Biocompatibility of mineral trioxide aggregate and biodentine as root-end filling materials: an in vivo study.

This study evaluated the biocompatibility of mineral trioxide aggregate (MTA) and Biodentine (BD) as root-end filling materials. Six mongrel dogs were divided into two equal groups according to the evaluation period; group A: one month and group B: three months. Three premolars of the same quadrant in each arch were used, summing up 36 teeth (6 teeth/dog). These teeth were randomly subdivided into three subgroups according to the root-end filling material used: MTA, BD and no root-end filling material (control). Endodontic access cavities were performed for induction of periapical pathosis. After the infection period, root canal instrumentation and obturation were accomplished. One day after root canal procedures, root-end surgery was performed. Surgical access was achieved and the root-end was resected approximately 3 mm above the apex. Root-end cavity was prepared ultrasonically and filled with the tested materials. All samples were evaluated by radiography and histopathology (Inflammation and new hard tissue formation). Data were collected and subjected to statistical analysis. In group A, MTA subgroup exhibited significant higher mean inflammatory score than BD subgroup (P < 0.05) while no significant difference was recorded between MTA and BD subgroups in group B (P > 0.05). Regarding mean mineralization score, there was no significant difference between all subgroups in both groups A and B (P > 0.05). Biodentine exhibited favorable biocompatibility in the initial stage of healing than MTA and comparable biomineralization. Clinical relevance: Biodentine could be considered as an acceptable alternative to MTA in peri-radicular surgeries.

Effect of Recombinant Human Platelet-Derived Growth Factor on Healing of Chronic Periapical Tissue Pathosis Following Apical Surgery in a Canine Model: A Histomorphometric and Microcomputed Tomography Analysis.

This canine in vivo study assessed the effect of recombinant human platelet-derived growth factor (rhPDGF) on the healing of periapical tissues following apical surgery. From a total of 96 premolar teeth, 64 teeth from six beagle dogs (2 years old) were classified as experimental and were randomly assigned to four experimental groups (16 teeth per group). After having the pulp extirpated, leaving teeth open to the oral cavity for 1 week, and sealing with an immediate restorative material for 8 weeks, nonsurgical endodontic treatment was performed. A split-mouth design was used, and intra-animal randomization of treatment sides was applied to the groups as follows: apical curettage + 1.5-mm root-end resection (Group 1); apicoectomy + mineral trioxide aggregate (MTA) root-end filling (Group 2); apicoectomy + MTA root-end filling + rhPDGF (Group 3); and apical curettage + rhPDGF (Group 4). The animals were sacrificed 24 months after apical surgery, and histologic and µCT analyses were performed for bone volume loss (BVL). Group 1 showed partial resolution of the periapical lesions without signs of tissue regeneration (BVL: 49.09 ± 10.97 mm3). Group 2 had minimal bone regeneration and showed cementum reformation in 9 teeth, with no direct attachment to the MTA (BVL: 35.34 ± 10.97 mm3). Group 3 showed regeneration of all damaged apical tissues without direct contact between the cementum and MTA (BLV: 4.51 ± 1.55 mm3). Group 4 showed regeneration of PDL, bone, and cementum and attachment of functional cementum fibers (BVL: 2.82 ± 2.3 mm3). The difference in BVL was statistically significant only for Groups 1 and 2 (P < .05). rhPDGF may help regenerate apical tissue structures following apical surgery.

Zinc oxide-ozonated olive oil as a new root canal filling material in primary molars: a clinical randomized controlled trial.

OBJECTIVES: The complex root canal anatomy of primary teeth keeps it very tricky to attain appropriate cleansing by biomechanical instrumentation, so obtaining an obturating material with excellent antimicrobial properties is a challenge in current clinical pulpectomy practice. So, this study aimed to assess the clinical and radiographic performance of zinc oxide-ozonated olive oil as a primary root canal filling material. MATERIALS AND METHODS: Ninety non-vital primary molars in children ranging from 4 to 8 years were allocated into three groups in which root canals were filled with zinc oxide-ozonated olive oil, zinc oxide-olive oil, and zinc oxide-eugenol (ZOE) according to each group after pulpectomy procedure. Clinical and radiographic evaluations were done at 3-, 6-, and 12-month follow-up periods. Statistical analysis was performed for the collected data. RESULTS: All study groups showed a significant improvement regarding clinical signs and symptoms during follow-up periods. Ozonated-olive oil group revealed a significant increase in furcation radiodensity and a decrease in periodontal ligament space at 3-, 6-, and 12-month follow-up intervals compared to other groups. CONCLUSION: Zinc oxide-ozonated olive oil and zinc oxide-olive oil paste had shown good clinical and radiographic success for primary teeth pulpectomy. CLINICAL RELEVANCE: The intricate torturous primary root canal anatomy, in addition to the child's negative behavior, interferes with the complete debridement, so the long-lasting antibacterial effect of the primary root canal filling material aids in the pulpectomy success.

