-way ANOVA and Tukey's test (p < 0.05). The incorporation of UT was associated with a decrease in flow, for both sealers, an increase in AH Plus setting time, increase in MTA Fillapex pH values, and solubility (after 14 days), for both sealers (p < 0.05). Regarding the antibacterial evaluation, bacterial reduction was reported after incorporation of UT into both AH Plus and MTA Fillapex, up to 7 days after handling of the material (P<0.05). UT incorporation decreased the cytotoxic effects of both AH Plus and MTA Fillapex sealers in a way directly proportional to their respective concentrations (p < 0.05). In conclusion, UT can be added to both sealers to reduce their cytotoxicity, and improve their antibacterial effects, without compromising their original physicochemical properties.

Investigation of a modified hydraulic calcium silicate-based material - Bio-C Pulpo

Abstract This study evaluated the physicochemical, biological, and antimicrobial properties of a new hydraulic calcium silicate-based modified material, and compared it with MTA Repair HP and MTA Angelus. The materials were assessed regarding color luminosity (L), color change, radiopacity, setting time, and ISO 6876:2012 linear flow. Volumetric filling and volume change were evaluated using microcomputed-tomography (µCT). Chemical characterization after 28 days in Hank's Balanced Salt Solution (HBSS) and pH analysis were also assessed. Biological characterization of cytotoxicity and microbiological assessment were also undertaken. Shapiro-Wilk, ANOVA, Levene and post hoc analyses with Bonferroni correction were performed, adopting a 5% significance level (p <0.05). Bio-C Pulpo exhibited the highest L values after 90 days. All tested materials demonstrated color change during the analyses, and had radiopacity above 5 mm Al. MTA Repair HP set faster than Bio-C Pulpo, whereas the latter had the highest linear flow. MTA Repair HP had the highest volumetric filling in µCT analysis. Bio-C Pulpo showed the highest alkalinity during all tested periods, and the highest volumetric loss (above 9%), in comparison with MTA Repair HP and MTA Angelus. Bio-C Pulpo did not form calcium hydroxide after hydration. MTA Repair HP demonstrated the highest cytocompatibility, and Bio-C Pulpo, the highest cytotoxicity. No inhibition halos were observed for any material, and similar higher turbidity values were seen after direct contact. Composition additives used in Bio-C Pulpo modified its properties, and both the absence of calcium hydroxide deposition after hydration, and the related cytotoxicity of this material are of particular concern.

Evaluación de la citotoxicidad de Biodentine, IRM y MTA en cultivos de fibroblastos del ligamento periodontal humano / An evaluation of the cytotoxicity of Biodentine, IRM and MTA in cultures of human periodontal ligament fibroblasts

Introducción: Los materiales para la obturación retrógrada son diversos. Actualmente, IRM y MTA son las alternativas clínicas más utilizadas, no obstante, es relativamente reciente la introducción de materiales a base de silicatos tricálcicos tal como Biodentine. Objetivo: Determinar la citotoxicidad de fibroblastos del ligamento periodontal humano expuestos a medios de cultivo condicionados con Biodentine, IRM y MTA. Material y métodos: 1 × 103 fibroblastos del ligamento periodontal humano fueron expuestos a medios DMEM/F12 condicionados con MTA, IRM y Biodentine en tres protocolos diferentes. Se realizó un ensayo de MTT para determinar la viabilidad celular a las cero, 24, 48, 72 horas, siete y 14 días. Se realizó una prueba ANOVA (p < 0.05). Resultados: En los tres protocolos con los diferentes medios de cultivo condicionados, la viabilidad de las células fue predominantemente proliferativa; sin embargo, las células expuestas a Biodentine mostraron una tendencia mayor que la MTA o la IRM. Conclusión: Las células expuestas a la Biodentine mostraron un comportamiento proliferativo a los 14 días de análisis. Se debe realizar más investigación a nivel in vivo y clínico para obtener más información sobre la conducta de estos materiales empleados para la obturación retrógrada (AU)

Introduction: The materials for retrograde filling are diverse. Currently, IRM and MTA are the most commonly used clinical alternatives, however, the introduction of materials based on tricalcium silicates such as Biodentine is relatively recent. Objective: To determine the cytotoxicity of human periodontal ligament fibroblasts exposed to culture media conditioned with Biodentine, IRM and MTA. Material and methods: 1 × 103 fibroblasts of the human periodontal ligament were exposed to DMEM/F12 media conditioned with MTA, IRM and Biodentine in 3 different protocols. An MTT assay was performed to determine cell viability at 0, 24, 48, 72 hours, seven and 14 days. An ANOVA test was performed (p < 0.05). Results: In the three protocols with the different conditioned culture media, the viability of the cells was predominantly proliferative, however, the cells exposed to Biodentine showed a higher tendency than the MTA or the IRM. Conclusion: The cells exposed to the Biodentine showed a proliferative behavior at 14 days of analysis. More research should be done at in vivo and clinical level to obtain more information about the behavior of these materials used for retrograde filling (AU)

Cytotoxicity and biocompatibility of a new bioceramic endodontic sealer containing calcium hydroxide

Abstract: This study evaluated the cytotoxicity and biocompatibility of a new bioceramic endodontic sealer (i.e., Sealer Plus BC) in comparison with those of MTA Fillapex and AH Plus. L929 fibroblasts were cultured and Alamar Blue was used to evaluate cell viability of diluted extracts (1:50, 1:100, and 1:200) from each sealer at 24 h. Polyethylene tubes that were filled with material or empty (as a control) were implanted in the subcutaneous tissue of rats. The rats were killed after 7 and 30 d (n = 8), and the tubes were removed for histological analysis. Parametric data was analyzed using a one-way ANOVA test, and nonparametric data was analyzed via the Kruskal-Wallis test followed by the Dunn test (p < 0.05). A reduction in cell viability was observed in the extracts that were more diluted for Sealer Plus BC when compared to that of Control and AH Plus (p < 0.05). However, the 1:50 dilution of the Sealer Plus BC was similar to that of the Control (p > 0.05). Conversely, more diluted extracts of MTA Fillapex (1:200) and AH Plus (1:100 and 1:200) were similar to the Control (p > 0.05). Histological analysis performed at 7 d did not indicate any significant difference between tissue response for all materials, and the fibrous capsule was thick (p > 0.05). At 30 d, Sealer Plus BC was similar to the Control (p > 0.05) and MTA Fillapex and AH Plus exhibited greater inflammation than the Control (p < 0.05). The fibrous capsule was thin for the Control and for most specimens of Sealer Plus BC and AH Plus. Thus, Sealer Plus BC is biocompatible when compared to MTA Fillapex and AH Plus, and it is less cytotoxic when less-diluted extracts are used.

