Effects of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide on adhesion of posts in teeth obturated with different sealers

Abstract The aim of this study was to evaluate the effect of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) on bond strength of fiberglass posts in root canals obturated with different endodontic sealers. Seventy-eight mandibular premolars were obturated with three sealers (n=26): Endofill (END), AH Plus (AHP), and Endosequence BC Sealer (EBS). After preparation of the post space, two subgroups were formed according to the cementation of the posts (n=13): with EDC (EDC), and without EDC (control - CON). The specimens were submitted to a pull-out test, failure mode classification, and root canal surface evaluation by scanning electron microscopy after post displacement. Regarding the bond strength, a significant difference between the EDC and CON subgroups occurred only in the END (p=0.001). No difference was detected among the CON subgroups (p=0.339). However, among the EDC subgroups, AHP presented significantly higher values (END versus AHP: p=0.001; AHP versus EBS: p=0.016). Upon classification of failure modes, score 1 (≥ 50% of cement) was the most commonly observed, except for the END + EDC. Remains of endodontic sealers and resin cements were found in the cervical third, but without statistical difference (p=0.269), while in the middle third, difference occurred (p=0.004). In conclusion, EDC decreases bond strength when associated with END sealer, without changing the failure mode between the resin cement and fiberglass post. The best performance was observed when EDC was combined with AHP sealer.

Resumo O objetivo deste estudo foi avaliar o efeito da 1-etil-3- (3-dimetilaminopropil) carbodiimida (EDC) na resistência de união de pinos de fibra de vidro em canais radiculares obturados com diferentes cimentos endodônticos. Setenta e oito pré-molares inferiores foram obturados com três cimentos endodônticos (n=26): Endofill (END), AH Plus (AHP) e Endosequence BC Sealer (EBS). Após o preparo do espaço para pino, dois subgrupos formaram-se conforme a cimentação dos pinos (n=13): com EDC e sem EDC (controle - CON). Os espécimes foram submetidos ao teste pull-out, classificação do modo de falha e avaliação da superfície do canal radicular por microscopia eletrônica de varredura após o deslocamento. Quanto à força de resistência de união, uma diferença estatisticamente significativa ocorreu entre os subgrupos EDC e CON apenas no END (p=0,001). Não foi detectada diferença entre os subgrupos CON (p=0,339). Contudo, no subgrupo EDC, o AHP apresentou maiores valores (END versus AHP: p=0,001; AHP versus EBS: p=0,016). Acerca da classificação dos modos de falha, o escore 1 (≥50% de cimento) foi o mais comumente observado, exceto para END + EDC. Restos de cimentos endodônticos e cimentos resinosos foram encontrados no terço cervical, mas sem diferença estatística (p=0,269), enquanto no terço médio, houve diferença (p=0,004). Em conclusão, o EDC diminui a resistência de união quando associado ao cimento END, sem alterar o modo de falha entre o cimento resinoso e o pino de fibra de vidro. O melhor desempenho foi observado quanto o EDC foi usado com o cimento AHP.

Citotoxicidade transdentinária e efeito da carbodiimida (EDC) na biomodificação do colágeno dentinário e na degradação da interface adesiva / Trandentinal cytotoxicity and effect of carbodiimide (EDC) on dentin collagen biomodification and adhesive interface degradation