The Effects of Calcium Silicate- and Calcium Hydroxide-based Root Canal Sealers on Postoperative Pain: A Randomized Clinical Trial.

INTRODUCTION: This study aimed to compare the postoperative pain level changes resulting from using calcium silicate- (EndoSeal MTA) and calcium hydroxide-based (Sealapex) root canal sealers in mandibular first and second molar teeth with symptomatic apical periodontitis. METHODS: A total of 60 patients with symptomatic apical periodontitis in their lower molar teeth were randomly allocated into 2 groups according to sealer type (n = 30). Demographic data, including gender, age, and smoking habit, and preoperative pain measures were recorded. Root canal treatments were performed in a single visit. Postoperative pain measurements and analgesic intake were measured at 6, 12, 24, and 48 hours and after 3, 5, and 7 days using the visual analog scale. The data were statistically analyzed using a chi-squared test (to compare gender, age, smoking habit, analgesic intake, and sealer extrusion), the Mann-Whitney U test (to compare pain levels), Friedman tests (for the evaluations of the reduction in pain levels over time), and Spearman's correlation test (to analyze the relationships of age, gender, smoking habit factors with postoperative pain) (P = .05). RESULTS: The statistical analysis showed no significant differences between the groups in postoperative pain and analgesic intake at any of the time intervals evaluated (P > .05). CONCLUSIONS: Patients treated with calcium silicate- and calcium hydroxide-based root canal sealers experienced similar postoperative pain and no statistically significant differences were observed in analgesic intake.

Inflammatory response to epoxy resin and calcium silicate sealers preheated with different temperatures: an in vivo study.

OBJECTIVE: To determine the impact of heat exposure of different sealers on inflammatory cytokine secretions and tissue response in vivo. MATERIALS AND METHODS: Silicone tubes were prefilled with epoxy resin (ER) or calcium silicate (CS) sealers, preheated at 37, 60, or 120 °C, and implanted in rat subcutaneous site. Peri-implant exudate and tissue were analyzed after 1 and 4 weeks for cytokine secretions and tissue organization. RESULTS: At 1 week, 120 °C-preheated CS and ER induced higher secretions of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6), respectively, as compared to sham/empty tube groups. At 4 weeks, whereas TNF-α secretion was reduced in CS, it increased in ER group, particularly for 120 °C. Both sealers revealed high IL-6 after 4 weeks as compared to sham/empty tube, and generally, higher IL-6 secretions were associated with ER. Histology at 1 week revealed lower degree of inflammatory infiltrate in the groups of the highest preheating temperature (120 °C). Nonetheless, at 4 weeks, whereas fibrous capsule area and inflammatory infiltrate remained low in the CS120 group, they were high in ER120. CONCLUSION: Preheating ER sealer to 120 °C induced high and prolonged secretion of proinflammatory cytokines (TNF-α and IL-6), whereas this effect was transient for the CS sealer. This was associated with increased fibrous capsule and inflammatory infiltrate in response to 120 °C-preheated ER. CLINICAL RELEVANCE: Heat-induced changes in sealer properties alter the inflammatory response in vivo, which may affect the clinical outcome. This will not only help appropriate selection of obturation technique for different sealers, but also for optimizing the properties of new generation of sealers.

Antibacterial activity and physicochemical properties of a sealer containing copaiba oil.

This study aimed to evaluate the antimicrobial activity and physicochemical properties of a novel dual-cure endodontic sealer containing copaiba oil. The copaiba oil was obtained and characterized by gas chromatography (GC), and the minimum inhibitory concentration (MIC) was performed. The experimental sealers were formulated with copaiba oil concentrations of 0, 0.5, 1, and 2%, and the RealSeal™ (Sybron endo, Orange, USA) and AH Plus (Dentsply De Trey Gmbh, Konstanz, Germany) were used as the commercial references. The antimicrobial activity of the sealers was evaluated by the direct contact test for 1h and 24h. To evaluate the physicochemical properties of the sealers, the degree of conversion, setting time, film thickness, dimensional stability, and radiopacity tests were performed. The data were statistically analyzed by one-way ANOVA and Tukey's test (α = 0.05). Concerning the results, the sealers containing copaiba oil showed antimicrobial activity without harming the physicochemical properties.

