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The objective of this study was to assess the cytotoxicity of EndoSeal, a novel mineral trioxide aggregate-based root canal sealer in comparison with two commonly used sealers, AH Plus and Sealapex. For cytotoxicity assay, MG-63 cells and human gingival fibroblasts were incubated in culture medium containing eluates of each sealer at 1, 3, and 7 days. Cell metabolism was evaluated by the WST-1 assay. For cell adhesion assay, disc specimens were fabricated from EndoSeal and AH Plus. MG-63 cells and human gingival fibroblasts were seeded on the discs, and after 1 day and 7 days of incubation, cell morphology and cell adhesion were examined by SEM. The order of cytotoxicity of root canal sealers was as follows: EndoSeal < AH Plus < Sealapex. Both types of cells seeded on the EndoSeal specimens were much larger, flat with rough margins compared to those on the AH Plus specimens. These results suggest that EndoSeal has a satisfactory cytocompatibility.

Bioceramic processing using rapid prototyping technique (RPT) results in a fragile device that requires thermal treatment to improve the mechanical properties. This investigation evaluates the effect of thermal treatment on the mechanical, porosity, and bioactivity properties as well as the cytotoxicity of a porous silica-calcium phosphate nanocomposite (SCPC) implant prepared by RPT. Porous SCPC implant was subject to 3-h treatment at 800°C, 850°C, or 900°C. The compressive strength (s) and modulus of elasticity (E) were doubled when the sintering temperature is raised from 850 to 900°C measuring (s = 15.326 ± 2.95 MPa and E = 1095 ± 164 MPa) after the later treatment. The significant increase in mechanical properties takes place with minimal changes in the surface area and the percentage of pores in the range 1-356 μm. The SCPC implant prepared at 900°C was loaded with rh-BMP-2 and grafted into a segmental defect in the rabbit ulna. Histology analyses showed highly vascularized bone formation inside the defect. Histopathological analyses of the liver, spleen, kidney, heart, and the lung of rabbits grafted with and without SCPC demonstrated healthy tissues with no signs of toxicity or morphology alterations. Results of the study suggest that it is possible to engineering the mechanical properties of the SCPC implant without compromising its bioactivity. The enhanced bone formation inside the porous SCPC facilitated cell-mediated graft resorption and prohibited any accumulation of the material in the body organs., (Copyright © 2013 Wiley Periodicals, Inc., a Wiley Company.)

The Cosmetic Ingredient Review Expert Panel (Panel) reviewed the safety of calcium sodium borosilicate, calcium aluminum borosilicate, calcium titanium borosilicate, silver borosilicate, and zinc borosilicate as used in cosmetics. These borosilicate glasses function mostly as bulking agents. Available animal and human data were considered along with data from a previous safety assessment of magnesium silicates. The similar structure, properties, functions, and uses of these ingredients enabled grouping them and using the available toxicological data to assess the safety of the entire group. Data submitted on calcium borosilicate, which is not a cosmetic ingredient, are also included as additional support for the safety of borosilicate glass ingredients. The Panel concluded that borosilicate glasses are safe as cosmetic ingredients in the practices of use and concentration as given in this safety assessment.

Aim: To investigate and compare the systemic toxic effect of DiaRoot BioAggregate and grey ProRoot Mineral trioxide aggregate (MTA) on the liver and kidney after 7 and 30 days., Methodology: Forty-two white albino rats were divided into two main groups. Group (1), considered the control group (n = 18), was further divided into two subgroups. The negative control subgroup (n = 6) received no treatment. The empty tube subgroup (n = 12) received empty sterile Teflon tubes. In Group (2), considered the experimental group (n = 24), the rats were divided equally into two subgroups. One subgroup received MTA, whilst the other received BioAggregate. The materials in the Teflon tubes were implanted subcutaneously in the dorsal side of the rats. Blood samples were taken to investigate the change of kidney and liver functions on day 7 and day 30. The liver and kidney organs were subjected to histopathological examination and calculation of the number of inflammatory cells. Data analysis was performed using one-way anova with post hoc multiple comparisons with the Tukey's test. Student's t-test was used to compare the changes in liver and kidney functions amongst the groups., Results: On day 7, a significantly more severe inflammatory reaction was observed in both experimental subgroups compared with the control (P < 0.05); the severity decreased after 30 days. The kidney functions were not affected after 7 days but had subsequently increased after 30 days (P < 0.001). Liver functions increased after 7 days and had decreased in the BioAggregate subgroup after 30 days, whilst in the MTA subgroup, a continuous increase in the level of liver function was observed., Conclusions: Mineral trioxide aggregate had adverse effects on the liver and kidney that were significantly more severe than BioAggregate but with no permanent damage., (© 2013 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Objectives: The purpose of this study was to evaluate and compare the cytotoxic effects of ProRoot MTA and DiaRoot BA, a bioceramic nanoparticulate cement, on subcutaneous rat tissue., Study Design: Fifty Sprouge Dawley rats were used in this study. Polyethylene tubes filled with ProRoot MTA and DiaRoot BioAggregate, along with a control group of empty, were implanted into dorsal connective tissue of rats for 7, 15, 30, 60, and 90 days. After estimated time intervals the rats were sacrificed. The specimens were fixed, stained with hematoxylin and eosin, and then evaluated under a light microscope for inflammatory reactions and mineralization., Results: All groups evoked a severe to moderate chronic inflammatory reaction at 7 and 15 days, which decreased with time. Both the MTA and BioAggregate groups showed similar inflammatory reactions, except at 90 days when MTA showed statistically significant greater inflammation (p>0.05). The MTA group showed foreign body reaction at all times. Compared to BioAggregate, MTA showed significantly more foreign body reaction at 60 and 90 days (p<0.0001). After 30 days foreign body reaction of BioAggregate decreased significantly. Both MTA and BioAggregate groups showed similar necrosis at 7 and 15 days (p=0.094 and p=0.186 respectively). No necrosis was observed after 15 days. Similarly there was no fibrosis after 30 days for both MTA and BioAggregate groups (p>0.05)., Conclusions: Since DiaRoot BioAggregate showed significantly better results than MTA, we can conclude that it is more biocompatible. However, further studies are required to confirm this result.

Objective: The aim of this study was to compare the cytotoxic effects of endodontic cements on human tooth germ stem cells (hTGSCs). MTA Fillapex, a mineral trioxide aggregate (MTA)-based, salicylate resin containing root canal sealer, was compared with iRoot SP, a bioceramic sealer, and AH Plus Jet, an epoxy resin-based root canal sealer., Material and Methods: To evaluate cytotoxicity, all materials were packed into Teflon rings (4 mmµ3 mm) and co-cultured with hTGSCs with the aid of 24-well Transwell permeable supports, which had a pore size of 0.4 µm. Coverslips were coated with MTA Fillapex, iRoot SP and AH Plus Jet and each coverslip was placed onto the bottom of one well of a six-well plate for scanning electron microscopy (SEM) analysis. Before the cytotoxicity and SEM analysis, all samples were stored at 37ºC and at 95% humidity and 5% CO2 for 24 hours to set. The cellular viability was analyzed using MTS test (3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium). The cytotoxic effects and SEM visualization of the tested materials were analyzed at 24-hour, 72-hour, one-week and two-week periods., Results: On the 1st day, only MTA Fillapex caused cytotoxicity compared to negative control (NC) group (p<0.008). No significant difference was observed between the other tested materials at this period (p>0.05). After 14 days of incubation with the test materials, MTA Fillapex exhibited significantly higher cytotoxicity compared with iRoot SP, AH Plus Jet and the NC group (P<0.008). In the SEM analysis, the highest levels of cell attachment were observed for iRoot SP and the control group. After 24 hours, MTA Fillapex reduced the number of cells attached to the surface., Conclusions: Within the limitations of this study, sealers exerted different cytotoxic effects on hTGSCs. Although all materials have exerted cellular toxicity, iRoot SP and AH Plus Jet may promote better attachment to hTGSCs.

