RESUMO
Enzymatic degradation of the hybrid layer can be accelerated by the activation of dentin metalloproteinases (MMP) during the bonding procedure. MMP inhibitors may be used to contain this process. Objective To evaluate the degree of conversion (DC%), dentin bond strength (µTBS) (immediate and after 1 year of storage in water), and nanoleakage of an experimental (EXP) and a commercial (SB) adhesive system, containing different concentrations of the MMP inhibitor GM1489: 0, 1 µM, 5 µM and 10 µM. Methodology DC% was evaluated by FT-IR spectroscopy. Dentin bond strength was evaluated by µTBS test. Half of beams were submitted to the µTBS test after 24 h and the other half, after storage for 1 year. From each tooth and storage time, 2 beams were reserved for nanoleakage testing. Data were analyzed using ANOVA and Tukey's test to compare means (α=0.05). Results All adhesive systems maintained the µTBS after 1 year of storage. Groups with higher concentrations of inhibitor (5 µM and 10 µM) showed higher µTBS values than groups without inhibitor or with 1 µM. The nanoleakage values of all groups showed no increase after 1 year of storage and values were similar for SB and EXP groups, in both storage periods. The inhibitor did not affect the DC% of the EXP groups, but the SB5 and SB10 groups showed higher DC% values than those of SB0 and SB1. Conclusions The incorporation of GM1489 in the adhesive systems had no detrimental effect on DC%. The concentrations of 5 µM GM1489 for SB and 5 µM or 10 µM for EXP provided higher µTBS than groups without GM1489, in the evaluation after 1 year of storage; whereas the concentration of inhibitor did not affect adhesive systems nanoleakage.
Assuntos
Cimentos Dentários/química , Dentina/química , Inibidores de Metaloproteinases de Matriz/química , Metacrilatos/química , Polietilenoglicóis/química , Ácidos Polimetacrílicos/química , Análise de Variância , Colagem Dentária/métodos , Corrosão Dentária/métodos , Infiltração Dentária , Dentina/efeitos dos fármacos , Humanos , Teste de Materiais , Valores de Referência , Reprodutibilidade dos Testes , Espectroscopia de Infravermelho com Transformada de Fourier , Estatísticas não Paramétricas , Propriedades de Superfície , Resistência à Tração , Fatores de TempoRESUMO
Abstract Enzymatic degradation of the hybrid layer can be accelerated by the activation of dentin metalloproteinases (MMP) during the bonding procedure. MMP inhibitors may be used to contain this process. Objective To evaluate the degree of conversion (DC%), dentin bond strength (µTBS) (immediate and after 1 year of storage in water), and nanoleakage of an experimental (EXP) and a commercial (SB) adhesive system, containing different concentrations of the MMP inhibitor GM1489: 0, 1 µM, 5 µM and 10 µM. Methodology DC% was evaluated by FT-IR spectroscopy. Dentin bond strength was evaluated by µTBS test. Half of beams were submitted to the µTBS test after 24 h and the other half, after storage for 1 year. From each tooth and storage time, 2 beams were reserved for nanoleakage testing. Data were analyzed using ANOVA and Tukey's test to compare means (α=0.05). Results All adhesive systems maintained the µTBS after 1 year of storage. Groups with higher concentrations of inhibitor (5 µM and 10 µM) showed higher µTBS values than groups without inhibitor or with 1 µM. The nanoleakage values of all groups showed no increase after 1 year of storage and values were similar for SB and EXP groups, in both storage periods. The inhibitor did not affect the DC% of the EXP groups, but the SB5 and SB10 groups showed higher DC% values than those of SB0 and SB1. Conclusions The incorporation of GM1489 in the adhesive systems had no detrimental effect on DC%. The concentrations of 5 µM GM1489 for SB and 5 µM or 10 µM for EXP provided higher μTBS than groups without GM1489, in the evaluation after 1 year of storage; whereas the concentration of inhibitor did not affect adhesive systems nanoleakage.
