Dose- and time-dependent effects of taxifolin on viability and mineralization markers of osteoblast-like cells.

The current study evaluated the effects of taxifolin treatments on the viability of osteoblast-like cells, and on the expression of early mineralization markers, as part of the ongoing search for new endodontic materials able to induce periapical healing without causing cytotoxicity. Saos-2 osteoblast-like cells were exposed to different concentrations of taxifolin (5 and 10 µM), applied as pretreatments either for 24h and 72h, or continuously throughout the experimental protocol. Cell viability using the methylthiazole tetrazolium (MTT) assay, alkaline phosphatase activity using thymolphthalein monophosphate assays, deposition of mineralized nodules using alizarin red staining, and expression of ALP and COL-1 by qPCR were determined after 6 and 13 days of treatment. The data were analyzed statistically (p<0.05). Taxifolin was not cytotoxic in the concentrations tested. Pretreatments with taxifolin for 24h and 72h at 10 µM stimulated ALP activity, and increased mineralized nodule deposition by Saos-2 cells. Continuous treatment with taxifolin was not effective in stimulating ALP activity and mineralization. ALP and COL-1 gene expression increased with taxifolin pretreatments, since the highest mRNA levels were observed after 72h of pretreatment with taxifolin at 10 µM on day 13. In conclusion, taxifolin was cytocompatible, and induced mineralization markers when applied for short periods in osteoblast-like cell culture.

Inhibitory activity of Varronia curassavica and Mikania laevigatafractions against pathogens associated with persistent dental infections

Herbal medicines have been studied as potential antimicrobial agents, emerging as treatments against oral diseases. The current study evaluated the antimicrobial activity of the crude extract and fractions of Mikania laevigata SchultzBip. ex Baker (ML) and Varronia curassavicaJacq (VC) against oral pathogens associated with persistent dental root infections, under planktonic and biofilm conditions. Minimal inhibitory concentrations and minimal bactericidal/fungicidal concentrations were determined for the ML and VC fractions/extracts against Enterococcus faecalis, Actinomyces israelii, Pseudomonas aeruginosa, and Candida albicans using the microdilution method. The best results were chosen for subsequent biofilm assays. All tested ML and VC extracts/fractions demonstrated inhibitory activity against E. faecalis and A. israelii. The ML ethyl acetate fraction affected the growth of all microorganisms tested. C. albicans and P. aeruginosawere not affected by any VC extract/fractions. The ML ethyl acetate fraction eliminated E. faecalis, A.israelii, and P. aeruginosa biofilms after 24h. A similar result was observed for ML crude hydroethanolic extract and its hexane fraction for A. israelii. The VC hexane fraction was able to eliminate A. israelli biofilms. None of the tested extracts or fractions eliminated C. albicans biofilm. The Mikania laevigata ethyl acetate fraction is an efficient antimicrobial agent against oral pathogens and could be indicated for the treatment of persistent dental infections.

Cytotoxicity and antimicrobial effects of citronella oil (Cymbopogon nardus) and commercial mouthwashes on S. aureus and C. albicans biofilms in prosthetic materials.

Although the effectiveness of some mouthwashes has been proven, phytotherapy is still a field to be explored as an alternative to commercial products. OBJECTIVE: To evaluate, in vitro, the cytotoxicity and efficacy of two solutions based on citronella oil (CN), on S. aureus and C. albicans biofilms (in formation-adhesion phase and 24 h-biofilm formation) on acrylic resin and nickel-chromium alloy samples (one trademark of each material), compared to two alcohol-free commercial mouthwashes. MATERIAL AND METHODS: Two solutions containing CN at concentrations of 5x and 10x the minimum bactericidal/fungicidal concentration (MBC/MFC) were prepared by microdilution. After contamination of the samples surfaces with these microorganisms, the mouthwashes (CN - 5x and 10x; CHX - 0,12% alcohol-free chlorhexidine and LT - alcohol-free essential oils) were evaluated. Mouthwash simulation was performed for 1 min at two moments, the first simulation after 4 h of microbial adhesion and 24 h-biofilm formation, and the second simulation, 6 h after the first simulation. For biofilm quantification, the number of cultured cells was evaluated by CFUs. The cytotoxicity assay was performed on HaCat epithelial cells and quantified by the MTT method. RESULTS: Tested solutions completely inhibited the growth of both microorganisms in the adhesion phase. All solutions showed inhibitory activity against 24 h-biofilm formation. However, CN led to greater microbial reduction, regardless of the surface of the sample. All solutions demonstrated a toxic effect. However, after serial dilution, CN presented the lowest cytotoxic effect. CONCLUSION: Citronella had a lower cytotoxic effect and a higher action compared to commercial solutions.

Development and characterization of p1025-loaded bioadhesive liquid-crystalline system for the prevention of Streptococcus mutans biofilms.