Cytotoxicity Evaluation of Root Canal Sealers Using an In Vitro Experimental Model with Roots

Abstract The aim of the present study was to evaluate the cytotoxicity of root canal sealers under conditions closely resembling a clinical reality. A primary human fibroblast cell line was seeded in 24-well acrylic plates with Dulbecco’s modified Eagle’s medium supplemented with 10% serum fetal bovine (SFB) and incubated for 24 h. Root canals from premolars were filled and individually attached to nylon devices to be stabilized in the wells with the already seeded cells. Specimens were divided into groups as follows: Control: gutta-percha cones (GPC); AH Plus+GPC; Sealapex+GPC; MTA Fillapex+GPC and Endofill+GPC. After 24 and 48 h, cell viability and morphology were evaluated by MTT assay and scanning electron microscopy (SEM), respectively. Statistical analysis was performed by Mann-Whitney test, complemented by Kruskal Wallis test (p<0.05). Only Endofill presented cytotoxicity after 24 h. MTA Fillapex and Endofill reduced the production of succinic desidrogenase after 48 h. AH Plus was non-toxic at any time point. SEM showed that the AH Plus and MTA Fillapex groups presented fibroblasts with morphology close to the control group, while the Endofill group presented few cells with thin extensions cells. The present study showed that good results were present in AH Plus and Sealapex, but not the Endofill group after 48 h. The method used enabled evaluation of the cytotoxicity of the studied sealers that diffused through the root apex.

Resumo O objetivo do presente estudo foi avaliar a citotoxicidade dos cimentos dos canais radiculares em condições próximas à realidade clinica. Uma linhagem primária de fibrolastos humanos foi semeada em placas acrílicas de 24-poços com meio de cultura Dulbecco’s modified Eagle’s medium suplementado com 10% de soro fetal bovino e incubados por 24 h. Os canais radiculares de pré-molares foram obturados e individualmente adaptados aos dispositivos de nylon para serem estabilizados nos poços com as células já semeadas. Amostras foram dividas de acordo com os grupos: Controle: cones de gutta-percha (CGP); AH Plus+CGP; Sealapex+CGP; MTA Fillapex+CGP e Endofill+CGP. Após 24 e 48 h, a viabilidade e a morfologia celular foram avaliadas pelo ensaio de MTT e microscopia eletrônica de varredura (MEV), respectivamente. Análises estatísticas foram realizadas pelo teste de Mann-Whitney, complementadas por Kruskal Wallis (p<0,05). Apenas o Endofill apresentou citotoxicidade após 24 h. MTA Fillapex e Endofill reduziram a produção da enzima desidrogenase succinica após 48 h. AH Plus não apresentou toxicidade em nenhum momento. MEV mostrou que os grupos AH Plus e o MTA Fillapex apresentaram fibroblastos com morfologia próxima ao grupo controle, enquanto que o grupo do Endofill apresentou poucas células com finos prolongamentos. O presente estudo demonstrou que resultados satisfatórios foram apresentados nos grupos AH Plus e Sealapex, mas não para o Endofill após 48 h. O método utilizado permitiu avaliar a citotoxicidade dos cimentos que se difundem pelo ápice radicular.

Physical-chemical properties and cytotoxicity analysis of 5 different endodontic sealers / Propriedades físico-químicas e análise da citotoxicidade de 5 diferentes cimentos endodônticos

The aim of this study was to analyze the radiopacity, setting time, flowability, pH, calcium ion release, solubility and cytotoxicity of bioceramic cements Totalfill BC Sealer and Totalfill BC RRM, and compare them with AH Plus, MTA Fillapex and MTA Angelus. The groups were divided and compared among them according to the filling and retro-filling cement functions. Totalfill BC Sealer was compared with AH Plus and MTA Fillapex; and Totalfill BC RRM retrofilling cement with MTA Angelus. For radiopacity analysis, specimens were placed in metal rings measuring 10x1 mm placed on occlusal film together with the aluminum scale. Digora 1.51 software was used to evaluate the digitized images and determine radiographic density. Setting time was tested in accordance with the American Society for Testing and Materials C266-08 standard specifications, but specimens were fabricated in accordance with the International Organization for Standardization 6876: 2001. Flow was tested in accordance with ANSI/ADA No.57/200 specifications. In total 30 acrylic teeth were filled with filling-cements and 20, with (retrograde cavity) retro-filling cements. All teeth were immersed in ultrapure water for pH and calcium ion release measurement (atomic absorption spectrophotometer) for time intervals of 1, 3, 24, 72, 168 and 360 hours. Solubility was tested by scanning and digitizing 50 acrylic teeth twice by Micro- CT, before and after immersion in ultrapure water for time intervals of 168, 360 and 720 hours. The images were reconstructed and volume (mm3) values of samples obtained by means of CTan software (CTan v1.11.10.0, SkyScan). The in vitro effects on cells were analyzed at concentrations of 100, 50, 10, 5, 1 mg/mL, and 0 mg / mLnegative control group and recorded in time intervals of 24, 48 and 72 hours by MTT reduction assay. The results were statistically analyzed by the ANOVA, Tukey, Kruskal-Wallis and Dunn tests (P<0.05). All radiopacity values according to ISO 6876/2001, AH Plus (7.86 mm Al) being the most radiopaque followed by Totalfill BC Sealer (4.84 mm Al), MTA Fillapex (3.41 mm Al), Totalfill BC RRM (6.8 mm Al), and MTA Angelus (6.7 mm Al). The following values were the initial and final setting time (in hours), respectively: AH Plus (8 and 15); Totalfill BC Sealer (11 and 24); MTA Fillapex (13 and 26); MTA Angelus (10 and 120 minutes) and Totalfill BC RRM (3 hours and 22 hours). In flow analysis, the cements behaved as follows: MTA Fillapex (47 mm), Totalfill BC Sealer (41.5 mm), Totalfill BC RRM (33.5 mm), AH Plus (33 mm) e MTA Angelus (17.5 mm) (p < 0.05). pH analysis showed in general the lowest values for AH Plus cement, followed by Totalfill BC RRM, MTA Angelus, MTA Fillapex and Totalfill BC Sealer. AH Plus showed the highest Ca2+ release in time interval 1 hour (1.38 mg/L); MTA Fillapex, in 360 hours (3.81 mg/L); MTA Angelus, 1 hour (1.38 mg/L); Totalfill BC Sealer, 360 hours (6.77 mg/L) and Totalfill BC RRM, 360 hours (3.81 mg/L). Almost all the sealers presented solubility lower than 3% in all periods, as recommended by ISO 6876/2001. Whereas, the MTA Fillapex solubility value was higher than 5% in all periods. Relative to cytotoxicity, all the cements were shown to be toxic at the concentration of 100 mg/mL, however, Totalfill BC Sealer and Totalfill BC RRM showed the best cell viability result compared with the other cements tested. We concluded that all root canal filling and root retro-filling complied with the requisites of radiopacity, setting time, flow, pH, calcium ion release, solubility and cytotoxicity. With the exception of the MTA Fillapex that not only fulfilled the requirement of solubility. Of the sealers, Totalfill BC Sealer was outstanding: it showed the highest pH and Ca2+ release, and lowest cytotoxicity. Among the retrofilling cements, Totalfill BC RRM maintained its high pH, higher Ca2+ release, and lower cytotoxicity. (AU)

