Physicochemical properties, cytotoxicity and bioactivity of a ready-to-use bioceramic repair material

Abstract The aim of this study was to evaluate the physicochemical properties, cytotoxicity and bioactivity of a ready-to-use bioceramic material, Bio-C Repair (Angelus), in comparison with White MTA (Angelus) and Biodentine (Septodont). The physicochemical properties of setting time, radiopacity, pH, solubility, dimensional and volumetric changes were evaluated. Biocompatibility and bioactivity were assessed in Saos-2 osteoblast cell cultures by the MTT assay 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), Neutral Red (NR), Alizarin Red (ARS), and cell migration tests. Statistical analysis was performed by ANOVA, Tukey or Bonferroni tests (α = 0.05). Bio-C Repair had the longest setting time (p < 0.05), but radiopacity and solubility were accordance with the ISO 6876/2012 standards, besides linear expansion. Bio-C Repair and MTA had similar volumetric change (p > 0.05); lower than Biodentine (p < 0.05). All the materials evaluated had an alkaline pH. Bio-C Repair was cytocompatible and promoted mineralized nodule deposition in 21 days and cell migration in 3 days. In conclusion, Bio-C Repair had adequate radiopacity above 3mm Al, solubility less than 3%, dimensional expansion, and low volumetric change. In addition, Bio-C Repair promoted an alkaline pH and presented bioactivity and biocompatibility similar to MTA and Biodentine, showing potential for use as a repair material.

Resumo O objetivo deste estudo foi avaliar as propriedades físico-químicas, citotoxicidade e bioatividade de um novo material biocerâmico pronto para uso, Bio-C Repair (Angelus), em comparação com MTA Branco (Angelus) e Biodentine (Septodont). Foram avaliadas as propriedades físico-químicas de tempo de presa, radiopacidade, pH, solubilidade, alterações dimensionais e volumétricas. A biocompatibilidade, bioatividade e capacidade de reparo foram avaliadas em culturas de células de osteoblastos Saos-2 pelo ensaio MTT brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazólio), vermelho neutro (NR), vermelho de alizarina ( ARS) e testes de migração celular. A análise estatística foi realizada pelos testes ANOVA, Tukey ou Bonferroni (α = 0,05). Bio-C Repair apresentou o maior tempo de presa (p < 0,05), mas radiopacidade e solubilidade de acordo com as normas ISO 6876/2012, além de expansão linear. Bio-C Repair e MTA tiveram variação volumétrica semelhante (p > 0,05), menor que Biodentine (p < 0,05). Todos os materiais avaliados apresentaram pH alcalino. Bio-C Repair foi citocompatível, além de promover deposição de nódulos mineralizados em 21 dias e migração celular em 3 dias. Em conclusão, o Bio-C Repair apresentou radiopacidade adequada acima de 3mmAl, solubilidade menor que 3%, expansão dimensional e baixa perda volumétrica.. Além disso, o Bio-C Repair promoveu um pH alcalino e apresentou bioatividade e biocompatibilidade semelhantes ao MTA e Biodentine, mostrando potencial para uso como material reparador

Physicochemical properties, cytocompatibility and antibiofilm activity of a new calcium silicate sealer

Abstract The aim of this study was to evaluate the physicochemical properties, cytocompatibility and antibiofilm activity of a new calcium silicate-based endodontic sealer, Sealer Plus BC (MK Life, Brazil), in comparison with TotalFill BC Sealer (FKG Dentaire SA, Switzerland) and AH Plus (Dentsply, Germany). Setting time and flow were evaluated based on ISO 6876 standard. The pH was evaluated after different periods, and radiopacity by radiographic analysis (mmAl). Solubility (% mass loss) and volumetric change (by micro-CT) were assessed after 30 days of immersion in distilled water. Cytocompatibility was assessed by methyltetrazolium (MTT) and neutral red (NR) assays, after exposure of Saos-2 cells to the sealer extract for 24 h. An additional analysis was performed by using MTT assay after 1, 3 and 7 days of exposure of Saos-2 to the sealers 1:8 dilution extracts. Antibiofilm activity against Enterococcus faecalis and/or Candida albicans was evaluated by crystal violet assay and modified direct contact test. The physicochemical properties were analyzed using ANOVA/Tukey tests; MTT and NR data were analyzed by ANOVA and Bonferroni tests; the antimicrobial tests were analyzed by Kruskal-Wallis and Dunn tests (α=0.05). Sealer Plus BC had proper setting time, radiopacity, flow and alkalization capacity. Sealer Plus BC was significantly more soluble than AH Plus (p<0.05) and presented volumetric change similar to AH Plus and TotalFill BC (p>0.05). Sealer Plus BC presented antibiofilm activity and no cytotoxic effect. In conclusion, although Sealer Plus BC had higher solubility, this sealer showed proper physicochemical properties, cytocompatibility, and antibiofilm activity.