O objetivo geral deste trabalho foi avaliar a citotoxicidade transdentinária da carbodiimida (EDC), bem como sua influência na degradação do colágeno dentinário e na estabilidade da união resina-dentina. No estudo 1, células MDPC23 foram plantadas na superfície pulpar de discos de dentina e a superfície oclusal foi tratada por 60s com uma das seguintes soluções: sem tratamento; EDC 0,1M; 0,3M ou 0,5M; glutaraldeído 5% (GA); tampão Sorensen ou H2O2 29%. A viabilidade e a morfologia celular foram analisadas pelos testes de MTT, Live/dead, produção de proteína total (PT), de colágeno e MEV. Os dados foram analisados pelos testes de Kruskal-Wallis e Mann-Whitney (p<0,05). O GA promoveu aumento do metabolismo celular. A morte por necrose e a morfologia celular não foram influenciadas pelos agentes cross-linkers. Não houve redução na produção de PT e colágeno após 7 dias. Para o estudo 2, espécimes de dentina foram completamente desmineralizados e a variação do módulo de elasticidade (E), inibição de MMP, perda de massa, liberação de hidroxiprolina (HYP) e degradação térmica do colágeno (DTC) foram analisados após tratamento com uma das seguintes soluções por 30s ou 60s: água deionizada (controle); EDC 0,5M; EDC 1M; EDC 2M e GA 10%. Os dados referentes ao E, atividade de MMP e liberação de HYP foram submetidos aos testes de Wilcoxon e KruskalWallis ou Mann-Whitney. Os valores de perda de massa e DT foram analisados pelos testes de ANOVA e Tukey (p<0,05). Os melhores resultados quanto ao E foram observados para o GA. Todos os cross-linkers reduziram a atividade de MMP e a liberação de HYP e aumentaram a temperatura de DT do colágeno. No estudo 3, sessenta palitos de dentina foram divididos em 6 grupos de acordo com a solução de tratamento: água deionizada (controle); EDC 0,1M; EDC 0,5M; EDC 0,5M + HEMA 35%; proantocianidina 5% (PA) ou clorexidina (CHX) 2%. Após condicionamento ácido os palitos foram tratados por 60s. A atividade de MMP foi analisada por ensaio colorimétrico. Os dados expressos em valores de absorbância e em equivalentes a atividade de MMP-9 foram submetidos aos testes de Kruskall-Wallis e Mann-Whitney (p<0.05). Todas as soluções inibiram a atividade de MMP. O EDC 0,5M e sua mistura com HEMA obtiveram os melhores resultados. Finalmente, no estudo 4, superfícies planas de dentina foram condicionadas e tratadas por 30 ou 60s com solução de EDC 0,5M ou água deionizada. Após o tratamento, o adesivo Single Bond 2 (SB2) foi aplicado e as coroas reconstruídas em resina composta. Espécimes foram produzidos e submetidos aos testes de microtração e nanoinfiltração após 24h, 6 ou 12 meses em saliva artificial. Os testes de ANOVA e Tukey (p<0.05) foram aplicados. O tratamento da dentina com EDC preveniu a redução da RU e o aumento da nanoinfiltração após 12 meses de armazenamento. Desta forma, pôde ser concluído que o pré-tratamento da dentina com agentes cross-linkers não exerceu efeito citotóxico trandentinário sobre células odontoblastóides, favoreceu as propriedades mecânicas do colágeno, foi capaz de inibir MMPs e prevenir a degradação da interface adesiva