O objetivo do presente estudo foi analisar a radiopacidade, tempo de presa, escoamento, pH, liberação de íons cálcio, solubilidade e citotoxicidade dos cimentos biocerâmicos Totalfill BC Sealer e Totalfill BC RRM e compará-los ao AH Plus, MTA Fillapex e MTA Angelus. Os grupos foram divididos e comparados entre si de acordo com as funções dos cimentos de obturação e retro-obturação. Comparamos o cimento obturador Totalfill BC Sealer com os cimentos AH Plus e MTA Filapex, e o cimento retrobturador Totalfill BC RRM com o cimento retrobturador MTA Angelus. Para análise da radiopacidade, os espécimes foram colocados em anéis metálicos medindo 10x1 mm, dispostos sobre um filme oclusal com uma escala de alumínio. O software Digora 1.51 foi utilizado para avaliar as imagens digitalizadas e determinar a densidade radiográfica. O tempo de presa foi realizado de acordo com as especificações da American Society for Testing and Materials C266-08 standard specifications, mas os espécimes foram feitos de acordo com a International Organization for Standardization 6876: 2001. O escoamento foi realizado de acordo com as especificações ANSI/ADA N0 57/2000. Trinta dentes acrílicos foram preenchidos com cimentos obturadores e vinte dentes de acrílico (com cavidade retrógrada) foram preenchidos com cimentos retro-obturadores e imersos em água ultrapura para mensuração do pH e liberação de íons cálcio (espectrofotômetro de absorção atômica) no período de 1, 3, 24, 72, 168 e 360 horas. Para o teste de solubilidade, foram escaneados 50 dentes acrílicos e digitalizados duas vezes pelo Micro-CT, antes e após a imersão em água ultrapura nos períodos de 168, 360 e 720 horas. As imagens foram reconstruídas e o volume (mm3) das amostras foi obtido usando o software CTan (CTan v1.11.10.0, SkyScan). Os efeitos celulares in vitro foram analisados nas concentrações de 100, 50, 10, 5, 1 mg/mL e 0 mg / mLgrupo controle negativo e registados nos períodos de 24, 48 e 72 horas através do ensaio de redução de MTT. Os resultados foram analisados estatisticamente pelos testes ANOVA, Tukey, Kruskal-Wallis e Dunn (p < 0.05). Todos os valores de radiopacidade estavam de acordo com a norma ISO 6876/2001, sendo o AH Plus (7.86 mm Al) o mais radiopaco seguido dos demais cimentos; Totalfill BC Sealer (4.84 mm Al), MTA Filapex (3.41 mm Al), Totalfill BC RRM (6,8 mm Al), MTA Angelus (6,7 mm Al). Os valores obtidos para o tempo de presa inicial e final foram respectivamente, AH Plus (8 e 15 horas), Totalfill BC Sealer (11 e 24 horas), MTA Filapex (13 e 26 horas), Totalfill BC RRM (3 horas e 22 horas) e MTA Angelus (10 e 120 minutos). Na análise de escoamento os cimentos se comportaram da seguinte forma: AH Plus (33 mm), MTA Filapex (47 mm), Totalfill BC Sealer (41,5 mm), Totalfill BC RRM (33,5 mm), e MTA Angelus (17,5 mm) (p < 0.05). A análise do pH mostrou que o cimento AH Plus de um modo geral, foi o que apresentou os menores valores, seguido do Totalfill BC RRM, MTA Angelus, MTA Filapex e Totalfill BC Sealer. A maior liberação de Ca2+ do AH Plus foi no período de 1 hora (1.38 mg/L), MTA Filapex foi em 360 horas (3.81 mg/L), Totalfill BC Sealer 360 horas (6.77 mg/L), Totalfill BC RRM 360 horas (3.81 mg/L) e MTA Angelus em 1 hora (1.38 mg/L). Todos os cimentos apresentaram solubilidade menor que 3% em todos os períodos, como recomendado pela ISO 6876/2001. Entretanto, os valores de solubilidade do MTA Fillapex excedeu mais que 5% em todos os períodos. Com relação à citotoxicidade, todos os cimentos mostraram-se tóxicos na concentração de 100 mg/mL, porém o Totalfill BC Sealer e Totalfill BC RRM apresentaram melhor resultado de viabilidade celular comparado aos demais cimentos testados. Concluiu-se que os cimentos de obturação e retro-obturação cumpriram os requisitos de radiopacidade, tempo de presa, escomento, pH, liberação de íons cálcio, solubilidade e citotoxicidade. Com exceção do MTA Fillapex que não cumpriu somente o requisito de solubilidade. Dos cimentos obturadores, o que melhor se portou foi o Totalfill BC Sealer, apresentando maior pH e liberação de íons cálcio e menor citotoxicidade. Dentre os cimentos retro-obturadores, Totalfill BC RRM foi o que melhor se destacou, mantendo seu pH elevado, possuindo maior liberação de Ca2+ e menor citotoxicidade. (AU)

Interaction of dental pulp stem cells with Biodentine and MTA after exposure to different environments

ABSTRACT Objective: The aim of the present study was to evaluate and compare the cytotoxic effects of Biodentine and MTA on dental pulp stem cells (DPSCs) and to assess cell viability and adherence after material exposure to an acidic environment. Material and Methods: DPSCs were cultured either alone or in contact with either: Biodentine; MTA set for 1 hour; or MTA set for 24 hours. After 4 and 7 days, cell viability was measured using the MTT assay. Biodentine and MTA were also prepared and packed into standardized bovine dentin disks and divided into three groups according to the storage media (n=6/group): freshly mixed materials without storage medium (Group A); materials stored in saline (Group B); materials stored in citric acid buffered at pH 5.4 (Group C). After 24 hours, DPSCs were introduced in the wells and cell adherence, viability, and cellular morphology were observed via confocal microscopy after three days of culture. Cell viability was analyzed using repeated-measures analysis of variance test with Tukey's post hoc tests (α=0.05). Results: Biodentine expressed significantly higher cell viability compared with all other groups after 4 days, with no differences after 7 days. Notably, cell viability was significantly greater in 24-hour set MTA compared with 1-hour set MTA and control groups after 7 days. Material exposure to an acidic environment showed an increase in cell adherence and viability in both groups. Conclusions: Biodentine induced a significantly accelerated cell proliferation compared with MTA. Setting of these materials in the presence of citric acid enhanced DPSC viability and adherence.

