Incorporating N-acetylcysteine and tricalcium phosphate into epoxy resin-based sealer improved its biocompatibility and adhesiveness to radicular dentine.

OBJECTIVE: This in vitro study was designed to evaluate the biocompatibility, adhesiveness, and antimicrobial activity of epoxy resin-based sealer associated with N-Acetylcysteine (NAC) or beta-tricalcium phosphate nanoparticles (ß-TCP) as an experimental retro-filling material. METHODS: Cytotoxicity was assessed using 2,3-Bis-(Methoxy-4-Nitro-5-Sulphophenyl)-2H-Tetrazolium-5-Carboxanilide (XTT) and Sulforhodamine B (SRB) assays after exposing human periodontal ligament fibroblasts to extracts of the materials for 1, 3, or 7 days. For the adhesive resistance test, root canals (48 single-root teeth) were instrumented with Reciproc #40 files (VDW GmbH, Germany) and obturated. After 7 days, the apices were sectioned and a retrograde cavity prepared and filled with the experimental materials (Mineral trioxide aggregate, Epoxy sealer, Epoxy sealer+NAC, and Epoxy sealer+ß-TCP). For the push-out test, one 2-mm thick slice was obtained from the apical third of each specimen. Antimicrobial activity was performed using agar diffusion method. Biofilms were grown in microplates and exposed to the extracts of retro-filled materials, followed by analysis of growth inhibition on agar plates. RESULTS: Epoxy sealer in association with ß-TCP or NAC showed better bond strength while Mineral trioxide aggregate allowed for the lowest adhesion. Mineral trioxide aggregate, Epoxy sealer+ß-TCP, and Epoxy sealer+NAC showed low cytotoxicity. Epoxy sealer was the most cytotoxic. In antimicrobial activity assays, all materials had no effect on Candida albicans. Addition of NAC improved the antimicrobial property of Epoxy sealer against Enterococcus faecalis compared to unmodified Epoxy sealer (P<0.05). SIGNIFICANCE: Incorporating ß-TCP or NAC with Epoxy sealer could improve the adhesiveness and biocompatibility for better use in endodontic therapy.

Effect of ethylenediaminetetraacetic acid and hyaluronic acid on the viability and cytokine expression of periodontal ligament fibroblasts.

BACKGROUND/AIMS: Dental root cell proliferation following tooth avulsion has not been well researched. Understanding the effects of dry time and dentin treatment influences on cell proliferation is essential to provide evidence-based guidelines for tooth replantation. The study evaluated the viability of periodontal ligament fibroblasts (PLF) in contact with roots, submitted to surface treatments with ethylenediaminetetraacetic acid (EDTA) and hyaluronic acid (HA) at different times, including to quantify inflammatory cytokines interleukin (IL)-6, IL-8, IL-1ß and TNF-α expressed by PLF. The adhesion of fibroblasts to treated root surfaces was also evaluated. MATERIAL AND METHODS: One hundred and eight cementum discs from bovine teeth were randomly divided into groups according to time periods of being dry (n = 12) as follows: (i) fresh discs that were not kept dry, (WD); (ii) dry for 1 hour (1 hd); and (iii) dry for 24 hours (24 hd). The discs were subdivided into 3 subgroups (n = 12) according to surface treatments: (iv) no treatment, (v) treatment with EDTA, (vi) treatment with HA. The discs were placed in 96-well plates, and PLF were seeded and kept in contact with the discs for 48 hours. Cell viability on the surface of the discs was assessed by XTT, and PLF adhesion was evaluated using scanning electron microscopy (SEM). Quantification of cytokines was performed using enzyme-linked immunosorbent assay (ELISA). Data were submitted to ANOVA and Tukey's test (α = .05). RESULTS: Surface treatment had a statistically significant effect on the cell viability in groups WD (P = .03), 1 hd (P = .01) and 24 hd (P = .048). PLF in contact with dried root surfaces expressed more cytokines without treatment with IL-6 decreasing the expression when treated with HA for 24 hours. SEM also showed adhesion of PLF to the surface of all discs at all time periods. CONCLUSION: EDTA + HA is an alternative treatment for cases of avulsed teeth as it promoted adhesion and increased viability of PLF.

