Preparation, properties and antibacterial applications of medical nano-metals and their oxides: a review / 生物工程学报

Antibiotics are playing an increasingly important role in clinical antibacterial applications. However, their abuse has also brought toxic and side effects, drug-resistant pathogens, decreased immunity and other problems. New antibacterial schemes in clinic are urgently needed. In recent years, nano-metals and their oxides have attracted wide attention due to their broad-spectrum antibacterial activity. Nano-silver, nano-copper, nano-zinc and their oxides are gradually applied in biomedical field. In this study, the classification and basic properties of nano-metallic materials such as conductivity, superplasticity, catalysis, and antibacterial activities were firstly introduced. Secondly, the common preparation techniques, including physical, chemical and biological methods, were summarized. Subsequently, four main antibacterial mechanisms, such as cell membrane, oxidative stress, DNA destruction and cell respiration reduction, were summarized. Finally, the effect of size, shape, concentration and surface chemical characteristics of nano-metals and their oxides on antibacterial effectiveness and the research status of biological safety such as cytotoxicity, genotoxicity and reproductive toxicity were reviewed. At present, although nano-metals and their oxides have been applied in medical antibacterial, cancer treatment and other clinical fields, some issues such as the development of green preparation technology, the understanding of antibacterial mechanism, the improvement of biosafety, and the expansion of application fields, require further exploration.

Impact of calcium aluminate cement with additives on dental pulp-derived cells

Abstract Calcium aluminate cement (CAC) has been highlighted as a promising alternative for endodontic use aiming at periapical tissue repair. However, its effects on dental pulp cells have been poorly explored. Objective: This study assessed the impact of calcium chloride (CaCl2) and bismuth oxide (Bi2O3) or zinc oxide (ZnO) additives on odontoblast cell response to CAC. Methodology: MDPC-23 cells were exposed for up to 14 d: 1) CAC with 2.8% CaCl2 and 25% ZnO (CACz); 2) CAC with 2.8% CaCl2 and 25% Bi2O3 (CACb); 3) CAC with 10% CaCl2 and 25% Bi2O3 (CACb+); or 4) mineral trioxide aggregate (MTA), placed on inserts. Non-exposed cultures served as control. Cell morphology, cell viability, gene expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), and dentin matrix protein 1 (DMP-1), ALP activity, and extracellular matrix mineralization were evaluated. Data were compared using ANOVA (α=5%). Results: Lower cell density was detected only for MTA and CACb+ compared with Control, with areas showing reduced cell spreading. Cell viability was similar among groups at days one and three (p>0.05). CACb+ and MTA showed the lowest cell viability values at day seven (p>0.05). CACb and CACb+ promoted higher ALP and BSP expression compared with CACz (p<0.05); despite that, all cements supported ALP activity. Matrix mineralization were enhanced in CACb+ and MTA. Conclusion: In conclusion, CAC with Bi2O3, but not with ZnO, supported the expression of odontoblastic phenotype, but only the composition with 10% CaCl2 promoted mineralized matrix formation, rendering it suitable for dentin-pulp complex repair.

Surface and vertical dimensional changes of mineral trioxide aggregate and biodentine in different environmental conditions

Abstract Surface changes in biological environments are critical for the evaluation of physical and biological activity of biomaterials. Objective: This study investigated surface alterations of calcium silicate-based cements after exposure to different environments. Material and Methods: Forty-eight cylindrical cavities were prepared on root surfaces. The cavities were filled using ProRoot MTA or Biodentine and assigned to four subgroups (n=6): dry, wet, acidic, and blood. Surface topographies were evaluated using an optical profilometer for 28 days, and the roughness of the material surfaces was quantified. Vertical dimensional change was measured by determining the height difference between the material surface and the flat tooth surface. Data were compared with a two-way repeated measures ANOVA and Bonferroni tests. Results: In dry condition, the surface roughness of MTA or Biodentine was constant up to 3 days (p>0.05) but decreased after 28 days (p<0.05). In dry condition, ProRoot MTA presented constant surface level through time, while Biodentine showed decreased surface level after 28 days. In wet condition, the roughness and the surface levels of both materials increased after 1 day (p<0.05). Neither the surface roughness nor the levels of the materials showed significant changes in acidic conditions (p>0.05). Both materials showed the highest roughness in blood conditions on the 1st day (p<0.05), while the surface roughness in blood decreased dramatically after 28 days. The roughness of Biodentine was higher in wet conditions up to 3 days compared with ProRoot MTA (p<0.05). Likewise, in blood condition, Biodentine showed higher roughness on the 28th day than ProRoot MTA (p<0.05). Conclusions: Dry, wet, and blood conditions had a time-dependent effect on the surface roughness and vertical dimensional changes of the materials. However, acidic conditions did not affect the roughness and the surface level of the materials.

Effect of sonic and ultrasonic activation on physicochemical properties of root canal sealers

Abstract Objective: To evaluate the effect of ultrasonic and sonic activation on physicochemical properties of AH Plus, MTA Fillapex, ADSeal, GuttaFlow Bioseal, and GuttaFlow 2 sealers. Methodology: Three experimental groups were formed: no activation (NA), ultrasonic activation (UA), and sonic activation (SA). The sealers were manipulated according to the manufacturers' instructions. A 3-mL syringe was adapted to receive 1 mL of sealer. Activation was performed with a 20/.01 ultrasonic insert (20 s/1W) in the UA group. A size 35.04 sonic tip was used (20 s/10,000 cycles/min-1) in the SA group. The molds for physicochemical analysis were filled and evaluated according to ANSI/ADA specification no. 57: setting time (ST), flow (FL), dimensional change (DC), solubility (SB), and radiopacity (RD). Statistical analysis was performed by Kruskal-Wallis, one-way ANOVA, and Tukey's tests (P<0.05). Results: Regarding ST, only AH Plus and GuttaFlow 2 in the NA group met the ANSI/ADA standards. All FL values were greater than 20 mm in diameter, as determined by ANSI/ADA. The tested sealers and protocols did not comply with the ANSI/ADA standards for DC. As for SB, only MTA Fillapex, regardless of the activation protocol, did not follow the ANSI/ADA standards. All of the investigated sealers, regardless of the activation protocol, presented radiographic density higher than 3 mm Al, as proposed by ANSI/ADA. Conclusions: UA and SA promoted changes in the physicochemical properties of the evaluated root canal sealers, mainly in ST and F. Thus, it is important to evaluate the physicochemical properties of endodontic sealers associated with activation techniques prior to clinical application in order to determine whether the properties follow the parameters set by ANSI/ADA, ensuring safety and quality of root canal filling.