Cytotoxic effects of new MTA-based cement formulations on fibroblast-like MDPL-20 cells

Abstract The present study aimed at evaluating the cytotoxic effects of a novel cement called CER on periodontal fibroblast-like cells of mice (MDPL-20), in comparison with different formulations of Mineral Trioxide Aggregate (MTA), by means of the cell viability test (MTT) and cell morphology analysis. Thirty-two round-shaped samples were fabricated with the following cements: white MTA, white and gray CER and experimental white MTA. The samples were immersed in serum-free culture medium for 24 hours or 7 days (n = 16). The extracts (culture medium + components released from the cements) were applied for 24 hours to previously cultured cells (40.000 cells/cm2) in the wells of 24-well plates. Cells seeded in complete culture medium were used as a negative control. Cell viability was assessed using the MTT assay. Two samples of each cement were used for cell morphology analysis by Scanning Electron Microscopy (SEM). The extracts obtained at the 7-day period presented higher cytotoxicity compared with the 24-hour period (p < 0.05). The gray CER obtained at 24 hours presented the highest cytotoxic effect, whereas the experimental white MTA presented the lowest, similar to the control (p > 0.05). However, at the 7-day period, the experimental white MTA presented no significant difference in comparison with the other cements (p > 0.05). At the 7-day period, CER cement presented cytotoxic effects on fibroblast-like cells, similar to different MTA formulations. However, the immersion period in the culture medium influenced the cytotoxicity of the cements, which was greater for CER cement at 24 hours.

Analysis of radiopacity, pH and cytotoxicity of a new bioceramic material

AbstractObjective RetroMTA® is a new hydraulic bioceramic indicated for pulp capping, perforations or root resorption repair, apexification and apical surgery. The aim of this study was to compare the radiopacity, pH variation and cytotoxicity of this material to ProRoot® MTA.Material and Methods Mixed cements were exposed to a digital x-ray along with an aluminum stepwedge for the radiopacity assay. pH values were verified after incubation period of 3, 24, 48, 72 and 168 hours. The cytotoxicity of each cement was tested on human periodontal ligament fibroblasts using a multiparametric assay. Data analysis was performed using ANOVA and Tukey’spost hoc in GraphPad Prism.Results ProRoot® MTA had higher radiopacity than RetroMTA®(p<0.001). No significant differences were observed for the pH of the materials throughout experimental periods (p>0.05) although pH levels of both materials reduced over time. Both ProRoot® MTA and RetroMTA® allowed for significantly higher cell viability when compared with the positive control (p<0.001). No statistical difference was observed between ProRoot® MTA and RetroMTA® cytotoxicity level in all test parameters, except for the ProRoot® MTA 48-hour extract media in the NR assay (p<0.05).Conclusion The current study provides new data about the physicochemical and biological properties of Retro® MTA concerning radiopacity, pH and cytotoxic effects on human periodontal ligaments cells. Based on our findings, RetroMTA® meets the radiopacity requirements standardized by ANSI/ADA number 572, and similar pH values and biocompatibility to ProRoot® MTA. Further studies should be performed to evaluate additional properties of this new material.

Evaluation of cytotoxicity, antimicrobial activity and physicochemical properties of a calcium aluminate-based endodontic material

A calcium aluminate-based endodontic material, EndoBinder, has been developed in order to reduce MTA negative characteristics, preserving its biological properties and clinical applications. Objectives: The aim of this study was to evaluate the cytotoxicity, antimicrobial activity, pH, solubility and water sorption of EndoBinder and to compare them with those of white MTA (WMTA). Material and Methods: Cytotoxicity was assessed through a multiparametric analysis employing 3T3 cells. Antimicrobial activity against Enterococcus faecalis (ATCC 29212), Staphylococcus aureus. (ATCC 25923) and Candida albicans (ATCC 10556) was determined by the agar diffusion method. pH was measured at periods of 3, 24, 72 and 168 hours. Solubility and water sorption evaluation were performed following ISO requirements. Data were statistically analyzed by ANOVA and Tukey`s test with a significance level of 5%. Results: EndoBinder and WMTA were non-cytotoxic in all tested periods and with the different cell viability parameters. There was no statistical differences between both materials (P>.05). All tested materials were inhibitory by direct contact against all microbial strains tested. EndoBinder and WMTA presented alkaline pH in all tested times with higher values of pH for WMTA (P<.05). Both materials showed values complying with the solubility minimum requirements. However, EndoBinder showed lower solubility than WMTA (P<.05). No statistical differences were observed regarding water sorption (P>.05). Conclusion: Under these experimental conditions, we concluded that the calcium aluminate-based endodontic material EndoBinder demonstrated suitable biological and physicochemical properties, so it can be suggested as a material of choice in root resorption, perforations and root-end filling. .

Human tooth germ stem cell response to calcium-silicate based endodontic cements

OBJECTIVE: The aim of this study was to compare the cytotoxic effects of endodontic cements on human tooth germ stem cells (hTGSCs). MTA Fillapex, a mineral trioxide aggregate (MTA)-based, salicylate resin containing root canal sealer, was compared with iRoot SP, a bioceramic sealer, and AH Plus Jet, an epoxy resin-based root canal sealer. MATERIAL AND METHODS: To evaluate cytotoxicity, all materials were packed into Teflon rings (4 mmµ3 mm) and co-cultured with hTGSCs with the aid of 24-well Transwell permeable supports, which had a pore size of 0.4 µm. Coverslips were coated with MTA Fillapex, iRoot SP and AH Plus Jet and each coverslip was placed onto the bottom of one well of a six-well plate for scanning electron microscopy (SEM) analysis. Before the cytotoxicity and SEM analysis, all samples were stored at 37ºC and at 95% humidity and 5% CO2 for 24 hours to set. The cellular viability was analyzed using MTS test (3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium). The cytotoxic effects and SEM visualization of the tested materials were analyzed at 24-hour, 72-hour, one-week and two-week periods. RESULTS: On the 1st day, only MTA Fillapex caused cytotoxicity compared to negative control (NC) group (p<0.008). No significant difference was observed between the other tested materials at this period (p>0.05). After 14 days of incubation with the test materials, MTA Fillapex exhibited significantly higher cytotoxicity compared with iRoot SP, AH Plus Jet and the NC group (P<0.008). In the SEM analysis, the highest levels of cell attachment were observed for iRoot SP and the control group. After 24 hours, MTA Fillapex reduced the number of cells attached to the surface. CONCLUSIONS: Within the limitations of this study, sealers exerted different cytotoxic effects on hTGSCs. Although all materials ...