Formation of a dental biofilm by Streptococcus mutans can cause dental caries, and remains a costly health problem worldwide. Recently, there has been a growing interest in the use of peptidic drugs, such as peptide p1025, analogous to the fragments 1025-1044 of S. mutans cellular adhesin, responsible for the adhesion and formation of dental biofilm. However, peptides have physicochemical characteristics that may affect their biological action, limiting their clinical performance. Therefore, drug-delivery systems, such as a bioadhesive liquid-crystalline system (LCS), may be attractive strategies for peptide delivery. Potentiation of the action of LCS can be achieved with the use of bioadhesive polymers to prolong their residence on the teeth. In line with this, three formulations - polyoxypropylene-(5)-polyoxyethylene-(20)-cetyl alcohol, oleic acid, and Carbopol C974P in different combinations (F1C, F2C, and F3C) were developed to observe the influence of water in the LCS, with the aim of achieving in situ gelling in the oral environment. These formulations were assessed by polarized light microscopy, small-angle X-ray scattering, rheological analysis, and in vitro bioadhesion analysis. Then, p1025 and a control (chlorhexidine) were incorporated into the aqueous phase of the formulation (F + p1025 and F + chlorhexidine), to determine their antibiofilm effect and toxicity on epithelial cells. Polarized light microscopy and small-angle X-ray scattering showed that F1C and F2C were LCS, whereas F3C was a microemulsion. F1C and F2C showed pseudoplastic behavior and F3C Newtonian behavior. F1C showed the highest elastic and bioadhesive characteristics compared to other formulations. Antibiofilm effects were observed for F + p1025 when applied in the surface-bound salivary phase. The p1025-loaded nanostructured LCS presented limited cytotoxicity and effectively reduced S. mutans biofilm formation, and could be a promising p1025-delivery strategy to prevent the formation of S. mutans dental biofilm.

Efeito de flavonoides sobre microrganismos de interesse endodôntico e sua influência na expressão de marcadores de mineralização / Effect of flavonoids on endodontic microorganisms and their influence on the expression of mineralization markers

O tratamento endodôntico de dentes permanentes jovens representa um grande desafio clínico devido a presença de paredes dentinárias finas, divergentes ou paralelas e ápice aberto, o que dificulta a desinfecção e a execução dos procedimentos convencionais. Terapias de regeneração endodôntica envolvem o uso de materiais capazes de promover uma desinfecção eficaz sem causar citotoxicidade, além de induzir a diferenciação de células-tronco ou bioestimular células remanescentes do tecido pulpar mesmo após a injúria. Nesse contexto, os flavonoides, polifenóis presentes em frutas e vegetais, poderiam ser agentes interessantes para o tratamento endodôntico de dentes imaturos devido a sua amplitude terapêutica. Dessa forma, o objetivo do presente trabalho foi avaliar o efeito antimicrobiano, citotóxico e indutor de mineralização de flavonoides com finalidade de aplicação endodôntica. Este trabalho de tese foi dividido em três capítulos. O capítulo 1 avaliou a toxicidade dos flavonoides taxifolina, crisina, pinocembrina e galangina sobre fibroblastos pelo ensaio de MTT, a atividade antimicrobiana pela determinação da concentração inibitória e bactericida mínima, e a ação antibiofilme do flavonoide com melhor efeito antimicrobiano, por meio de ensaios em placas de poliestireno e em dentina radicular bovina por meio da análise por microscopia confocal. Os resultados mostraram que o flavonoide taxifolina não foi tóxico para os fibroblastos em nenhuma das concentrações analisadas, enquanto que os flavonoides crisina, pinocembrina e galangina apresentaram efeitos citotóxicos. Crisina, pinocembrina e galangina não apresentaram efeito antimicrobiano frente E. faecalis and S. mutans nas concentrações testadas. A taxifolina foi capaz de inibir todas as bactérias testadas, eliminar biofilmes de E. faecalis e S. mutans em placas de poliestireno e reduzir significantemente o biofilme de E. faecalis em túbulos dentinários. O capítulo 2 avaliou a citotoxicidade do flavonoide taxifolina e o seu potencial sobre a indução de marcadores de mineralização dentinária (produção de fosfatase alcalina - ALP, nódulos de mineralização ­ NM e expressão dos genes DSPP ­ sialofosfoproteína dentinária e DMP-1 ­ proteína da matriz dentinária - 1) em células semelhantes a odontoblastos MDPC-23, após tratamentos de 24, 72h e contínuo. A taxifolina não apresentou citotoxicidade em nenhum dos três tipos de tratamento analisados. Todas as concentrações do tratamento de 24h e as concentrações de 10 e 5µM do tratamento de 72h aumentaram a atividade de ALP. A formação de NM aumentou com os tratamentos de taxifolina à 10µM em ambos os tratamentos de 24 e 72h, e à 5µM no tratamento de 24h. A expressão de DMP-1 aumentou com o tratamento de taxifolina em ambos os tratamentos de 24 e 72h, enquanto que a de DSPP aumentou apenas com o tratamento de 72h na concentração de 5µM. O capítulo 3 avaliou a citotoxicidade da taxifolina, e o seu potencial sobre a indução de marcadores de mineralização óssea (ALP, NM e expressão dos genes ALP e colágeno 1 - Col-1) em células semelhantes a osteoblastos Saos-2, após tratamentos de 24, 72h e contínuo. Os resultados mostraram que os tratamentos com taxifolina nas concentrações de 10, 5 e 1µM não foram citotóxicos em nenhum dos períodos analisados. O tratamento de 72h da taxifolina à 10µM foi capaz de aumentar a atividade de ALP e a formação de NM, além de aumentar a expressão de Col-1 após 13 dias. O tratamento de 24h da taxifolina na concentração de 10µM aumentou a expressão de ALP após 6 dias. Conclui-se que a taxifolina é um flavonoide com potencial uso para o tratamento endodôntico de dentes permanentes jovens, devido à sua ação antimicrobiana/antibiofilme, baixa citotoxicidade e capacidade de estimular a mineralização em odontoblastos e osteoblastos(AU)