Penetration of bioceramic and epoxy-resin endodontic cements into lateral canals

Abstract: The aim of the present study was to assess the penetration capacity of two endodontic cements, Endosequence BC Sealer and AH Plus, in artificial lateral canals. Twenty-six two-rooted, maxillary first premolars were instrumented to size 40.06 using K3 files. In each root, six lateral canals of two diameters (0.06 and 0.10 mm) were created with a working length of 2, 4, and 6 mm. The specimens were randomly divided into two groups according to the endodontic cement to be used (Endosequence BC Sealer and AH Plus) and obturated by the single-cone technique. The specimens were imaged by digital periapical radiography and scores from 0 to 4 were attributed according to the degree of penetration by sealers into the lateral canals. Data were analyzed statistically by Kruskal-Wallis and Student-Newman-Keuls tests, and a significance level of p < 0.05 was adopted. No significant difference was observed between the two endodontic cements used to fill the simulated lateral canals (p > 0.05). The diameter of lateral canals only influenced the capacity of the Endosequence BC Sealer in filling the canals, and presented greater penetration in the lateral canals of diameter 0.10 mm (p < 0.05). We concluded that the bioceramic endodontic cement Endosequence BC Sealer presented similar ability as AH Plus to fill simulated lateral canals.

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.

Ytterbium oxide as radiopacifier of calcium silicate-based cements. physicochemical and biological properties

Abstract This study evaluated physicochemical properties, cytotoxicity and bioactivity of MTA Angelus (MTA), calcium silicate-based cement (CSC) and CSC with 30% Ytterbium oxide (CSC/Yb2O3). Setting time was evaluated using Gilmore needles. Compressive strength was evaluated in a mechanical machine. Radiopacity was evaluated using radiographs of materials and an aluminum scale. Solubility was evaluated after immersion in water. Cell viability was evaluated by means of MTT assay and neutral red staining, and the mineralization activity by using alkaline phosphatase activity and Alizarin Red staining. The data were submitted to ANOVA, Tukey and Bonferroni tests (5% significance). The bioactive potential was evaluated by scanning electron microscopy. The materials presented similar setting time. MTA showed the lowest compressive strength. MTA and CSC/Yb2O3 presented similar radiopacity. CSC/Yb2O3 showed low solubility. Saos-2 cell viability tests showed no cytotoxic effect, except to 1:1 dilution in NR assay which had lower cell viability when compared to the control. ALP at 1 and 7 days was similar to the control. MTA and CSC had greater ALP activity at 3 days when compared to control. All the materials present higher mineralized nodules when compared with the control. SEM analysis showed structures suggesting the presence of calcium phosphate on the surface of materials demonstrating bioactivity. Ytterbium oxide proved to be a properly radiopacifying agent for calcium silicate-based cement since it did not affected the physicochemical and biological properties besides preserving the bioactive potential of this material.

Resumo Este estudo avaliou as propriedades físico-químicas, citotoxicidade e bioatividade do MTA Angelus (MTA), cimento à base de silicato de cálcio (CSC) e CSC com 30% de óxido de itérbio (CSC/Yb2O3). O tempo de presa foi avaliado usando agulhas Gilmore. A resistência à compressão foi avaliada em uma máquina mecânica. A radiopacidade foi avaliada utilizando radiografias dos materiais e uma escala de alumínio. A solubilidade foi avaliada após imersão em água. A viabilidade celular foi avaliada por meio do ensaio MTT e coloração de vermelho neutro (NR), e a atividade de mineralização por meio da atividade da fosfatase alcalina e a coloração com Vermelho de Alizarina. Os dados foram submetidos aos testes ANOVA, Tukey e Bonferroni (5% de significância). O potencial bioativo foi avaliado por microscopia eletrônica de varredura. Os materiais apresentaram tempo de presa semelhante. O MTA mostrou menor resistência à compressão. MTA e CSC/Yb2O3 apresentaram radiopacidade semelhante. CSC/Yb2O3 apresentou menores valores de solubilidade. A viabilidade celular realizada pelos ensaios de MTT e NR não revelaram efeitos ctotóxicos em todas as diluições, exceto na diluição 1:1 no NR, o qual mostrou baixa viabilidade celular (p<0.05) em todos materiais testados quando comparado ao controle. A atividade de ALP em 1 e 7 dias foi similar ao controle (p>0.05). MTA e CSC tiveram significante aumento na atividade de ALP aos 3 dias quando comarados ao controle (p>0.05). Todos os materiais apresentaram grande produção de nódulos mineralizados quando comparados ao controle (P<0.05). A análise da SEM mostrou estruturas que sugerem a presença de depósitos de fosfato de cálcio na superfície dos materiais demonstrando bioatividade. O Yb2O3 mostrou ser um agente radiopacificador adequado em cimentos à base de silicato de cálcio uma vez que não afetou as propriedades físico-químicas e biológicas e ainda preservou o potencial bioativo desse material.