Assuntos
Humanos , Polietilenoglicóis/química , Ácidos Polimetacrílicos/química , Cimentos Dentários/química , Dentina/química , Inibidores de Metaloproteinases de Matriz/química , Metacrilatos/química , Valores de Referência , Propriedades de Superfície , Resistência à Tração , Fatores de Tempo , Teste de Materiais , Reprodutibilidade dos Testes , Análise de Variância , Colagem Dentária/métodos , Espectroscopia de Infravermelho com Transformada de Fourier , Estatísticas não Paramétricas , Infiltração Dentária , Dentina/efeitos dos fármacos , Corrosão Dentária/métodosRESUMO
PURPOSE: To evaluate the microshear bond strength (µSBS) to deep (DD) or superficial (SD) dentin (µSBS) overtime, nanoleakage (AG%), degree of conversion (DC%), water sorption (WSp), and solubility (WSl) of an adhesive system [Adper Single Bond 2(ASB)] containing matrix metalloproteinases (MMP) inhibitors [GM1489 (ASB-GM), Batimastat (ASB-BAT), or Chlorhexidine diacetate (ASB-CHX)]. ASB without inhibitor was used as control (CONTROL). MATERIALS AND METHODS: WSp and WSL were calculated based on ISO4049. DC% was analyzed using FT-IR spectroscopy. Dentin discs were used for µSBS evaluation. For AG%, resin-dentin beams were analyzed under scanning electronic microscopy. Data were analyzed using three-way ANOVA (AG% and µSBS) or ANOVA (DC%, WSp, WSl) and Tukey's HSD test. RESULTS: ASB-CHX presented the lowest DC%, lowest WSp, and highest WSl. ASB-GM reached the highest immediate µSBS in SD, only different from ASB-CHX. In DD, ASB-BAT and ASB-GM had the highest µSBS, statistically different from ASB-CHX. After twelve months, ASB-GM and ASB-BAT presented higher µSBS in SD when compared to CONTROL and ASB-CHX. In DD, ASB-GM reached the highest value, which was statistically different from CONTROL and ASB-CHX. CONTROL at both dentin depths and ASB-CHX at DD did not maintain bond stability. In SD after 12 months, ASB-BAT and ASB-GM decreased AG%. In DD, only ASB-GM reduced AG%. CONCLUSION: The ASB containing Batimastat and GM1489 maintained resin-dentin bond stability after 12 months for both dentin depths, without jeopardizing WSp, WSl, or DC% of the adhesive system. The ASB-GM presented greater µSBS after 12 months when compared with ASB. CLINICAL SIGNIFICANCE: Batimastat and GM1489 could be suitable for inclusion as an MMP-inhibitor into Single Bond to improve the bond stability to superficial and deep dentin, without jeopardize the physic-mechanical proprieties.
Assuntos
Adesivos Dentinários/química , Dentina/química , Inibidores de Metaloproteinases de Matriz/química , Cimentos de Resina/química , Condicionamento Ácido do Dente , Adesivos/análise , Adolescente , Adulto , Clorexidina/química , Resinas Compostas/química , Colagem Dentária , Humanos , Técnicas In Vitro , Modelos Lineares , Teste de Materiais , Fenilalanina/análogos & derivados , Fenilalanina/química , Solubilidade , Estresse Mecânico , Propriedades de Superfície , Resistência à Tração , Tiofenos/química , Triptofano/análogos & derivados , Triptofano/química , Adulto JovemRESUMO
A new collagenase producing a strain of Bacillus cereus, isolated from the pollen of a bee of Amazon Region (Brazil), had its enzyme characterized and the production medium composition and culture conditions enhanced. A two-level design on three factors, namely initial medium pH, the substrate (gelatin) concentration and agitation intensity, allowed identifying the first two variables as the most significant ones, while a central composite design (CCD) was subsequently used to identify their optimal levels. Statistics highlighted maximized collagenolytic activity when substrate concentration and initial medium pH were selected at their highest levels (positive effects), whereas agitation intensity at the lowest (negative effect). Triplicate runs performed under predicted optimal conditions (pH 7.8 and 1.7% gelatin concentration) yielded a collagenolytic activity (305.39 ± 5.15 U) 4.6- to 15-fold those obtained with the preliminary design. The enzyme displayed optimum activity at 45 °C and pH 7.2, was stable over wide ranges of pH values and temperatures (7.2-11.0 and 25-50 °C, respectively) and was strongly inhibited by 10 mM phenylmethylsulphonyl fluoride. The zymogram showed two prominent bands at 50 and 76 kDa. These results are a first attempt to elucidate the features of this new collagenase, its production conditions, and possible scale-up.