The endodontic treatment of young permanent teeth represents a great clinical challenge due to the presence of thin, divergent or parallel dentin walls and the open apex that makes it difficult to disinfect and perform conventional endodontic procedures. Endodontic regeneration therapies involve the use of materials capable of promoting effective disinfection without causing cytotoxicity, in addition to inducing differentiation of stem cells or biostimulating remaining pulp tissue cells even after injury. In this context, the flavonoids, polyphenols present in fruits and vegetables, could be interesting agents for the endodontic treatment of immature teeth due to the wide therapeutic use. Thus, the objective of the present study was to evaluate the antimicrobial, cytotoxic effects and capacity of mineralization induction of flavonoids for endodontic application. This thesis was divided into three chapters. The chapter 1 evaluated the toxicity of taxifolin, chrysin, pinocembrin and galangin flavonoids on fibroblasts by the MTT method, antimicrobial activity by determining the minimum inhibitory and bactericidal concentrations and analyzed the antibiofilm action of the flavonoid with the best antimicrobial effect, by means of the biofilm assays in polystyrene plates and in bovine root dentin and confocal microscopy analysis. The results showed that the flavonoid taxifolin was not toxic on fibroblasts in any tested concentration, while chrysin, pinocembrin and galangin flavonoids showed cytotoxic effects. Chrysin, pinocembrin and galangin showed no antimicrobial effect against E. faecalis and S. mutans in any tested concentrations. Taxifolin was able to inhibit all tested bacteria, to eliminate E. faecalis and S. mutans biofilms on polystyrene plates and significantly reduce E. faecalis biofilms from the dentin tubules. The chapter 2 evaluated the cytotoxicity of taxifolin and its potential on the induction of dentin mineralization markers (alkaline phosphatase production - ALP, mineralization nodules - MN and expression of genes DSPP - dentin sialophosphoprotein and DMP-1 - dentin matrix protein - 1) on odontoblast-like cells MDPC-23, after treatments of 24, 72h and continuous. Taxifolin did not present cytotoxicity at any of the three types of treatments analyzed. All concentrations of 24h-treatment and 10 and 5µM of 72htreatment increased ALP activity. NM formation increased with taxifolin treatments at 10µM in both 24 e 72h treatments, and at 5µM in the 24h-treatment. Expression of DMP-1 increased with taxifolin in both 24 e 72h-treatments, whereas DSPP expression increased only with 72h-treatment at 5µM. The chapter 3 evaluated the cytotoxicity of taxifolin and its potential on the induction of bone mineralization markers (ALP, NM and expression of ALP and collagen 1 -Col-1 genes) on Saos-2 osteoblast-like cells, after treatments of 24, 72h and continuous. The results showed that taxifolin treatments at 10, 5 and 1µM were not cytotoxic in any of the periods analyzed. The 72h-treatment of taxifolin at 10µM was able to increase ALP activity and NM formation, in addition to increasing Col-1 expression after 13 days. The 24h-treatment of taxifolin at 10µM increased ALP expression after 6 days. It is concluded that taxifolin is a flavonoid with potential use for endodontic treatment of young permanent teeth due to its antimicrobial/antibiofilm action, low cytotoxicity and ability to stimulate mineralization in odontoblasts and osteoblasts(AU)

Evaluation of the Cytotoxicity and Biocompatibility of New Resin Epoxy-based Endodontic Sealer Containing Calcium Hydroxide.