In vitro inflammatory modulation of bioceramic endodontic sealer in macrophages stimulated by bacterial lipopolysaccharide.

AIM: To evaluate the effects of AH Plus (Dentsply), Sealer 26 (Dentsply), and Sealer Plus BC (Produtos Médicos e Odontológicos) on cytotoxicity and inflammation in macrophage cultures exposed to bacterial lipopolysaccharide (LPS). METHODOLOGY: After initial setting, the sealers were conditioned with serum-free culture medium for 24 h (1 ml/cm2 ). Macrophages from the RAW 264.7 strain were exposed to sealer extracts in a 1:16 ratio in a culture medium with or without LPS. Cell morphology, viability, mitochondrial activity, oxidative stress and gene expression of interleukin 1ß (IL-1ß) and tumour necrosis factor-alpha (TNF-α) were evaluated. Data on mitochondrial activity, oxidative stress and TNF-α were analysed using a two-way analysis of variance (anova) test, followed by the Student-Newman-Keuls post-test. IL-1ß data were analysed using one-way anova, followed by SNK, and the t-test was used for intragroup comparison. The significance level was set at 5%. RESULTS: In the absence of LPS, only AH Plus and Sealer 26 showed a reduction in cell density, while in the presence of LPS, Sealer 26 had the lowest density compared to the other groups. In terms of mitochondrial activity, at 24 and 48 h, Sealer Plus BC had significantly higher mean values than Sealer 26 and AH Plus (p < .05). Sealer 26 exhibited the lowest levels of oxidative stress and IL-1ß and TNF-α expression, regardless of the presence of LPS (p < .05). CONCLUSIONS: Although all sealers interfere with the response of macrophages to LPS, contact with epoxy resin-based sealers can impair cell activity in vitro, while bioceramic sealer seems to favour the inflammatory functions of these cells.

Interleukin-6, tumor necrosis factor-α, and CD5 immunolabeling of new experimental endodontic sealer and repair material.

The aim of this study was to evaluate the biocompatibility and immunoinflammatory response of the Sealepox and Sealepox-RP, based on interleukin (IL)-6, tumor necrosis factor (TNF)-α, and CD5 immunolabelling. The ProRoot MTA (PRMTA) was used for comparison. Polyethylene tubes (1.0-mm internal, 1.6-mm external diameter, and 10.0-mm length; ISO 10993) with or without (control) materials were randomly implanted in the dorsum of 35 rats (4 per rat). After 7, 15, 30, 60, and 90 days (n = 7), the tubes were removed for histological and immunohistochemical analysis. The Kruskal-Wallis and Dunn's test for non-parametric data and, ANOVA and Tukey test for parametric data were used (P < 0.05). Hematoxylin and eosin staining revealed that the concentration of inflammatory cells decreased over time with no differences between groups in all periods (P > 0.05). Regarding IL-6 immunostaining, there was no difference at 7 days (P > 0.05); all groups decreased over time, being faster for the PRMTA group and also, with no differences between groups in the last period (P > 0.05). For TNF-α, at 7 days there was no difference between groups (P > 0.05); there was an increase at 15 days for PRMTA and, at 30 and 60 days, for PRMTA and Sealepox compared to the control (P < 0.05). At 90 days, Sealepox RP showed the lowest immunostaining being similar to the control (P > 0.05). Regarding CD5 cells, at 7 days, there was high immunostaining for PRMTA compared to the control (P < 0.05); and significant reduction over time with difference for all groups at 30 and 60 days. (P < 0.05); Sealepox was similar to the control in all periods (P > 0.05). Sealepox RP showed the highest immunostaining at 15 days, being different from the control and PRMTA (P < 0.05); in the other periods it was similar to the control (P > 0.05). It can be concluded that Sealepox and Sealepox-RP were biocompatible and demonstrated similar immunoinflammatory response regarding IL-6, TNF-α, and CD5 compared to PRMTA.

Laboratory study of tissue repair of resin-based endodontic sealers in critical surgical defects.