Aim: To characterize and investigate the genotoxic effect of a new endodontic cement based on dicalcium- and tricalcium-silicate (CS) with hydroxyapatite (HA) on human lymphocytes., Methodology: Hydrothermal treatment was applied for synthesis of CS and HA. The final mixture HA-CS, with potential to be used in endodontic practice, is composed of CS (34%) and HA (66%). Human lymphocytes were incubated with HA, HA-CS and CS for 1 h, at 37 °C and 5% CO2. Cell viability was determined using the trypan blue exclusion assay. To evaluate the level of DNA damage comet assay (single cell gel electrophoresis) was performed. For the statistical analysis anova and Duncan's Post Hoc Test were used., Results: The SEM analysis indicated that CS consisted mostly of agglomerates of several micrometers in size, built up from smaller particles, with dimensions between 117 and 477 nm. This is promising because dimensions of agglomerates are not comparable with channels inside the cell membranes, whereas their nano-elements provide evident activity, important for faster setting of these mixtures compared to MTA. Values of DNA damage obtained in the comet assay indicated low genotoxic risk of the new endodontic materials., Conclusions: The significantly improved setting characteristics and low genotoxic risk of the new material support further research., (© 2012 International Endodontic Journal.)

Rationale: Occupational coexposure to asbestos and other fibers or particles could modify the carcinogenicity of asbestos with regard to pleural mesothelioma., Objectives: To estimate associations between pleural mesothelioma and occupational mineral wool and silica exposure and to study the impact of occupational coexposure on the risk of pleural mesothelioma., Methods: A total of 1,199 male cases and 2,379 control subjects were included in a French pooled case-control study. Complete job histories were collected, and occupational exposure to asbestos, mineral wool (MW), and silica were assessed by three French job exposure matrices. Unconditional logistic regression models adjusted for age, birth date, and occupational asbestos exposure were used to estimate odds ratios (OR) and 95% confidence intervals (CIs)., Measurements and Main Results: A significant association between mesothelioma and MW exposure was observed after adjustment for occupational asbestos exposure. OR for subjects exposed to less than 0.01 fibers·ml(-1)·yr(-1) was 1.6 (95% CI, 1.2-2.1) and increased to 2.5 (95% CI, 1.8-3.4) for subjects exposed to more than 0.32 fibers·ml(-1)·yr(-1). All ORs for silica exposure were around the null. Coexposure to either asbestos and MW or asbestos and silica seemed to increase the risk of pleural mesothelioma. ORs were 17.6 (95% CI, 11.8-26.2) and 9.8 (95% CI, 4.2-23.2) for subjects exposed to both asbestos and MW and for subjects exposed to both asbestos and silica, respectively, compared with 4.3 (95% CI, 1.9-9.8) for occupational asbestos exposure alone., Conclusions: Our results are in favor of an increased risk of pleural mesothelioma for subjects exposed to both asbestos and MW or asbestos and silica.

Introduction: This study examined the effect of a new bioactive dentin substitute material (Biodentine) on the viability of human gingival fibroblasts., Methods: Biodentine, White ProRoot mineral trioxide aggregate (MTA), and glass ionomer cement were evaluated. Human gingival fibroblasts were incubated for 1, 3, and 7 days both in the extracts from immersion of set materials in culture medium and directly on the surface of the set materials immersed in culture medium. Fibroblasts cultured in Dulbecco modified Eagle medium were used as a control group. Cytotoxicity was evaluated by flow cytometry, and the adhesion of human gingival fibroblasts to the surface of the set materials was assessed by using scanning electron microscopy. The data of cell cytotoxicity were analyzed statistically by using a one-way analysis of variance test at a significance level of P< .05., Results: Cells exposed to extracts from Biodentine and MTA showed the highest viabilities at all extract concentrations, whereas cells exposed to glass ionomer cement extracts displayed the lowest viabilities (P< .05). There was no significant difference in cell viabilities between Biodentine and MTA during the entire experimental period (P> .05). Human gingival fibroblasts in contact with Biodentine and MTA attached to and spread over the material surface after an overnight culture and increased in numbers after 3 and 7 days of culture., Conclusions: Biodentine caused gingival fibroblast reaction similar to that by MTA. Both materials were less cytotoxic than glass ionomer cement., (Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

This study aimed to evaluate sublethal effects of a short-term exposure (96 h) to titanium silicate nanoparticles (TiSiO(4)-NP) on Pelophylax perezi tadpoles. Tadpoles were exposed to five concentrations of TiSiO(4)-NP (8.2, 10.2, 12.8, 16 and 20 mg/L) plus a control. Effect criteria were: mortality, cholinesterases, glutathione S-transferases, lactate dehydrogenase, and catalase activities, and alanine and lactate contents. Light scattering was used for physical characterization of TiSiO(4)-NP suspensions, revealing a high aggregation state of the NP, consistent with low z-potential values (<30 mV). Mortality among TiSiO(4)-NP treatments was <11%. Significant differences relatively to the control were observed at the biochemical level (for CAT and LDH) and in lactate and alanine contents, which may end-up in increased oxidative stress. Overall, some of the monitored endpoints suggest metabolic alterations in TiSiO(4)-NP exposed tadpoles, highlighting the potential of TiSiO(4)-NP long-term effects on these organisms., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Introduction: The 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., Methods: Cytotoxicity, 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%., Results: In 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)., Conclusions: Although MTA Fillapex was more cytotoxic than AH Plus, it showed suitable physicochemical properties for an endodontic sealer., (Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

The aim of this study was to compare the in vitro cytotoxicity of white mineral trioxide aggregate (MTA), MTA Fillapex® and Portland cement (PC) on human cultured periodontal ligament fibroblasts. Periodontal ligament fibroblast culture was established and the cells were used for cytotoxic tests after the fourth passage. Cell density was set at 1.25 X10 4 cells/well in 96-well plates. Endodontic material extracts were prepared by placing sealer/cement specimens (5x3mm) in 1mL of culture medium for 72 h. The extracts were then serially two-fold diluted and inserted into the cell-seeded wells for 24, 48 and 72 h. MTT assay was employed for analysis of cell viability. Cell supernatants were tested for nitric oxide using the Griess reagent system. MTA presented cytotoxic effect in undiluted extracts at 24 and 72 h. MTA Fillapex® presented the highest cytotoxic levels with important cell viability reduction for pure extracts and at ½ and ¼ dilutions. In this study, PC did not induce alterations in fibroblast viability. Nitric oxide was detected in extract-treated cell supernatants and also in the extracts only, suggesting presence of nitrite in the soluble content of the tested materials. In the present study, MTA Fillapex displayed the highest cytotoxic effect on periodontal ligament fibroblasts followed by white MTA and PC.