Effect of blood contamination on the push-out bond strength of calcium silicate cements

Abstract This study investigated the effect of blood-contamination on the push-out bond strength of BiodentineTM (BD) and MTA Angelus® (MTA-A) to root dentin over time. Twenty-five teeth were sectioned horizontally to obtain 120 root slices. The lumens were filled with MTA-A or BD: 60 for each cement (30 uncontaminated and 30 blood contaminated). Push out bond strength to dentin was assessed at 24 h (n=10), 7 days (n=10) and 28 days (n=10). Failure modes were classified as: cohesive, adhesive or mixed failure. Two-way ANOVA was used to investigate the interaction between blood contamination vs. hydration period. Mann Whitney test compared different materials in each period, and it also compared the contaminated versus uncontaminated material for each period. Friedman, followed by Dunn`s test, compared periods of hydration for each material, regardless of blood contamination. Failure modes were reported descriptively. The interaction hydration period vs. blood contamination was highly significant for MTA-A (P=0.001) and it was not significant for BD (P=0.474). There were no differences between bond strength of uncontaminated and contaminated BD in any of the periods. Bond strength of uncontaminated MTA-A increased at each time of hydration; but it remained stable over time for blood-contaminated samples. BD had higher bond strength than MTA-A in all periods of hydration. Cohesive failure predominated. Only for MTA-A, the over time bond strength to dentin was affected by blood contamination.

Resumo Este estudo investigou o efeito da contaminação sanguínea na resistência de união do BiodentineTM (BD) e do MTA Angelus® (MTA-A) à dentina, em diferentes períodos. Vinte e cinco dentes foram seccionados para obter 120 fatias de dentina. Os lúmens das fatias foram preenchidos com MTA-A ou BD: 60 para cada cimento (30 não-contaminados e 30 contaminados com sangue). A resistência de união à dentina foi medida por teste push-out em 24 horas (n=10), 7 dias (n=10) e 28 dias (n=10). Os tipos de falha foram classificados como: falha coesiva, adesiva ou mista. Two-way ANOVA foi usado para investigar a interação entre contaminação sanguínea vs. período de hidratação. O teste de Mann Whitney comparou os diferentes materiais em cada período, e comparou as amostras contaminadas e não contaminadas de cada material em cada tempo. O teste de Friedman, seguido pelo teste de Dunn, comparou os períodos de hidratação de cada material, independentemente da contaminação. A análise estatística mostrou a interação entre contaminação sanguínea vs. período de hidratação. Os tipos de falha foram reportados de maneira descritiva. A interação entre contaminação sanguínea vs. período de hidratação foi altamente significativa para o MTA-A (P=0,001), e não foi significativa para o BD (P=0,474). Não houve diferenças entre a resistência de união entre o BD contaminado e não-contaminado independente do período. A resistência de união do MTA-A não-contaminado aumentou a cada tempo de hidratação; mas, permaneceu estável ao longo do tempo para as amostras contaminadas com sangue. BD obteve maior resistência de união que o MTA-A em todos os períodos de hidratação. Falhas coesivas predominaram. A contaminação ao longo do tempo influenciou a resistência de união no grupo MTA-A.

Bond strength of experimental root canal sealers based on mta and butyl ethylene glycol disalicylate

Abstract This study investigated the bond strength of two experimental root canal sealers based on MTA and butyl ethylene glycol disalicylate: MTAe and MTAe-HA. The reference materials used for comparison were AH Plus and MTA Fillapex. Twenty human upper incisors were selected and one 1 mm slice was obtained from the cervical third of each root. On the coronal surface of each slice, four 0.9 mm wide holes were drilled through the dentine. Standardized irrigation was performed and holes were filled with one of the four tested sealers: AH Plus, MTA Fillapex, MTAe, and MTAe-HA. The filled slices were stored in a PBS solution (pH 7.2) for 7 days at 37 °C. A push-out assessment was performed with a 0.7 mm plunger tip. Load was applied at a crosshead speed of 0.5 mm/min until sealer displacement. The results were expressed in MPa. The Kruskal-Wallis test was applied to assess the effect of each sealer on the push-out bond strength. Mann-Whitney with Bonferroni correction was used to isolate the differences. The alpha-type error was set at 0.05. Significant differences among medians values obtained by materials were observed (p<0.001). AH Plus displayed the highest value of bond strength (p<0.001). In contrast, MTA Fillapex presented the lowest bond strength among all tested sealers (p<0.001). Experimental sealers showed intermediary bond strength values, with no statistical differences between them (p>0.05). In conclusion, experimental root canal sealers presented suitable bond strength outcomes when compared to MTA Fillapex.

Resumo Esse estudo investigou a resistência de união de dois cimentos endodônticos experimentais à base de MTA e butiletilenoglicol dissalicilato: MTAe e MTAe. Os materiais de referência utilizados para comparação foram os cimentos endodônticos MTA Fillapex e AH Plus. Vinte incisivos superiores humanos foram selecionados e um slice dentinário de 1 mm de espessura foi obtido do terço cervical de cada raiz. Na superfície coronária de cada slice, quatro orifícios com 0,9 mm de diâmetro foram confeccionados através da dentina. Uma irrigação padronizada foi realizada e os orifícios foram preenchidos com um dos quatro cimentos endodônticos avaliados: AH Plus, MTA Fillapex, MTAe, e MTAe-HA. Os slices preenchidos foram armazenados em solução PBS (pH 7,2) durante 7 dias a 37°C. O ensaio de push-out foi realizado por meio de um dispositivo com 0,7 mm de diâmetro. A carga foi aplicada com a velocidade de 0,5 mm/min até a obtenção de deslocamento do material obturador. Os resultados foram expressos em MPa. O teste de Kruskal-Wallis foi aplicado para avaliar o efeito da resistência de união de cada cimento endodôntico. O teste de Mann-Whitney com correção de Bonferroni foi utilizado para isolamento das diferenças. O erro do tipo-alfa foi fixado em 0,05. Diferenças significantes entre os valores de medianas obtidos pelos materiais foram observados (p<0,001). O AH Plus demonstrou os maiores valores de resistência de união (p<0,001). Em contraste, o MTA Fillapex apresentou a menor resistência de união entre todos os cimentos testados (p<0,001). Os cimentos experimentais demonstraram valores intermediários, com ausência de diferenças estatísticas entre si (p>0,05). Em conclusão, os cimentos endodônticos experimentais à base de MTA e butiletilenoglicol dissalicilato apresentaram resultados adequados de resistência de união quando comparados ao MTA Fillapex.