Assuntos
Bacillus cereus/enzimologia , Colagenases/química , Animais , Bacillus cereus/genética , Técnicas de Tipagem Bacteriana , Abelhas , Brasil , Colagenases/isolamento & purificação , Meios de Cultura , Precursores Enzimáticos/química , Precursores Enzimáticos/isolamento & purificação , Gelatina/metabolismo , Concentração de Íons de Hidrogênio , Inibidores de Metaloproteinases de Matriz/química , Pólen/microbiologia , RNA Ribossômico 16S/genética , TemperaturaRESUMO
Objetivo: realizar uma revisão de literatura acerca da eficácia de utilização da clorexidina (CHX) e de outros tipos de inibidores de metaloproteinases (MMPs) na resistência de união da camada híbrida. Métodos: a busca bibliográfica foi realizada na base de dados PubMed, nos meses de novembro e dezembro de 2018. A pesquisa ocorreu em três fases, com os descritores previamente selecionados. Foram incluídas publicações dos últimos 10 anos no formato de pesquisas científicas realizadas in vitro ou in vivo. Após análise, obedecendo aos critérios de inclusão e exclusão, foram incluídos sete estudos na presente revisão. Resultados/Revisão de literatura: na interface adesiva, os estudos mostram que as MMPs são ativadas durante a etapa de ataque ácido realizada nos protocolos de aplicação de sistemas adesivos, podendo ser ativada tanto por procedimentos adesivos com condicionamento ácido prévio como por sistemas adesivos autocondicionantes. Além da CHX, outras substâncias foram pesquisadas e se mostraram eficazes na inibição de MMPs. Considerações finais: por meio da inibição da atividade das MMPs, é possível obter uma maior durabilidade da interface adesiva e uma menor degradação hidrolítica do colágeno presente na camada híbrida. (AU)
Objective: to perform a literature review on the efficacy of chlorhexidine (CHX) and other types of metalloproteinase inhibitors (MMPs) on hybrid layer bond strength. Methods: the bibliographic search was performed in PubMed, in the months of november and december of 2018. The research was carried out in three phases with the previously selected descriptors. Publications have been included in the last 10 years in the form of scientific research conducted in vitro or in vivo. After analysis, following the inclusion and exclusion criteria, 7 studies were included in the present review. Results / Literature review: in the adhesive interface, the studies show that the MMPs are activated during the acid attack stage carried out in the application protocols of adhesive systems, and can be activated either by adhesive procedures with prior acid conditioning or self-etching adhesive systems. In addition to CHX, other substances were investigated and shown to be effective in inhibiting MMPs. Final considerations: through the inhibition of the MMPs activity it is possible to obtain a greater durability of the adhesive interface and lower hydrolytic degradation of the collagen present in the hybrid layer. (AU)
Assuntos
Humanos , Clorexidina/química , Adesivos Dentinários/química , Dentina/química , Inibidores de Metaloproteinases de Matriz/química , Compostos de Benzalcônio/química , Colágenos Fibrilares/efeitos dos fármacos , Proantocianidinas/química , Dentina/efeitos dos fármacosRESUMO
The adhesive process to dentin substrate depends on the condition determined by the combined action of the mineral loss and the endogenous enzymes activity. Thus, considering a more complete therapeutic approach, sodium trimetaphosphate (STMP) may be a novel strategy that conciliates the remineralization potential to the promotion of dentin strengthening and its stability, possibly directing mineral nucleation and controlling the rate of biodegradation. In this study, the effect of STMP was evaluated in 2 studies. In study 1, different concentrations of STMP (0.5, 1.5, 3.5 and 5%) were investigated to assess their anti-proteolytic capacity on human purified MMPs-2 and -9 by zymography. Afterwards, only the concentrations (1.5, 3.5 and 5%) that showed total inhibition of both MMPs were used to evaluate their remineralizing capacity in dentin substrate submitted to artificial cariogenic challenge, through surface hardness (SH) and cross-sectional