The aim of this study was to evaluate the response of rat subcutaneous tissue to MTA Fillapex® (Angelus), an experimental root canal filling material based on Portland cement and propylene glycol (PCPG), and a zinc oxide, eugenol and iodoform (ZOEI) paste. These materials were placed in polyethylene tubes and implanted into the dorsal connective tissue of Wistar rats for 7 and 15 days. The specimens were stained with hematoxylin and eosin, and evaluated regarding inflammatory reaction parameters by optical microscopy. The intensity of inflammatory response against the sealers was analyzed by two blinded and previously calibrated examiners for all experimental periods (kappa=0.96). The histological evaluation showed that all materials caused a moderate inflammatory reaction at 7 days, which subsided with time. A greater inflammatory reaction was observed at 7 days in the tubes filled with ZOEI paste. Tubes filled with MTA Fillapex presented some giant cells, macrophages and lymphocytes after 7 days. At 15 days, the presence of fibroblasts and collagen fibers was observed indicating normal tissue healing. The tubes filled with PCPG showed similar results to those observed in MTA Fillapex. At 15 days, the inflammatory reaction was almost absent at the tissue, with several collagen fibers indicating normal tissue healing. Data were analyzed by the nonparametric Kruskal-Wallis test (α=0.05). Statistically significant difference (p<0.05) was found only between PCPG at 15 days and ZOEI at 7 days groups. No significant differences were observed among the other groups/periods (p>0.05). MTA Fillapex and Portland cement added with propylene glycol had greater tissue compatibility than the PCPG paste.

Introduction: Various materials have been advocated for use as root-end filling materials. The purpose of the present in vitro study was to compare the cytotoxicity of 4 root-end filling materials: glass ionomer cement (GIC; Fuji II, GC Corp, Tokyo, Japan), reinforced zinc oxide-eugenol cement (IRM; Dentsply Tulsa Dental, Tulsa, OK), and 2 types of mineral trioxide aggregate., Methods: This study used MG-63 cells derived from a human osteosarcoma. To quantitatively evaluate the cytotoxicity of test materials, the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Each specimen was examined by scanning electron microscopy for the observation of cell morphology., Results: The XTT assay showed that the cell viability of ProRoot MTA (Dentsply Tulsa Dental) was higher than that of GIC and Ortho MTA (BioMTA, Seoul, Republic of Korea) at all time points. IRM showed significantly lower cell viability than the other groups. The scanning electron microscopic analysis revealed that elongated, dense, and almost confluent cells were observed in the cultures of GIC, Ortho MTA, and ProRoot MTA specimens. In contrast, cells on the surface of IRM were rounded in shape, and the numbers and the density of the cells were smaller than that in the other groups., Conclusions: ProRoot MTA and GIC showed good biocompatibility in this study. However, Ortho MTA showed lower biocompatibility compared with ProRoot MTA and GIC., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Intensive study of the natural asbestiform minerals that cause human diseases, and the consequent understanding of their hazardous characteristics, has enabled the development of manufactured fibres whose physical and/or chemical properties, in particular as they relate to biopersistence, have been adjusted to minimize possible harm to health. A strong driver for the developmentof new high temperature insulation materials wasthe perception of the toxicity of refractory ceramic fibres (RCF)and their classification in the EU as a category 2 carcinogen under Directive 67/548/EEC. Such classification carries with it the requirement for substitution by less hazardous materials. This paper focuses on the development of alkaline earth silicate (AES) wools as a new class of high temperature insulation with the capability of such substitution in a number of applications. These wools have only a low potential to cause harm because they do not persist in lung tissue once deposited, and have produced minimal effects in experimental test systems. AES wools are increasingly being used in a wide range of high temperature applications., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Introduction: The purpose of this study was to evaluate the subcutaneous connective tissue reaction in rats to a mineral trioxide aggregate (MTA)-based endodontic sealer Fillapex (Angelus, Londrina, PR, Brazil) and compare it with Grossman sealer (Farmadental, Buenos Aires, Argentina)., Methods: Sterile medical-grade silicone tubes containing the test materials were implanted in 24 Wistar rats. After 10, 30, and 90 days, the animals (n = 8 per period) were euthanized, and the implants along with their surrounding tissues were dissected, fixed, and processed for histologic evaluation. A 4-category evaluation system was used to evaluate the microscopic observations. The tissue response on the lateral walls of the silicone tubes was used as the negative control. The data were analyzed for statistical significance using the Wilcoxon signed rank, Kruskal-Wallis, and Dunn tests., Results: Fillapex showed a severe tissue reaction for all 3 observation periods. Grossman sealer showed similar features after 10 and 30 days, but the reaction decreased slightly after 90 days. In contrast, the negative controls did not show adverse reactions in any sample of the 3 time periods. After 10 and 30 days, no statistically significant differences were found between Fillapex and Grossman sealer (P > .05); however, the difference was significant after 90 days (P < .05). For all experimental periods, there were statistically significant differences between both Fillapex and Grossman sealer and the negative controls (P < .05)., Conclusions: It was concluded that both MTA-Fillapex and Grossman sealer remained toxic to subcutaneous tissues in rats after 90 days., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: To investigate the effects of root repair materials on the cytotoxicity and gelatinolytic activity of matrix metalloproteinases (MMPs) in 3T3 fibroblasts., Methodology: Fibroblasts (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 24 h to determine the levels of MMP-2 and MMP-9 gelatinolytic activity by gelatin zymography. Data were analysed using anova and Tukey's test., Results: Cells 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)., Conclusions: Calcium 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., (© 2012 International Endodontic Journal.)

Introduction: The purpose of this in vitro study was to evaluate the cytotoxicity and alkaline phosphatase (ALP) activity of a new bioceramic root repair material, EndoSequence Root Repair Material (ESRRM; Brasseler USA, Savannah, GA), and to compare these characteristics with those of ProRoot MTA (Dentsply Tulsa Dental, Tulsa, OK) and Geristore (GR; Den-Mat LLC, Santa Maria, CA)., Methods: Human Saos-2 osteoblast-like cells were exposed to 1-, 3-, and 7-day elutes of the materials (100% and 50% strength) for 24 hours after which the bioactivity and ALP activity of the cells were evaluated using a methylthiazol sulfophenyl (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay and para-Nitrophenylphosphate colorimetric assay, respectively. In the positive control group, Triton X-100 (Boehringer Mannheim Corp, Indianapolis, IN) was used to lyse the cells, representing 100% cytotoxicity, and in the negative control group cells received fresh culture medium only. Data were statistically analyzed using the unpaired t test and 1-way analysis of variance., Results: The results revealed that the bioactivity of the cells as well as ALP activity were significantly decreased after exposure to ESRRM elutes in almost all time periods, both in 100% and 50% concentrations, with the exception of ALP activity of day 1 elutes of ESRRM at 50% concentration. MTA did not change the bioactivity or ALP activity of the cells. GR elutes of 100% concentration reduced the bioactivity on days 1 and 3, whereas GR elutes of 50% concentration affected the cells only on day 1. None of the GR elutes had any effect on ALP activity of the cells., Conclusions: It was concluded that ESRRM elutes of all time periods in general reduced the bioactivity and ALP activity of osteoblast-like cells. GR reduced bioactivity only, whereas MTA had no effect on the cells., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: To investigate the cytotoxicity of three flowable resin composites that potentially useful as retrograde filling materials, compared with mineral trioxide aggregate (MTA)., Methods: Ten standard cylinder discs were used for each of the tested materials: Tetric Flow, Filtex Flow, Aeliteflo, and MTA, which were prepared under aseptic conditions. Cytotoxicity of eluates from all materials after 1-4 days' immersion in culture medium and direct contact cytotoxicity were evaluated using cultured human periodontal ligament cells (PDLC). The colorimetric (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and scanning electron microscope of cell morphology (direct contact only) were used., Results: No eluates of set materials demonstrated cytotoxicity at any concentration or elution time. Freshly-mixed MTA was cytotoxic in direct contact, but not set MTA. Freshly-mixed Aeliteflo was also cytotoxic, as was set material up to 2 days' elution. With morphological assessment, some differences were seen among resin composites, but all changes were rated as slight using the International Standard Organization criteria., Conclusions: Of all of the materials tested, Tetric Flow showed the least cytotoxic effects on PDLC. Further research is needed to determine the clinical usefulness of flowable composites as retrograde filling materials., (© 2012 Blackwell Publishing Asia Pty Ltd.)