In Vitro Cytotoxicity of White MTA, MTA Fillapex® and Portland Cement on Human Periodontal Ligament Fibroblasts

The aim of this study was to compare the in vitro cytotoxicity of white mineral trioxide aggregate (MTA), MTA Fillapex® and Portland cement (PC) on human cultured periodontal ligament fibroblasts. Periodontal ligament fibroblast culture was established and the cells were used for cytotoxic tests after the fourth passage. Cell density was set at 1.25 X10 4 cells/well in 96-well plates. Endodontic material extracts were prepared by placing sealer/cement specimens (5X3mm) in 1mL of culture medium for 72 h. The extracts were then serially two-fold diluted and inserted into the cell-seeded wells for 24, 48 and 72 h. MTT assay was employed for analysis of cell viability. Cell supernatants were tested for nitric oxide using the Griess reagent system. MTA presented cytotoxic effect in undiluted extracts at 24 and 72 h. MTA Fillapex® presented the highest cytotoxic levels with important cell viability reduction for pure extracts and at ½ and » dilutions. In this study, PC did not induce alterations in fibroblast viability. Nitric oxide was detected in extract-treated cell supernatants and also in the extracts only, suggesting presence of nitrite in the soluble content of the tested materials. In the present study, MTA Fillapex displayed the highest cytotoxic effect on periodontal ligament fibroblasts followed by white MTA and PC.

Resumo O objetivo deste estudo foi comparar a citotoxicidade in vitro de agregado trióxido mineral (MTA) branco, MTA Fillapex® e cimento Portland (PC) em cultura de fibroblastos de ligamento periodontal humano. A cultura de fibroblastos de ligamento periodontal foi estabelecida e as células foram utilizadas para os testes citotóxicos após a quarta passagem. A densidade celular foi ajustada em 1,25X10 4 células/poço em placas de 96 poços. Extratos dos materiais endodônticos foram preparados por meio da inserção de corpos de prova dos cimentos (5 X 3 mm) em 1 mL de meio de cultura durante 72 h. Os extratos foram diluídos serialmente na razão de ½ e inseridos aos poços contendo as células por 24, 48 e 72 h. Ensaio de MTT foi realizado para a avaliação da viabilidade celular. O sobrenadante das células foi testado em relação à presença de óxido nítrico utilizando o sistema de reagentes de Griess. O MTA apresentou efeito citotóxico quando o extrato era aplicado sem diluição durante 24 e 72 h. O MTA Fillapex apresentou os maiores níveis de citotoxicidade com importante redução da viabilidade celular quando o extrato foi aplicado puro e em diluições de ½ e ». Neste estudo, PC não induziu alterações na viabilidade de fibroblastos. Óxido nítrico foi detectado no sobrenadante de células tratadas com os extratos e ainda nos extratos somente, o que sugere a presença de nitrito no conteúdo solúvel dos materiais testados. No presente estudo, MTA Fillapex foi o material que demonstrou o maior efeito citotóxico sobre fibroblastos de ligamento periodontal seguido do MTA branco e do PC. .

Cytotoxicity of root canal sealers on endothelial cell cultures

This study evaluated, in vitro, the cytotoxicity of six root canal sealers after 12, 24 and 72 h of contact time, using an endothelial ECV-304 cell line. The MTT assay was used for analysis of cell viability. Twelve specimens of each sealer were prepared and randomly assigned to 6 groups according to the commercial brands (n=4/time). A control group was also formed, which was not subjected to the contact with sealers. To assess the effects of sealers on endothelial cells, the specimens were placed in culture plate wells and incubated at 37°C with 5% CO2 and 100% humidity. MTT assays were performed in quadruplicate after 12, 24 and 72 h of contact of the sealer specimens with monolayers. Statistical analysis was performed by two-way ANOVA with Bonferroni post-hoc test at a significance level of 5%. Analysis of absorbance in the experimental groups showed that GuttaFlow presented the lowest cytotoxicity, with a mean absorbance of 0.048, followed by Pulp Canal Sealer (0.038), Sealer 26 (0.038), Endo Densell (0.036) and Pulp Fill (0.035). The control group had a mean absorbance of 0.098. Based on the results, Endofill and GuttaFlow were the most and the least cytotoxic sealers, respectively.

Este estudo avaliou, in vitro, a citotoxicidade de 6 cimentos endodônticos após 12, 24 e 72 h de tempo de contato, utilizando-se uma linhagem de células endoteliais ECV-304. Para a avaliação da viabilidade celular, utilizou-se o teste de citotoxicidade MTT. Para cada cimento foram preparados 12 corpos de prova que foram distribuídos em 6 grupos experimentais de acordo com as marcas comerciais, sendo 4 para cada tempo. Foi criado um grupo controle que não foi submetido à ação de cimento. Para avaliação do efeito dos cimentos sobre as células endoteliais, os corpos de prova foram inseridos nos poços da placa cultura, incubados a 37°C em presença de 5% de CO2 e 100% de umidade. Os testes MTT foram realizados em quadruplicata, após 12, 24 e 72 h de contato das amostras com o tapete celular. Foi utilizada a prova two-way Anova com o teste post hoc de Bonferroni com nível de significância de 5%. Quando analisadas as médias gerais de absorbância dos grupos analisados observou-se que o cimento GuttaFlow se apresentou como o cimento com menor índice de citotoxicidade, apresentando média de absorbência de 0,048. Logo após, apresentando médias de absorbância iguais (0,038) encontraram-se os cimentos Pulp Canal Sealer e Sealer 26; seguidos do Densell Endo e do Pulp Fill, com 0,036 e 0,035, respectivamente. O grupo controle apresentou média de absorbância de 0,098. Portanto, tendo como base os resultados obtidos, pôde-se concluir que o cimento Endofill foi o que apresentou maior citotoxicidade e o cimento GuttaFlow, o menos citotóxico.

