Emmanuel João Nogueira Leal da Silva

Evaluation of Cytotoxicity and Physicochemical Properties of Calcium Silicate-based Endodontic Sealer MTA Fillapex

INTRODUCTIONnThe aim of the study was to evaluate the cytotoxicity, radiopacity, pH, and flow of a calcium silicate-based and an epoxy resin-based endodontic sealer, MTA Fillapex (Angelus, Londrina, PR, Brazil) and AH Plus (Dentsply, Konstanz, Germany), respectively.nnnMETHODSnCytotoxicity, radiopacity, and flow evaluation were performed following ISO requirements. The pH level was measured at periods of 3, 24, 72, and 168 hours. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay to check the Balb/c 3T3 cells viability at 1- to 4-week periods. Data were statistically analyzed by analysis of variance and the Tukey test with a significance level of 5%.nnnRESULTSnIn all tested periods, MTA Fillapex was more cytotoxic than AH Plus (P < .05). Although AH Plus presented higher radiopacity than MTA Fillapex (P < .05), both sealers showed minimum required values. MTA Fillapex presented alkaline pH in all experimental times, whereas AH Plus cement showed a slightly neutral pH and a flow significantly lower than that of MTA Fillapex (P < .05).nnnCONCLUSIONSnAlthough MTA Fillapex was more cytotoxic than AH Plus, it showed suitable physicochemical properties for an endodontic sealer.

Evaluation of cytotoxicity and up-regulation of gelatinases in human fibroblast cells by four root canal sealers

AIMnu2002 To investigate the effects of root canal sealers on the cytotoxicity and gelatinolytic activity of matrix metalloproteinases (MMPs) in human fibroblasts.nnnMETHODOLOGYnu2002 Human fibroblasts (MRC5, 3×10(5) cells per well) were incubated directly or indirectly with AH Plus, Endomethasone N, Pulp Canal Sealer EWT or Sealapex for 30u2003min, 1, 4 or 24u2003h (time-points). The cytotoxicity of all root canal sealers was determined by counting viable cells using the trypan blue exclusion assay. Supernatants of cell cultures incubated with root sealers directly or indirectly were collected after each time-point to determine the levels of MMP-2 and MMP-9 gelatinolytic activity by gelatin zymography. Data were analysed using anova and the Tukeys tests.nnnRESULTSnu2002 Cells secreted MMP-2 after periods of 4 and 24u2003h; however, there were no significant differences between the sealers. Secretion of gelatinases was elevated by root canal sealers in direct contact with the cell monolayer when compared to indirect contact (Pu2003<u20030.05). At the time-points tested, no gelatinolytic activity could be detected in the control group without the sealers. The cytotoxicity results revealed that all sealers were cytotoxic in both contact forms. Sealapex had the lowest cytotoxicity and AH Plus the most cytotoxicity.nnnCONCLUSIONSnu2002 All root canal sealers induced the expression of MMP-2 in MRC5 fibroblasts. AH Plus had the highest cytotoxicity amongst the tested sealers, but all were associated with cytotoxic effects.

Long-term cytotoxic effects of contemporary root canal sealers

Objectives: The aim of the present study was to investigate the effects of root canal sealers on the cytotoxicity of 3T3 fibroblasts during a period of 5 weeks. Material and Methods: Fibroblasts (3T3, 1x105 cells per well) were incubated with elutes of fresh specimens from eight root canal sealers (AH Plus, Epiphany, Endomethasone N, EndoReZ, MTA Fillapex, Pulp Canal Sealer EWT, RoekoSeal and Sealapex) and with elutes of the same specimens for 5 succeeding weeks after immersing in simulated body fluid. The cytotoxicity of all root canal sealers was determined using the MTT assay. Data were analyzed using ANOVA and Tukeys test. Results: RoekoSeal was the only sealer that did not show any cytotoxic effects (p<0.05). All the other tested sealers exhibited severe toxicity initially (week 0). MTA Fillapex remained moderately cytotoxic after the end of experimental period. Toxicity of the other tested sealers decreased gradually over time. The evaluated root canal sealers presented varying degrees of cytotoxicity, mainly in fresh mode. Conclusions: RoekoSeal had no cytotoxic effect both freshly mixed and in the other tested time points. MTA Fillapex was associated with significantly less cell viability when compared to the other tested root canal sealers.

Evaluation of cytotoxicity and up-regulation of gelatinases in fibroblast cells by three root repair materials.