Nanotechnology creates new possibilities to control and improve material properties for civil infrastructure. Special focus in this area is put on Portland cement and gypsum. Together their annual production is by far larger than for any other material worldwide. Nanomodification of these materials can be done during the few hours between dissolution and hardening, especially by nucleation of the re-crystallization with suitable colloids. Here we report first results in homogeneous seeding of the precipitation of calcium silicate hydrates within a real Portland cement composition. The occupational safety during the production phase and during mixing of concrete paste is addressed in detail by in vivo testing. We perform 5-day inhalation with 21-day recovery in rats and analyze organ-specific toxicity and 71 endpoints from bronchoalveolar lavage (BALF) and blood. In BALF parameters, no test-related changes were observed, indicating the generally low toxicity of the test material. Some mild lesions were observed in larynx level. In the lungs, all animals of the 50 mg/m³ concentration group revealed a minimal to mild increase in alveolar macrophages, which recovered back to control level.

Introduction: Quick-setting calcium aluminosilicate cement with improved washout resistance is a potential substitute for calcium silicate cements in endodontics. This study examined the effect of an experimental calcium aluminosilicate cement (Quick-Set; Primus Consulting, Bradenton, FL) on the viability of odontoblast-like cells., Methods: The biocompatibility of Quick-Set and white ProRoot MTA (WMTA; Dentsply Tulsa Dental Specialties, Tulsa, OK) cements and their eluents was evaluated using a murine dental papilla-derived odontoblast-like cell line (MDPC-23); 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to examine the effects of the 2 hydraulic cements on mitochondrial metabolic activity. Flow cytometry and confocal laser scanning microscopy were used to identify the effects of the 2 cements on cell death-induced plasma membrane permeability to fluorescent dyes and DNA stains., Results: After the first week of immersion in culture medium, Quick-Set and WMTA were more cytotoxic than the Teflon-negative control (P < .05), and the cells exhibited more apoptosis/necrosis than Teflon (P < .05). After the second week of immersion, the 2 cements were as biocompatible as Teflon (P > .05), with cells exhibiting minimal apoptosis/necrosis. Eluents from the set cements at 1:1 dilution were significantly more cytotoxic that eluents at 1:10 or 1:100 dilution (P < .05)., Conclusions: Quick-Set and WMTA exhibited similar cytotoxicity profiles. They possess negligible in vitro toxicologic risks after time-dependent elution of toxic components., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: The main purpose of this study was to evaluate the biocompatibility and bioactivity of a new mineral trioxide aggregate (MTA)-based endodontic sealer, MTA Fillapex (MTA-F; Angelus, Londrina, Brazil), in human cell culture., Methods: Human osteoblast-like cells (Saos-2) were exposed for 1, 2, 3, and 7 days to MTA-F, Epiphany SE (EP-SE; SybronEndo, Orange, CA), and zinc oxide-eugenol sealer (ZOE). Unexposed cultures were the control group (CT). The viability of the cells was assessed by MTT assay and the morphology by scanning electron microscopy (SEM). The bioactivity of MTA-F was evaluated by alkaline phosphatase activity (ALP) and the detection of calcium deposits in the culture with alizarin red stain (ARS). Energy-dispersive X-ray spectroscopy (EDS) was used to chemically characterize the hydroxyapatite crystallites (HAP). Saos-2 cells were cultured for 21 days for ARS and SEM/EDS. ARS results were expressed as the number of stained nodules per area. Statistical analysis was performed with analysis of variance and Bonferroni tests (P < .01)., Results: MTA-F exposure for 1, 2, and 3 days resulted in increased cytotoxicity. In contrast, viability increased after 7 days of exposure to MTA-F. Exposure to EP-SE and ZOE was cytotoxic at all time points. At day 7, ALP activity increase was significant in the MTA-F group. MTA-F presented the highest percentage of ARS-stained nodules (MTA-F > CT > EP-SE > ZOE). SEM/EDS analysis showed hydroxyapatite crystals only in the MTA-F and CT groups. In the MTA-F group, crystallite morphology and chemical composition were different from CT., Conclusions: After setting, the cytotoxicity of MTA-F decreases and the sealer presents suitable bioactivity to stimulate HAP crystal nucleation., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Context: Inhalation of asbestos or silica is associated with chronic and progressive diseases, including fibrosis, cancer, and increased risk of systemic autoimmunity. Because there is a need for treatment options for these diseases, a better understanding of their mechanistic etiologies is essential. While oxidative stress in macrophages is an early consequence of these exposures, it may also serve as a signaling mechanism involved in downstream immune dysregulation. The system x(c)(-) exchange protein is induced by oxidative stress, and exchanges equimolor levels of extracellular cystine for intracellular glutamate. Cystine is subsequently reduced to cysteine, the rate-limiting precursor for glutathione synthesis., Objective: As the primary transporter responsible for cystine/glutamate exchange on macrophages, system x(c)- was hypothesized to be inducible in response to asbestos and silica, and to increase viability through protection from oxidative stress., Results: When challenged with amphibole asbestos, but not crystalline silica, RAW 264.7 macrophages increased expression of xCT and the rate of cystine/glutamate exchange in sodium-free conditions. This upregulation was prevented with N-acetylcysteine, implicating oxidative stress. Cystine protected the macrophages from asbestos-induced oxidative stress and cell death, supporting the hypothesis that imported cystine was used for synthesis of cellular antioxidants. System x(c)(-) inhibitors, glutamate and S-4-carboxyphenylglycine ((S)-4-CPG), significantly increased oxidative stress and cell death of asbestos-treated macrophages., Conclusion: System x(c)(-) plays a critical role in survival of macrophages exposed to asbestos, but not silica. These data demonstrate a very early difference in the cellular response to these silicates that may have important downstream implications in the pathologic outcome of exposure.

Exposure to some manufactured beryllium compounds via skin contact or inhalation can cause sensitization. A portion of sensitized persons who inhale beryllium may develop chronic beryllium disease (CBD). Little is understood about exposures to naturally occurring beryllium minerals. The purpose of this study was to assess the bioaccessibility of beryllium from bertrandite ore. Dissolution of bertrandite from two mine pits (Monitor and Blue Chalk) was evaluated for both the dermal and inhalation exposure pathways by determining bioaccessibility in artificial sweat (pH 5.3 and pH 6.5), airway lining fluid (SUF, pH 7.3), and alveolar macrophage phagolysosomal fluid (PSF, pH 4.5). Significantly more beryllium was released from Monitor pit ore than Blue Chalk pit ore in artificial sweat buffered to pH 5.3 (0.88 ± 0.01% vs. 0.36 ± 0.00%) and pH 6.5 (0.09 ± 0.00% vs. 0.03 ± 0.01%). Rates of beryllium released from the ores in artificial sweat were faster than previously measured for manufactured forms of beryllium (e.g., beryllium oxide), known to induce sensitization in mice. In SUF, levels of beryllium were below the analytical limit of detection. In PSF, beryllium dissolution was biphasic (initial rapid diffusion followed by latter slower surface reactions). During the latter phase, dissolution half-times were 1,400 to 2,000 days, and rate constants were ~7 × 10(-10) g/(cm(2)·day), indicating that bertrandite is persistent in the lung. These data indicate that it is prudent to control skin and inhalation exposures to bertrandite dusts.