Influência do ressecamento da superfície radicular, do tratamento com EDTA e ácido Hialurônico na viabilidade e expressão de citocinas inflamatórias de fibroblastos do ligamento periodontal / Influence of root surface dryness and treatment with EDTA and Hyaluronic acid on the viability and expression of inflammatory cytokines of periodontal ligament fibroblasts

O momento ideal para o reimplante de um dente avulsionado é imediatamente após a avulsão, no entanto, isso nem sempre é possível. Uma série de fatores influenciam na viabilidade das células do ligamento periodontal (PDL) contribuindo para acelerar ou minimizar a ocorrência da reabsorção radicular ou anquilose, consequências mais frequentes dos reimplantes. Um destes fatores é o período extra-oral e o tratamento da superfície radicular. O EDTA (ácido etilenodiamino tetracético) a 17% e o ácido hialurônico (AH) são utilizados para tratar a superfície radicular, visando menor ocorrência de reabsorção inflamatória e por substituição. Sendo assim, os objetivos deste estudo foram: a) avaliar a viabilidade de fibroblastos do ligamento periodontal (PDLF) em contato com discos radiculares submetidos ou não ao ressecamento de superfície por diferentes tempos e tratados com EDTA e/ou AH; b) quantificar as citocinas inflamatórias IL-6, IL-8, IL-1ß e TNF-α expressas por PDLF; c) observar a adesão de PDLF na superfície do discos radiculares tratados. Foram obtidos 108 discos de dentina e cemento da superfície radicular de dentes bovinos, com 4,5 mm de diâmetro, que foram previamente submetidos a dissolução do ligamento periodontal em solução de hipoclorito de sódio 1% por 15 min. Em seguida os discos foram esterilizados em concentrações decrescentes de álcool (100%,90%,80% e 70%). Os espécimes foram submetidos ou não ao ressecamento radicular por 1h ou 24h e tratados com EDTA associado ou não ao ácido hialurônico, colocados em placas de 96 poços onde foram semeadas células de culturas primárias de PDLF, que ficaram em contato com os discos por 48 h. A viabilidade celular na superfície dos discos foi avaliada através do ensaio de XTT. A microscopia eletrônica de varredura foi utilizada para verificar a adesão de PDLF à superfície dos discos. A detecção e quantificação das citocinas foi realizada pelo teste ELISA. Os dados foram submetidos à análise estatística pelo teste ANOVA e Tukey (p < 0,05). A maior média foi apresentada no grupo sem ressecamento para o tratamento EDTA+AH (148,39), que diferiu significativamente dos grupos controle e EDTA. No grupo ressecamento 1 h EDTA+AH (144,91) foi diferente dos demais. Para ressecamento 24h, verificou-se que o grupo EDTA+AH diferiu do grupo controle. Não houve modificações na expressão das citocinas IL-6, IL-8, IL-1ß e TNF-α quando foi acrescentado os tratamentos propostos. A IL-6 mostrou uma diminuição quando em contato com AH no periodo de 24h. Foi observada pelo MEV adesão de PDLF na superfície de todos os discos tratados e em todos os períodos analisados. Conclui-se que o ácido hialurônico é uma alternativa de tratamento para casos de dentes avulsionados já que mostrou seu papel promovendo adesão e aumento da viabilidade(AU)

The ideal time for reimplantation of an avulsed tooth is immediately after avulsion, however, this is not always possible. A number of factors influence the viability of the cells of the periodontal ligament (PDL) contributing to accelerate or minimize the occurrence of root resorption or ankylosis, more frequent consequences of reimplantation. One of these factors is the extra-oral period and root surface treatment. EDTA (17% ethylenediaminetetraacetic acid) and hyaluronic acid (HA) are used to treat the root surface, aiming for a lower occurrence of inflammatory resorption and replacement. Thus, the objectives of this study were: a) to evaluate the viability of periodontal ligament fibroblasts (PDLF) in contact with root disks submitted to surface dryness at different times and treated with EDTA and / or AH; b) quantify the inflammatory cytokines IL-6, IL-8, IL-1ß and TNF-α expressed by PDLF; c) observe the adhesion of PDLF on the surface of the treated root discs. 108 dentin and cementum disks were obtained from the root surface of bovine teeth, 4.5 mm in diameter, which were previously submitted to dissolution of the periodontal ligament in 1% sodium hypochlorite solution for 15 min. Then the disks were sterilized in decreasing concentrations of alcohol (100%, 90%, 80% and 70%). The specimens were submitted to root resection for 1 h or 24 h and treated with EDTA, whether or not associated with hyaluronic acid, placed in 96-well plates where cells from PDLF primary cultures were seeded and left in contact with the discs for 48 h. Cell viability at disc surfaces was assessed by the XTT assay. Scanning electron microscopy was used to verify the adhesion of PDLF to the disc surface. Detection and quantification of cytokines was performed by ELISA. Data were submitted to statistical analysis by the ANOVA and Tukey test (p <0.05). The highest mean was presented in the non-dry group for EDTA + AH treatment (148,39), which differed significantly from the control and EDTA groups. In the dryness group 1 h EDTA + AH (144.91) was different from the others. For dryness 24 h, it was found that the EDTA + AH group differed from the control group. There were no changes in the expression of cytokines IL-6, IL-8, IL-1ß and TNFα when the proposed treatments were added. IL-6 showed a decrease when in contact with HA in the 24-hour period. It was observed by the MEV adhesion of PDLF on the surface of all treated discs and in all periods analyzed. It is concluded that hyaluronic acid is an alternative treatment for cases of avulsed teeth since it showed its role promoting adhesion and increased viability(AU)

