Alleviation of calcium hydroxide nanoparticles induced genotoxicity and gastritis by coadministration of calcium titanate and yttrium oxide nanoparticles in mice.

Diverse applications of nanoparticles due to their unique properties has rapidly increased human exposure to numerous nanoparticles such as calcium hydroxide (Ca(OH)2), calcium titanate (CaTiO3), and yttrium oxide (Y2O3) nanoparticles almost in all aspect of daily life. However, very limited data are available on the effect of these nanoparticles on genomic DNA integrity and inflammation induction in the gastric tissues. Hence, this study estimated the effect of Ca(OH)2, CaTiO3, or/and Y2O3 nanoparticles multiple oral administration on the genomic DNA damage and inflammation induction in the mice gastric tissues. A suspension containing 50 mg/kg b.w of Ca(OH)2, CaTiO3, or Y2O3 nanoparticles were given orally to male mice separately or together simultaneously three times a week for two consecutive weeks. Multiple oral administration of Ca(OH)2 nanoparticles led to significant elevations in DNA damage induction and ROS generation, in contrast to the non-significant changes observed in the level of induced DNA damage and generated ROS after administration of CaTiO3 or Y2O3 nanoparticles separately or in combination with Ca(OH)2 nanoparticles. Oral administration of Ca(OH)2 nanoparticles alone also highly upregulated INOS and COX-2 genes expression and extremely decreased eNOS gene expression. However, high elevations in eNOS gene expression were detected after multiple administration of CaTiO3 and Y2O3 nanoparticles separately or together simultaneously with Ca(OH)2 nanoparticles. Meanwhile, non-remarkable changes were noticed in the expression level of INOS and COX-2 genes after administration of CaTiO3 and Y2O3 nanoparticles separately or simultaneously together with Ca(OH)2 nanoparticles. In conclusion: genomic DNA damage and inflammation induced by administration of Ca(OH)2 nanoparticles alone at a dose of 50 mg/kg were mitigated by about 100% when CaTiO3 and Y2O3 nanoparticles were coadministered with Ca(OH)2 nanoparticles until they reached the negative control level through altering the expression level of eNOS, INOS and COX-2 genes and scavenging gastric ROS. Therefore, further studies are recommended to investigate the toxicological properties of Ca(OH)2, CaTiO3 and Y2O3 nanoparticles and possibility of using CaTiO3 and Y2O3 nanoparticles to mitigate genotoxicity and inflammation induction by Ca(OH)2 nanoparticles.

Genotoxicity and oxidative stress induction by calcium hydroxide, calcium titanate or/and yttrium oxide nanoparticles in mice.

Intensive uses of Calcium hydroxide (Ca(OH)2NPs), calcium titanate (CaTiO3NPs) and yttrium oxide (Y2O3NPs) nanoparticles increase their environmental release and human exposure separately or together through contaminated air, water and food. However, too limited data are available on their genotoxicity. Therefore, this study explored the effect of Ca(OH)2NPs, CaTiO3NPs or/and Y2O3NPs administration on the genotoxicityand oxidative stress induction in mice hepatic tissue. Mice were orally administered Ca(OH)2NPs, CaTiO3NPs and Y2O3NPs separately or simultaneously together at a dose level of 50 mg/kg b.w. for two successive weeks (3 days per week). Marked induction of DNA damage noticed after oral administration of Ca(OH)2NPs or CaTiO3NPs alone together with high Ca(OH)2NPs induced reactive oxygen species (ROS) generation and a slight CaTiO3NPs induced ROS production were highly decreased after simultaneous coadministration of administration of Y2O3NPs with Ca(OH)2NPs and CaTiO3NPs up to the negative control level. Oral administration of Y2O3NPs alone also did not cause observable changes in the genomic DNA integrity and the ROS generation level compared to the negative control levels. Similarly, significant elevations in P53 gene expression and high reductions in Kras and HSP-70 genes expression were observed only after administration of Ca(OH)2NPs alone, while, remarkable increases in the Kras and HSP-70 genes expression and non-significant changes in p53 gene expression were noticed after administration of CaTiO3NPs and Y2O3NPs separately or simultaneously together with Ca(OH)2NPs. Conclusion: Ca(OH)2NPs exhibited the highest genotoxic effect through oxidative stress induction and disruption of apoptotic (p53 and Kras) and protective (HSP-70) genes expression. Slight DNA damage was noticed after CaTiO3NPs administration. However, administration of Y2O3NPs alone was non-genotoxic and coadministration of Y2O3NPs with Ca(OH)2NPs and CaTiO3NPs restored genomic DNA integrity and normal expression of apoptotic p53 and protective HSP-70 genes disrupted by Ca(OH)2NPs and CaTiO3NPs. Thus co-administration of Y2O3NPs with Ca(OH)2NPs and CaTiO3NPs is recommended to counter Ca(OH)2NPs and CaTiO3NPs induced genotoxicity and oxidative stress.