OBJECTIVE: Filling materials should be restricted to the root canal space. However, sometimes it is impossible to control the apical extrusion, in this case, the fate of the filling material and the result of the treatment will depend on its physicochemical properties and biocompatibility. To evaluate the tissue response and bone repair capacity of endodontic sealers that were implanted in the calvaria of Wistar rats, forming the groups (n=16): AH Plus and Sealer Plus, compared to the clot group. METHODOLOGY: On days 30 and 60, the animals were euthanized, the calvaria was removed and processed for hematoxylin-eosin, immunohistochemistry for collagen type I, Picrosirus red and microtomographic analysis. Data were subjected to ANOVA and Tuckey tests (p<0.05). RESULTS: At 30 days, all groups showed an intense inflammatory reaction (p>0.05). At 60 days, the AH Plus and Sealer Plus maintained an intense inflammatory infiltrate compared to the clot group (p<0.05). We observed immunopositive areas for type I collagen in all groups at 30 days and 60 days (p>0.05). We observed more red collagen fibers for the Sealer Plus compared to the clot group at 30 days (p<0.05). Considering the total fibers, the clot group at 30 days compared to 60 days after surgery showed an increase in the amount of matrix (p<0.05). There were no statistical differences between groups for green and yellow fibers (p>0.05). Regarding morphometric parameters, at 30 days, the newly formed bone volume and number of bone trabeculae were higher in the groups with sealers compared to the clot group (p<0.05). At 60 days, AH Plus and Sealer Plus showed greater bone neoformation compared to the clot group (p<0.05). CONCLUSIONS: Despite AH Plus and Sealer Plus induced an intense inflammatory reaction, they can be considered biocompatible materials, since they allowed bone repair.

Influence of supplement administration of omega-3 on the subcutaneous tissue response of endodontic sealers in Wistar rats.

AIM: Natural substances such as omega-3 have been used in the medical field due to their numerous properties and, in particular, modulating effect on the systemic and local inflammatory processes. Thus, this study evaluated the influence of omega-3 supplementation on the subcutaneous tissue response of endodontic sealers in Wistar Rats. METHODOLOGY: Polyethylene tubes were implanted in the subcutaneous tissue of 48 animals (one empty for control and three filled with Sealapex, AH Plus or Endofill). The animals were treated with omega-3 (TO) or water (TW). Treatments started 15 days before implantation until euthanasia. After 5, 15 and 30 days (n = 8), animals were euthanized and polyethylene tubes and surrounding tissue were removed and processed for histological analysis. The inflammatory reaction was analysed by Haematoxylin and Eosin stain and immunolabelling for IL-6 and TNF-α. The collagen maturity was analysed by picrosirius red stain and calcium deposition by von Kossa stain and polarized light. Results were statistically analysed (p < .05). RESULTS: Amongst TW sealer groups, Endofill evoked a more intense inflammatory infiltrate compared with AH Plus and control in the 30-day period (p = .009). However, in TO sealer groups, there was no difference amongst the sealers and control in all periods (p > .05). Comparing each sealer as a function of the supplementation with water or omega-3, there are differences for Endofill (p = .001) and Sealapex (p = .005) in the 30-day period, presenting lower inflammatory infiltrate in the animals treated with omega-3. A higher percentage of immature fibres was observed at 15 and 30 days in the TO group, compared with the TW group (p < .05). The deposition of calcium particles was observed only by Sealapex in all periods, despite the supplementation procedure. CONCLUSIONS: Omega-3 supplementation influence the tissue reactions of endodontic sealers, modulating inflammation, the immunolabelling of IL-6 and TNF-α, the repair process and it does not interfere with calcium deposition.

A comprehensive in vitro comparison of the biological and physicochemical properties of bioactive root canal sealers.

OBJECTIVES: To evaluate the biological and physicochemical features of bioactive root canal sealers. MATERIALS AND METHODS: Human periodontal ligament fibroblasts (hPDLF) and human osteoblasts (hOB) were exposed to eluates of three bioactive root canal sealers, GuttaFlow® bioseal (GF), BioRoot™ RCS (BR), and TotalFill® BC Sealer (TF), and the epoxy resin-based sealer AH plus® (AH). Cytotoxicity and cellular inflammatory response were evaluated. The osteogenic potential was examined using human mesenchymal stem cells (hMSC). Film thickness, flowability, and pH were assessed. Root canal treatment was performed on human extracted teeth to evaluate the sealers' tightness towards bacterial penetration. The antibacterial activity against common pathogens in primary root canal infections was tested. RESULTS: AH was severely cytotoxic to hPDLF and hOB (p < 0.001). The bioactive sealers were generally less cytotoxic. IL-6 levels in hPDLF were elevated in the presence of AH (p < 0.05). AH and GF suppressed IL-6 production in hOB (p < 0.05). AH and BR stimulated the PGE2 production in hPDLF and hOB (p < 0.05). BR was the only sealer that led to calcium deposits in hMSC (p < 0.05). TF and AH showed the lowest film thickness and the highest flowability. Bacterial tightness was best in teeth filled with AH and BR. All sealers showed similar antimicrobial activity, but the overall antimicrobial efficacy was moderate as the bacteria were reduced by just one log scale (p < 0.05). CONCLUSIONS: This study revealed favorable in vitro results regarding the biocompatibility of the bioactive root canal sealers. CLINICAL RELEVANCE: Bioactive root canal sealers may be a useful alternative to epoxy resin-based sealers.