Influência do tempo de endurecimento no comportamento físico e biológico de sete cimentos endodônticos / Setting time of influence on the physical and biological behavior seven sealers

Objetivo: O objetivo deste estudo foi avaliar a influência do tempo de endurecimento sobre as propriedades físicas (teste de push-out) e biológicas (XTT, MNT) de sete cimentos endodônticos: Apexit Plus, Real Seal, Endo Rez, Roeko Seal, AH Plus, Endomethasone N e o cimento biocerâmico iRoot SP. Material e Métodos: Para a análise da citotoxicidade (XTT), foram utilizados fibroblastos do ligamento periodontal humano (PDLF). Para o número de micronúcleos (MNT), este estudo avaliou culturas de células expostas a diluições dos cimentos testados sobre células V79 No teste de push-out foram usados cento e quarenta dentes humanos unirradiculares que tiveram suas coroas removidas. Os canais radiculares foram preparados biomecanicamente com o sistema rotatório Mtwo (VDW GmbH, München, Alemanha) até o número de série 702 a 25 mm (25-40). Durante todo o preparo biomecânico, os canais foram irrigados com 2 ml de hipoclorito de sódio. Os espécimes foram seccionados em fatias de 2 mm nos terços cervical, médio e apical. O teste de Push-out foi realizado a uma velocidade de 1 mm / min e célula de carga de 50 Kgf. Os testes biológicos foram avaliadas após 24 h, 72 h, 1 semana, 1 mês, 3 meses, 6 meses e um ano após a manipulação e o teste de push-out 15 dias e 1 ano.Os dados foram analisados pelo teste de Kruskal-Wallis e teste de Dunn (p = 0,05). Resultados: IRooot SP, Roeko seal, Apexit Plus, AH Plus e Real Seal não mostraram citotoxicidade nas células PDLF. AH Plus, Real Seal, IRooot SP e Endomethasone N não apresentaram genotoxicidade. Roeko Seal e Apexit Plus mostraram ser genotóxico e EndoRez apresentou alta genotoxicidade. Roeko Seal, IRooot SP e EndoRez tiveram menor resistência à tração nos terços apical, médio e cervical, tanto em 15 dias e 1 ano de análise. Além disso, Endomethasone N, Real Seal e AH Plus apresentaram uma força de adesão satisfatória após 1 ano em todos os terços. Conclusão: Conclui-se que AH Plus e Real Seal foram os cimentos com menor ...

Objective: The aim of this study was to evaluate the influence of curing time on physical (push-out test) and biological properties (XTT, MNT) of seven sealers: Apexit Plus, Royal Seal, Endo Rez, Roeko Seal, AH Plus, Endomethasone, and bioceramic cement iRoot SP. Material and Methods: To the citotoxity analysis (XTT), fibroblasts of the human periodontal ligament were used (PDLF). To the number of micronuclei (MNT), this study evaluated cell cultures exposed to dilutions of the tested sealers, among them from causing excessive cytotoxic effects on V79 cells. To push-out test was used one hundred and forty single-rooted teeth which had the crowns removed. The root canals were prepared biomechanically with the rotary system Mtwo (VDW GmbH, München, Alemanha) until to series number 702 to file 25 mm (25-40). Throughout the biomechanical preparation, the canals were irrigated with 2 ml of sodium hypochlorite. The specimens were sectioned in slices of 2 mm in cervical, middle and apical. The push-out test was performed at speed of 1 mm/min and 50 kgf load cell. The tests were evaluated after 24 h, 72 h, 1 week, 1 month, 3 months, 6 months and one year after manipulation. Data were analyzed by Kruskal-Wallis and Dunn test (p = 0.05). Results: IRooot SP, Roeko Seal, Apexit Plus, AH plus and Real Seal showed no cytotoxicity in PDLF cells. AH plus, Real Seal, IRooot SP and Endomethasone N showed no genotoxicity. Roeko Seal and Apexit Plus have shown to be genotoxic and EndoRez showed high genotoxicity. Roeko Seal, IRooot SP and EndoRez had lower bond strength in the apical, middle and cervical thirds in both the 15-day and 1 year analysis. Apexit Plus, Endomethasone N, Real Seal and AH plus had a satisfactory bond strength after 1 year in all thirds. Conclusion: It is conclude that AH plus and Real Seal were the sealer with lower cytotoxicity and genotoxicity, with good values of bond strength in all third, keeping as the material of choice for endodontic therapy