Comparison of the Physicochemical Properties, Antimicrobial Action, and Cytotoxicity of Ambroxol Hydrochloride, N-acetylcysteine, and Calcium Hydroxide Pastes.

OBJECTIVE: To evaluate the solubility, pH, antimicrobial action, and cytotoxicity of ambroxol hydrochloride (AMB), N-acetylcysteine (NAC), and calcium hydroxide (CH) pastes for use as intracanal medications. METHODS: Solubility was determined by micro-CT, based on the paste volume remaining after immersion in water for 7 days. pH was measured by immersing acrylic tubes containing the pastes in ultrapure water and then measuring pH after 3 hours, 3 days, and 7 days. Antimicrobial action against Enterococcus faecalis was assessed based on the percentage of living cells, using the live/dead staining method under confocal microscopy. Cytotoxicity was assessed based on the cell viability of L929 fibroblast-like cells after 6, 24, and 48 hours. Cytotoxicity data were compared using the ANOVA and Tukey tests, and the antimicrobial data were compared using the Kruskal-Wallis and Dunn tests. The significance level used was 5% (α=0.05). RESULTS: The solubility values for all the study groups were significantly different (P<0.05), where the highest values were for NAC, followed by AMB, and then CH. Likewise, the pH levels were all significantly different (P<0.05), where NAC and AMB levels were acidic, and CH levels were alkaline. The antimicrobial action of AMB was significantly higher than that of CH (P<0.05), and that of NAC was also higher than that of CH, albeit not significantly. AMB and NAC were more cytotoxic than CH, and higher dilutions of CH promoted higher cell viability levels than lower dilutions of the same paste (P<0.05). CONCLUSION: The NAC and AMB pastes were more soluble and cytotoxic than the CH paste and had acidic pH levels. The AMB paste displayed the highest antimicrobial action against Enterococcus faecalis biofilm.

Cytotoxicity and Genotoxicity of Calcium Hydroxide and Two Antibiotic Pastes on Human Stem Cells of The Apical Papilla.

OBJECTIVE: This study aimed to assess the cytotoxicity and genotoxicity of triple antibiotic paste, double antibiotic paste and calcium hydroxide medicaments on human stem cells of the apical papilla. METHODS: In this experimental study, stem cells were isolated from the apical papilla and cultured. They are treated with different concentrations 0.1, 0.5, 1, 10 and 100 mg/ml of medicaments for 24, 48 and 72 hours. The cytotoxicity and genotoxicity of the medicaments were determined using methyl thiazolyl tetrazolium assay and Comet test, respectively. RESULTS: Results showed all tested concentrations of the calcium hydroxide had no significant effect on stem cells at any time point. Triple antibiotic paste showed cytotoxicity in 10 and 100 mg/mL concentrations at all-time points and in 1, 10 and 100 mg/ml concentrations at 72 hours. In addition, its genotoxicity was significantly higher than that of other groups (P<0.05). Double antibiotic paste showed cytotoxic effects only in 100 mg/ml concentration at 24 hours and 10 and 100 mg/ml concentrations at 48 and 72 hours. And also, its genotoxicity in these concentrations was significantly higher than that of control and calcium hydroxide groups (P<0.05). CONCLUSION: In contrast to calcium hydroxide, triple antibiotic paste and double antibiotic paste, especially in their higher concentrations, induced cytotoxicity and genotoxicity on human stem cells of the apical papilla.

Induction of genotoxicity and differential alterations of p53 and inflammatory cytokines expression by acute oral exposure to bulk- or nano-calcium hydroxide particles in mice "Genotoxicity of normal- and nano-calcium hydroxide".

With the high increases in the uses of calcium hydroxide in various applications due its distinctive properties, human exposure has increased to normal- and nano-calcium hydroxide. However, its impact on the DNA integrity, expression of inflammatory cytokines, and induction of oxidative stress has not been clearly studied. Therefore, here we estimate the induction of DNA damage, inflammation, and oxidative stress in mice orally administrated a single dose (100 mg/kg) of normal- or nano-sized calcium hydroxide for 24 hour. Comet, Diphenylamine and laddered DNA fragmentation assays were done to assess DNA damage induction. Acute oral administration of normal- or nano-calcium hydroxide particles disrupted the DNA integrity, caused generation of ROS and also concurrent increases in both the nitric oxide concentration and inducible nitric oxide synthase gene expression in a reverse proportional to the calcium hydroxide particles' size. Increases in the concentration of calcium ions as well as alterations in the expression level of p53 and proinflammatory cytokines were also observed in calcium hydroxide administrated groups. Moreover, administration of normal- or nano-calcium hydroxide particles suspension elevated the level of malondialdehyde and decreased both the glutathione peroxidase activity and the reduced glutathione level, as well as caused tissue injuries (e.g. renal tube degeneration, congested blood vessels, atrophied lymphoid follicles, interstitial inflammatory reaction, and hyalinosis of myocardial muscles). Thus, we conclude that calcium hydroxide acutely orally administrated in its ordinary or nano-particulate form causes DNA damage induction by generating free radicals and altering the expression levels of p53 gene and proinflammatory cytokines.

