Efficacy of a thermally treated single file compared with rotary systems in endodontic retreatment of curved canals: a micro-CT study.

OBJECTIVES: This study aimed to compare WaveOne Gold with ProTaper and RaCe systems regarding remaining filling material, apical transportation (AT), and working time (WT) after (i) filling removal and (ii) shaping of curved canals. METHODS: Thirty mesiobuccal canals of maxillary molars were prepared and filled. After 30 days, they were randomly assigned into three groups (n = 10), according to the instruments used for filling removal and shaping, respectively: WOG-WaveOne Gold Primary and Medium; PTG-ProTaper Retreatment and ProTaper Next; RCG-D-RaCe and RaCe. Micro-CT analysis assessed the residual filling material and AT. WT was recorded. Data were statistically analyzed (α = .05). RESULTS: There was no significant difference between groups in the amount of filling material at any evaluated moment (P > .05). All groups presented low AT values. The WT was similar in all groups in filling removal (P > .05), and in shaping step WOG was faster than PTG and RCG (P < .05). CONCLUSIONS: Neither system could completely remove the filling material. The instruments evaluated were safe and the reciprocating system was faster than the rotary systems in shaping the canals. CLINICAL RELEVANCE: This study provided consistent information on filling material removal capacity of WaveOne Gold. Considering that all tested systems were safe, WaveOne Gold may be an alternative with cost-effectiveness and shorter learning curve for endodontic retreatment.

Evaluation of n-butyl cyanoacrylate adhesive in rat subcutaneous tissue.

BACKGROUND: Tissue adhesives have been widely used for wound closure, especially in children, because they are painless, fast, and easy to use and result in minimal scarring. OBJECTIVE: To analyze the biocompatibility of an adhesive based on n-butyl-cyanoacrylate in the subcutaneous tissue of rats. MATERIALS AND METHODS: Two surgical sites were prepared (approximately 3 cm apart): one on the left side of the animal and the other on the right side); polyethylene tubes were implanted in each surgical site. The tube on the left was filled with n-butyl-cyanoacrylate (treated group) and the tube on the right side was unfilled (control group). After 7, 30, and 120 days, the animals were killed, and the specimens were processed for histologic analysis. RESULTS: No significant inflammatory reaction occurred in the treated group, showing results similar to the control group. CONCLUSION: This adhesive based on n-butyl-cyanoacrylate is biocompatible in the subcutaneous tissue of rats.

Cytotoxicity and Bioactivity of Calcium Silicate-based Cements in a Culture of Stem Cells from the Apical Papilla.

Introduction: The present in vitro study evaluated the cytotoxicity and bioactivity of commonly-used calcium silicate-based cements in a culture of stem cells from the apical papilla (SCAPs). Materials and Methods: NeoMTA Plus (Avalon Biomed), BiodentineTM (Septodont) and MTA HP Repair (Angelus) cements were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and sulphorhodamine-B (SRB) viability assays. Cells were seeded (1*104 cells mL-1) in 96-well plates and exposed to 1:4 diluted extract in 24 h and 72 h. For the analysis of bioactivity, alkaline phosphatase (ALP) enzyme activity and Alizarin Red S (AZR) were assessed after 24 h of cell culture in 12-well plates (1*104 cells mL-1), where cells were exposed to 1:4 diluted extract on days 1 and 7. Minimum Essential Eagle's Medium alpha modification was used as control. ANOVA and Tukey's post hoc test were used to compare the different cements at each experimental time point. Results: No significant differences were found between the cements and the control specimens on MTT at 24 h and 72 h (P>0.05); however, the calcium silicate-based cement materials showed higher cell viability compared to the control group (P<0.05). In the 24-h SRB, NeoMTA Plus showed lower cell viability than BiodentineTM and MTA HP Repair (P<0.05), with all groups similar to the control group (P>0.05). Compared to 24-h results, only NeoMTA Plus presented increased cell viability at 72 h (P<0.05). ALP activity was similar across the materials at 1 day (P>0.05). ALP activity was higher for BiodentineTM when compared to NeoMTA Plus (P<0.05), nevertheless, it was similar to MTA HP Repair and control groups (P>0.05) at 7 days. At 1- and 7-day periods of AZR assay, BiodentineTM presented higher levels of mineralized nodule formation (P<0.05). Conclusion: All evaluated calcium silicate-based cements demonstrated cell viability and bioactivity, suggesting that these (bio)materials may be indicated for use in regenerative dentine-pulp complex procedures.

Biological Properties of Experimental Methacrylate-Based Sealers Containing Calcium Phosphates.