Evaluation of the sealing ability of different root canal sealers: a combined SEM and micro-CT study

Abstract Objective: The purpose of this study was to analyze the ability of multiple compounds to seal the dental tubules using scanning electron microscopy (SEM) and micro-computed tomogra-phy (micro-CT). Material and Methods: Twenty-four single-root human mandibular premolars were selected and instrumented with nickel-titanium rotary file and the final file size was #40/06. They were then randomly allocated into 2 groups, and all samples were filled with single cone gutta-percha (#40/06) and one of the tested sealers (AH Plus and EndoSequence BC sealers). All specimens were scanned using micro-CT and then three from each group were randomly selected for SEM analysis. Results: According to SEM, both root canal sealers showed sufficient adaptation to dentin along the whole length of the root canal, though the coronal sections presented superior sealing than the apical sections. Micro porosity analyses revealed that the volume of closed pores and the surface of closed pores had the largest values in the coronal sections, followed by the middle and the apical sections for both sealants (p<0.05). However, no significant difference was observed for those two parameters between AH Plus and EndoSequence BC sealers in any of the three sections (p>0.05), whereas they were larger in the apical section when the AH Plus sealer was used. Conclusions: By using the single cone technique, neither EndoSequence or AH Plus pro-vides a porosity-free root canal filling. The EndoSequence BC sealer may have similar sealing abilities regarding the whole root canal as the AH Plus sealer. A better sealing effect could be obtained in the coronal and middle sections of a root canal than the apical part by using the tested sealers.

Solubility, porosity and fluid uptake of calcium silicate-based cements

Abstract Objective: To evaluate the absorption/fluid uptake, solubility and porosity of White mineral trioxide aggregate (MTA) Angelus, Biodentine (BIO), and zinc oxide-eugenol (ZOE). Material and Methods: Solubility was evaluated after immersion in distilled water for 7 and 30 days. Porosity was evaluated using digital inverted microscope, scanning electron microscope (SEM) and micro-computed tomography (micro-CT). For the fluid uptake test, specimens were immersed in Hank's balanced salt solution (HBSS) for 1, 7, 14 and 28 days. Fluid absorption, solubility and porosity of the materials were measured after each period. Statistical evaluation was performed using one-way analysis of variance (ANOVA) and Tukey tests, with a significance level at 5%. Results: After 7 and 30 days, BIO showed the highest solubility (p<0.05). All methods demonstrated that MTA had total porosity higher than BIO and ZOE (p<0.05). Micro-CT analysis showed that MTA had the highest porosity at the initial period, after its setting time (p<0.05). After 7 and 30 days, ZOE had porosity lower than MTA and BIO (p<0.05). Absorption was similar among the materials (p>0.05), and higher fluid uptake and solubility were observed for MTA in the fluid uptake test (p<0.05). Conclusions: BIO had the highest solubility in the conventional test and MTA had higher porosity and fluid uptake. ZOE had lower values of solubility, porosity and fluid uptake. Solubility, porosity and fluid uptake are related, and the tests used provided complementary data.

Tricalcium silicate-based cements: properties and modifications

Abstract: Mineral trioxide aggregate (MTA) has been widely used for different reparative procedures in endodontics. The extensive use of this cement for pulp capping, apexifications, apical surgeries, and revascularization is related to its ability to induce tissue repair and to stimulate mineralization. Several research studies have tested modifications in the composition of MTA-based cements in order to enhance their clinical performance. Novel formulations have been introduced in the market with the aim of increasing flowability. Important properties such as appropriate radiopacity and setting time, color stability, alkaline pH, release of calcium ions, and biocompatibility have to be considered in these new formulations. The latest research studies on the physical, chemical, and biological properties of tricalcium silicate-based cements are discussed in this critical review.

Physicochemical properties of calcium silicate-based formulations MTA Repair HP and MTA Vitalcem

Abstract Objective This study aimed to analyze the following physicochemical properties: radiopacity, final setting time, calcium release, pH change, solubility, water sorption, porosity, surface morphology, and apatite-forming ability of two calcium silicate-based materials. Material and methods We tested MTA Repair HP and MTA Vitalcem in comparison with conventional MTA, analyzing radiopacity and final setting time. Water absorption, interconnected pores and apparent porosity were measured after 24-h immersion in deionized water at 37°C. Calcium and pH were tested up to 28 d in deionized water. We analyzed data using two-way ANOVA with Student-Newman-Keuls tests (p<0.05). We performed morphological and chemical analyses of the material surfaces using ESEM/EDX after 28 d in HBSS. Results MTA Repair HP showed similar radiopacity to that of conventional MTA. All materials showed a marked alkalinizing activity within 3 h, which continued for 28 d. MTA Repair HP showed the highest calcium release at 28 d (p<0.05). MTA Vitalcem showed statistically higher water sorption and solubility values (p<0.05). All materials showed the ability to nucleate calcium phosphate on their surface after 28 d in HBSS. Conclusions MTA Repair HP and MTA Vitalcem had extended alkalinizing activity and calcium release that favored calcium phosphate nucleation. The presence of the plasticizer in MTA HP might increase its solubility and porosity. The radiopacifier calcium tungstate can be used to replace bismuth oxide.