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.

The presence of smear layer affects the antimicrobial action of root canal sealers.

AIM: To assess the chemical and microstructural characteristics of dentine after the use of two irrigation protocols and correlate this with the antimicrobial properties of hydraulic calcium silicate cement (HCSC) sealers and changes to the dentine structure/chemistry after sealer placement. METHODOLOGY: Two irrigation protocols - Protocol A using 2% NaOCl used 5 mL/5 min and Protocol B with 2% NaOCl (5 mL/5 min) followed by 17% EDTA (5 mL/3 min) - were used to prepare dentine. The chemical and microstructural changes following irrigation were assessed by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and Fourier transform infrared (FT-IR) spectroscopy (n = 5) on dentine obtained from the mid-root and coronal parts of extracted human teeth. Four sealers (AH Plus, BioRoot, MTA Fillapex, TotalFill) were characterized by SEM/EDS (n = 3). The ability of the sealers to eradicate intratubular Enterococcus faecalis biofilms was assessed by live/dead dye and confocal laser scanning microscopy to measure the percentage of living cells. The effect of combined irrigation and root filling on dentine was assessed by SEM and EDS analysis (n = 5). Statistical analysis was undertaken using one-way anova and a number of post hoc tests to detect intergroup differences. The F-test was used for comparison of variances in the microbiology testing. RESULTS: The use of NaOCl alone left the smear layer intact, with traces of chlorine remaining on dentine. The use of BioRoot sealer restored the calcium levels of dentine which had been depleted by the irrigation with EDTA. BioRoot exhibited antimicrobial properties against intratubular bacteria even in the presence of smear layer (Protocol A). Smear layer removal improved the bactericidal effect of all sealers and Ca2+ leaching. The use of a chelating agent was important for the intratubular sealer penetration for AH Plus but not the other sealers. CONCLUSION: The removal of smear layer was necessary for penetration of AH Plus into the dentinal tubules. BioRoot was a more effective sealer in reducing the bacterial load in the dentinal tubules than the other materials tested and the presence of smear layer did not affect its activity.

Cytocompatibility of filling pastes by primary teeth root simulating model.

Evaluate the cytocompatibility of Calen®/ZO, Calcicur®, Vitapex®, Endoflas®, and zinc oxide/eugenol-based (ZOE) root canal pastes (RCP) to human primary osteoblasts (HPO) through a simplified model for primary teeth. The model employed pipette tips filled with 0.037 g of paste, exposed to 185 µL of culture medium for 24 h (n = 6). Release of components was analysed by Proton Nuclear Magnetic Resonance Spectroscopy (1H-NMR). HPO were exposed to conditioned media for 24 h. Cell viability was assessed by cell density and metabolic activity, and release of interleukin 6 (IL-6), vascular endothelial growth factor (VEGF) and fibroblast growth factor (bFGF) by immunological assay. Physicochemical properties and antimicrobial efficacy were also evaluated. 1H-NMR spectra analysis showed similarity between ZOE, Endoflas®, Calcicur®, and Vitapex® compared to Calen®/ZO and positive control, which showed distinct released components. Calen®/ZO and Calcicur® exhibited high alkaline pH in all periods and showed similar solubility. Calen®/ZO, ZOE, and Vitapex® showed similar flow rate. Calen®/ZO, Calcicur®, and Vitapex® did not exhibit antimicrobial efficacy. Calen®/ZO presented cytotoxicity (p < 0.05). Pastes did not increase IL-6 release compared to control. Apart from Vitapex®, all pastes significantly induced VEGF/bFGF release. Interactive effects among released products may affect biological response to filling pastes. Calcicur®, ZOE, Endoflas® and Calen®/ZO presented good to moderate cytocompatibility, with low impact on pro-inflammatory cytokine release and induction of growth factors of interest to tissue repair. This simplified model, specific for the evaluation of the cytocompatibility of RCPs on primary teeth, suggests how these pastes might contribute to bone repair in clinical situations of apical periodontitis in children.