A New Calcium Silicate-Based Root Canal Dressing: Physical and Chemical Properties, Cytotoxicity and Dentinal Tubule Penetration.

The aims of this study were to evaluate the physical and chemical properties, cytotoxicity and dentinal tubule penetration of a new calcium silicate-based root canal dressing. For pH and calcium ion release evaluation (1, 24, 72 and 168 h) were used a pH meter and colorimetric spectrophotometer, respectively. Radiopacity evaluation followed the ISO 6876:2012. Cytotoxicity was evaluated by the percentage of cell viability using MTT assay. Illustrative images of dentinal tubule penetration were obtained using confocal laser scanning microscopy (CLSM). Data from pH and calcium ion release were statistically analyzed by two-way analysis of variance and Tukey test. Radiopacity was analyzed using the Student t-test. The statistical tests for cytotoxicity results were the one-way analysis of variance and Tukey test. Both materials showed alkaline pH in all experimental times. The pH values for calcium hydroxide paste were higher than bioceramic paste at 1, 24, and 72 h (p<0.05). The calcium ion release of bioceramic was lower than the calcium hydroxide paste only at 24 h (p<0.05). The bioceramic was more radiopaque than the calcium hydroxide paste (p<0.05). Bioceramic paste presented a dose and time-dependent cytotoxic effect after MTT assay. CLSM images showed absence of tubule penetration for both pastes. The new calcium silicate-based canal dressing presented alkaline pH, high calcium release, and acceptable radiopacity. Bio C Temp showed a dose and time-dependent cytotoxic and absence of dentinal tubule penetration.

A new calcium silicate-based root canal dressing: physical and chemical properties, cytotoxicity and dentinal tubule penetration

Abstract The aims of this study were to evaluate the physical and chemical properties, cytotoxicity and dentinal tubule penetration of a new calcium silicate-based root canal dressing. For pH and calcium ion release evaluation (1, 24, 72 and 168 h) were used a pH meter and colorimetric spectrophotometer, respectively. Radiopacity evaluation followed the ISO 6876:2012. Cytotoxicity was evaluated by the percentage of cell viability using MTT assay. Illustrative images of dentinal tubule penetration were obtained using confocal laser scanning microscopy (CLSM). Data from pH and calcium ion release were statistically analyzed by two-way analysis of variance and Tukey test. Radiopacity was analyzed using the Student t-test. The statistical tests for cytotoxicity results were the one-way analysis of variance and Tukey test. Both materials showed alkaline pH in all experimental times. The pH values for calcium hydroxide paste were higher than bioceramic paste at 1, 24, and 72 h (p<0.05). The calcium ion release of bioceramic was lower than the calcium hydroxide paste only at 24 h (p<0.05). The bioceramic was more radiopaque than the calcium hydroxide paste (p<0.05). Bioceramic paste presented a dose and time-dependent cytotoxic effect after MTT assay. CLSM images showed absence of tubule penetration for both pastes. The new calcium silicate-based canal dressing presented alkaline pH, high calcium release, and acceptable radiopacity. Bio C Temp showed a dose and time-dependent cytotoxic and absence of dentinal tubule penetration.

Resumo Os objetivos deste estudo foram avaliar as propriedades físicas e químicas, citototoxidade e penetração tubular de uma nova medicação à base de silicato de cálcio. Para o teste de pH, e liberação de íons cálcio (1, 24, 72 e 168 h) foi usado medidor de pH e espectofotômetro colorimétrico, respectivamente. Avaliação da radiopacidade, seguiu a ISO 6876:2012). A citotoxicidade foi avaliada pela porcentagem de células viáveis usando o ensaio MTT. Imagens ilustrativas de penetração tubular foram obtidas usando microscopia confocal de varredura a laser (CLSM). Os dados de pH e liberação de cálcio foram analisados através do teste de Análise de Variância de duas vias e teste de Tukey. A radiopacidade foi avaliada usando o teste T de Student. Para a citotoxicidade foi empregada a Análise de Variância de uma via e teste de Tukey. Ambos os materiais apresentaram pH alcalino em todos os tempos experimentais. Os valores de pH da pasta de hidróxido de cálcio foram superiores à pasta biocerâmica em 1, 24 e 72 h (p<0,05). A liberação de cálcio da pasta biocerâmica foi inferior à pasta de hidróxido de cálcio apenas em 24 h (p<0,05). Bio-C Temp foi mais radiopaco que o Ultracal XS (p<0,05). A pasta biocerâmica apresentou efeito citotóxico dependente da dose e do tempo de exposição. Imagens de CLSM mostraram ausência de penetração intratubular para ambas as pastas. A nova medicação à base de silicato de cálcio apresentou pH alcalino, alta liberação de cálcio e boa radiopacidade. Bio C Temp apresentou um efeito citotóxico dependente da dose e do tempo de exposição e ausência de penetração tubular.