Push-out bond strength of different tricalcium silicate-based filling materials to root dentin

Abstract: The aim of this study was to evaluate the bond strength of different triccalcium silicate cements to retrograde cavity using a push out test. Thirty maxillary central incisors were shaped using #80 hand files and sectioned transversally. Root slices were obtained from the apical 4 mm after eliminating the apical extremity. The specimens were embedded in acrylic resin and positioned at 45° to the horizontal plane for preparation of root-end cavities with a diamond ultrasonic retrotip. The samples were divided into three groups according to the root-end filling material (n = 10): MTA Angelus, ProRoot MTA and Biodentine. A gutta-percha cone (#80) was tugged-back at the limit between the canal and the root-end cavity. The root-end cavity was filled and the gutta-percha cone was removed after complete setting of the materials. The specimens were placed in an Instron machine with the root-end filling turned downwards. The push-out shaft was inserted in the space previously occupied by the gutta-percha cone and push out testing was performed at a crosshead speed of 1.0 mm/min. There was no statistically significant difference in resistance to push out by the materials tested (p > 0.01). MTA Angelus and ProRoot MTA showed predominantly mixed failure while Biodentine exhibited mixed and cohesive failures. The tricalcium silicate-based root-end filling materials showed similar bond strength retrograde cavity.

Physical properties and biological effects of mineral trioxide aggregate mixed with methylcellulose and calcium chloride

Abstract Objectives: Methylcellulose (MC) is a chemical compound derived from cellulose. MTA mixed with MC reduces setting time and increases plasticity. This study assessed the influence of MC as an anti-washout ingredient and CaCl2 as a setting time accelerator on the physical and biological properties of MTA. Material and Methods: Test materials were divided into 3 groups; Group 1(control): distilled water; Group 2: 1% MC/CaCl2; Group 3: 2% MC/CaCl2. Compressive strength, pH, flowability and cell viability were tested. The gene expression of bone sialoprotein (BSP) was detected by RT-PCR and real­ time PCR. The expression of alkaline phosphatase (ALP) and mineralization behavior were evaluated using an ALP staining and an alizarin red staining. Results: Compressive strength, pH, and cell viability of MTA mixed with MC/CaCl2 were not significantly different compared to the control group. The flowability of MTA with MC/CaCI2 has decreased significantly when compared to the control (p<.05). The mRNA level of BSP has increased significantly in MTA with MC/CaCl2 compared to the control (p<.05). This study revealed higher expression of ALP and mineralization in cells exposed to MTA mixed with water and MTA mixed with MC/CaCl2 compared to the control (p<.05). Conclusions: MC decreased the flowability of MTA and did not interrupt the physical and biological effect of MTA. It suggests that these cements may be useful as a root-end filling material.

Effect of photodynamic therapy and non-thermal plasma on root canal filling: analysis of adhesion and sealer penetration

Abstract Objective The aim of this study was to evaluate the effect of photodynamic therapy (PDT) and non-thermal plasma (NTP) on adhesion and sealer penetration in root canals. Material and Methods Sixty single-rooted premolars were used. The teeth were prepared using a crown-down technique. NaOCl and EDTA were used for irrigation and smear layer removal, respectively. The root canals were divided into three groups: control, PDT, and NTP. After treatments, the roots were filled using gutta-percha and either AH Plus (AHP) or MTA Fillapex (MTAF) sealers. Samples were sectioned at 4, 8, and 12 mm from the apex (1-mm slices)and analyzed by the push-out bond strength test (adhesion) and confocal laser scanning microscopy (sealer penetration). Data were statistically evaluated using Kruskal-Wallis, Dunn's, and Spearman's tests. Results Regarding AHP, bond strength was similar in the NTP group and in the control group, but significantly lower in the PDT group. As to MTAF, both therapies showed lower values than the control group. In the confocal analysis of AHP, maximum and mean penetration, and penetrated area were statistically higher in the control group than in the PDT and NTP groups. Penetrated perimeter was similar among groups. Regarding MTAF, all parameters yielded better results in the NTP than in the control group. The PDT and control groups showed similar results except for penetrated area. Conclusion PDT and plasma therapy affected the adhesion and sealer penetration of root canals filled with AH Plus and MTA Fillapex and there is no positive correlation between adhesion and sealer penetration.

Evaluation of physicochemical properties of root-end filling materials using conventional and Micro-CT tests

Abstract Objective To evaluate solubility, dimensional stability, filling ability and volumetric change of root-end filling materials using conventional tests and new Micro-CT-based methods. Material and Methods Solubility (loss of mass) after 7 and 30 days, and dimensional stability (in mm) were evaluated in accordance with Carvalho-Junior, et al. 7 (2007). The filling ability and volumetric change (in mm3) were evaluated by Micro-CT (Bruker-MicroCT, Kontich, Belgium) using resin models with cavities 3 mm deep and 1 mm in diameter. The cavities were filled with materials to evaluate filling ability, and then scanned by Micro-CT. After 7 and 30 days immersed in distilled water, the filled cavities were scanned again to evaluate the volumetric change. MTA Angelus (MTA), Biodentine (BIO) and zinc oxide-eugenol cement (ZOE) were evaluated. Data were submitted to analysis of variance (ANOVA) and Tukey's test with 5% significance level. Results The results suggested correlated or complementary data between the proposed tests. At 7 days, BIO showed higher solubility and at 30 days, showed higher volumetric change in comparison with MTA (p<0.05). With regard to volumetric change, the tested materials were similar (p>0.05) at 7 days. At 30 days, they presented similar solubility. BIO and MTA showed higher dimensional stability than ZOE (p<0.05). ZOE and BIO showed higher filling ability (p<0.05). Conclusions ZOE presented a higher dimensional change, and BIO had greater solubility after 7 days. BIO presented filling ability and dimensional stability, but greater volumetric change than MTA after 30 days. Micro-CT can provide important data on the physicochemical properties of materials complementing conventional tests.