Antibacterial Activity of a New Ready-To-Use Calcium Silicate-Based Sealer

Abstract The aim of this study was to evaluate the antibacterial potential of a calcium silicate-based sealer (Bio-C Sealer, Angelus) against common bacteria in primary and secondary endodontic infections. Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus mutans were exposed to fresh Bio-C Sealer for 24 h by the agar diffusion method (n=5). Additionally, the antibacterial activity was investigated against E. faecalis and S. mutans biofilms (48 h old) grown in discs with 4 mm in diameter and 2 mm in height. (n=3) of set discs of Bio-C Sealer (Angelus), EndoFill (Dentsply-Mallefer), Sealer 26 (Dentsply), AH Plus (Dentsply), Sealapex (Sybron-Endo) and EndoSequence BC Sealer (Brasseler). The antibacterial activity was evaluated by colony forming unity (CFU) counting using ImageJ software. Data were compared by one-way ANOVA followed by Holm-Sidak test (a=5%). Fresh Bio-C Sealer exhibited antimicrobial activity against all bacteria evaluated by agar diffusion method, except for S. mutans. Set discs of all endodontic sealers tested showed similar CFU values for E. faecalis (p>0.05). S. mutans in biofilms showed higher susceptibility to EndoFill compared with the other sealers (p<0.05). In conclusion, the results indicate that fresh Bio-C Sealer does not inhibit S. mutans growth, but exhibits antibacterial activity against E. faecalis, S. aureus, P. aeruginosa and E. coli. After setting, the Bio-C Sealer exhibits an antimicrobial potential comparable to that of the other sealers evaluated in E. faecalis biofilm, but lower than that of EndoFill for S. mutans biofilm.

Resumo O objetivo deste estudo foi avaliar o potencial antibacteriano do novo cimento biocerâmico (Bio-C Sealer, Angelus) contra bactérias comuns em infecções endodônticas primárias e secundárias. Culturas de Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus e Streptococcus mutans foram expostos a amostras frescas do Bio-C sealer durante 24 h pelo método de difusão em agar (n=5). A atividade antibacteriana de amostras dos cimentos Bio-C Sealer (Angelus), EndoFill (Dentsply-Mallefer), Sealer 26 (Dentsply), AH Plus (Dentsply), Sealapex (Sybron-Endo) e EndoSequence BC Sealer (Brasseler) após a presa também foi investigada em biofilmes de 48 h das bactérias E. faecalis e S. mutans, crescidos em discos com 4 mm de diâmetro e 2 mm de altura. A atividade antibacteriana foi avaliada por contagem das unidades formadoras de colônias (UFC) utilizando o software ImageJ. Os dados foram comparados por ANOVA a um critério seguido pelo pós-teste Holm-Sidak (a=5%). Amostras frescas do Bio-C Sealer exibiram atividade antimicrobiana contra todas as bactérias avaliadas pelo método de difusão em ágar, exceto para S. mutans. A análise da formação de biofilme mostrou que todos os cimentos endodônticos testados apresentaram valores similares de UFC para E. faecalis (p> 0,05), enquanto biofilmes de S. mutans foram mais suscetíveis ao EndoFill em comparação com os demais cimentos (p<0,05). Conclui-se que o cimento Bio-C Sealer fresco exibe atividade antibacteriana para E. faecalis, S. aureus, P. aeruginosa e E. coli, mas não inibe o crescimento de S. mutans. Após a presa, o cimento Bio-C Sealer exibe potencial antimicrobiano similar ao dos demais cimentos avaliados em biofilme de E. faecalis, mas inferior ao do EndoFill para S. mutans.

Immunoinflammatory response and bioactive potential of GuttaFlow bioseal and MTA Fillapex in the rat subcutaneous tissue.