Maximum contaminant levels are used to control potential health hazards posed by chemicals in drinking water, but no primary national or international limits for aluminum (Al) have been adopted. Given the differences in toxicological profiles, the present evaluation derives total allowable concentrations for certain water-soluble inorganic Al compounds (including chloride, hydroxide, oxide, phosphate and sulfate) and for the hydrated Al silicates (including attapulgite, bentonite/montmorillonite, illite, kaolinite) in drinking water. The chemistry, toxicology and clinical experience with Al materials are extensive and depend upon the particular physical and chemical form. In general, the water solubility of the monomeric Al materials depends on pH and their water solubility and gastrointestinal bioavailability are much greater than that of the hydrated Al silicates. Other than Al-containing antacids and buffered aspirin, food is the primary source of Al exposure for most healthy people. Systemic uptake of Al after ingestion of the monomeric salts is somewhat greater from drinking water (0.28%) than from food (0.1%). Once absorbed, Al accumulates in bone, brain, liver and kidney, with bone as the major site for Al deposition in humans. Oral Al hydroxide is used routinely to bind phosphate salts in the gut to control hyperphosphatemia in people with compromised renal function. Signs of chronic Al toxicity in the musculoskeletal system include a vitamin D-resistant osteomalacia (deranged membranous bone formation characterized by accumulation of the osteoid matrix and reduced mineralization, reduced numbers of osteoblasts and osteoclasts, decreased lamellar and osteoid bands with elevated Al concentrations) presenting as bone pain and proximal myopathy. Aluminum-induced bone disease can progress to stress fractures of the ribs, femur, vertebrae, humerus and metatarsals. Serum Al ≥100 µg/L has a 75-88% positive predictive value for Al bone disease. Chronic Al toxicity is also manifest in the hematopoietic system as an erythropoietin-resistant microcytic hypochromic anemia. Signs of Al toxicity in the central nervous system (speech difficulty to total mutism to facial grimacing to multifacial seizures and dyspraxia) are related to Al accumulation in the brain and these symptoms can progress to frank encephalopathy. There are four groups of people at elevated risk of systemic Al intoxication after repeated ingestion of monomeric Al salts: the preterm infant, the infant with congenital uremia and children and adults with kidney disease. There is a dose-dependent increase in serum and urinary Al in people with compromised renal function, and restoration of renal function permits normal handling of systemically absorbed Al and resolution of Al bone disease. Clinical experience with 960 mg/day of Al(OH)(3) (~5 mg Al/kg-day) given by mouth over 3 months to men and women with compromised renal function found subclinical reductions in hemoglobin, hematocrit and serum ferritin. Following adult males and females with reduced kidney function found that ingestion of Al(OH)(3) at 2.85 g/day (~40 mg/kg-day Al) over 7 years increased bone Al, but failed to elicit significant bone toxicity. There was one report of DNA damage in cultured lymphocytes after high AlCl(3) exposure, but there is no evidence that ingestion of common inorganic Al compounds presents an increased carcinogenic risk or increases the risk for adverse reproductive or developmental outcomes. A number of studies of Al exposure in relation to memory in rodents have been published, but the results are inconsistent. At present, there is no evidence to substantiate the hypothesis that the pathogenesis of Alzheimer's Disease is caused by Al found in food and drinking water at the levels consumed by people living in North America and Western Europe. Attapulgite (palygorskite) has been used for decades at oral doses (recommended not to exceed two consecutive days) of 2,100 mg/day in children of 3-6 years, 4,200 mg/day in children of 6-12 years, and 9,000 mg/day in adults. Chronic ingestion of insoluble hydrated Al silicates (in kg) can result in disturbances in iron and potassium status, primarily as a result of clay binding to intestinal contents and enhanced fecal iron and zinc elimination. Sufficiently high doses of ingested Al silicates (≥50 g/day) over prolonged periods of time can elicit a deficiency anemia that can be corrected with oral Fe supplements. There is essentially no systemic Al uptake after ingestion of the hydrated Al silicates. Rats fed up to 20,000 ppm Ca montmorillonite (equivalent to 1,860 ppm total Al as the hydrated Al silicate) for 28 weeks failed to develop any adverse signs. The results of dietary Phase I and II clinical trials conducted in healthy adult volunteers over 14 days and 90 days with montmorillonite found no adverse effects after feeding up to 40 mg/kg-day as Al. Since the Al associated with ingestion of hydrated Al silicates is not absorbed into the systemic circulation, the hydrated Al silicates seldom cause medical problems unless the daily doses consumed are substantially greater than those used clinically or as dietary supplements. A no-observable-adverse-effect-level (NOAEL) of 13 mg/kg-day as total Al can be identified based on histologic osteomalacia seen in adult hemodialysis patients given Al hydroxide for up to 7 years as a phosphate binder. Following U.S. EPA methods for calculation of an oral reference dose (RfD), an intraspecies uncertainty factor of 10x was applied to that value results in a chronic oral reference dose (RfD) of 1.3 mg Al/kg-day; assuming a 70-kg adult consumes 2 L of drinking water per day and adjusting for a default 20% relative source contribution that value corresponds to a drinking water maximum concentration of 9 mg/L measured as total Al. A chronic NOAEL for montmorillonite as representative of the hydrated Al silicates was identified from the highest dietary concentration (20,000 ppm) fed in a 28-week bioassay with male and female Sprague-Dawley rats. Since young rats consume standard laboratory chow at ~23 g/day, this concentration corresponds to 56 mg Al/kg-day. Application of 3x interspecies uncertainty factor and a 3x factor to account for study duration results in a chronic oral RfD of 6 mg Al/kg-day. Of note, this RfD is 5-10 fold less than oral doses of Al silicates consumed by people who practice clay geophagy and it corresponds to a maximum drinking water concentration of 40 mg Al/L. To utilize the values derived here, the risk manager must recognize the particular product (e.g., alum) or source (e.g., groundwater, river water, clay or cement pipe) of the Al found in tap water, apply the appropriate analytical methods (atomic absorption, energy dispersive X-ray diffraction, infrared spectral analysis and/or scanning transmission electron microscopy) and compare the results to the most relevant standard. The drinking water concentrations derived here are greater than the U.S. EPA secondary maximum contaminant level (MCL) for total Al of 0.05-0.2 mg/L [40 CFR 143.3]. As such, domestic use of water with these concentrations is likely self-limiting given that its cloudy appearance will be greater than the maximum permitted (0.5-5.0 nephalometric turbidity units; 40 CFR Parts 141 and 142). Therefore, the organoleptic properties of Al materials in water determine public acceptance of potable water as contrast to any potential health hazard at the concentrations ordinarily present in municipal drinking water.

Introduction: MTA has good biological properties, and it is a mineralization-inducing material with different indications in endodontics. Initially this material was not recommended as root canal sealer. However, a resin sealer based on mineral trioxide aggregate (MTA Fillapex) was recently released with this indication. Because MTA is in contact with the periodontal tissues, bone, and pulp, it is important to know its cytotoxic and genotoxic effects. The purpose of this study was to evaluate the cytotoxicity and genotoxicity of MTA canal sealer (Fillapex) compared with white MTA cement and AH Plus., Methods: Chinese hamster fibroblasts (V79) were placed in contact with different dilutions of culture media previously exposed to such materials. Cytotoxicity was evaluated by methol-thiazol-diphenyl tetrazolium assay in spectrophotometer to check the viability rate and cell survival. The genotoxicity was accessed by the micronucleus formation assay. Cell survival rate and micronuclei number were assessed before and after exposure to cement extracts, and the results were statistically analyzed by Kruskal-Wallis and Dunn tests (P < .05)., Results: The results showed that the cell viability remained above 50% in white MTA group for all dilutions. AH Plus induced an intermediate cytotoxicity in a dilution-dependent manner, followed by Fillapex MTA., Conclusions: White MTA group was the less cytotoxic material in this study. Both AH Plus and Fillapex MTA sealer showed the lowest cell viability rates and caused an increased micronucleus formation when compared with control untreated group., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