Rat subcutaneous tissue response to MTA Fillapex® and Portland cement

The aim of this study was to evaluate the response of rat subcutaneous tissue to MTA Fillapex® (Angelus), an experimental root canal filling material based on Portland cement and propylene glycol (PCPG), and a zinc oxide, eugenol and iodoform (ZOEI) paste. These materials were placed in polyethylene tubes and implanted into the dorsal connective tissue of Wistar rats for 7 and 15 days. The specimens were stained with hematoxylin and eosin, and evaluated regarding inflammatory reaction parameters by optical microscopy. The intensity of inflammatory response against the sealers was analyzed by two blinded and previously calibrated examiners for all experimental periods (kappa=0.96). The histological evaluation showed that all materials caused a moderate inflammatory reaction at 7 days, which subsided with time. A greater inflammatory reaction was observed at 7 days in the tubes filled with ZOEI paste. Tubes filled with MTA Fillapex presented some giant cells, macrophages and lymphocytes after 7 days. At 15 days, the presence of fibroblasts and collagen fibers was observed indicating normal tissue healing. The tubes filled with PCPG showed similar results to those observed in MTA Fillapex. At 15 days, the inflammatory reaction was almost absent at the tissue, with several collagen fibers indicating normal tissue healing. Data were analyzed by the nonparametric Kruskal-Wallis test (α=0.05). Statistically significant difference (p<0.05) was found only between PCPG at 15 days and ZOEI at 7 days groups. No significant differences were observed among the other groups/periods (p>0.05). MTA Fillapex and Portland cement added with propylene glycol had greater tissue compatibility than the PCPG paste.

O objetivo deste estudo foi avaliar a resposta do tecido subcutâneo de rato ao MTA Fillapex® (Angelus), a um cimento endodôntico experimental à base de cimento Portland e propilenoglicol, e à pasta de óxido de zinco e eugenol com iodofórmio. Estes materiais foram colocados em tubos de polietileno e implantados no tecido conjuntivo do dorso de ratos Wistar, por 7 e 15 dias. Os espécimes foram corados com hematoxilina e eosina e os parâmetros de reação inflamatória foram avaliados em microscópio óptico. A intensidade da resposta inflamatória provocada pelos cimentos foi analisada em todos os períodos por dois observadores previamente calibrados (kappa 0,96) e sem conhecimento dos grupos experimentais. O exame histológico mostrou que todos os materiais provocaram reação inflamatória moderada aos 7 dias que regrediu com o tempo. A maior resposta inflamatória do tecido foi observada aos 7 dias, nos tubos preenchidos com pasta de Óxido de Zinco e Eugenol com Iodofórmio. Os tubos com MTA Fillapex apresentaram algumas células gigantes, macrófagos e linfócitos após 7 dias. Aos 15 dias, a presença de fibroblastos e fibras de colágenas foi observada, indicando processo de cicatrização do tecido. Os tubos com o cimento Portland mostraram resultados semelhantes aos observados no grupo MTA Fillapex. Aos 15 dias, a reação inflamatória apresentada foi praticamente ausente, com muitas fibras colágenas, indicando cicatrização normal do tecido. A análise estatística mostrou diferença estatisticamente significante entre o grupo de cimento Portland (15 dias) e óxido de zinco eugenol com Iodofórmio (7 dias) (p<0,05). Nos outros grupos não houve diferença estatística significante. MTA Fillapex e cimento Portland são mais biocompatíveis do que os outros cimentos testados.

Long-term cytotoxic effects of contemporary root canal sealers

Objectives: The aim of the present study was to investigate the effects of root canal sealers on the cytotoxicity of 3T3 fibroblasts during a period of 5 weeks. Material and Methods: Fibroblasts (3T3, 1×105 cells per well) were incubated with elutes of fresh specimens from eight root canal sealers (AH Plus, Epiphany, Endomethasone N, EndoREZ, MTA Fillapex, Pulp Canal Sealer EWT, RoekoSeal and Sealapex) and with elutes of the same specimens for 5 succeeding weeks after immersing in simulated body fluid. The cytotoxicity of all root canal sealers was determined using the MTT assay. Data were analyzed using ANOVA and Tukey's test. Results: RoekoSeal was the only sealer that did not show any cytotoxic effects (p<0.05). All the other tested sealers exhibited severe toxicity initially (week 0). MTA Fillapex remained moderately cytotoxic after the end of experimental period. Toxicity of the other tested sealers decreased gradually over time. The evaluated root canal sealers presented varying degrees of cytotoxicity, mainly in fresh mode.Conclusions: RoekoSeal had no cytotoxic effect both freshly mixed and in the other tested time points. MTA Fillapex was associated with significantly less cell viability when compared to the other tested root canal sealers.

Effect of time of extraction on the biocompatibility of endodontic sealers with primary human fibroblasts

The aim of this work was to evaluate the effects of different times of extraction on the cytotoxicity of six representatives of different root canal sealer groups-Real Seal SE, AH Plus, GuttaFlow, Sealapex, Roth 801, and ThermaSeal Plus-with human gingival fibroblasts. The materials were prepared according to manufacturers' specifications, and were incubated in culture medium (DMEM) at 37ºC for 1, 7, 14, 21, and 28 days, with daily washing, to simulate periodontal ligament clearance. Human fibroblasts were exposed to the final extracts at 24 hours, and cell viability was determined by MTT assay, with exposure to unconditioned DMEM as a negative control. Statistical analysis comparing cytotoxicities at each exposure time was performed by ANOVA with Scheffé adjustment for multiple comparisons at a 95% confidence level. Results indicated that GuttaFlow was significantly less cytotoxic than all other sealers (p < 0.05) at 1 day of extraction. After 7 days of extraction, cell viability for GuttaFlow was significantly increased as compared with that of all groups except sealer AH Plus. At day 14, cytotoxicity of Sealapex was significantly higher than that of all other sealers (p < 0.05). At days 21 and 28, there were no significant differences in cytotoxicity among sealer groups. All materials presented some level of cytotoxicity to fibroblasts, while GuttaFlow was the least cytotoxic sealer tested. However, the cytotoxicity of all materials seemed to decrease similarly in a time-dependent manner.