Effect of preheating on cytotoxicity and physicochemical properties of light-cured calcium-based cements.

The aim of this study was to evaluate the degree of conversion, cytotoxicity, solubility and pH of photopolymerizable calciumbased cements submitted to preheating. The degree of conversion was analyzed by Fourier transform infrared, cytotoxicity by the MTT test and solubility through loss of mass. The data were subjected to statistical tests (ANOVA / Tukey's, p<0.05). The photopolymerizable materials showed a low degree of conversion, regardless of preheating. All materials caused a reduction in cell viability at 24 hours and 7 days, with the Dycal (control) being more cytotoxic. Heat had a positive effect on Biocal at 7 days. Dycal is the most soluble material. Heat had no effect on the solubility or pH of the polymerizable materials. It is concluded that photopolymerizable calcium-based cements have a low degree of conversion and are soluble, which results in mild to moderate cytotoxicity.

O objetivo do presente estudo foi avaliar o grau de conversão, citotoxicidade, solubilidade e pH de cimentos à base de cálcio fotopolimerizáveis submetidos a pré-aquecimento. O grau de conversão foi analisado por espectroscopia no infravermelho com transformada de Fourier, a citotoxicidade pelo teste de MTT e a solubilidade através da perda de massa. Os dados foram submetidos a testes estatísticos (ANOVA/Tukey, p<0,05). Os materiais fotopolimerizáveis apresentaram baixo grau de conversão, independente do pré-aquecimento. Todos os materiais causaram redução da viabilidade celular nas análises de 24 horas e 7 dias, sendo que o Dycal (controle) apresentouse mais citotóxico e o calor apresentou efeito positivo sobre o Biocal na análise de 7 dias. O Dycal é o material mais solúvel e o calor não causou efeito na solubilidade e pH dos materiais polimerizáveis. Assim, conclui-se que os cimentos à base de cálcio fotopolimerizáveis apresentam baixo grau de conversão e são solúveis, que resulta em citotoxicidade suave e moderada.

Effect of preheating on cytotoxicity and physicochemical properties of light-cured calcium-based cements / Efeito do pré-aquecimento na citotoxicidade e propriedades físico-químicas de cimentos à base de cálcio fotopolimerizáveis

ABSTRACT The aim of this study was to evaluate the degree of conversion, cytotoxicity, solubility and pH of photopolymerizable calciumbased cements submitted to preheating. The degree of conversion was analyzed by Fourier transform infrared, cytotoxicity by the MTT test and solubility through loss of mass. The data were subjected to statistical tests (ANOVA / Tukey's, p<0.05). The photopolymerizable materials showed a low degree of conversion, regardless of preheating. All materials caused a reduction in cell viability at 24 hours and 7 days, with the Dycal (control) being more cytotoxic. Heat had a positive effect on Biocal at 7 days. Dycal is the most soluble material. Heat had no effect on the solubility or pH of the polymerizable materials. It is concluded that photopolymerizable calcium-based cements have a low degree of conversion and are soluble, which results in mild to moderate cytotoxicity.

RESUMO O objetivo do presente estudo foi avaliar o grau de conversão, citotoxicidade, solubilidade e pH de cimentos à base de cálcio fotopolimerizáveis submetidos a pré-aquecimento. O grau de conversão foi analisado por espectroscopia no infravermelho com transformada de Fourier, a citotoxicidade pelo teste de MTT e a solubilidade através da perda de massa. Os dados foram submetidos a testes estatísticos (ANOVA/Tukey, p<0,05). Os materiais fotopolimerizáveis apresentaram baixo grau de conversão, independente do pré-aquecimento. Todos os materiais causaram redução da viabilidade celular nas análises de 24 horas e 7 dias, sendo que o Dycal (controle) apresentouse mais citotóxico e o calor apresentou efeito positivo sobre o Biocal na análise de 7 dias. O Dycal é o material mais solúvel e o calor não causou efeito na solubilidade e pH dos materiais polimerizáveis. Assim, conclui-se que os cimentos à base de cálcio fotopolimerizáveis apresentam baixo grau de conversão e são solúveis, que resulta em citotoxicidade suave e moderada.