INTRODUCTION: Many endodontic sealers are available, but the search for the ideal sealer continues. This study evaluated the cytotoxicity and biocompatibility of Sealer Plus, a new resin epoxy-based endodontic sealer containing calcium hydroxide. AH Plus, Endofill, and SimpliSeal endodontics sealers were used for comparison. METHODS: L929 fibroblasts were cultured, and an MTT assay was used to determine the cytotoxicity of the sealer extracts at 6, 24, 48, and 72 hours. Tubes containing materials or empty tubes for control were inserted into the subcutaneous tissues of 20 rats. After 7 and 30 days, the rats were killed, and the tubes were removed with the surrounding tissues for histologic analysis. The data were submitted to statistical tests (P < .05). RESULTS: Undiluted Sealer Plus exhibited less cytotoxicity compared with other undiluted extracts at 6 hours (P < .05), and cell viability was higher for all Sealer Plus extracts after 24 hours (P < .05). At 48 hours, the undiluted and ½ Sealer Plus dilution were the extracts with less cytotoxicity (P < .05). At 72 hours, cell viability was higher for the undiluted and ½ Sealer Plus dilution compared with the other sealers (P < .05). At 7 days, Endofill and SimpliSeal had higher inflammation compared with the control and Sealer Plus (P < .05); AH Plus had moderate inflammation (P > .05). At 30 days, control, Sealer Plus, and AH Plus had less inflammation (P < .05). The fibrous capsule was thick at 7 days and thin at 30 days, except for SimpliSeal. CONCLUSIONS: In general, Sealer Plus promoted greater cell viability and was more biocompatible compared with the other sealers.

KR-12-a5 is a non-cytotoxic agent with potent antimicrobial effects against oral pathogens.

This study evaluated the cytotoxicity and antimicrobial activity of analogs of cationic peptides against microorganisms associated with endodontic infections. L-929 fibroblasts were exposed to LL-37, KR-12-a5 and hBD-3-1CV and chlorhexidine (CHX, control), and cell metabolism was evaluated with MTT. The minimal inhibitory concentration (MIC) and the minimal bactericidal/fungicidal concentration (MBC/MFC) of the peptides and CHX were determined against oral pathogens associated with endodontic infections. Enterococcus faecalis and Streptococcus mutans biofilms were cultivated in bovine dentin blocks, exposed to different concentrations of the most efficient antimicrobial peptide and analyzed by confocal laser scanning microscopy. CHX and peptides affected the metabolism of L-929 at concentrations > 31.25 and 500 µg ml-1, respectively. Among the peptides, KR-12-a5 inhibited growth of both the microorganisms tested with the lowest MIC/MBC/MFC values. In addition, KR-12-a5 significantly reduced E. faecalis and S. mutans biofilms inside dentin tubules. In conclusion, KR-12-a5 is a non-cytotoxic agent with potent antimicrobial and anti-biofilm activity against oral pathogens associated with endodontic infections.

Antimicrobial Activity and Biocompatibility of the Psidium cattleianum Extracts for Endodontic Purposes

Abstract Psidium cattleianum (PC) has been displaying inhibitory effect against a variety of microorganisms, but this effect has not yet been tested against endodontic pathogens. The aim of this study was to evaluate the antimicrobial activity and biocompatibility of the aqueous (PCAE) and hydroethanolic (PCHE) extracts from Psidium cattleianum (PC) leaves. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were determined using the microdilution broth method in order to analyze the antimicrobial effect against Enterococcus faecalis, Pseudomonas aeruginosa, Actinomyces israelii and Candida albicans in planktonic conditions. Biofilm assays were conducted only with the extracts that were able to determine the MLC for microorganisms in planktonic conditions. Immediate and late tissue reactions against PC extracts were evaluated using edemogenic test and histological analysis of subcutaneous implants in Wistar rats. The results showed that the MIC and MLC values ranged between 0.25 and 4 mg/mL. The MLC obtained for PCHE inhibited 100% growth of all the tested strains, except for C. albicans. PCAE had the same effect for E. faecalis and P. aeruginosa. Both PC extracts were able to eliminate E. faecalis biofilms and only the PCHE eliminated P. aeruginosa biofilms. The positive controls inhibited the growth of all tested strains in MIC and MLC essays, but no CHX tested concentrations were able to eliminate A. israelii biofilm. PCAE caused a discrete increase in the edema over time, while PCHE caused a higher initial edema, which decreased progressively. Both PCAE and PCHE extracts were biocompatible, but PCHE showed better results with slight levels of inflammation at 28 days. In conclusion, PCHE was biocompatible and presented better antimicrobial effect against important pathogens associated with persistent endodontic infections