AIMnTo investigate the effects of root repair materials on the cytotoxicity and gelatinolytic activity of matrix metalloproteinases (MMPs) in 3T3 fibroblasts.nnnMETHODOLOGYnFibroblasts (3T3, 3 × 10(5) cells per well) were incubated with elutes of calcium hydroxide (Biodinâmica, Ibiporã, PR, Brazil), EndoBinder (Binderware, São Carlos, SP, Brazil) and mineral trioxide aggregate (MTA) (Angelus, Londrina, PR, Brazil) for 24 h. The cytotoxicity of all root repair materials was determined using the MTT assay. Supernatants of cell cultures incubated with materials were collected after 24u2003h to determine the levels of MMP-2 and MMP-9 gelatinolytic activity by gelatin zymography. Data were analysed using anova and Tukeys test.nnnRESULTSnCells secreted MMP-2 after 24 h with calcium hydroxide inducing significantly greater MMP-2 expression in relation to the control and the other root repair materials (P < 0.05). The cytotoxicity results revealed that there was no significant difference in the cell viability of MTA, EndoBinder and the control group. However, there was a significantly reduced cell viability of 3T3 fibroblasts in association with calcium hydroxide (P < 0.05).nnnCONCLUSIONSnCalcium hydroxide was associated with significantly less cell viability when compared with EndoBinder and MTA. All materials had gelatinolytic activity for MMP-2 with calcium hydroxide being associated with the greatest activity.

A multiparametric assay to compare the cytotoxicity of soy milk with different storage media.

BACKGROUND/AIMnThe aim of this study was to evaluate the cytotoxicity of soy milk compared with several other storage media [coconut water, Hanks Balanced Salt Solution (HBSS) and whole milk], assessed through a multiparametric analysis employing 3T3 cells.nnnMATERIALS AND METHODSnPlates containing confluent 3T3 fibroblasts were exposed to the various media for 24 h, at 37°C with 5% CO₂, and cell viability was evaluated by a multiparametric assay assessing sequentially, on the same cells, mitochondrial activity (XTT), membrane integrity (neutral red test) and total cell density (crystal violet dye exclusion test). Results from each test were compared by two-way analysis of variance (ANOVA).nnnRESULTSnStatistical analysis showed that whole milk, HBSS and soy milk were the most effective media in maintaining cell viability at all tested times (P < 0.05). The least amount of viable cells was observed when using coconut water.nnnCONCLUSIONSnThis study shows that the efficacy of soy milk in maintaining the viability of 3T3 fibroblasts is similar to that of HBSS and milk, as shown by three different cell viability tests.

Influence of the curing mode on the degree of conversion of a dual-cured self-adhesive resin luting cement beneath ceramic

Abstract Objective. To evaluate the effect of the delayed photoactivation and ceramic barrier on the degree of conversion (DC) of self-adhesive resin cement. Materials and methods. Circular specimens (5 mm in diameter × 1 mm in thickness) of the RelyX U-100 resin cement were made using the following curing protocols (n = 10): (G1) 40 s beneath a IPS Empress II ceramic; (G2) 40 s of direct photocuring; (G3) 80 s beneath the ceramic; (G4) 80 s of direct photocuring; (G5) self-curing; (G6) 5 min in the absence of light (self-curing) followed by transceramic photocuring for 40 s; (G7) 5 min in the absence of light (self-curing) followed by transceramic photocuring for 80 s. All the specimens were photoactivated by LED (800 mW/cm2). After 24 h of dry storage, the DC was measured by FTIR, on the top surface of the specimens. Data were submitted to one-way ANOVA and Tukey test (p ≤ 0.05). Results. Direct photocuring with no ceramic interposition, regardless of the curing time (40 s and 80 s) promoted the highest conversion mean (56.79 ± 1.19 and 59.98 ± 2.93, respectively) and the 5 min delay time for the transceramic photocuring presented a similar mean compared to the immediate transceramic photocuring. The DC was influenced by the ceramic barrier, decreasing the conversion values (49.72 ± 1.91 for 40 s and 52.36 ± 2.50 for 80 s), with no statistical difference from the groups with the previous 5 min of photoactivation delay. The self-cure only showed the worst DC values. Conclusion. Direct photocuring provided a higher degree of conversion for the self-adhesive resin cement. The delayed light activation did not influence the degree of conversion for the resin cement tested.