From the archives of the National Toxicology Program, National Institutes of Health, kidney sections from twenty-four carcinogenicity studies (representing twenty-three chemicals) in male and female F344 rats were histopathologically re-evaluated to grade the severity of chronic progressive nephropathy (CPN) on an expanded scale of 0-8, and to record the presence of renal tubule tumors (RTT) and their precursor, atypical tubule hyperplasia (ATH). The data were statistically analyzed using SAS software for logistic regression analysis. This histopathological survey of 2,436 F344 rats showed clear evidence of a qualitative and statistically significant association between advanced stages of CPN severity and the development of low-grade RTT and ATH. Advanced CPN severity therefore represents a risk factor for the development of RTT and appears to be an underlying basis for spontaneous occurrence of RTT in the F344 rat. The difference in incidence and severity of CPN between the sexes also explains the 9:1 male-to-female sex difference in the spontaneous occurrence of ATH and RTT observed here. The regulatory significance of this finding is that chemicals exacerbating CPN as their only renal effect are likely to show a numerical increase in RTT with dose, which does not represent a direct tumorigenic effect of the chemical.

Introduction: The purpose of this study was to compare the cytotoxicity and cytokine expression profiles of EndoSequence Root Repair Material (ERRM; Brasseler, Savannah, GA) putty, ERRM flowable, and ProRoot mineral trioxide aggregate (MTA; Dentsply Tulsa Dental, Johnson City, TN) using osteoblast cells (MG-63)., Methods: Four millimeters in diameter of each material was placed in the center of a 6-well culture plate, and a 2-mL suspension (10(5) cells/mL) of human osteoblasts was seeded in each well. Photomicrograph images were used to evaluate cytotoxicity as evidenced by the lack of osteoblast cell growth in relation to the materials with AH-26 (Dentsply Tulsa Dental) as the positive control. In addition, reverse-transcriptase polymerase chain reaction (RT-PCR) was used to evaluate the expression of interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor-α (TNF-α). Cytokine expression of MG-63 cells upon lipopolysaccharide treatment was used as controls. RT-PCR results were normalized by the expression of the housekeeping gene β-actin and were used to measure cytokine expression. Statistical analysis was performed using analysis of variance., Results: Results showed that ERRM putty and MTA exhibited minimal levels of cytotoxicity; however, ERRM was slightly more cytotoxic although not statistically significant. The expression of IL-1β, IL-6, and IL-8 was detected in all samples with minimal TNF-α expression., Conclusions: We concluded that ERRM and MTA showed similar cytotoxicity and cytokine expressions., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: The purpose of this in vitro study was to compare the cytotoxicity of white mineral trioxide aggregate cement (AMTA, MTA-Angelus), Brasseler Endosequence Root Repair Putty (ERRM), Dycal, and Ultra-blend Plus (UBP) by using human dermal fibroblasts and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay., Methods: Cultured adult human dermal fibroblasts were exposed to multiple concentrations of material elutes. The test material samples were immersed and incubated in the culture medium for 2, 5, or 8 days at 37°C. The cytotoxic effects were recorded by using an MTT-based colorimetric assay. Positive and negative controls were used. The results were statistically examined by one-way analysis of variance and Tukey post tests., Results: The cell viability of cultures exposed to all dilutions of AMTA, ERRM, and UBP was statistically similar to the negative control at 2 and 5 days. Only the Dycal-exposed specimens exhibited a statistically significant increase in cytotoxicity at the 2 initial evaluation periods. After exposure to the 8-day elutes, the respective percentage of cell survivability was 91% (Brasseler), 88% (MTA-Angelus), 76% (Ultra-blend Plus), and 37% (Dycal)., Conclusions: From the data in this in vitro study, AMTA, ERRM, and UBP had statistically similar adult human dermal fibroblast cytotoxicity levels. Relative to the negative control, only Dycal was shown to have a statistically significant cytotoxic effect to adult human dermal fibroblasts at all tested intervals., (Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

In this ex-vivo study, we assessed the adhesion of human gingival fibroblast (HGF) to mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) cement using a scanning electronic microscope (SEM). Test biomaterials were prepared and inserted in polyethylene wells (n = 18). Glass coverslips were used for control groups. HGFs were seeded in the glass coverslips, MTA and CEM. For the positive-control group, distilled water was added to the medium; the samples were observed with SEM at one, three and seven-day intervals. Normal cell morphology was observed in the negative controls. In the positive controls, round cells with rougher surfaces were seen; these cells showed few attachments to the glass coverslip. HGFs spread and adhered similarly on MTA and CEM by forming a monolayer of flat cells; there were no statistical differences between these two experimental groups. HGF cells displayed a favorable biologic response in contact with MTA and CEM. This demonstrates non-cytotoxicity of CEM as a novel endodontic biomaterial.

Aim:   To compare the cytotoxicity of four endodontic sealers (Sealapex, Pulp Canal Sealer EWT, Real Seal and MTA Fillapex) either 1 or 7 days after mixing, when assessed through a multiparametric analysis employing human primary cells closely related to periapical tissues., Methodology:   Extracts of each sealer were prepared following 24-h exposure to culture media, at either 24 h or 7 days after mixing. Primary human osteoblasts were exposed to extracts for 24 h, at 37 °C with 5% CO(2) , 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 and experimental time were compared by 2-way analysis of variance (anova)., Results:   All endodontic sealers had strong cytotoxicity 24 h after mixing, according to all parameters evaluated. At a longer setting period (7 days), viability for Sealapex was significantly increased (P < 0.05) and Pulp Canal Sealer achieved levels of cytocompatibility similar to the control group. The anova indicated a general correlation between the cytotoxicity of the materials and the time after mixing, with some level of dependence on the cell viability assay employed., Conclusions:   All materials had high cytotoxic levels for human primary cells, mostly on a time-dependent basis, as shown by three different cell viability tests., (© 2011 International Endodontic Journal.)

This aim of this study was to evaluate the physicochemical and biological properties of novel experimental cements (Hybrid, Paste and Resin) based on synergistic combinations of existing materials, including pH, diametral tensile strength (DTS) and cytotoxicity comparing them with mineral trioxide aggregate (MTA - Angelus®) and a glass ionomer cement (GIC) developed at our laboratory. For the physicochemical and biological tests, specimens with standard dimensions were produced. pH measurements were performed with digital pH meter at the following time intervals: 3, 24, 48 and 72 h. For the DTS test, cylindrical specimens were subjected to compressive load until fracture. The MTT assay was performed for cytotoxicity evaluation. Data were analyzed by ANOVA and Tukey's test (α=0.05). Paste group showed pH values similar to MTA, and Hybrid group presented pH values similar to GIC (p>0.05). The tested materials showed pH values ranging from alkaline to near neutrality at the evaluated times. MTA and GIC showed similar DTS values. The lowest and highest DTS values were seen in the Paste and Resin groups, respectively (p<0.05). Cell viability for MTA and experimental Hybrid, Paste and Resin groups was 49%, 93%, 90% and 86%, respectively, when compared with the control group. The photo-cured experimental resin cement showed similar or superior performance compared with the current commercial or other tested experimental materials.