To evaluate the effect of GuttaFlow bioseal (GFB) and MTA Fillapex (MTAF) in comparison with Endofill (EF) in the subcutaneous tissue. Polyethylene tubes with GFB, MTAF, EF or empty tubes (control group; CG) were implanted into subcutaneous of rats. After 7, 15, 30 and 60 days, the capsule thickness, inflammatory reaction, interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), caspase-3, TUNEL-positive cells, von Kossa and ultrastructural features were evaluated. The data were statistically analyzed (p ≤ 0.05). At all periods, the number of IL-6- and VEGF-immunolabelled cells, and capsule thickness were lower in GFB than MTAF, which was lower than EF (p < 0.0001). At 60 days, the number of inflammatory cells was similar in GFB and MTAF (p = 0.58). Significant differences in the number of TUNEL- and caspase-3-positive cells were not observed among GFB, MTAF and CG whereas the highest values were found in EF specimens. The EF specimens exhibited several cells with condensed chromatin, typical of apoptosis. von Kossa-positive and birefringent structures were only observed in GFB and MTAF, suggesting the presence of calcite crystals. Taken together, these results show that cellular and structural damage induced by GFB and MTAF sealers were recovery over time. Moreover, these sealers express bioactive potential in subcutaneous tissue.

Effect of a novel bioceramic root canal sealer on the angiogenesis-enhancing potential of assorted human odontogenic stem cells compared with principal tricalcium silicate-based cements.

OBJECTIVE: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). METHODOLOGY: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers' instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6-8 h. Statistical analyses included Kruskal-Wallis, Mann-Whitney U, and Friedman and Wilcoxon signed rank tests. RESULTS: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). CONCLUSION: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.

Avaliação da citotoxicidade in vitro de diferentes pastas endodônticas utilizadas em odontopediatria / In vitro cytotoxic effects of different endodontic pastes used in pediatric dentistry

Objetivo: O presente estudo teve como objetivo avaliar o potencial citotóxico de três diferentes pastas endodônticas em cultura celular de osteoblastos humanos. Material e Métodos: As pastas endodônticas Calen PMCC (SSWhite Artigos Dentários LTDA, Rio de Janeiro, Brasil), Feapex (Fórmula e Ação, São Paulo, Brasil) e CTZ (Lenzafarm, Belo Horizonte, Brasil) foram preparadas e eluídas em meio de cultura celular durante 24 horas em estufa a 37°C e 5% de CO2. Foram realizadas quatro diluições distintas desses meios nas concentrações 1: 1, 1: 2, 1: 4 e 1: 8. Culturas de células de Osteoblastos Humanos da linhagem Saos-2 foram expostas a estas diluições durante 24 horas. O controle negativo foi realizado expondo as células ao meio de cultura sem contato com nenhuma pasta endodôntica. A citotoxicidade desses meios foi avaliada utilizando o ensaio MTT e os resultados foram transformados em porcentagens de células viáveis em relação ao grupo controle negativo. A análise estatística foi realizada com nível de significância de 5%, utilizando ANOVA seguido do teste de Tukey. Resultados: A viabilidade celular foi significativamente alterada de acordo com o material testado (p <0,05) e sua concentração (p <0,05). Em todas as concentrações testadas, a pasta Feapex apresentou maior viabilidade celular comparada aos demais materiais (p <0,05). Embora não tenha sido observada diferença estatisticamente significante entre a pasta Calen PMCC e a pasta CTZ nas concentrações de 1: 1 e 1: 2 (p> 0,05), a pasta CTZ apresentou maior citotoxicidade nas concentrações de 1: 4 e 1: 8 (p <0,05 ). De forma geral, a citotoxicidade diminuiu com o aumento da diluição do material. Conclusão: De acordo com os resultados do presente estudo, a pasta endodôntica Feapex parece ser a melhor opção para utilização entre as pastas analisadas, pois apresentou menor citotoxicidade que as pastas Calen PMCC e CTZ

Objective: The aim of the present study was to evaluate the cytotoxic potential of three different endodontic filling materials on human osteoblast cell cultures. Material and Methods: The endodontic pastes Calen PMCC (SSWhite Dental Articles LTDA, Rio de Janeiro, Brazil), Feapex (Formula and Action, São Paulo, Brazil) and CTZ (Lenzafarm, Belo Horizonte, Brazil) were eluted in cell culture medium during 24 hours in an oven at 37°C and 5% CO2. Four distinct dilutions of these media were performed at the concentrations 1:1, 1:2, 1:4 and 1:8. Cell cultures of Saos-2 Human Osteoblast-like were exposed to these dilutions for 24 hours. The negative control group was performed by exposing the cells to culture medium without contact with any endodontic paste. The cytotoxic potential of these media was evaluated using the MTT assay and the results were transformed into viable cell percentages in relation to the negative control group. Statistical analysis was submitted with a significance level of 5%, using univariate Analysis of Variance (ANOVA) followed by Tukey's test. Results: Cell viability was significantly altered according to the material tested (p<0.05) and its concentration (p<0.05). Feapex samples presented higher cell viability than the other materials in all concentrations tested (p<0.05). Although no statistically significant difference was observed between Calen PMCC paste and CTZ paste at concentrations of 1: 1 and 1: 2 (p>0.05), CTZ paste showed a higher cytotoxicity at concentrations of 1: 4 and 1: 8 (p<0.05). In general, cytotoxicity decreases with increasing material dilution. Conclusion: According to the results of the present study, Feapex endodontic paste seems to be the better option for use among the analyzed pastes, since it presented lower cytotoxicity than Calen PMCC and CTZ pastes