Crown discoloration promoted by materials used in regenerative endodontic procedures and effect of dental bleaching: spectrophotometric analysis

Abstract Regenerative endodontic procedure (REP) has been proposed as a new approach to treat immature permanent teeth. However, materials used in REP for root canal disinfection or cervical sealing may induce tooth discoloration. Objectives To assess tooth crown’s color after intracanal treatment with triple antibiotic paste (TAP) or calcium hydroxide (CH); cervical sealing with glass ionomer cement (GIC) or mineral trioxide aggregate (MTA); and bleaching with carbamide peroxide. Material and Methods After pulp removal and color spectrophotometer measurement, 50 bovine incisors were divided into 4 experimental groups and one control (untreated). Experiments were performed in phases (Ph). Ph1: TAP (ciprofloxacin, metronidazole, minocycline), TAPM (ciprofloxacin, metronidazole, amoxicillin), DAP (ciprofloxacin, metronidazole), or CH treatment groups. After 1 and 3 days (d); 1, 2, 3 weeks (w); and 1, 2, 3 and 4 months (m), color was measured and medications were removed. Ph2: GIC or MTA cervical sealing, each using half of the specimens from each group. Color was assessed after 1d, 3d; 1w, 2w, 3w; 1m and 2m. Ph3: Two bleaching sessions, each followed by color measurement. Data were analyzed with ANOVA and post-hoc Holm-Sidak method. Results Ph1: Specimens of TAP group presented higher color alteration (ΔE) mean than those of TAPM group. No significant difference was found among TAP or TAPM and CH, DAP or Control groups. Ph2: cervical sealing materials showed no influence on color alteration. Ph3: Different ΔE means (from different groups), prior to bleaching, became equivalent after one bleaching session. Conclusions TAP induces higher color alteration than TAPM; color alteration increases over time; cervical sealing material has no influence on color alteration; and, dental bleaching was able to recover, at least partially, the tooth crown’s color.

Evaluation of sealing ability, color alteration and surface morphology of MTA and Portland cement associated with different radiopacifiers and liquids / Avaliação da capacidade seladora, alteração de cor e de componentes superficiais do MTA e cimento Portland associado à diferentes radiopacificadores e diferentes líquidos

This study aimed to analyze the influence of two different vehicles for MTA manipulation on color alteration, chemical characteristics, and radiopacity after immersion in blood or distilled water (DW). In addition, some physical-chemical properties, volumetric solubility and bioactivity of new brands of calcium silicate cements such as MTA Repair HP, MTA Vitalcem and MTA Flow were compared with MTA Angelus. MTA Angelus mixed with 100% Distilled Water (DW) or 80% DW/20% propylene glycol (PG) as vehicles were placed into rubber rings and incubated at 37°C and 100% relative humidity. Color assessment and scanning electron microscopy/energy-dispersive spectroscopy (SEM/EDX) analysis were performed after setting and repeated after 7, 15, and 30 days after immersion in blood and DW. The data were analyzed by using Kruskal-Wallis followed by the Dunn test. The association of 80% DW/20% PG used as vehicle presented significantly lower color alterations compared with 100% DW, when immersed in blood (p < 0.05). All samples surfaces displayed loss of bismuth after immersion in both media and consequently a decrease in radiopacity was observed with significant results after 30 days. This study showed that ratio of 80% DW/20% PG as a vehicle for MTA Angelus results in a lower color alteration when in contact with blood. The physical-chemical properties such as radiopacity, calcium release, the pH change, solubility, water sorption, porosity, surface morphology, and bioactivity of MTA Repair HP and MTA Vitalcem were compared with MTA Angelus. Water absorption, interconnected pores and apparent porosity were measured after 24h-immersion in DW at 37ºC. Calcium and pH were tested up to 28d in DW. Morphological and chemical analyses of the material surfaces were performed by using ESEM-EDX after 28d in HBSS. All the materials showed good bioactivity after 28d in HBSS. The data were analyzed by using two-way ANOVA followed by the RM Student-Newman-Keuls test (p<0.05). MTA Repair HP and MTA Vitalcem had some positives properties, such as extended alkalinizing activity and calcium release that favored the bioactivity. However, MTA Vitalcem showed scant radiopacity, long setting time and high solubility. The radiopacifier Calcium Tungstate, present in MTA Repair HP can be used to replace the traditional Bismuth Oxide. In relation to the comparison between MTA Angelus and MTA Flow, it was analyzed the pH, volumetric change, radiopacity and bioactivity. In the volumetric change test, the root-end fillings of 20 acrylic teeth with both materials were scanned twice by a Micro- CT, before and after immersion in DW for 168 h. The morphological and bioactivity of the materials were performed by using SEM-EDX after 28d in HBSS. The data were analyzed by using two-way ANOVA followed by the RM Student-Newman-Keuls test (p<0.05). MTA Angelus showed statistically higher radiopacity values (p<0.05). the results pointed that the new cement MTA Flow showed good physical-chemical characteristics and bioactivity, showing values similar to those of MTA Angelus. (AU)