Resumo Psidium cattleianum (PC) tem apresentado atividade inibitória frente diversos microrganismos, entretanto esse efeito ainda não foi testado contra microrganismos de interesse endodôntico. O objetivo desse estudo foi avaliar a atividade antimicrobiana e a biocompatibilidade dos extratos aquoso (EAPC) e hidroetanólico (EHPC) das folhas de Psidium cattleianum. As concentrações inibitória mínima (CIM) e letal mínima (CLM) foram determinadas pelo método de microdiluição em caldo, com o objetivo de analisar o efeito antimicrobiano frente Enterococcus faecalis, Pseudomonas aeruginosa, Actinomyces israelii e Candida albicans em condições planctônicas. Os ensaios de biofilme foram realizados somente com os extratos em que se determinou a CLM frente os microrganismos em condições planctônicas. Respostas teciduais imediata e tardia frente aos extratos de Psidium cattleianum foram avaliadas por teste edemogênico e análise histológica de implantes subcutâneos em ratos Wistar. Os resultados mostraram que CIM e CLM variaram entre 0,25 e 4 mg/mL. As CLMs determinadas pelo EHPC inibiram 100% do crescimento de todas as cepas testadas, exceto Candida albicans. EAPC apresentou o mesmo efeito para E. faecalis e P. aeruginosa. Ambos os extratos de PC conseguiram eliminar o biofilme de E. faecalis, e somente o EHPC eliminou o biofilme de P. aeruginosa. Os controles positivos inibiram o crescimento de todos os microrganismos testados nos ensaios de CIM e CLM, mas nenhuma das concentrações de clorexidina testadas foi capaz de eliminar o biofilme de A. israelii. O EAPC provocou um discreto aumento de edema com o tempo, enquanto EHPC provocou um edema inicial severo, que diminuiu progressivamente. Ambos os extratos EAPC e EHPC foram biocompatíveis, entretanto, EHPC apresentou melhores resultados com baixos níveis de inflamação em 28 dias. Pode-se concluir que EHPC foi biocompatível e apresentou melhor efeito antimicrobiano frente importantes patógenos associados a infecções endodônticas persistentes.

Antimicrobial Activity and Biocompatibility of the Psidium cattleianum Extracts for Endodontic Purposes.

Psidium cattleianum (PC) has been displaying inhibitory effect against a variety of microorganisms, but this effect has not yet been tested against endodontic pathogens. The aim of this study was to evaluate the antimicrobial activity and biocompatibility of the aqueous (PCAE) and hydroethanolic (PCHE) extracts from Psidium cattleianum (PC) leaves. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were determined using the microdilution broth method in order to analyze the antimicrobial effect against Enterococcus faecalis, Pseudomonas aeruginosa, Actinomyces israelii and Candida albicans in planktonic conditions. Biofilm assays were conducted only with the extracts that were able to determine the MLC for microorganisms in planktonic conditions. Immediate and late tissue reactions against PC extracts were evaluated using edemogenic test and histological analysis of subcutaneous implants in Wistar rats. The results showed that the MIC and MLC values ranged between 0.25 and 4 mg/mL. The MLC obtained for PCHE inhibited 100% growth of all the tested strains, except for C. albicans. PCAE had the same effect for E. faecalis and P. aeruginosa. Both PC extracts were able to eliminate E. faecalis biofilms and only the PCHE eliminated P. aeruginosa biofilms. The positive controls inhibited the growth of all tested strains in MIC and MLC essays, but no CHX tested concentrations were able to eliminate A. israelii biofilm. PCAE caused a discrete increase in the edema over time, while PCHE caused a higher initial edema, which decreased progressively. Both PCAE and PCHE extracts were biocompatible, but PCHE showed better results with slight levels of inflammation at 28 days. In conclusion, PCHE was biocompatible and presented better antimicrobial effect against important pathogens associated with persistent endodontic infections.

Cytotoxicity and the effect of cationic peptide fragments against cariogenic bacteria under planktonic and biofilm conditions.

This study evaluated the cytotoxicity and effect of fragments derived from three oral cationic peptides (CP): LL-37, D6-17 and D1-23 against cariogenic bacteria under planktonic and biofilm conditions. For cytotoxicity analysis, two epithelial cell lines were used. The minimum inhibitory concentration and the minimal bactericidal concentration were determined for the CP fragments and the control (chlorhexidine-CHX) against cariogenic bacteria. The fractional inhibitory concentration was obtained for the combinations of CP fragments on Streptococcus mutans. Biofilm assays were conducted with the best antimicrobial CP fragment against S. mutans. The results indicated that D6-17 was not cytotoxic. D1-23, LL-37 and CHX were not cytotoxic in low concentrations. D1-23 presented the best bactericidal activity against S. mutans, S. mitis and S. salivarius. Combinations of CP fragments did not show a synergic effect. D1-23 presented a higher activity against S. mutans biofilm than CHX. It was concluded that D1-23 showed a substantial effect against cariogenic bacteria and low cytotoxicity.

Antimicrobial action of calcium hydroxide-based endodontic sealers after setting, against E. faecalis biofilm.