Resumo O objetivo deste estudo foi avaliar as propriedades físico-químicas, a citocompatibilidade e a atividade antibiofilme de um novo cimento endodôntico à base de silicato de cálcio, Sealer Plus BC (MK Life, Brasil), em comparação com TotalFill BC Sealer (FKG Dentaire SA, Suíça) e AH Plus (Dentsply, Alemanha). O tempo de presa e o escoamento foram avaliados com base nas normas ISO 6876. O pH foi avaliado após diferentes períodos, e a radiopacidade por análise radiográfica (mmAl). A solubilidade (% de perda de massa) e alteração volumétrica (por micro-CT) foram avaliadas após 30 dias de imersão em água destilada. Citocompatibilidade foi avaliada pelos ensaios metiltetrazólio (MTT) e vermelho neutro (NR), após exposição das células Saos-2 ao extrato de cimento por 24 horas. Análise adicional foi realizada através do ensaio MTT após 1, 3 e 7 dias de exposição das células Saos-2 aos extratos dos cimentos na diluição de 1:8. Atividade antibiofilme contra Enterococcus faecalis e/ou Candida albicans foi avaliada pelos ensaios cristal violeta e contato direto modificado. As propriedades físico-químicas foram analisadas utilizando os testes ANOVA e Tukey; MTT e NR foram analisados pelos testes ANOVA e Bonferroni; os ensaios antimicrobianos foram analisados pelos testes Kruskal-Wallis e Dunn (α=0.05). Sealer Plus BC apresentou tempo de presa, radiopacidade e escoamento adequados, além de capacidade de alcalinização. Sealer Plus BC foi significantemente mais solúvel que AH Plus (p<0.05) e apresentou alteração volumétrica similar à de AH Plus e TotalFill BC (p>0.05). Sealer Plus BC apresentou atividade antibiofilme, sem efeito citotóxico. Como conclusão, embora Sealer Plus BC apresente maior solubilidade, este cimento apresentou propriedades físico-químicas adequadas, citocompatibilidade e atividade antibiofilme.

Influência da agulha e fluxo de irrigação na limpeza do canal radicular e extrusão apical de irrigante: análise em micro-CT / Influence of needle and irrigation flow rate on root canal cleaning and apical extrusion of irrigant: Micro-CT analysis

Introdução: O tipo de agulha e fluxo de irrigação é importante para a limpeza adequada no terço apical do canal radicular. Objetivo: Avaliar a influência do tipo de agulha e fluxo de irrigação na limpeza do canal radicular e canais laterais simulados e na extrusão apical do irrigante. Métodos: Trinta e dois dentes de resina foram utiliza- dos. Após a instrumentação do canal radicular, foram feitos quatro canais laterais a 2 e 7 mm do ápice. Os canais radiculares foram preenchidos com solução de contraste. Os canais foram irrigados com dois tipos de agulha, 29G e 31G, com diferentes designs (abertura lateral e apical) e dois fluxos (2 ou 5 mL/min), a 1 mm aquém do comprimento de trabalho. O volume da solução de contraste nos canais principal e laterais após irrigação e a extrusão apical do irrigante (mm3 ) foi avaliado por micro-CT, em comparação com a análise inicial. Os dados em porcentagem foram analisados pelos testes ANOVA e Tukey (α=0,05). Resultados: não houve diferença entre os protocolos de irrigação para limpeza da solução de contraste. Maior volume de extrusão apical com agulha 29G com abertura apical e 5 mL/min foi observado, em comparação ao mesmo tipo de agulha e 2 mL/min (p<0,05). Conclusão: Todos os protocolos de irrigação foram associados à limpeza dos canais radiculares. Maior extrusão apical foi observada com agulha 29G ­ abertura apical com maior fluxo de irrigação (5 mL/min). Implicações clínicas: Irrigação endodôntica com agulha com abertura apical e maior fluxo da solução pode favorecer a extrusão dos irrigantes para os tecidos periapicais (AU).