hardness (CSH). In study 2, based on the previous results, the capacity of the 1.5% STMP associated or not with NaF or Ca(OH)2 solutions in improving the dentin bond strength of a universal adhesive system was evaluated by the microtensile test . Thus, these studies suggest that 1.5% STMP is an effective inhibitor of collagen degradation mediated by purified human MMPs-2 and -9. In addition, demineralized and treated dentin with 1.5% STMP supplemented with Ca(OH)2 may induce remineralization. Thus, the use of STMP can be introduced as a new strategy that combines enzymatic inhibition and remineralization potential, reestablishing favorable conditions to affected dentin. These evidences support perspectives of therapies to restructure dentin and propose feasible and promising clinical strategies.(AU)
O processo adesivo ao substrato dentinário depende da condição determinada pela ação combinada da perda mineral e atividade de enzimas endógenas. Deste modo, considerando uma abordagem terapêutica mais completa, o trimetafosfato de sódio (STMP) pode ser uma estratégia inovadora que concilia o potencial remineralizador à promoção do fortalecimento da dentina e sua estabilidade, possivelmente direcionando a nucleação mineral e controlando a taxa de biodegradação. Neste trabalho, o efeito do STMP foi avaliado em 2 estudos. No estudo 1, diferentes concentrações de STMP (0,5; 1,5; 3,5 e 5%) foram investigadas para avaliar sua capacidade anti-proteolítica sobre as MMPs-2 e -9 purificadas humanas, por zimografia. Posteriormente, somente as concentrações (1,5; 3,5 e 5%) que apresentaram capacidade de inibição total de ambas MMPs foram utilizadas para avaliar sua capacidade remineralizadora em substrato dentinário submetido ao desafio cariogênico artificial, através da dureza de superfície (DS) e longitudinal (DL). No estudo 2, baseado nos resultados anteriores, foi avaliada a capacidade do STMP à 1,5% associado ou não a soluções de NaF ou Ca(OH)2 em melhorar a resistência de união à dentina de um sistema adesivo universal pelo teste de microtração. Desta forma, estes estudos sugerem que o STMP à 1,5% apresenta-se como um inibidor eficaz da degradação do colágeno mediada por MMPs-2 e -9 humanas purificadas. Além disso, a dentina humana desmineralizada e tratada com STMP à 1,5% suplementada com Ca(OH)2 pode induzir à remineralização. Assim, o uso de STMP pode ser introduzido como uma nova estratégia que combina inibição enzimática e potencial de remineralização, reestabelecendo condições favoráveis a partir de uma dentina afetada. Estas evidências sustentam perspectivas de terapias para reestruturar a dentina e propor estratégias clínicas factíveis e promissoras.(AU)
Assuntos
Humanos , Cariostáticos/química , Dentina/efeitos dos fármacos , Metaloproteinases da Matriz/efeitos dos fármacos , Polifosfatos/química , Remineralização Dentária/métodos , Testes de Dureza , Inibidores de Metaloproteinases de Matriz/química , Reprodutibilidade dos Testes , Fluoreto de Sódio/química , Resistência à TraçãoRESUMO
A receptor-independent (RI) four-dimensional structure-activity relationship (4D-QSAR) formalism was applied to a set of sixty-four ß-N-biaryl ether sulfonamide hydroxamate derivatives, previously reported as potent inhibitors against matrix metalloproteinase subtype 9 (MMP-9). MMP-9 belongs to a group of enzymes related to the cleavage of several extracellular matrix components and has been associated to cancer invasiveness/metastasis. The best RI 4D-QSAR model was statistically significant (N=47; r(2) =0.91; q(2) =0.83; LSE=0.09; LOF=0.35; outliers=0). Leave-N-out (LNO) and y-randomization approaches indicated the QSAR model was robust and presented no chance correlation, respectively. Furthermore, it also had good external predictability (82 %) regarding the test set (N=17). In addition, the grid cell occupancy descriptors (GCOD) of the predicted bioactive conformation for the most potent inhibitor were successfully interpreted when docked into the MMP-9 active site. The 3D-pharmacophore findings were used to predict novel ligands and exploit the MMP-9 calculated binding affinity through molecular docking procedure.