Enterococcus faecalis are gram positive bacteria that can mostly resist endodontic therapy, inducing persistent infection in the root canal system. Endodontic sealers with antimicrobial activity may help eliminate residual microorganisms that survive endodontic treatment. The present study aimed at comparing the antimicrobial activity of Acroseal, Sealapex and AH Plus endodontic sealers in an in vitro biofilm model. Bovine dentin specimens (144) were prepared, and twelve blocks for each sealer and each experimental time point (2, 7 and 14 days) were placed and left in contact with plates containing inoculum of E. faecalis (ATCC 51299), to induce biofilm formation. After 14 days, the samples were transferred to another plate with test sealers and kept at 37°C and 5% CO2 for 2, 7 and 14 days. The specimens without sealers were used as a control for each period. The samples were agitated in a sonicator after each experiment. The suspensions were agitated in a vortex mixer, serially diluted in saline, and triple plated onto m-Enterococcus agar. Colonyforming units were counted, and the data were statistically analyzed using ANOVA, Shapiro-Wilk and Kruskal-Wallis one-way tests (p < 0.05) to determine antimicrobial potential. Sealapex showed significant differences at all the experimental time points, in comparison with all the other groups. AH Plus and Acroseal showed antimicrobial activity only on the 14th experimental day. Neither of the sealers tested were able to completely eliminate the biofilm. Sealapex showed the highest antimicrobial activity in all the experimental periods. The antimicrobial activity of all the sealers analyzed increased over time.

Antimicrobial action of calcium hydroxide-based endodontic sealers after setting, against E. faecalis biofilm

Abstract Enterococcus faecalis are gram positive bacteria that can mostly resist endodontic therapy, inducing persistent infection in the root canal system. Endodontic sealers with antimicrobial activity may help eliminate residual microorganisms that survive endodontic treatment. The present study aimed at comparing the antimicrobial activity of Acroseal, Sealapex and AH Plus endodontic sealers in an in vitro biofilm model. Bovine dentin specimens (144) were prepared, and twelve blocks for each sealer and each experimental time point (2, 7 and 14 days) were placed and left in contact with plates containing inoculum of E. faecalis (ATCC 51299), to induce biofilm formation. After 14 days, the samples were transferred to another plate with test sealers and kept at 37°C and 5% CO2 for 2, 7 and 14 days. The specimens without sealers were used as a control for each period. The samples were agitated in a sonicator after each experiment. The suspensions were agitated in a vortex mixer, serially diluted in saline, and triple plated onto m-Enterococcus agar. Colonyforming units were counted, and the data were statistically analyzed using ANOVA, Shapiro-Wilk and Kruskal-Wallis one-way tests (p < 0.05) to determine antimicrobial potential. Sealapex showed significant differences at all the experimental time points, in comparison with all the other groups. AH Plus and Acroseal showed antimicrobial activity only on the 14th experimental day. Neither of the sealers tested were able to completely eliminate the biofilm. Sealapex showed the highest antimicrobial activity in all the experimental periods. The antimicrobial activity of all the sealers analyzed increased over time.

Análise da atividade antimicrobiana dos extratos de Cordia verbenacea DC, Mikania laevigata Sch. Bip. ex Baker e Psidium Cattleianum frente a microrganismos endodônticos / Antimicrobial analysis of the Cordia verbenacea DC, Mikania laevigata Sch. Bip. ex Baker e Psidium Cattleianum plant extracts against endodontic microorganisms

Na busca por fontes alternativas de antimicrobianos, diversos autores tem explorado o uso de plantas medicinais para o tratamento de várias doenças. A atividade antimicrobiana de várias espécies de plantas como, Cordia verbenacea DC, Mikania laevigata Schultz Bip. Ex Baker e Psidium cattleianum, popularmente chamadas de erva-baleeira, guaco e de araçá, respectivamente, tem sido relatada contra diversos microrganismos orais. O objetivo do presente estudo foi avaliar o efeito antimicrobiano dos extratos hidroetanólico bruto e as suas respectivas frações aquosa, butanol, hexano e acetato de etila de Cordia verbenacea (CV) e Mikania laevigata (ML) e extratos aquoso e hidroetanólico de Psidium cattleianum (PC) contra Enterococcus faecalis, Pseudomonas aeruginosa, Actinomyces israelii e Candida albicans, sob condições planctônicas e de biofilme. Os microrganismos foram analisados em condições planctônicas por meio de ensaios para a determinação da concentração inibitória mínima (CIM) e concentração letal mínima (CLM), pelo método de microdiluição em caldo. Posteriormente, foram selecionados somente os extratos capazes de eliminar completamente cada microrganismo (CLM 100%), para a análise da atividade antimicrobiana em ensaios de biofilme. Os resultados mostraram que os valores de CIM e CLM variaram de 250 à 4000μg/ml. O biofilme formado por E. faecalis foi eliminado frente à ação dos extratos aquoso e hidroetanólico de PC e à fração acetato de etila de ML. A mesma fração de ML e o extrato hidroetanólico de PC também foram capazes de eliminar biofilme de P. aeruginosa. O mesmo ocorreu com as frações hexânicas de CV e ML e o extrato hidroetanólico de ML frente ao biofilme de A. israelii. Nenhum extrato/fração foi capaz de eliminar biofilme de C. albicans. Este estudo demonstrou o potencial antimicrobiano de extratos ou frações de Cordia verbenacea, Mikania laevigata e Psidium cattleianum, sugerindo seu emprego com finalidade endodôntica devido à sua efetividade...