O objetivo do estudo foi analisar a influência de dois diferentes veículos utilizados na manipulação do MTA em relação a alteração de cor, características superficiais e radiopacidade, antes de depois do contato com sangue ou Água Destilada (AD). Adicionalmente, foram analisadas as propriedades físico-quimicas, solubilidade volumétrica e a bioatividade de novas marcas comerciais de cimentos de silicato de cálcio como MTA Repair HP, MTA Vitalcem e MTA Flow comparados com o MTA Angelus. Foram realizadas associações do MTA Angelus com 100% de Água Destilada (AD) e outra com 80% de AD e 20% de Propilenoglicol (PG) como veículos e, após isso, estas foram inseridas em anéis de borracha e encubadas a 37ºC e 100% de humidade. A análise de cor e realização do escaneamento da amostra em microscópio eletrônico de varredura (MEV) juntamente com a espectroscopia de energia dispersiva (EDX) foram realizadas após a presa e depois de 7, 15 e 30 dias após a imersão em sangue ovino e água destilada. Os dados foram submetidos aos testes não paramétricos de Kruskal-Wallis e Dunn. A associação com o veículo de 80% AD com 20% de PG apresentou alteração de cor significativamente menor quando comparada com o veículo de 100% de AD quando imersos em sangue (p < 0.05). Em todas as amostras a análise superficial comprovou a perda de partículas de bismuto durante a imersão nos diferentes líquidos e consequentemente a redução da radiopacidade apresentando resultados significativos após 30 dias. Esse estudo demonstrou que o uso do veículo 80% de AD com 20% de PG no MTA Angelus apresentou menor alterações de cor quando em contato com sangue. As propriedades físico químicas como radiopacidade, liberação de cálcio, pH, solubilidade, absorção de água, porosidade, análise das características superficiais e bioatividade dos cimentos MTA Repair HP e MTA Vitalcem foram comparadas com as do MTA Angelus. A absorção de água e a porosidade aparente foram analisadas 24h após a imersão dos espécimes em AD a 37ºC. A liberação e cácio e o pH foram anaisados após 28d de imersão em AD. As características superficiais e a Bioatividade foram analisadas em MEV/EDX após 28 dias de imersão em HBSS. Os dados foram submetidos aos testes de two-way ANOVA seguido por RM Student-Newman-Keuls (p<0.05). Todos os materiais apresentaram boa bioatividade após 28 dias de imersão em HBSS. O MTA Repair HP e o MTA Vitalcem apresentaram boa atividade alcalinizante e liberação de ions cálcio favorecendo a bioatividade. Entretanto o MTA Vitalcem mostrou baixa radiopacidade, longo tempo de presa e alta solubilidade. O radiopacificador Tungstato de Cálcio, utilizado no MTA Repair HP pode ser usado como substituto do tradicional óxido de bismuto. Em relação a comparação entre o MTA Angelus e o MTA Flow, foram realizados as analises de pH, alteração volumétrica, radiopacidade e bioatividade. Em relação a alteração volumétrica, 20 retro-cavidades de dentes de acrílico preenchidas com os materiais foram escaneadas duas vezes em um aparelho de Micro-CT antes e após a imersão por 168h em AD. As características superficiais e a bioatividade foram realizadas por MEV/EDX depois de 28d de imersão em HBSS. Os dados foram submetidos aos testes de two-way ANOVA seguido por RM Student-Newman-Keuls (p<0.05). O MTA Angelus apresentou estatisticamente maior radiopacidade (p<0.05). Os resultados apontaram que o novo cimento MTA Flow apresentou boas características físico-quimicas e bioatividade similares ao MTA Angelus. (AU)

Physical-chemical properties and cytotoxicity analysis of 5 different endodontic sealers / Propriedades físico-químicas e análise da citotoxicidade de 5 diferentes cimentos endodônticos

The aim of this study was to analyze the radiopacity, setting time, flowability, pH, calcium ion release, solubility and cytotoxicity of bioceramic cements Totalfill BC Sealer and Totalfill BC RRM, and compare them with AH Plus, MTA Fillapex and MTA Angelus. The groups were divided and compared among them according to the filling and retro-filling cement functions. Totalfill BC Sealer was compared with AH Plus and MTA Fillapex; and Totalfill BC RRM retrofilling cement with MTA Angelus. For radiopacity analysis, specimens were placed in metal rings measuring 10x1 mm placed on occlusal film together with the aluminum scale. Digora 1.51 software was used to evaluate the digitized images and determine radiographic density. Setting time was tested in accordance with the American Society for Testing and Materials C266-08 standard specifications, but specimens were fabricated in accordance with the International Organization for Standardization 6876: 2001. Flow was tested in accordance with ANSI/ADA No.57/200 specifications. In total 30 acrylic teeth were filled with filling-cements and 20, with (retrograde cavity) retro-filling cements. All teeth were immersed in ultrapure water for pH and calcium ion release measurement (atomic absorption spectrophotometer) for time intervals of 1, 3, 24, 72, 168 and 360 hours. Solubility was tested by scanning and digitizing 50 acrylic teeth twice by Micro- CT, before and after immersion in ultrapure water for time intervals of 168, 360 and 720 hours. The images were reconstructed and volume (mm3) values of samples obtained by means of CTan software (CTan v1.11.10.0, SkyScan). The in vitro effects on cells were analyzed at concentrations of 100, 50, 10, 5, 1 mg/mL, and 0 mg / mLnegative control group and recorded in time intervals of 24, 48 and 72 hours by MTT reduction assay. The results were statistically analyzed by the ANOVA, Tukey, Kruskal-Wallis and Dunn tests (P<0.05). All radiopacity values according to ISO 6876/2001, AH Plus (7.86 mm Al) being the most radiopaque followed by Totalfill BC Sealer (4.84 mm Al), MTA Fillapex (3.41 mm Al), Totalfill BC RRM (6.8 mm Al), and MTA Angelus (6.7 mm Al). The following values were the initial and final setting time (in hours), respectively: AH Plus (8 and 15); Totalfill BC Sealer (11 and 24); MTA Fillapex (13 and 26); MTA Angelus (10 and 120 minutes) and Totalfill BC RRM (3 hours and 22 hours). In flow analysis, the cements behaved as follows: MTA Fillapex (47 mm), Totalfill BC Sealer (41.5 mm), Totalfill BC RRM (33.5 mm), AH Plus (33 mm) e MTA Angelus (17.5 mm) (p < 0.05). pH analysis showed in general the lowest values for AH Plus cement, followed by Totalfill BC RRM, MTA Angelus, MTA Fillapex and Totalfill BC Sealer. AH Plus showed the highest Ca2+ release in time interval 1 hour (1.38 mg/L); MTA Fillapex, in 360 hours (3.81 mg/L); MTA Angelus, 1 hour (1.38 mg/L); Totalfill BC Sealer, 360 hours (6.77 mg/L) and Totalfill BC RRM, 360 hours (3.81 mg/L). Almost all the sealers presented solubility lower than 3% in all periods, as recommended by ISO 6876/2001. Whereas, the MTA Fillapex solubility value was higher than 5% in all periods. Relative to cytotoxicity, all the cements were shown to be toxic at the concentration of 100 mg/mL, however, Totalfill BC Sealer and Totalfill BC RRM showed the best cell viability result compared with the other cements tested. We concluded that all root canal filling and root retro-filling complied with the requisites of radiopacity, setting time, flow, pH, calcium ion release, solubility and cytotoxicity. With the exception of the MTA Fillapex that not only fulfilled the requirement of solubility. Of the sealers, Totalfill BC Sealer was outstanding: it showed the highest pH and Ca2+ release, and lowest cytotoxicity. Among the retrofilling cements, Totalfill BC RRM maintained its high pH, higher Ca2+ release, and lower cytotoxicity. (AU)