Introduction: needle and irrigation flow rate are important for proper cleaning of the root canal. Aim: to evaluate the influence of type of needle and irrigation flow rate on cleaning of root canal and simulated lateral canals and the apical extrusion of irrigant. Methods: Thirty-two resin teeth were used. After root canal instrumentation, four lateral canals were made at 2 and 7 mm from the apex. Root canals were filled with contrast solution. The root canals were irrigated with two types of needle, 29G and 31G, with different designs (side and apical opening) and two flow rates (2 or 5 mL/min), at 1 mm short of the working length. The volume of the contrast solution in the main and lateral canals after irrigation and apical extrusion of the irrigant (mm3) were evaluated by Micro-CT, in comparison with the initial analysis. Data in percentage were analyzed by ANOVA and Tukeys tests (α = 0.05). Results: no difference among the irrigation protocols for contrast solution cleaning was observed. Higher volume of apical extrusion using needle 29G-apical opening and 5 mL/min in comparison with the same type of needle and 2 mL/min was observed (p < 0.05). Conclusion: all irrigation protocols were associated with root canals cleaning. Greater apical extrusion was observed for needle 29G-apical opening with higher irrigation flow rate (5 mL/min). Clinical implications: endodontic irrigation using a needle with apical opening and higher flow rate of solution may favor extrusion of irrigant to the periapical tissues (AU).

Influence of voxel size on dentinal microcrack detection by micro-CT after root canal preparation

Abstract The aim of this study was to assess the influence of micro-computed tomography (micro-CT) voxel size on dentinal microcrack detection after root canal preparation using rotary heat-treated nickel-titanium files. Curved mesial root canals (n = 24) of mandibular molars were prepared using ProDesign Logic 30/.05 (PDL) or HyFlex EDM 25/.08 (HEDM). The specimens were scanned by micro-CT at 5 μm voxel size before and after root canal preparation. The percentage of microcracks was evaluated in images at 5, 10 and 20 μm voxel size, by two examiners at two moments. The Kappa and McNemar tests (α = 0.05) were used. The percentage of dentinal microcracks was similar before and after PDL and HEDM preparations, at 10 and 20 μm (p > 0.05). HEDM showed a higher percentage of dentinal microcracks in the middle third at 5 μm after preparation (p < 0.05). The detection of dentinal microcracks before and after instrumentation using PDL was more accurate at 5 μm than at 20 μm, in all thirds (p < 0.05). Within the limitations of this ex vivo study, as expected, the results showed that different resolutions influence the micro-CT analysis of microcracks. The highest accuracy in detecting microcracks was observed for analyses performed at 5 μm voxel size. HyFlex EDM caused even more microcracks to develop in the middle third, detectable only by visualization of images made at 5 μm voxel size.

Evaluation of flow and filling of root canal sealers using different methodologies / Avaliação do escoamento e preenchimento de cimentos obturadores utilizando diferentes metodologias

Abstract Introduction Flow and filling ability of root canal sealers are indispensable for hermetic sealing of the root canal. Micro-computed tomography (micro-CT) can be used as a complementary methodology to evaluate such properties. Objective To evaluate the flow and filling ability of AH Plus, Endofill and Sealapex by conventional methodology and micro-CT. Material and method The flow of the sealers was analyzed according to ISO 6876/2012 and complemented by the area evaluation. Glass plates were manufactured with diameters of 1×1×2 mm and 1×1×1 mm (length, width and height), with a central cavity and four grooves in the horizontal and vertical directions. Each material was placed in the central cavity. Another glass plate and a metal weight were placed on the cement and kept for 10 minutes. The glass plate/sealer set was scanned using micro-CT. The flow was calculated by linear measurement of the material in the grooves. The central filling (mm3) was calculated in the central cavity and the lateral filling was measured up to 2 mm from the central cavity. Data were submitted to ANOVA/Tukey tests (α=0.05). Result All evaluated sealers presented flow according to ISO 6876 standards. The materials showed central cavity filling capacity higher than 80% and lateral filling greater than 75%. There was no difference in flow (mm and mm2) and in the filling ability (mm3) provided by the materials (p>0.05). Conclusion All evaluated root canal sealers showed adequate flow and filling capacity, suggesting their clinical application.