The purpose of this study was to evaluate the trandentinal cytotoxicity of carbodiimide (EDC), as well as its influence on dentinal collagen degradation and stability of resin-dentin bonds. In the first experiment, MDPC-23 cells were seeded on the pulp surface of the disks and one of the following solutions was applied on the occlusal surface for 60s: no treatment (negative control), 0.1M, 0.3M or 0.5M EDC; 5% glutaraldehyde (GA); Sorensen buffer or 29% hydrogen peroxide (positive control). Cell viability and morphology were analyzed by MTT, Live/Dead assays, total protein (TP) and collagen production and SEM. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (p<0.05). Only GA increased cellular metabolism. Cell death by necrosis and cell morphology were not affected by the cross-linker agents. There was no reduction in TP and collagen production after 7 days. For the second experiment, dentin beams were completely demineralized and the variation in elastic modulus (E), MMP activity, dry mass loss, hydroxyproline release (HYP) and collagen thermal degradation (CTD) were analyzed after the dentin treatment for 30s or 60s with the following solutions: water; 0.5M; 1M or 2M EDC and 10% GA. Data from E and MMP activity and HYP release were submitted to Wilcoxon and Kruskal-Wallis or Mann-Whitney tests. Dry mass loss and CTD data were analyzed by ANOVA and Tukey's tests (p>0.05). GA group obtained the highest E values. All cross-linking agents decreased MMP activity and HYP release and increased CTD. In the third experiment, sixty dentin beams were randomly divided into 6 groups according to the treatment solution: deionized water (control), 0.1M EDC, 0.5M EDC, 0.5M EDC+35% HEMA, 5% proanthocyanidin (PA) or 2% chlorhexidine (CHX). The beams were acid etched and treated for 60s. The total MMP activity was analyzed by a colorimetric assay (Sensolyte®). Data were expressed as absorbance values at 412nm and MMP-9 equivalents and subjected to Kruskal-Wallis and MannWhitney tests (p<0.05). All cross-liking solutions inhibited MMPs. The 0.5 M EDC solution and its mixture with HEMA had the highest inhibition values. Finally, in experiment 4, flat dentin surfaces were etched and treated for 30 or 60s with 0.5M EDC or deionized water. After treatment, Single Bond 2 (SB2) was applied and the crowns were reconstructed with composite resin. Dentin specimens were produced and submitted to microtensile and nanoleakage tests after 24h, 6 or 12 months in artificial saliva. Bond strength (BS) data (MPa) were analyzed by ANOVA and Tukey tests (p<0.05). The dentin treatment with EDC did not affect SB2 immediate BS and prevented the degradation of the adhesive interface, even after 12 months of saliva storage. Thus, the dentin treatment with cross-linking agents did not exert transdentinal cytotoxic effects on odontoblastlike cells, increased collagen mechanical properties, was able to inhibit MMPs and prevent the degradation of the adhesive interface

In Vitro and In Vivo Imaging of Prostate Cancer Angiogenesis Using Anti-Vascular Endothelial Growth Factor Receptor 2 Antibody-Conjugated Quantum Dot

OBJECTIVE: Authors aimed to determine the targeting ability of vascular endothelial growth factor receptor 2 (VEGFR2)-conjugated quantum dots (QDs) in vitro, and apply it for a xenograft prostate cancer mouse model. MATERIALS AND METHODS: Conjugation reaction of QDs was performed by using the N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) and sulfo-(N-hydroxysulfosuccinimide) (Sulfo-NHS). The human umbilical vein cord endothelial cells (HUVECs) were incubated with QDs, conjugated with antiVGFR2, to see a specific binding in vitro. Fluorescent cell images were taken by a confocal microscope. The human prostate cancer cells (PC3) were injected to five nude mice on hind limbs to make the xenograft tumor model. QD-antiVEGFR2 antibody complex was injected into the tumor model and fluorescence measurements were performed at 1, 4, 9, 12, 15, and 24 hours after the injection. RESULTS: The specific interaction between HUVECs and QD-antiVEGFR2 antibody was clearly shown in vitro. The in vivo fluorescence image disclosed that there was an increased signal of tumor, 12 hours after the injection of QDs. CONCLUSION: By showing endothelial cells binding with QDs-antiVEGFR2 antibodyand an experimental application of the antibody for VEGFR2 imaging in the prostate cancer xenograft mouse model, we suggests that the antibody-conjugated QDs can be a potential imaging tool for angiogenesis of the cancer.

Biocompatibility of decellularized canine carotid artery allograft cross-linked by carbodiimide / 中华整形外科杂志

<p><b>UNLABELLED</b>OBJECTIVE Crosslink decellularized canine carotid artery allograft by EDC [1-3-(dimethylamino)propyl-3-ethylcarbodiimide methiodide] and evaluate the biocompatibility of it.</p><p><b>METHODS</b>Use the multi-step detergent-enzyme method to construct decellularized canine carotid artery allograft and cross-link it by EDC with the weight ratio of decellularized artery to EDC 1:1 and 1:2. Evaluate the biocompatibility of it by the cytotoxical MTT test and the rat subdermal bury test.</p><p><b>RESULTS</b>Decellularized canine carotid artery cross-linked by EDC has a lower degradation rate treated by collagenase type II, the result of MTT test show that the EDC cross-linked decellularized artery has no cytotoxity and the rat subdermal bury test show that crosslinking greatly enhance the ability of decellularize artery to resist the enzyme degradation and lower the immune reaction. The more the artery was cross-linked , the more effects it has.</p><p><b>CONCLUSIONS</b>Decellularized canine carotid artery cross-linked by EDC has fairly good biocompatibility and ability to resist the collagenase degradation.</p>