Cytotoxicity and biocompatibility of a new bioceramic endodontic sealer containing calcium hydroxide.

This study evaluated the cytotoxicity and biocompatibility of a new bioceramic endodontic sealer (i.e., Sealer Plus BC) in comparison with those of MTA Fillapex and AH Plus. L929 fibroblasts were cultured and Alamar Blue was used to evaluate cell viability of diluted extracts (1:50, 1:100, and 1:200) from each sealer at 24 h. Polyethylene tubes that were filled with material or empty (as a control) were implanted in the subcutaneous tissue of rats. The rats were killed after 7 and 30 d (n = 8), and the tubes were removed for histological analysis. Parametric data was analyzed using a one-way ANOVA test, and nonparametric data was analyzed via the Kruskal-Wallis test followed by the Dunn test (p < 0.05). A reduction in cell viability was observed in the extracts that were more diluted for Sealer Plus BC when compared to that of Control and AH Plus (p < 0.05). However, the 1:50 dilution of the Sealer Plus BC was similar to that of the Control (p > 0.05). Conversely, more diluted extracts of MTA Fillapex (1:200) and AH Plus (1:100 and 1:200) were similar to the Control (p > 0.05). Histological analysis performed at 7 d did not indicate any significant difference between tissue response for all materials, and the fibrous capsule was thick (p > 0.05). At 30 d, Sealer Plus BC was similar to the Control (p > 0.05) and MTA Fillapex and AH Plus exhibited greater inflammation than the Control (p < 0.05). The fibrous capsule was thin for the Control and for most specimens of Sealer Plus BC and AH Plus. Thus, Sealer Plus BC is biocompatible when compared to MTA Fillapex and AH Plus, and it is less cytotoxic when less-diluted extracts are used.

Cytotoxicity and Genotoxicity of a New Intracanal Medicament, 2-hydroxyisocaproic Acid-An In Vitro Study.

INTRODUCTION: A successful outcome of root canal therapy relies on effective disinfection of the root canal system, including the use of intracanal medicaments, which vary in their bactericidal and cytotoxic properties. Assessing the benefits and risks associated with the use of these medicaments is of extreme importance, especially in regenerative endodontic procedures, because residual stem cells may be harmed. In this study, we tested the cytotoxicity and genotoxicity of a novel agent, 2-hydroxyisocaproic acid (HICA), and compared its properties with those of a well-established medicament, calcium hydroxide. METHODS: Human periodontal ligament fibroblasts were exposed to varying concentrations of HICA (1, 2.5, 5, 10, 20, and 40 mg/mL) for 24 hours, and a dose-response curve was generated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Immunofluorescence for 2 markers of DNA double-strand breaks, phosphorylated γH2AX and 53BP1, was used to establish the genotoxicity of HICA at various half maximal effective concentration (EC50) fractions. Cytotoxicity and genotoxicity of HICA and calcium hydroxide at 1 mg/mL were compared at 24 and 48 hours using the same methods. RESULTS: At 10 mg/mL and higher, HICA was significantly more cytotoxic and genotoxic than the control (P < .05 and P < .0001, respectively). Calcium hydroxide at 1 mg/mL was more cytotoxic than HICA at 1 mg/mL at 24 and 48 hours (P < .05 for both), whereas no difference in the accumulated DNA damage was observed. CONCLUSIONS: HICA is not cytotoxic and genotoxic at concentrations <10 mg/mL. At the concentration of 1 mg/mL, HICA is significantly less cytotoxic than calcium hydroxide.

Cytotoxicity of intracanal dressings on apical papilla cells differ upon activation with E. faecalis LTA.

OBJECTIVE: The aim of this study was to investigate the cytotoxic effects of modified triple antibiotic paste and an experimental composition using calcium hydroxide on lipoteichoic acid (LTA)-primed apical papilla cells (APC). MATERIAL AND METHODS: Human APC were tested for in vitro cytotoxicity of modified Triple Antibiotic Paste (mTAP - Ciprofloxacin, Metronidazole and Cefaclor at 1:1:1) and of a paste of Ciprofloxacin, Metronidazole and Calcium hydroxide (CMC - 1:1:2) and modified CMC (mCMC - 2:2:1) by using MTT assay. The substances were reconstituted in DMEM at 1,000 µg/mL and » serially diluted before being kept in contact with cells for 1, 3, 5 and 7 days. Further, cells were primed with 1 µg/mL of Enterococcus faecalis LTA for 7 days prior to the viability test with 1,000 µg/mL of each substance. Statistical analysis was performed using one-way analysis of variance (ANOVA) and two-way ANOVA respectively followed by Tukey's post-test. Significance levels were set at p<0.05. RESULTS: In the first assay, the higher cytotoxic rates were reached by mTAP for all experimental periods. CMC was found toxic for APC at 5 and 7 days, whereas mCMC did not affect the cell viability. Only CMC and mCMC were able to induce some cellular proliferation. In the second assay, when considering the condition with medium only, LTA-primed cells significantly proliferated in comparison to LTA-untreated ones. At this context, mTAP and CMC showed similar cytotoxicity than the observed for LTA-untreated cells, while mCMC was shown cytotoxic at 7 days only for LTA-primed APC. Comparing the medications, mTAP was more cytotoxic than CMC and mCMC. CONCLUSION: mTAP showed higher cytotoxicity than CMC and mCMC and the effect of topic antimicrobials might differ when tested against apical papilla cells under physiological or activated conditions.