Resumo Introdução Escoamento e capacidade de preenchimento de cimentos obturadores são indispensáveis para um selamento hermético do canal radicular. Microtomografia computadorizada (micro-CT) pode ser utilizada como uma metodologia complementar para avaliação de tais propriedades. Objetivo Avaliar escoamento e capacidade de preenchimento de AH Plus, Endofill e Sealapex, por meio de metodologia convencional e micro-CT. Material e método O escoamento dos cimentos foi analisado de acordo com as normas ISO 6876/2012 e complementado pela avaliação em área. Placas de vidro foram confeccionadas nos diâmetros de 1×1×2 mm e 1×1×1 mm (comprimento, largura e altura), com uma cavidade central e quatro canaletas nas direções horizontal e vertical. Cada material foi colocado na cavidade central. Outra placa de vidro e um peso de metal foram colocados sobre o cimento e mantidos por 10 minutos. O conjunto placa de vidro/cimento foi escaneado usando micro-CT. O escoamento foi calculado por medição linear do material nas canaletas. O preenchimento (mm3) central foi calculado na cavidade central e o preenchimento lateral foi medido até 2 mm a partir da cavidade central. Os dados foram submetidos aos testes ANOVA/Tukey (α=0.05). Resultado Todos os cimentos avaliados apresentaram escoamento de acordo com as normas ISO 6876. Os materiais mostraram capacidade de preenchimento da cavidade central superior a 80% e preenchimento lateral superior a 75%. Não houve diferença no escoamento (mm e mm2) e na capacidade de preenchimento (mm3) proporcionada pelos materiais (p>0.05). Conclusão Todos os cimentos obturadores avaliados mostraram adequado escoamento e capacidade de preenchimento, sugerindo a aplicação clínica dos mesmos.

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.

Micro-CT analysis of filling ability and porosity of root-end filling materials / Análise em micro-CT da capacidade de preenchimento e porosidade de materiais retrobturadores

Introduction: Filling ability of retrograde cavity and porosity are important properties for root-end filling materials and may be evaluated by using microcomputed tomography (micro-CT). Objective: To evaluate filling ability and porosity of root-end filling materials using microcomputed tomography (micro-CT). Material and method: Cavities with 1 mm internal diameter and 3 mm depth were prepared in bovine dentin sections by using ultrasonic tips (CVD No. 6.1107-6), and filled by Mineral Trioxide Aggregate (MTA); Sealer 26 (S26) and zinc oxide and eugenol cement (ZOE). Before and after filling, cavities were scanned by using micro-CT (SkyScan 1176). Filling and porosity were analyzed by using CTAn software. Filling ability was calculated based on volumetric percentage of the filled cavity. The number and percentage of closed pores were measured throughout entire extension of the filled cavity (total) and in each third (cervical, middle and apical), by using bi and tridimensional analyses. The filling data were submitted to ANOVA and Tukey statistical tests, and porosity data to Kruskall-Wallis and Dunn tests, at a 5% significance level. Result: S26 and ZOE presented higher filling ability than MTA (p<0.05). S26 showed the highest total porosity (number and percentage) (p<0.05). In all thirds after 2D and 3D analyses, porosity was higher for S26 in comparison to MTA and ZOE (p<0.05). Conclusion: Although Sealer 26 presented more porosity, the material was associated with a great filling ability. Micro-CT analysis showed no correlation between filling ability and porosity.

Introdução: Capacidade de preenchimento da cavidade retrógrada e porosidade são propriedades importantes de materiais retrobturadores e podem ser avaliadas por meio de microtomografia computadorizada (micro-CT). Objetivo: Avaliar a capacidade de preenchimento e porosidade de materiais retrobturadores por meio de micro-CT. Material e método: Cavidades com 1 mm de diâmetro e 3 mm de altura foram preparadas em dentina bovina utilizando pontas ultrassônicas (CVD No. 6.1107-6) e foram preenchidas com Mineral Trióxido Agregado (MTA), Sealer 26 (S26) e cimento de óxido de zinco e eugenol (OZE). As cavidades foram escaneadas em micro-CT antes e após o preenchimento. A capacidade de preenchimento foi calculada com base na porcentagem em volume, das cavidades preenchidas. O número e porcentagem dos poros fechados foram avaliados em toda extensão da cavidade preenchida (total) e por terços (cervical, médio e apical) por meio de análises bi e tridimensionais. Os dados de preenchimento foram submetidos aos testes estatísticos ANOVA e Tukey e a porosidade aos testes de Kruskall-Wallis e Dunn, com nível de significância de 5%. Resultado: S26 e OZE apresentaram maior capacidade de preenchimento que o MTA (p<0,05). S26 mostrou maior porosidade total (em número e porcentagem) (p<0,05). Em todos os terços, após as análises 2D e 3D, a porosidade foi maior para S26 em comparação ao MTA e OZE (p<0,05). Conclusão: Embora Sealer 26 tenha apresentado maior porosidade, o material foi associado a uma adequada capacidade de preenchimento. A análise em micro-CT mostrou ausência de correlação entre capacidade de preenchimento e porosidade.

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.