Estudo comparativo da citotoxicidade de cimentos obturadores sobre culturas de células endoteliais da linhagem ECV 304 / Comparative study of cytotoxicity of sealers on cultured endothelial cells line ECV 304

Este estudo teve como objetivo avaliar, in vitro, a citotoxicidade dos cimentos endodônticos Densell Endo®, Pulp-Fill®, Endofill®, Sealer 26®, Pulp Canal Sealer® e GuttaFlow® após 12, 24 e 72 horas de tempo de contato, utilizando-se uma linhagem de células endoteliais ECV-304. Para a avaliação da viabilidade celular, utilizou-se o teste de citotoxicidade MTT. Para cada cimento foram preparados 12 corpos de prova que foram distribuídos em seis grupos experimentais de acordo com as marcas comerciais, sendo quatro para cada tempo. Foi criado um grupo controle que não foi submetido à ação de cimento. Para avaliação do efeito dos cimentos sobre as células endoteliais, os corpos de prova foram inseridos nos poços da placa cultura, incubados a 37ºC em presença de 5% de CO2 e 100% de umidade. Os testes MTT foram realizados, em quadruplicata, após 12, 24 e 72 horas de contato das amostras com o tapete celular. Foi utilizada a prova Two-Way Anova com o teste Post Hoc de Bonferroni com nível de significância de 5%. Na análise de 12 horas, foi possível observar que o cimento GuttaFlow® apresentou média de absorbância de 0,055, seguido do Sealer 26® (média = 0,038). Os cimentos Pulp Canal Sealer® e Densell Endo® apresentaram a mesma média de absorbância (0,031). O Pulp Fill® e o Endofill® foram os cimentos que apresentaram maior citotoxicidade (média de absorbância = 0,024 e 0,021, respectivamente). O grupo controle apresentou média de absorbância de 0,158. Em 24 horas observou-se que os cimentos GuttaFlow® e Sealer 26® apresentaram as maiores médias de absorbância (0,041 e 0,037, respectivamente), seguidos pelo cimento Pulp Canal Sealer® que apresentou média de absorbância de 0,035. Já os cimentos Densell Endo® e Pulp Fill® apresentaram médias de absorbância de 0,033 e 0,032, respectivamente. O cimento Endofill® apresentou uma média de 0,026 e o grupo controle de 0,086. Quando analisados em 72 horas, o cimento Pulp Canal Sealer® obteve média de absorbância ...

This study aimed to evaluate, in vitro, cytotoxicity of root canal sealers Densell Endo®, Pulp-Fill®, Endofill®, Sealer 26®, Pulp Canal Sealer® and GuttaFlow® after 12, 24 and 72 hours of contact time, using a endothelial cell line ECV-304. For the assessment of cell viability, used the MTT cytotoxicity test. For any cement were prepared 12 specimens that were divided into six groups according to the trademarks, four for each time. Created a control group that was not subjected to action of cement. To assess the effects of cements on endothelial cells, the specimens were placed in culture plate wells, incubated at 37°C with 5% CO2 and 100% humidity. MTT tests were performed, in quadruplicate, after 12, 24 and 72 contact hours of the samples with monolayers. Was used to test Two-Way Anova with Bonferroni Post Hoc test with significance level of 5%. In the analysis of 12 hours, was observed that the cement GuttaFlow® had a mean absorbance of 0,055, followed by Sealer 26® (average = 0,038). Cements Pulp Canal Sealer® and Densell Endo® had the same mean absorbance (0,031). The Pulp Fill® and Enfofill® were the cements with higher cytotoxicity (mean absorbance = 0,024 and 0,021, respectively). The control group had a mean absorbance of 0,158. In 24 hours it was observed that the cements Sealer 26® and GuttaFlow® had the highest average absorbance (0,041 and 0,037, respectively), followed by cement Pulp Canal Sealer® that had a mean absorbance of 0.035. Already cements Densell Endo® and Pulp Fill® showed means absorbance of 0,033 and 0,032, respectively. Cement Endofill showed an average of 0,026 and the control group, 0,086. When analyzed at 72 hours, the cement Pulp Canal Sealer® had an average absorbance of 0,049, followed by cement GuttaFlow® and Pulp Fill®, both with 0,048. Cements Densell Endo®, Sealer 26® and Endofill® showed respectively, averaging 0,044, 0,040 and 0,036. The control group differed significantly ...(AU)

Estudo comparativo da citotoxicidade de cimentos obturadores sobre culturas de células endoteliais da linhagem ECV 304 / Comparative study of cytotoxicity of sealers on cultured endothelial cells line ECV 304

Este estudo teve como objetivo avaliar, in vitro, a citotoxicidade dos cimentos endodônticos Densell Endo®, Pulp-Fill®, Endofill®, Sealer 26®, Pulp Canal Sealer® e GuttaFlow® após 12, 24 e 72 horas de tempo de contato, utilizando-se uma linhagem de células endoteliais ECV-304. Para a avaliação da viabilidade celular, utilizou-se o teste de citotoxicidade MTT. Para cada cimento foram preparados 12 corpos de prova que foram distribuídos em seis grupos experimentais de acordo com as marcas comerciais, sendo quatro para cada tempo. Foi criado um grupo controle que não foi submetido à ação de cimento. Para avaliação do efeito dos cimentos sobre as células endoteliais, os corpos de prova foram inseridos nos poços da placa cultura, incubados a 37ºC em presença de 5% de CO2 e 100% de umidade. Os testes MTT foram realizados, em quadruplicata, após 12, 24 e 72 horas de contato das amostras com o tapete celular. Foi utilizada a prova Two-Way Anova com o teste Post Hoc de Bonferroni com nível de significância de 5%. Na análise de 12 horas, foi possível observar que o cimento GuttaFlow® apresentou média de absorbância de 0,055, seguido do Sealer 26® (média = 0,038). Os cimentos Pulp Canal Sealer® e Densell Endo® apresentaram a mesma média de absorbância (0,031). O Pulp Fill® e o Endofill® foram os cimentos que apresentaram maior citotoxicidade (média de absorbância = 0,024 e 0,021, respectivamente). O grupo controle apresentou média de absorbância de 0,158. Em 24 horas observou-se que os cimentos GuttaFlow® e Sealer 26® apresentaram as maiores médias de absorbância (0,041 e 0,037, respectivamente), seguidos pelo cimento Pulp Canal Sealer® que apresentou média de absorbância de 0,035. Já os cimentos Densell Endo® e Pulp Fill® apresentaram médias de absorbância de 0,033 e 0,032, respectivamente. O cimento Endofill® apresentou uma média de 0,026 e o grupo controle de 0,086. Quando analisados em 72 horas, o cimento Pulp Canal Sealer® obteve média de absorbância...