Estimation of genomic instability and mitochondrial DNA damage induction by acute oral administration of calcium hydroxide normal- and nano- particles in mice.

Nowadays, there is increasing interest in using calcium hydroxide (Ca(OH)2) nanoparticles rather than normal-sized Ca(OH)2 because of its higher antimicrobial activities. However, the genotoxicity of Ca(OH)2 nanoparticles has not been well investigated. Therefore, this study was performed to estimate the possible genomic instability and mitochondrial DNA damage induction by normal and nano-sized Ca(OH)2 particles in mice. Oral administration of normal or nano-sized Ca(OH)2 particles induced DNA breakages and apoptosis causing genomic instability as a result of increased Calcium content, ROS and MDA levels and decreased SOD and Gpx activities reversely proportional to Ca(OH)2 particles size in the liver, brain and bone marrow tissues. However, decreases in mitochondrial membrane potential concurrently with downregulated expression of POLG, POLG1 and TFAM genes were only observed in the brain and bone marrow tissues confirmed mitochondrial DNA damage. In contrast, the expressions of POLG, POLG1 and TFAM genes have been improved and mitochondrial membrane potential has not unchanged in liver tissues. Conclusion: single oral administration of normal or nano-sized Ca(OH)2 particles induced genomic instability through ROS generation that exhausted the antioxidant defense system in the liver, brain and bone marrow tissues but impaired the mitochondrial DNA only in the brain and bone marrow tissues.

Cytotoxicity and biocompatibility of a new bioceramic endodontic sealer containing calcium hydroxide

Abstract: This study evaluated the cytotoxicity and biocompatibility of a new bioceramic endodontic sealer (i.e., Sealer Plus BC) in comparison with those of MTA Fillapex and AH Plus. L929 fibroblasts were cultured and Alamar Blue was used to evaluate cell viability of diluted extracts (1:50, 1:100, and 1:200) from each sealer at 24 h. Polyethylene tubes that were filled with material or empty (as a control) were implanted in the subcutaneous tissue of rats. The rats were killed after 7 and 30 d (n = 8), and the tubes were removed for histological analysis. Parametric data was analyzed using a one-way ANOVA test, and nonparametric data was analyzed via the Kruskal-Wallis test followed by the Dunn test (p < 0.05). A reduction in cell viability was observed in the extracts that were more diluted for Sealer Plus BC when compared to that of Control and AH Plus (p < 0.05). However, the 1:50 dilution of the Sealer Plus BC was similar to that of the Control (p > 0.05). Conversely, more diluted extracts of MTA Fillapex (1:200) and AH Plus (1:100 and 1:200) were similar to the Control (p > 0.05). Histological analysis performed at 7 d did not indicate any significant difference between tissue response for all materials, and the fibrous capsule was thick (p > 0.05). At 30 d, Sealer Plus BC was similar to the Control (p > 0.05) and MTA Fillapex and AH Plus exhibited greater inflammation than the Control (p < 0.05). The fibrous capsule was thin for the Control and for most specimens of Sealer Plus BC and AH Plus. Thus, Sealer Plus BC is biocompatible when compared to MTA Fillapex and AH Plus, and it is less cytotoxic when less-diluted extracts are used.

Cytotoxicity of intracanal dressings on apical papilla cells differ upon activation with E. faecalis LTA