The current study was conducted in order to investigate the short-term effects (6, 12, and 24 h) of silicon (Si) on the endogenous hormonal composition of rice (Oryza sativa L. cv. Dongjin-beyo), with and without wounding stress. Si applied in different concentrations (0.5, 1.0, and 2.0 mM) significantly promoted shoot length, plant biomass, and chlorophyll content of rice plants. Plants treated with different concentrations of sole Si for 6, 12, and 24 h had higher endogenous jasmonic acid contents than control. However, a combined application of wounding stress and Si induced a significantly small quantity of endogenous jasmonic acid as compared with control. On the contrary, endogenous salicylic acid level was significantly higher in sole Si-treated plants, while after wounding stress, a similar trend was observed yet again. After 6, 12, and 24 h of Si applications, with and without wounding stress, ethylene levels were significantly lower in comparison to their respective controls. The findings of the present study perpetrate the beneficial role of Si on the growth and development of rice plant by relieving physical injury and stress. Si also affects endogenous jasmonic acid and ethylene levels, while an inverse correlation exists between jasmonic acid and salicylic acid under wounding stress conditions.

Objective: This study evaluated the cytotoxicity of GuttaFlow and EndoSequence BC sealers and compared them with AH Plus and Tubli-Seal sealers., Study Design: Samples (0.5 mg) of freshly mixed or set BC, GuttaFlow, AH Plus, and Tubli-Seal sealers were eluted with 300, 600, and 1,000 μL cell culture medium for 24 and 72 hours. L929 cells were seeded into 96-well plates at 3 × 10(4) cells/well and cultured with 100 μL eluate from each eluate group. Cells cultured only with culture medium served as control. After 24 hours' incubation, the cytotoxicity was evaluated by MTT assay. Cell viability was calculated as the percentage of the control group, and the results were analyzed with a one-way analysis of variance., Results: For the freshly mixed sealer, cell viability in the AH Plus group was less than in all of the other 3 sealer groups. The Tubli-Seal sealer group had less cell viability than the EndoSequence BC and GuttaFlow sealer groups. For the set sealer, the Tubli-Seal and AH Plus groups had less cell viability than the EndoSequence BC and GuttaFlow sealer groups. There was no cell viability difference between the EndoSequence BC and GuttaFlow sealer groups in the either freshly mixed or set sealer group., Conclusions: The GuttaFlow and EndoSequence BC sealers have lower cytotoxicity than the AH Plus and Tubli-Seal sealers., (Copyright © 2011 Mosby, Inc. All rights reserved.)

Due to the toxic effect of asbestos, other materials with similar chemical-physical characteristics have been introduced to substitute it. We evaluate the angiogenic effect of certain asbestos substitute fibres such as glass fibres (GFs), ceramic fibres (CFs) and wollastonite fibres (WFs) and then compare angiogenic responses to those induced by crocidolite asbestos fibres (AFs). An in vitro model using human endothelial cells in small islands within a culture matrix of fibroblasts (Angio-Kit) was used to evaluate vessel formation. The release of IL-6, sIL-R6, IL-8, VEGF-A and their soluble receptors, sVEGFR-1, sVEGFR-2, was determined in the conditioning medium of Angio-Kit system after fibre treatment. ROS formation and cell viability were evaluated in cultured endothelial cells (HUVEC). To evaluate the involvement of intracellular mechanisms, EGFR signalling, ROS formation and nuclear factor-κB (NFκB) pathway were then inhibited by incubating HUVEC cells with AG1478, NAC and PDTC respectively, and the cytokine and growth factor release was analyzed in the culture medium after 7 days of fibre incubation. Among the mineral fibres tested, WFs markedly induced blood vessel formation which was associated with release of IL-6 and IL-8, VEGF-A and their soluble receptors. ROS production was observed in HUVEC after WFs treatment which was associated with cell cytotoxicity. The EGFR-induced ERK phosphorylation and ROS-mediated NFκB activation were involved in the cytokine and angiogenic factor release. However, only the EGFR activation was able to induce angiogenesis. The WFs are potential angiogenic agents that can induce regenerative cytokine and angiogenic factor production resulting in the formation of new blood vessels., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Introduction: Generex A and Generex B (calcium silicate based), Capasio (calcium-phospho-alumino silicate based) along with Ceramicrete-D (magnesium phosphate based) are being introduced as a new generation of endodontic materials with the potential to facilitate bone healing. The aim of this study was to evaluate the biocompatibility and osteogenic potential of these new materials by using primary osteoblasts., Methods: Primary osteoblasts were prepared from rat calvaria and exposed to mineral trioxide aggregate (MTA), Generex A, Generex B, Capasio, and Ceramicrete-D prepared to standardized size and shape (n = 5). Trypan blue staining was used to evaluate cell viability from 1-6 days. Mineralization potential was evaluated by scanning electron microscopy for the presence of mineralized nodules. Data were analyzed by Kruskal-Wallis and Mann-Whitney U tests., Results: Only Generex A and MTA allowed cell growth and proliferation throughout the experiment. There were statistically significant differences between groups throughout the experiment beginning on day 1. The greatest amount of cell growth was consistently observed with Generex A and MTA. There was no difference in mineralized nodule formation between any test materials., Conclusions: Generex A was the only new generation endodontic material that supported primary osteoblast growth; no material besides MTA facilitated nodule formation., (Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Objective: To study the cytotoxicity induced by chrysotile asbestos (CA), rock wool (RW) and wollastonite (WS)., Methods: V79 cells were divided into 4 groups. i.e. CA group, WS group, RW group and control group (200 microl PBS). The exposure concentration of dusts was 100 mg/L, The cell viability was detected by MTT and lactate dehydrogenase (LDH) activity assays. The technique of scanning electron microscopy was used to examine the change of V79 cells., Results: SiO2 was main constituent for 3 kinds of dusts. In MTT assay, the cell viability of RW and WS groups was 64.8% and 65.7%, respectively, which were significantly higher than that (54.5%) of CA group (P < 0.01). In LDH assay, the LDH activity of RW and WS groups [(15.7 +/- 50.9), (12.3 +/- 3.7) U/L, respectively] was significantly lower than that [(20.2 +/- 0.9) U/L] of CA group (P < 0.05). In scanning electron microscopy examination, it was found that the two ends of V79 cells in CA group contained a great deal of fibers remaining bodies, but the V79 cell appearance in RW and WS groups was normal., Conclusion: The cytotoxicity induced by RW and WS is significantly lower than that induced by CA for V79 cell.