This study aimed to evaluate, in vitro, cytotoxicity of root canal sealers Densell Endo®, Pulp-Fill®, Endofill®, Sealer 26®, Pulp Canal Sealer® and GuttaFlow® after 12, 24 and 72 hours of contact time, using a endothelial cell line ECV-304. For the assessment of cell viability, used the MTT cytotoxicity test. For any cement were prepared 12 specimens that were divided into six groups according to the trademarks, four for each time. Created a control group that was not subjected to action of cement. To assess the effects of cements on endothelial cells, the specimens were placed in culture plate wells, incubated at 37°C with 5% CO2 and 100% humidity. MTT tests were performed, in quadruplicate, after 12, 24 and 72 contact hours of the samples with monolayers. Was used to test Two-Way Anova with Bonferroni Post Hoc test with significance level of 5%. In the analysis of 12 hours, was observed that the cement GuttaFlow® had a mean absorbance of 0,055, followed by Sealer 26® (average = 0,038). Cements Pulp Canal Sealer® and Densell Endo® had the same mean absorbance (0,031). The Pulp Fill® and Enfofill® were the cements with higher cytotoxicity (mean absorbance = 0,024 and 0,021, respectively). The control group had a mean absorbance of 0,158. In 24 hours it was observed that the cements Sealer 26® and GuttaFlow® had the highest average absorbance (0,041 and 0,037, respectively), followed by cement Pulp Canal Sealer® that had a mean absorbance of 0.035. Already cements Densell Endo® and Pulp Fill® showed means absorbance of 0,033 and 0,032, respectively. Cement Endofill showed an average of 0,026 and the control group, 0,086. When analyzed at 72 hours, the cement Pulp Canal Sealer® had an average absorbance of 0,049, followed by cement GuttaFlow® and Pulp Fill®, both with 0,048. Cements Densell Endo®, Sealer 26® and Endofill® showed respectively, averaging 0,044, 0,040 and 0,036. The control group differed significantly...

Biocompatibility of four root canal sealers: a histopathological evaluation in rat subcutaneous connective tissue

The aim of this study was to evaluate the subcutaneous biocompatibility of: Epiphany, AH Plus, Pulp Canal Sealer and Sealapex root canal sealers. Sixty rats were randomly assigned to 4 groups, according to the sealer. Polyethylene tubes containing the tested materials were inserted into the connective tissue. The implants were removed after 7, 15 and 30 days, and the tissue samples were processed, stained and examined by light microscopy. The descriptive analysis considered: thickness of the fibrous capsule, severity of the inflammatory reaction, and presence of giant cells. After 7 days, all sealers induced moderate to severe inflammatory reaction. After 15 days, Epiphany and AH Plus sealers showed a moderate inflammatory reaction, while Pulp Canal Sealer and Sealapex induced severe and mild inflammatory reactions, respectively. After 30 days, mild inflammatory reactions were observed for Epiphany, Sealapex and AH Plus. Sealapex induced the lowest inflammatory response at all evaluation periods, and only Pulp Canal Sealer did not show a decreased in the inflammatory reaction over time.

O objetivo deste estudo foi avaliar a biocompatibilidade subcutânea de cimentos endodônticos radiculares: Epiphany, AH Plus, Pulp Canal Sealer e Sealapex. Sessenta ratos foram divididos aleatoriamente em 4 grupos, de acordo com o cimento. Tubos de polietileno contendo os materiais testados foram inseridos no tecido conjuntivo. Os implantes foram removidos após 7, 15 e 30 dias, e as amostras de tecido foram processadas, coradas e examinadas por microscopia de luz. A análise descritiva considerou: espessura da cápsula fibrosa, severidade da reação inflamatória e presença de células gigantes. Após 7 dias, todos os cimentos induziram moderada e severa reação inflamatória. Após 15 dias, os cimentos Epiphany e AH Plus apresentaram uma reação inflamatória moderada, enquanto Pulp Canal Sealer e Sealapex induziram severa e leve reações inflamatórias, respectivamente. Após 30 dias, leve reação inflamatória foi observada para o Epiphany, Sealapex e AH Plus. Sealapex induziu a menor resposta inflamatória em todos os períodos de avaliação, e somente Pulp Canal Sealer não apresentou uma diminuição da reação inflamatória ao longo do tempo.

An endodontic sealer induces a pathological condition when associated with persistent tissue toxicity and presence of myofibroblasts

The aims of this study were to evaluate the ratio between inflammatory reactions induced by four endodontic sealers and the occurrence of fibrosis and the number of myofibroblasts with positivity to α-smooth-actin muscle (α-SMA). Polyethylene tubes were filled with a root canal sealer (Endofill, AH Plus, Acroseal and Epiphany) and inserted into 4 site at the dorsal region of 24 Wistar rats; 2 empty tubes (control) were grafted in 6 rats. After 7, 21, and 45 days, 8 animals were euthanized, providing 6 specimens per test group and 2 specimens from the control group. The fragments were subjected to histological processing and immunohistochemical analysis for anti α-SMA protein. All specimens, except those from the control group, presented severe inflammatory reaction on the 7th postoperative day, which also coincided with a large number of myofibroblasts. On the 21st and 45th days post-surgery, the inflammatory reaction induced by Endofill, AH Plus and Acroseal decreased significantly, which coincided with reduced presence of myofibroblasts and usual collagen deposition. In contrast, in the group filled with Epiphany, significant inflammatory cell infiltrate was present in all analyzed periods. The persistence of an inflammatory reaction induced by endodontic sealer may also induce the development of fibrosis in combination with presence of myofibroblasts.

O objetivo deste estudo foi avaliar a relação entre reação inflamatória induzida por quatro cimentos endodônticos e a presença de fibrose e quantidade de miofibroblastos que apresentam positividade para α-SMA. Tubos de polietileno foram preenchidos com o cimento (I: Endofill; II: AH Plus; III: Acroseal; IV: Epiphany) e inseridos em 4 regiões do dorso de 24 ratos Wistar, enquanto 2 tubos vazios (V - controle) foram inseridos em 6 ratos. Após 7, 21 e 45 dias, oito animais foram sacrificados obtendo 6 indivíduos por grupo e 2 para o grupo controle. Os fragmentos foram submetidos ao processamento histológico e à análise imuno-histoquímica para a proteína anti-α-SMA. Todos os grupos, exceto o controle, demonstraram notável reação inflamatória no 7º dia pós-operatório, que também coincidiu com uma grande quantidade de miofibroblastos. No 21º e 45º dia pós-operatório, a reação inflamatória induzida pelo Endofill, AH Plus e Acroseal diminuiu significativamente, o que coincidiu com reduzida presença de miofibroblastos e deposição de colágeno normal. Em contraste, no grupo Epiphany, infiltrado inflamatório significativo esteve presente em todos os períodos analisados. A persistência do infiltrado inflamatório induzido por cimento endodôntico pode também provocar uma fibrose associada com a presença de miofibroblastos.