Abstract Objective The aim of this study was to investigate the cytotoxic effects of modified triple antibiotic paste and an experimental composition using calcium hydroxide on lipoteichoic acid (LTA)-primed apical papilla cells (APC). Material and Methods Human APC were tested for in vitro cytotoxicity of modified Triple Antibiotic Paste (mTAP - Ciprofloxacin, Metronidazole and Cefaclor at 1:1:1) and of a paste of Ciprofloxacin, Metronidazole and Calcium hydroxide (CMC - 1:1:2) and modified CMC (mCMC - 2:2:1) by using MTT assay. The substances were reconstituted in DMEM at 1,000 µg/mL and » serially diluted before being kept in contact with cells for 1, 3, 5 and 7 days. Further, cells were primed with 1 µg/mL of Enterococcus faecalis LTA for 7 days prior to the viability test with 1,000 µg/mL of each substance. Statistical analysis was performed using one-way analysis of variance (ANOVA) and two-way ANOVA respectively followed by Tukey's post-test. Significance levels were set at p<0.05. Results In the first assay, the higher cytotoxic rates were reached by mTAP for all experimental periods. CMC was found toxic for APC at 5 and 7 days, whereas mCMC did not affect the cell viability. Only CMC and mCMC were able to induce some cellular proliferation. In the second assay, when considering the condition with medium only, LTA-primed cells significantly proliferated in comparison to LTA-untreated ones. At this context, mTAP and CMC showed similar cytotoxicity than the observed for LTA-untreated cells, while mCMC was shown cytotoxic at 7 days only for LTA-primed APC. Comparing the medications, mTAP was more cytotoxic than CMC and mCMC. Conclusion mTAP showed higher cytotoxicity than CMC and mCMC and the effect of topic antimicrobials might differ when tested against apical papilla cells under physiological or activated conditions.

Novel endodontic sealers induced satisfactory tissue response in mice.

OBJECTIVES: The aim of this study was to evaluate the subcutaneous response induced by Roeko Guttaflow2 (RG), Sealapex Xpress (SX), AH Plus (AHP) sealers. METHODS: 100 BALB/c mice received implants in the subcutaneous tissue with the tested materials (10 animals per period for each evaluated sealer) and were evaluated after different experimental periods (7, 21 and 63 days), in each animal was placed a tube, the control group was an empty tube. Histological analysis evaluated semi-quantitatively the inflammatory infiltration, collagen fiber formation and tissue thickness. In addition, immunohistochemistry was performed for interleukin-6 (IL-6). Data were statistically analyzed (α = 0.05). RESULTS: RG promoted a greater collagen fiber formation at 7 days and 63 days compared to the CG (p = 0.004) and AHP (p = 0.005) respectively, while at 21 days, the SX promoted a greater reaction (p = 0.021). For the tissue thickness, there was a greater reaction at 7 days with CG (p = 0.0156) and with RG at 63 days (p = 0.03). Regarding the inflammatory infiltrate, there was no difference at 7 days and 63 days (p = 0.5; p = 0.27), while at 21 days, a statistically difference was found between SX, CG (p = 0.04) and RG (p = 0.027). In addition, the presence of IL-6 was observed in almost all groups, with a more intense marking at 7days. SIGNIFICANCE: All cements evaluated presented a satisfactory tissue response, however, RG was the one that presented a more satisfactory tissue response.

Comparative Evaluation of Cytotoxicity of Root Canal Sealers on Cultured Human Periodontal Fibroblasts: In vitro Study.

AIM: To evaluate and compare the cytotoxic effects of different types of root canal. MATERIALS AND METHODS: The sealers were eluted with culture medium for 1 hour, 7 days, and 14 days. Cell viability was estimated by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and trypan blue exclusion method on human periodontal ligament (PDL) fibroblast cells. Sealers used are mineral trioxide aggregate (MTA)-based sealer (MTA Fillapex, Angelus), calcium hydroxide-based sealer (Apexit Plus, Ivoclar Vivadent), resin-based sealer (AH Plus, Dentsply), and zinc oxide eugenol-based sealer (Tubli Seal, SybronEndo). RESULTS: The order of cytotoxicity through MTT assay, at the end of the second week, was observed as MTA Fillapex> Tubli Seal> Apexit Plus > AH Plus. The percentage cell viability obtained after trypan blue exclusion method decreased in the order of Apexit Plus> Tubli Seal> AH Plus> MTA Fillapex, which was similar to the reported cytotoxicity from the MTT assay after 1 hour. CONCLUSION: Each type of sealer showed moderate-to-severe cytotoxic response when compared with the control. The MTA Fillapex was found to be the most cytotoxic sealer. Use of resin-based material as a root canal sealer may result in a more favorable response to PDL fibroblasts. CLINICAL SIGNIFICANCE: Having knowledge of the cytotoxicity of various sealers will help in increasing patient's comfort.

Analysis of Aloe vera cytotoxicity and genotoxicity associated with endodontic medication and laser photobiomodulation.