Seeking for alternative sources of the antimicrobials, several authors have explored the use of herbal medicines for the treatment of diseases. The antimicrobial activity of various species of plants, such as Cordia verbenacea DC, Mikania laevigata Schultz Bip. Ex Baker and Psidium cattleianum, known as erva-baleeira, guaco and araçá, respectively, has been reported against several oral microorganisms. The present study aimed to evaluate the antimicrobial effect of Cordia verbenacea (CV) and Mikania laevigata (ML) hydroethanolic crude extracts and their fractions aqueous, butanol, hexane and ethyl acetate and aqueous and hydroethanolic extracts of Psidium cattleianum (PC) against Enterococcus faecalis, Pseudomonas aeruginosa, Actinomyces israelii and Candida albicans, under planktonic and biofilm conditions. The microorganisms in planktonic conditions were analyzed by determination of minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) assays using the microdilution broth method. Posteriorly, only the extracts that were able to totally eliminate each microorganism (MLC 100%) were selected for the antimicrobial activity analysis in the biofilm assay. The results showed that the MIC and MLC values ranged between 250 and 4000μg/ml. E. faecalis biofilm was eliminated by PC aqueous and hydroethanolic extracts and the ML ethyl acetate fraction. The same ML fraction and PC hydroethanolic extract were also able to eliminate the P. aeruginosa biofilm. The same occurred with the CV and ML hexane fractions and ML hydroethanolic extract against the A. israelii biofilm. None of the extracts/fractions were able to eliminate the C. albicans biofilm. This study confirmed the antimicrobial activity potential of Cordia verbenacea, Mikania laevigata and Psidium cattleianum, suggesting their use for endodontic purposes due to their effectiveness against pathogens associated with persistent or secondary endodontic infections...

Análise da atividade antimicrobiana dos extratos de Cordia verbenacea DC, Mikania laevigata Sch. Bip. ex Baker e Psidium Cattleianum frente a microrganismos endodônticos / Antimicrobial analysis of the Cordia verbenacea DC, Mikania laevigata Sch. Bip. ex Baker e Psidium Cattleianum plant extracts against endodontic microorganisms

Na busca por fontes alternativas de antimicrobianos, diversos autores tem explorado o uso de plantas medicinais para o tratamento de várias doenças. A atividade antimicrobiana de várias espécies de plantas como, Cordia verbenacea DC, Mikania laevigata Schultz Bip. Ex Baker e Psidium cattleianum, popularmente chamadas de erva-baleeira, guaco e de araçá, respectivamente, tem sido relatada contra diversos microrganismos orais. O objetivo do presente estudo foi avaliar o efeito antimicrobiano dos extratos hidroetanólico bruto e as suas respectivas frações aquosa, butanol, hexano e acetato de etila de Cordia verbenacea (CV) e Mikania laevigata (ML) e extratos aquoso e hidroetanólico de Psidium cattleianum (PC) contra Enterococcus faecalis, Pseudomonas aeruginosa, Actinomyces israelii e Candida albicans, sob condições planctônicas e de biofilme. Os microrganismos foram analisados em condições planctônicas por meio de ensaios para a determinação da concentração inibitória mínima (CIM) e concentração letal mínima (CLM), pelo método de microdiluição em caldo. Posteriormente, foram selecionados somente os extratos capazes de eliminar completamente cada microrganismo (CLM 100%), para a análise da atividade antimicrobiana em ensaios de biofilme. Os resultados mostraram que os valores de CIM e CLM variaram de 250 à 4000μg/ml. O biofilme formado por E. faecalis foi eliminado frente à ação dos extratos aquoso e hidroetanólico de PC e à fração acetato de etila de ML. A mesma fração de ML e o extrato hidroetanólico de PC também foram capazes de eliminar biofilme de P. aeruginosa. O mesmo ocorreu com as frações hexânicas de CV e ML e o extrato hidroetanólico de ML frente ao biofilme de A. israelii. Nenhum extrato/fração foi capaz de eliminar biofilme de C. albicans. Este estudo demonstrou o potencial antimicrobiano de extratos ou frações de Cordia verbenacea, Mikania laevigata e Psidium cattleianum, sugerindo seu emprego com finalidade endodôntica devido à sua efetividade...