O objetivo do presente estudo foi analisar a radiopacidade, tempo de presa, escoamento, pH, liberação de íons cálcio, solubilidade e citotoxicidade dos cimentos biocerâmicos Totalfill BC Sealer e Totalfill BC RRM e compará-los ao AH Plus, MTA Fillapex e MTA Angelus. Os grupos foram divididos e comparados entre si de acordo com as funções dos cimentos de obturação e retro-obturação. Comparamos o cimento obturador Totalfill BC Sealer com os cimentos AH Plus e MTA Filapex, e o cimento retrobturador Totalfill BC RRM com o cimento retrobturador MTA Angelus. Para análise da radiopacidade, os espécimes foram colocados em anéis metálicos medindo 10x1 mm, dispostos sobre um filme oclusal com uma escala de alumínio. O software Digora 1.51 foi utilizado para avaliar as imagens digitalizadas e determinar a densidade radiográfica. O tempo de presa foi realizado de acordo com as especificações da American Society for Testing and Materials C266-08 standard specifications, mas os espécimes foram feitos de acordo com a International Organization for Standardization 6876: 2001. O escoamento foi realizado de acordo com as especificações ANSI/ADA N0 57/2000. Trinta dentes acrílicos foram preenchidos com cimentos obturadores e vinte dentes de acrílico (com cavidade retrógrada) foram preenchidos com cimentos retro-obturadores e imersos em água ultrapura para mensuração do pH e liberação de íons cálcio (espectrofotômetro de absorção atômica) no período de 1, 3, 24, 72, 168 e 360 horas. Para o teste de solubilidade, foram escaneados 50 dentes acrílicos e digitalizados duas vezes pelo Micro-CT, antes e após a imersão em água ultrapura nos períodos de 168, 360 e 720 horas. As imagens foram reconstruídas e o volume (mm3) das amostras foi obtido usando o software CTan (CTan v1.11.10.0, SkyScan). Os efeitos celulares in vitro foram analisados nas concentrações de 100, 50, 10, 5, 1 mg/mL e 0 mg / mLgrupo controle negativo e registados nos períodos de 24, 48 e 72 horas através do ensaio de redução de MTT. Os resultados foram analisados estatisticamente pelos testes ANOVA, Tukey, Kruskal-Wallis e Dunn (p < 0.05). Todos os valores de radiopacidade estavam de acordo com a norma ISO 6876/2001, sendo o AH Plus (7.86 mm Al) o mais radiopaco seguido dos demais cimentos; Totalfill BC Sealer (4.84 mm Al), MTA Filapex (3.41 mm Al), Totalfill BC RRM (6,8 mm Al), MTA Angelus (6,7 mm Al). Os valores obtidos para o tempo de presa inicial e final foram respectivamente, AH Plus (8 e 15 horas), Totalfill BC Sealer (11 e 24 horas), MTA Filapex (13 e 26 horas), Totalfill BC RRM (3 horas e 22 horas) e MTA Angelus (10 e 120 minutos). Na análise de escoamento os cimentos se comportaram da seguinte forma: AH Plus (33 mm), MTA Filapex (47 mm), Totalfill BC Sealer (41,5 mm), Totalfill BC RRM (33,5 mm), e MTA Angelus (17,5 mm) (p < 0.05). A análise do pH mostrou que o cimento AH Plus de um modo geral, foi o que apresentou os menores valores, seguido do Totalfill BC RRM, MTA Angelus, MTA Filapex e Totalfill BC Sealer. A maior liberação de Ca2+ do AH Plus foi no período de 1 hora (1.38 mg/L), MTA Filapex foi em 360 horas (3.81 mg/L), Totalfill BC Sealer 360 horas (6.77 mg/L), Totalfill BC RRM 360 horas (3.81 mg/L) e MTA Angelus em 1 hora (1.38 mg/L). Todos os cimentos apresentaram solubilidade menor que 3% em todos os períodos, como recomendado pela ISO 6876/2001. Entretanto, os valores de solubilidade do MTA Fillapex excedeu mais que 5% em todos os períodos. Com relação à citotoxicidade, todos os cimentos mostraram-se tóxicos na concentração de 100 mg/mL, porém o Totalfill BC Sealer e Totalfill BC RRM apresentaram melhor resultado de viabilidade celular comparado aos demais cimentos testados. Concluiu-se que os cimentos de obturação e retro-obturação cumpriram os requisitos de radiopacidade, tempo de presa, escomento, pH, liberação de íons cálcio, solubilidade e citotoxicidade. Com exceção do MTA Fillapex que não cumpriu somente o requisito de solubilidade. Dos cimentos obturadores, o que melhor se portou foi o Totalfill BC Sealer, apresentando maior pH e liberação de íons cálcio e menor citotoxicidade. Dentre os cimentos retro-obturadores, Totalfill BC RRM foi o que melhor se destacou, mantendo seu pH elevado, possuindo maior liberação de Ca2+ e menor citotoxicidade. (AU)