Effect of a novel bioceramic root canal sealer on the angiogenesis-enhancing potential of assorted human odontogenic stem cells compared with principal tricalcium silicate-based cements

Abstract Objective: This study evaluated the angiogenesis-enhancing potential of a tricalcium silicate-based mineral trioxide aggregate (ProRoot MTA), Biodentine, and a novel bioceramic root canal sealer (Well-Root ST) in human dental pulp stem cells (hDPSCs), human periodontal ligament stem cells (hPLSCs), and human tooth germ stem cells (hTGSCs). Methodology: Dulbecco's modified Eagle's medium was conditioned for 24 h by exposure to ProRoot MTA, Biodentine, or Well-Root ST specimens (prepared according to the manufacturers' instructions). The cells were cultured in these conditioned media and their viability was assessed with 3-(4,5-dimethyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H tetrazolium (MTS) on days 1, 3, 7, 10, and 14. Angiogenic growth factors [platelet-derived growth factor (PDGF), basic fibroblast growth factor (FGF-2), and vascular endothelial growth factor (VEGF)] were assayed by sandwich enzyme-linked immunosorbent assay (ELISA) on days 1, 7, and 14. Human umbilical vein endothelial cell (HUVEC) migration assays were used to evaluate the vascular effects of the tested materials at 6-8 h. Statistical analyses included Kruskal-Wallis, Mann-Whitney U, and Friedman and Wilcoxon signed rank tests. Results: None of tricalcium silicate-based materials were cytotoxic and all induced a similar release of angiogenic growth factors (PDGF, FGF-2, and VEGF) (p>0.05). The best cell viability was observed for hDPSCs (p<0.05) with all tricalcium silicate-based materials at day 14. Tube formation by HUVECs showed a significant increase with all tested materials (p<0.05). Conclusion: The tricalcium silicate-based materials showed potential for angiogenic stimulation of all stem cell types and significantly enhanced tube formation by HUVECs.

Design, Evaluation And Antimicrobial Activity Of Egyptian Propolis-Loaded Nanoparticles: Intrinsic Role As A Novel And Naturally Based Root Canal Nanosealer.

BACKGROUND: Propolis is a unique natural adhesive product collected by honeybees. It contains a diversity of bioactive compounds with reported functional properties such as antioxidants, antibacterial, antifungal, anti-inflammatory, antiviral and anticancer activity. Dental caries is a worldwide problem that caused by microbial growth usually progress from tooth enamel to the underlying pulpal tissues and root canal. This situation could be controlled by a sequence of steps to remove microorganisms and fill root canal with a suitable long-lasting root canal sealer. Unfortunately, leachable and degradation products of the currently used sealers compromised their antimicrobial activity by inflammatory modulation associated with irritation and toxicity of periapical tissues. MATERIALS AND METHODS: Hence, propolis was selected to be designed as a natural root canal sealer due to its amazing functional properties. Moreover, its handling properties were enhanced and potentiated by its incorporation in polymeric nanoparticles (NPs). Frist, propolis was collected, extracted and analyzed for its bioactive compounds. After that, propolis-loaded NPs of PLGA (ProE-loaded NPs) were developed and fully characterized regarding physicochemical properties, in vitro release and in vitro cytotoxicity. Then, root canal sealers were fabricated and assayed for their antimicrobial activity. Both cytotoxicity and antimicrobial activity were compared to those of a model sealer; AH Plus®. RESULTS: The results revealed that spherical nanoscopic NPs with narrow size distribution were obtained. ProE-loaded NPs exhibited accepted entrapment efficiency (>80) and prolonged release. In vitro cytotoxicity study confirmed the safety of ProE-loaded NPs. Also, the developed sealers showed antimicrobial activity versus bacterial strains of Enterococcus faecalis and Streptococcus mutans and antifungal activity against Candida albicans. CONCLUSION: ProE-loaded NPs could be incorporated in and represented as a root canal sealer with prolonged release and enhanced cytocompatibility as well as antimicrobial activities.