Seeking for alternative sources of the antimicrobials, several authors have explored the use of herbal medicines for the treatment of diseases. The antimicrobial activity of various species of plants, such as Cordia verbenacea DC, Mikania laevigata Schultz Bip. Ex Baker and Psidium cattleianum, known as erva-baleeira, guaco and araçá, respectively, has been reported against several oral microorganisms. The present study aimed to evaluate the antimicrobial effect of Cordia verbenacea (CV) and Mikania laevigata (ML) hydroethanolic crude extracts and their fractions aqueous, butanol, hexane and ethyl acetate and aqueous and hydroethanolic extracts of Psidium cattleianum (PC) against Enterococcus faecalis, Pseudomonas aeruginosa, Actinomyces israelii and Candida albicans, under planktonic and biofilm conditions. The microorganisms in planktonic conditions were analyzed by determination of minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) assays using the microdilution broth method. Posteriorly, only the extracts that were able to totally eliminate each microorganism (MLC 100%) were selected for the antimicrobial activity analysis in the biofilm assay. The results showed that the MIC and MLC values ranged between 250 and 4000μg/ml. E. faecalis biofilm was eliminated by PC aqueous and hydroethanolic extracts and the ML ethyl acetate fraction. The same ML fraction and PC hydroethanolic extract were also able to eliminate the P. aeruginosa biofilm. The same occurred with the CV and ML hexane fractions and ML hydroethanolic extract against the A. israelii biofilm. None of the extracts/fractions were able to eliminate the C. albicans biofilm. This study confirmed the antimicrobial activity potential of Cordia verbenacea, Mikania laevigata and Psidium cattleianum, suggesting their use for endodontic purposes due to their effectiveness against pathogens associated with persistent or secondary endodontic infections

Alteração cromática de resinas compostas laboratoriais submetidas à imersão em diferentes soluções / Chromatic alteration of composite resins laboratory submitted to immersion in different solutions

A pigmentação das restaurações indiretas de resina composta ainda continua sendo um dos motivos que determina o sucesso ou insucesso do tratamento restaurador. O propósito deste estudo foi investigar o efeito de diferentes soluções sobre a mudança de cor de resinas compostas indiretas. Cinco marcas diferentes de resinas foram avaliadas: Adoro, Resilab, Cristobal, Sinfony e Epricord. As amostras foram submetidas a onze diferentes tipos de soluções (n = 10): soluções de bebida (coca, vinho, café e suco de laranja), colutórios (Listerine, Oral-B, Plax, Periogard) e agentes de clareamento dental (peróxido de carbamida 16%, 7,5% e peróxido de hidrogênio 38%), e a saliva artificial (controle). A cor foi analisada por espectrofotômetro, antes, e após 7, 14 e 21 dias de imersão nas bebidas; após 12, 24, 36 e 60 horas de imersão em colutórios; e após 7 e 14 dias de imersão em agentes clareadores. As resinas Cristobal e Adoro apresentaram os maiores valores de ΔE, estatisticamente significante, em relação aos valores obtidos pelas outras marcas de resina. Os valores de ΔE da resina Adoro alteraram significativamente após imersão em soluções de vinho e café e entre os períodos mensurados. As resinas Cristobal e Sinfony apresentaram os maiores valores de ΔE após imersão em Listerine, com diferença significativa em relação ao controle. Além disso, houve diferença significativa dos valores de ΔE para a resina Cristobal após imersão em hidrogénio a 7,5% em comparação ao controle. Pode-se concluir que todas as soluções promoveram alteração de cor das resinas indiretas, no entanto os valores ΔE estão dentro de limites clínicos aceitáveis

The pigmentation of indirect composite resins has been one of the most common reasons that lead the restorative treatment to failure. The purpose of this study was to investigate the effect of different solutions on color stability of indirect composite resins. Five brands of indirect composite resins were tested: Adoro, Resilab, Cristobal, Sinfony and Epricord. The samples were immerged in eleven solutions (n=10): common liquid foods (coke soft drink, red wine, coffee and orange juice), mounthrinses (Listerine, Oral-B, Colgate Plax and Periogard) and bleaching agents (carbamide peroxide 16%, 7.5% and hydrogen peroxide 38%) and artificial saliva (control group). The color was measured by a spectrophotometer before and after 7, 14 and 21 days of immersion in common liquid foods, after 12, 24, 36 and 60 hours of immersion in mounthrinses and after 7 and 14 days of immersion in bleaching agents. The Cristobal and Adoro resins showed highest values of ΔE statistically significant compared to the others resin brands. Adoro’s ΔE values changed significantly after the immersion process in red wine and coffee and also between periods measured. The resins Cristobal and Sinfony showed the highest values of ΔE after the immersion process in Listerine with difference statistically significant in comparison to control group. Besides, there was difference statistically significant of the ΔE values for Cristobal after immersion in hydrogen peroxide 7,5% in comparison to control group. It can be conclude that all the solutions promoted color change on the indirect composite resins. However, ΔE values are whitin the values clinically acceptable

Upper lip prosthesis retained to a palatal obturator.

In cases of total or partial maxillectomies, the prosthetic rehabilitation is an effective alternative to minimize the sequelae left by surgical resection. The present study reports a clinical case of a 52-year-old patient who underwent partial maxillectomy, with upper lip involvement. The oronasal communication, resultant from surgical resection, did not allow the patient to return to her normal social life. Besides, the upper lip partial resection damaged her face's aesthetics. The proposed treatment was the confection of an upper lip prosthesis retained by a palatal obturator. The prosthesis insertion restored the patient's facial aesthetics, contributing not only to function, but also to psychosocial adaptation.