Efficacy of different final irrigant activation protocols on smear layer removal by EDTA and citric acid

The aim of this study was to evaluate the influence of different activation protocols for chelating agents used after chemo‐mechanical preparation (CMP), for smear layer (SL) removal. Forty‐five single‐rooted human premolars with straight canals and fully formed apex were selected. The specimens were randomly divided into three groups depending on the chelating agent used for smear layer removal: distilled water (DW, control group); 17% ethylenediaminetetraacetic acid (EDTA); and 10% citric acid (CA). Each group was further divided into three subgroups according to the activation protocol used: no‐activation (NA), manual dynamic activation (MDA), or sonic activation (SA). After CMP, all specimens were sectioned and processed for observation of the apical thirds by using scanning electron microscopy (SEM). Two calibrated evaluators attributed scores to each specimen. The differences between activation protocols were analyzed with Kruskal‐Wallis and Mann‐Whitney U tests. Friedman and Wilcoxon signed rank tests were used for comparison between each root canal third. When chelating agents were activated, either by MDA or SA, it was obtained the best cleaning results with no significant difference between EDTA and CA (P > 0.05). Sonic activation showed the best results when root canal thirds were analyzed, in comparison to MDA and NA groups (P < 0.05). The activation of chelating agents, independent of the protocol used, benefits smear layer removal from root canals. Microsc. Res. Tech. 76:364–369, 2013.

Evaluation of Gelatinases, Tissue Inhibitor of Matrix Metalloproteinase-2, and Myeloperoxidase Protein in Healthy and Inflamed Human Dental Pulp Tissue

INTRODUCTIONnThe aim of this study was to compare the gelatinolytic activity of matrix metalloproteinase (MMP)-2 and MMP-9 and the expression of tissue inhibitor of matrix metalloproteinase (TIMP)-2 and myeloperoxidase protein (MPO) in clinically healthy human pulp and inflamed pulp tissue specimens.nnnMETHODSnTwenty dental pulps clinically diagnosed as inflammatory tissues and 20 healthy pulp tissues from enclosed third molars were harvested and evaluated. The gelatinolytic activity for MMP-2 and MMP-9 was assessed by using the zymography technique, TIMP-2 gene expression was evaluated using the enzyme-linked immunosorbent assay, and MPO was determined using the MPO assay.nnnRESULTSnData showed increased levels of MMP-9, active MMP-2, TIMP-2, and MPO in inflammatory pulp tissues compared with healthy tissues (P < .05). No statistical difference could be observed for pro-MMP-2 (P > .05).nnnCONCLUSIONSnAlthough all samples were associated with MMP-2 expression, the active form of this MMP was observed only in inflamed pulps. Inflamed pulps showed an up-regulation of MMP-9, TIMP-2, and MPO.

Furcal-perforation repair with mineral trioxide aggregate: Two years follow-up.

Furcal perforations are significant iatrogenic complications of endodontic treatment and could lead to endodontic failure. Mineral trioxide aggregate (MTA) has been regarded as an ideal material for perforation repair, retrograde filling, pulp capping, and apexification. This case report describes a furcal perforation in a maxillary first molar, which was repaired using MTA. The tooth was endodontically treated and coronally restored with resin composite. After 2 years, the absence of periradicular radiolucent lesions, pain, and swelling along with functional tooth stability indicated a successful outcome of sealing the perforation using MTA.

Response of mice connective tissue to three different endodontic materials

The aim of this study was to evaluate the biocompatibility of mineral trioxide aggregate (MTA) Bio and Portland cement (PC) and compare with those of ProRoot MTA. Polyethylene tubes were filled with materials and placed into dorsal subcutaneous connective tissue of Wistar albino rats. After 7, 30, and 60 days after the surgical procedure, the implants with the surrounding tissue were removed. Tissue samples were subjected to histological processing, and sections were stained with hematoxylin and eosin. Sections were evaluated for the intensity of inflammation, predominant cell type, presence of fibrous capsule and granulation tissue. Data were submitted to Kruskal‐Wallis test at a significant level of P ≤ 0.05. No statistical differences were observed at any evaluated condition among tested materials (P > 0.05). Statistically significant differences were observed between mean inflammatory scores, cell types and granulation tissue of the same material in different experimental periods (P < 0.05). Can be concluded that biocompatibility of MTA bio and PC were comparable with that of ProRoot MTA. Microsc. Res. Tech. 76:311–315, 2013.