Assuntos
Antineoplásicos/química , Metaloproteinase 9 da Matriz/química , Inibidores de Metaloproteinases de Matriz/química , Melanoma/tratamento farmacológico , Neoplasias Cutâneas/tratamento farmacológico , Domínio Catalítico , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Ligação de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Ácidos Hidroxâmicos/química , Melanoma/enzimologia , Simulação de Acoplamento Molecular , Ligação Proteica , Relação Quantitativa Estrutura-Atividade , Neoplasias Cutâneas/enzimologia , Sulfonamidas/químicaRESUMO
UNLABELLED: The use of gels and mouthrinses with MMP inhibitors (chlorhexidine, and green tea extract) was shown to prevent erosive wear. The aim of this study was to analyze the protective effect of toothpastes containing MMP inhibitors on dentine loss induced by erosion in vitro. MATERIAL AND METHODS: Five groups each containing 12 specimens of human root dentine were prepared. The specimens were subjected to 1 min erosion by immersion in a cola drink, 4 times a day, for 5 d. Each day, after the first and last erosive challenges, the specimens were brushed for 15 s with a slurry of dentifrice and water (1:3) containing placebo, 1,100 ppm fluoride, 0.61% green tea extract, 0.12% chlorhexidine or 0.004% chlorhexidine (commercial toothpaste). Between the acid challenges, the specimens were stored in artificial saliva with remineralizing potential until the next treatment. Dentine loss was determined using profilometry. Data were analyzed using one-way ANOVA after log transform (p<0.05). RESULTS: The mean wear values (µm) were as follows: placebo 1.83±0.53; 0.61% green tea extract 1.00±0.21; fluoride 1.27±0.43; 0.12% chlorhexidine 1.19±0.30; and 0.004% chlorhexidine 1.22±0.46. There was a significant difference in wear between placebo and all the treatment toothpastes, which did not differ from each other. CONCLUSION: The results suggest that toothpastes containing MMP inhibitors are as effective as those based on NaF in preventing dentine erosion and abrasion.
Assuntos
Dentina/efeitos dos fármacos , Inibidores de Metaloproteinases de Matriz/química , Abrasão Dentária/prevenção & controle , Erosão Dentária/prevenção & controle , Cremes Dentais/química , Análise de Variância , Bebidas Gaseificadas , Clorexidina/química , Humanos , Teste de Materiais , Distribuição Aleatória , Saliva Artificial/química , Propriedades de Superfície/efeitos dos fármacos , Fatores de Tempo , Escovação DentáriaRESUMO
ABSTRACT The use of gels and mouthrinses with MMP inhibitors (chlorhexidine, and green tea extract) was shown to prevent erosive wear. The aim of this study was to analyze the protective effect of toothpastes containing MMP inhibitors on dentine loss induced by erosion in vitro. Material and Methods Five groups each containing 12 specimens of human root dentine were prepared. The specimens were subjected to 1 min erosion by immersion in a cola drink, 4 times a day, for 5 d. Each day, after the first and last erosive challenges, the specimens were brushed for 15 s with a slurry of dentifrice and water (1:3) containing placebo, 1,100 ppm fluoride, 0.61% green tea extract, 0.12% chlorhexidine or 0.004% chlorhexidine (commercial toothpaste). Between the acid challenges, the specimens were stored in artificial saliva with remineralizing potential until the next treatment. Dentine loss was determined using profilometry. Data were analyzed using one-way ANOVA after log transform (p<0.05). Results The mean wear values (μm) were as follows: placebo 1.83±0.53; 0.61% green tea extract 1.00±0.21; fluoride 1.27±0.43; 0.12% chlorhexidine 1.19±0.30; and 0.004% chlorhexidine 1.22±0.46. There was a significant difference in wear between placebo and all the treatment toothpastes, which did not differ from each other. Conclusion The results suggest that toothpastes containing MMP inhibitors are as effective as those based on NaF in preventing dentine erosion and abrasion.
Assuntos
Humanos , Abrasão Dentária/prevenção & controle , Erosão Dentária/prevenção & controle , Cremes Dentais/química , Dentina/efeitos dos fármacos , Inibidores de Metaloproteinases de Matriz/química , Saliva Artificial/química , Propriedades de Superfície/efeitos dos fármacos , Fatores de Tempo , Escovação Dentária , Teste de Materiais , Bebidas Gaseificadas , Distribuição Aleatória , Clorexidina/química , Análise de VariânciaRESUMO
A highly regiospecific synthesis of a series of indenoindoles is reported, together with X-ray studies and their activity against human prostate cancer cells PC-3 and LNCaP in vitro. The most effective compound 7,7-dimethyl-5-[(3,4-dichlorophenyl)]-(4bRS,9bRS)-dihydroxy-4b,5,6,7,8,9bhexahydro-indeno[1,2-b]indole-9,10-dione 7q reduced the viability in both cell lines in a time and dose-dependent manner. Inhibitory effects were also observed on the adhesion, migration, and invasion of the prostate cancer cells as well as on clonogenic possibly by inhibition of MMP-9 activity. Molecular docking of 7q and 6k into MMP-9 human active site was also performed to determine the probable binding mode.