This study aims to evaluate, in vitro, the effect of Aloe vera associated with endodontic medication, with or without laser photobiomodulation (FTL) irradiation in FP6 human pulp fibroblasts. The materials were divided into eight groups: CTR - control; CL - FTL alone; AA - Aloe vera with distilled water; AL - Aloe vera with distilled water and FTL; HA - calcium hydroxide P.A. with distilled water; HL - calcium hydroxide P.A. with distilled water and FTL; HAA - calcium hydroxide P.A. with Aloe vera and distilled water; HAL - calcium hydroxide P.A. with Aloe vera, distilled water, and FTL. The cytotoxicity was evaluated by MTT assay at 24, 48, and 72h and the genotoxicity by micronucleus test assay. This study was performed in triplicate. Data obtained in both tests were statistically analyzed by ANOVA and Tukey's tests (p≤0.05). Group AA presented high genotoxicity and low cytotoxicity. After 24, 48, and 72h, the group HAA significantly reduced the cell viability. Interaction with FTL showed slightly increase cell viability after 24 and 48h in groups CL and HL (p<0.001), despite the high genotoxicity in group CL and low genotoxicity in group HL. Group AL showed higher cell survival rate at 72h (p<0.05) and high genotoxicity (p<0.001). It was concluded that Aloe vera allowed higher cell viability in human pulp fibroblasts in the presence of calcium hydroxide or with FTL separately, but genotoxicity increased in these associations.

A Comparative Evaluation of the Effect of the Addition of Pachymic Acid on the Cytotoxicity of 4 Different Root Canal Sealers-An In Vitro Study.

INTRODUCTION: Root canal sealers exhibit varying degrees of cytotoxicity to periapical tissues. This in turn results in inflammation, delayed wound healing, and even bone resorption. This study aimed to explore the effect of the addition of an antioxidant like pachymic acid on the cytotoxicity of 4 root canal sealers, namely, Tubliseal (Kerr, Romulus, MI), a zinc oxide eugenol-based sealer; AH Plus (Dentsply De Trey GmbH, Konstanz, Germany), an epoxy resin-based sealer; Sealapex (Kerr), a calcium hydroxide-based sealer; and EndoREZ (Ultradent Products, South Jordan, UT), a methacrylate resin-based sealer. METHODS: Sealers mixed according to the manufacturers' instructions formed the experimental groups. Subgroups were determined based on the absence (subgroup A) or addition (subgroup B) of pachymic acid. The experimental sealers were added to L929 mouse fibroblast cells immediately after mixing. Cell viability was evaluated by methylthiazoletetrazolium assay after 24 hours. Data were analyzed using 1-way analysis of variance. Intergroup comparisons were performed using the Kruskal-Wallis test, and intragroup comparisons were done using independent t and post hoc tests. RESULTS: All 4 sealers were cytotoxic but to varying degrees. In both the subgroups, Sealapex exhibited the lowest cytotoxicity followed by AH Plus, Tubliseal, and EndoREZ (P < .05). The addition of pachymic acid reduced the cytotoxicity of all the sealers except that of EndoREZ (P > .05). CONCLUSIONS: Calcium hydroxide-based Sealapex showed the least cytotoxicity compared with the other sealers. Pachymic acid could be a viable therapeutic agent to overcome the potential adverse effects associated with root canal sealers.

Evaluation of the genotoxicity and cytotoxicity of filling pastes used for pulp therapy on deciduous teeth using the micronucleus test on bone marrow from mice (Mus musculus).

Pulp therapy is the last resort for preserving deciduous teeth. However, the genotoxic and cytotoxic effects of many products used in this therapy are not well established. The aim of this study was to use the micronucleus test on bone marrow from mice to evaluate the genotoxic and cytotoxic effects of four filling pastes: zinc oxide, calcium hydroxide P.A., mineral trioxide aggregate and an iodoform paste (iodoform + camphorated + paramonochlorophenol + rifamycin + prednisolone). Male Swiss mice were divided into 4 groups of 10 animals, each exposed to one of the pastes, and were subdivided according to the dilutions tested: 1/10, 1/50, 1/500 and 1/1000 administered intraperitoneally (0.1ml/10g of weight). Cyclophosphamide was the positive control. The negative controls were dimethylsulfoxide and buffered saline solution. Five animals were killed 24h and five 48h after the treatment. The material was processed in accordance with Schmid (1976) and micronuclei were counted in 1000 polychromatic erythrocytes (PCE), under an optical microscope in a blinded test. Cytotoxicity was evaluated using the PCE/normochromatic erythrocyte (NCE) ratio in 200 erythrocytes. The micronucleus analysis results were evaluated using the conditional test for comparing proportions in situations of rare events. Analysis of variance and Tukey's test were used to evaluate the PCE/NCE ratio. There was significantly greater occurrence of micronuclei in the animals treated with iodoform paste at all the dilutions tested, at both sacrifice times. Greater occurrence of micronuclei was observed among the animals treated with zinc oxide and sacrificed 48h after the treatment, at the dilutions 1:50; 1:500 and 1:1000. Calcium hydroxide P.A. and mineral trioxide aggregate did not present any genotoxic or cytotoxic effects. The genotoxicity and cytotoxicity of zinc oxide and iodoform paste revealed here constitute an initial step towards their contraindication, but additional studies will be necessary in order to securely establish the risks involved in their use.