Introduction: The purpose of the present study was to evaluate the biocompatibility of 2 root-end filling materials, Endosequence Root Repair Material Putty (ERRM Putty) and Paste (ERRM Paste) and compare them with gray mineral trioxide aggregate (MTA)., Methods: ERRM Putty, ERRM Paste, MTA, intermediate restorative material (IRM), and Cavit G were tested. For cytotoxicity assay, human gingival fibroblasts were incubated for 1, 3, and 7 days with extracts of varying concentrations from materials set for 2 days or 7 days. Cell viability was evaluated by methyl-thiazol-tetrazolium (MTT) assay. For cell adhesion assay, materials set for 7 days were examined under scanning electron microscope directly after setting, after incubation in cell culture medium for 7 days, and after incubation in gingival fibroblast suspension at a density of 5 × 10(4) cells/well for 2 and 7 days. The constituents of crystals formed on surface of materials were determined by energy dispersive analysis by x-ray., Results: Cell viability was significantly correlated with the type of material, setting time, and incubation time (P < .001 for all parameters). ERRM Putty and ERRM Paste displayed similar cell viabilities to MTA at all experimental conditions, except that fresh samples of ERRM Paste showed slightly lower cell viabilities than MTA. Cell viabilities with IRM and Cavit G were significantly lower than with the other 3 materials (P < .001). Similar surface crystallographic features and cell adhesion were observed on ERRM Paste, ERRM Putty, and MTA., Conclusions: ERRM Putty and ERRM Paste displayed similar in vitro biocompatibility to MTA., (Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: The present in vitro study was conducted with the aim of evaluating and comparing the cytotoxic effects of three root-end filling materials, ProRoot mineral trioxide aggregate (ProRoot MTA; Dentsply Tulsa Dental, Memphis, TN), MTA Angelus (Angelus, Londrina, Brazil), and a modified zinc oxide-eugenol cement (Super-EBA; Bosworth Co, Skokie, IL) on human periodontal ligament (PDL) fibroblasts., Methods: PDL cells were cultured in an mineral trioxide aggregate (MTA)- or a Super-EBA-conditioned medium to assess the viability as determined by the trypan blue exclusion assay. The proliferation of the cells was recorded, and the cellular morphology was observed by confocal microscopy. Moreover, PDL cell aggregates were cultured on the substrate surfaces to assess cell adhesion., Results: ProRoot MTA was found to be the most biocompatible material, whereas Super-EBA was found to be the most cytotoxic material because it significantly inhibited the cell growth and adherence on its. In the presence of ProRoot MTA, the PDL cell proliferation was almost unaltered. MTA Angelus was found to be more cytotoxic than ProRoot MTA, offering, however, excellent scaffold properties for the adhesion of cell aggregates., Conclusions: Under the conditions of the present study, it seems that commercially available forms of MTA may behave in different ways regarding their proliferative effect on human PDL fibroblasts. ProRoot MTA appears to be the most biocompatible of the three tested materials when considering use for root-end endodontic microsurgery., (Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Background: The aim of this study was to compare the reaction of rat connective tissue to two root-end filling materials: white Mineral Trioxide Aggregate (WMTA) and Diaket., Methods: Each of the materials was placed in dentine tubes and implanted subcutaneously in the dorsal connective tissue of 21 Wistar albino rats. Tissue biopsies were collected 7, 30, and 60 days after the implantation procedure. The specimens were processed and stained with hematoxylin and eosin and examined microscopically. After determining inflammatory cell numbers in sections from each specimen, inflammatory reaction scores were defined as follows: 0; no or few inflammatory cells (no reaction), 1; less than 25 cells (mild reaction), 2; 25 to 125 cells, (moderate reaction), and 3; 125 or more cells (severe reaction). Statistical analysis was performed using the Kruskal-Wallis and Mann-Whitney tests., Results: There were statistically significant differences in the median inflammatory cell numbers throughout the three test periods, with the most severe degree of inflammation observed at the one-week period. Few cases of necrosis were observed with WMTA. Diaket exhibited the most severe degree of inflammation and necrosis. After 30 days, both materials provoked moderate inflammatory reaction. The eight-week period showed the least severe degree of inflammation in all groups., Conclusions: It was concluded that WMTA exhibits a more favourable tissue response compared with Diaket which induced more severe inflammatory reaction than WMTA and the control., (© 2011 Al-Omari et al; licensee BioMed Central Ltd.)

Silica hazard is a growing occupational problem and has been reported to be associated with scleroderma via case reports and occupational studies. The aim of this study is to demonstrate whether oral or subcutaneous silicate exposure can induce an autoimmunity and scleroderma susceptibility in immunosensitive rats. Sodium silicate in a dose of 3 mg in 0.2 ml NS was administered through oral and subcutaneous routes to 20 brown Norway rats. Autoantibodies including ANA, anti-RNP, anti-SCL70 and anti-centromere were measured and compared with pre- and post-challenge serum samples. Serum ANA and anti-RNP were high in significant number of rats (P < 0.05) of only the subcutaneous silicate group. There is an increase in the number of positive readings of autoantibodies at 14th week in comparison with the number of positive readings of autoantibodies at 7th week but P values were not significant. It may be concluded that silicate might induce autoimmunity and scleroderma and it seems to be that the longer the duration of exposure the greater the risk. This is probably the first experimental animal study demonstrating the induction of scleroderma-related autoantibodies after challenge with silicate.

Introduction: This study investigated the setting time and micohardness of a premixed calcium phosphate silicate-based sealer (EndoSequence BC Sealer; Brasseler USA, Savannah, GA) in the presence of different moisture contents (0-9 wt%). The moisture content that produced the most optimal setting properties was used to prepare set EndoSequence BC Sealer for cytotoxicity comparison with an epoxy resin-based sealer (AH Plus; Dentsply Caulk, Milford, DE)., Methods: Standardized disks were created with BC Sealer, AH Plus, Pulp Canal Sealer EWT (positive control) (SybronEndo, Orange CA), and Teflon (Small Parts Inc., Miami Lakes, FL; negative control). Disks were placed in Transwell Inserts, providing indirect contact with MC3T3-E1 cells. Succinate dehydrogenase activity of the cells was evaluated over a 6-week period using MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Cytotoxicity profiles of BC Sealer and AH Plus were fitted with polynomial regression models. The time for 50% of the cells to survive (T(0.5)) was analyzed using the Wald statistic with a two-tailed significance level of 0.05., Results: BC Sealer required at least 168 hours to reach the final setting using the Gilmore needle method, and its microhardeness significantly declined when water was included in the sealer (P = .004). All set sealers exhibited severe cytotoxicity at 24 hours. The cytotoxicity of AH Plus gradually decreased and became noncytotoxic, whereas BC Sealer remained moderately cytotoxic over the 6-week period. A significant difference (P < .001) was detected between T(0.5) of BC Sealer (5.10 weeks; 95% confidence interval [CI], 4.69-5.42, standard error [SE] = 0.09) and T(0.5) of AH Plus (0.86 weeks; 95% CI, 0.68-1.05; SE = 0.18)., Conclusions: Further studies are required to evaluate the correlation between the length of setting time of BC Sealer and its degree of cytotoxicity., (Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Natural clays have been used in ancient and modern medicine, but the mechanism(s) that make certain clays lethal against bacterial pathogens has not been identified. We have compared the depositional environments, mineralogies, and chemistries of clays that exhibit antibacterial effects on a broad spectrum of human pathogens including antibiotic resistant strains. Natural antibacterial clays contain nanoscale (<200 nm), illite-smectite and reduced iron phases. The role of clay minerals in the bactericidal process is to buffer the aqueous pH and oxidation state to conditions that promote Fe(2+) solubility. Chemical analyses of E. coli killed by aqueous leachates of an antibacterial clay show that intracellular concentrations of Fe and P are elevated relative to controls. Phosphorus uptake by the cells supports a regulatory role of polyphosphate or phospholipids in controlling Fe(2+). Fenton reaction products can degrade critical cell components, but we deduce that extracellular processes do not cause cell death. Rather, Fe(2+) overwhelms outer membrane regulatory proteins and is oxidized when it enters the cell, precipitating Fe(3+) and producing lethal hydroxyl radicals.

Aim: To compare the subcutaneous tissue response to grey mineral trioxide aggregate (GMTA), white mineral trioxide aggregate (WMTA) and a new experimental cement (calcium enriched cement, CEM)., Methodology: Thirty-six Wistar male albino rats each received three implants, containing one of the tested materials, and an empty tube as a control. Seven, 30 and 60 days after implantation, the animals were sacrificed. After histological preparation and H&E staining, the specimens were evaluated for capsule thickness, necrosis, and for the type, the severity, and the extent of inflammation. Kruskal Wallis and Chi-square tests were used for data analysis., Results: After 1 week, CEM produced no necrosis compared to both types of WMTA and GMTA (P = 0.007). After 30 days, GMTA specimens had significantly less inflammation compared with WMTA and CEM (P = 0.011). After 60 days, less inflammation was associated with CEM specimens (P = 0.0001) compared to the other materials. Dystrophic calcifications in the connective tissue adjacent to all experimental material were detected., Conclusion: Histological observation illustrated that all materials were well tolerated by the subcutaneous tissues., (© 2010 International Endodontic Journal.)