Immobilization of lipase by chemical modification of chitosan / 生物工程学报

Lipase (EC3.1.1.3) from Candida sp. 99-125 was immobilized on chitosan by chemical covalence. Lipase was first immobilized to chitosan beads by activating its hydroxyl groups with carbodiimide followed by cross-linking more lipase to the amino groups with glutaraldehyde. In this article, different factors that influenced the immobilization were investigated, and the optimum conditions were ascertained. Comparative studies of organic solvent and thermal stability between free lipase and immobilized lipase were conducted. Immobilization enhanced the lipase stability against changes of temperature and organic solvent. Immobilization lipase can be reused in the synthesis system of palmitate hexadecyl. Operational stability tests indicated that the immobilized lipase occurs after 16 consecutive batches, the conversion rate remained 85%. Such results revealed good potential for recycling under esterification system.

Further Studies on Effects of Some Local Anesthetics on Calcium Binding to Lipid-extracted RBC Membrane Fragments: Effect of Modification of Carboxyl Group with Carbodiimide

The interaction of calcium and local anesthetics was investigated with the lipid extracted human RBC membrane fragments treated with carbodiimide in order to titrate carboxyl groups. A water soluble carbodiimide [1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide methotoluene-p-sulfonate], referred to as a carbodiimide reagent, and glycine methylester were used for this purpose. About 76% of carboxyl groups of the fragments were modified at a concentration of 0.05M carbodiimide reagent. The interaction of calcium and local anesthetics such as procaine and lidocaine with these fragments still showed typical competition. However, when the calcium binding was decreased to 8% at a higher concentration of carbodiimide reagent (0.08M), the local anesthetics still inhibited the calcium binding, but were not competitive in nature. In other words, if concentrations of the carbodiimide reagent were raised, the degree of inhibition by the local anesthetics was gradually decreased and was not competitive in nature. Finally, no inhibition was demonstrated when the concentration of the reagent was 0.1 to 0.4M. The above findings, seem to suggest that local anesthetics such as procaine and lidocaine interact with carboxyl groups, in addition to phosphodiester groups of phospholipids as previously reported, and inhibited competitively calcium binding to carboxyl groups of the membrane fragments.

Further Studies on Effects of Some Local Anesthetics on Calcium Binding to Lipid-extracted RBC Membrane Fragments: Effect of Modification of Carboxyl Group with Carbodiimide

The interaction of calcium and local anesthetics was investigated with the lipid extracted human RBC membrane fragments treated with carbodiimide in order to titrate carboxyl groups. A water soluble carbodiimide [1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide methotoluene-p-sulfonate], referred to as a carbodiimide reagent, and glycine methylester were used for this purpose. About 76% of carboxyl groups of the fragments were modified at a concentration of 0.05M carbodiimide reagent. The interaction of calcium and local anesthetics such as procaine and lidocaine with these fragments still showed typical competition. However, when the calcium binding was decreased to 8% at a higher concentration of carbodiimide reagent (0.08M), the local anesthetics still inhibited the calcium binding, but were not competitive in nature. In other words, if concentrations of the carbodiimide reagent were raised, the degree of inhibition by the local anesthetics was gradually decreased and was not competitive in nature. Finally, no inhibition was demonstrated when the concentration of the reagent was 0.1 to 0.4M. The above findings, seem to suggest that local anesthetics such as procaine and lidocaine interact with carboxyl groups, in addition to phosphodiester groups of phospholipids as previously reported, and inhibited competitively calcium binding to carboxyl groups of the membrane fragments.