This study aimed to evaluate, in vitro and in vivo, the biocompatibility of experimental methacrylate-based endodontic sealers containing α-tricalcium phosphate (α-TCP) or nanostructured hydroxyapatite (HAp). Experimental methacrylate-based dual-cure sealers with the addition of α-TCP or HAp, at 10%wt were formulated and compared to AH Plus (AHP). Cell viability was assessed by 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT), and sulforhodamine B (SRB). Sealers were implanted in rats' subcutaneous tissue and histologically evaluated. Bioactivity was assessed by alkaline phosphatase enzyme activity (ALP) and Alizarin Red (AR), using apical papillary cells (SCAPs), and by the bone deposition measured in surgical cavities on rats' femur filled with AH Plus or α-TCP. In both viability assays, HAp and AHP sealers were similar, and α-TCP presented lower viability compared to the others at MTT assay (p<0.05). A gradual decrease of the inflammatory response according to the periods was observed and AHP was the only that presented giant cells (7-day period). Collagen fibers condensation increased according to the periods, with no differences among sealers. There was an increase at ALP activity and mineralized nodules deposition according to periods. HAp and α-TCP presented higher values for ALP activity at 5 days and at 5, 10, and 15 days for AR and were different from AHP (p<0.05). α-TCP presented superior values at 10 and 15 days compared to HAp and AHP for AR (p<0.05). At 90 days, α-TCP and control (empty cavity) showed high bone deposition compared to AHP (p<0.05). α-TCP and HAp, in a methacrylate-based sealer, presented biocompatibility and bioactivity, with the potential to be used as endodontic sealers in clinical practice. Further investigations are required to gain information on the physicochemical properties of these sealers formulation before its clinical implementation.

Cytocompatibility and cell proliferation evaluation of calcium phosphate-based root canal sealers.

OBJECTIVES: This study aimed to evaluate the cell viability and migration of Endosequence Bioceramic Root Canal Sealer (BC Sealer) compared to MTA Fillapex and AH Plus. MATERIALS AND METHODS: BC Sealer, MTA Fillapex, and AH Plus were placed in contact with culture medium to obtain sealers extracts in dilution 1:1, 1:2 and 1:4. 3T3 cells were plated and exposed to the extracts. Cell viability and migration were assessed by 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) and Scratch assay, respectively. Data were analyzed by Kruskal-Wallis and Dunn's test (p < 0.05). RESULTS: The MTT assay revealed greater cytotoxicity for AH Plus and MTA Fillapex at 1:1 dilution when compared to control (p < 0.05). At 1:2 and 1:4 dilutions, all sealers were similar to control (p > 0.05) and MTA Fillapex was more cytotoxic than BC Sealer (p < 0.05). Scratch assay demonstrated the continuous closure of the wound according to time. At 30 hours, the control group presented closure of the wound (p < 0.05). At 36 hours, only BC Sealer presented the closure when compared to AH Plus and MTA Fillapex (p < 0.05). At 42 hours, AH Plus and MTA Fillapex showed a wound healing (p > 0.05). CONCLUSIONS: All tested sealers demonstrated cell viability highlighting BC Sealer, which showed increased cell migration capacity suggesting that this sealer may achieve better tissue repair when compared to other tested sealers.

Influence of photobiomodulation therapy on root development of rat molars with open apex and pulp necrosis.

This study aimed to evaluate the role of photobiomodulation (PBM) in apexification and apexogenesis of necrotic rat molars with an open apex. Rat molars were exposed to the oral environment for 3 weeks. Canals were rinsed with 2.5% NaOCl and 17% EDTA, filled with antibiotic paste and sealed. After 7 days, canals were rinsed and divided into six groups (n=6): mineral trioxide aggregate (MTA); blood clot (BC); human dental pulp stem cells (hDPSC); MTA+PBM; BC+PBM; and hDPSC+PBM. In hDPSC groups, a 1% agarose gel scaffold was used. Two groups were not exposed: healthy tooth+PBM (n = 6), healthy tooth (n = 3); and one was exposed throughout the experiment: necrotic tooth (n = 3). In PBM groups, irradiation was performed with aluminum gallium indium phosphide (InGaAlP) diode laser for 30 days within 24-h intervals. After that, the specimens were processed for histological and immunohistochemical analyses. Necrotic tooth showed greater neutrophil infiltrate (p < 0.05). Necrotic tooth, healthy tooth, and healthy tooth+PBM groups showed absence of a thin layer of fibrous condensation in the periapical area. All the other groups stimulated the formation of a thicker layer of fibers (p < 0.05). All groups formed more mineralized tissue than necrotic tooth (p < 0.05). PBM associated with MTA, BC, or hDPSC formed more mineralized tissue (p < 0.05). MTA+PBM induced apexification (p < 0.05). Rabbit polyclonal anti-bone sialoprotein (BSP) antibody confirmed the histological findings of mineralized tissue formation, and hDPSC groups exhibited higher percentage of BSP-positive cells. It can be concluded that PBM improved apexification and favored apexogenesis in necrotic rat molars with an open apex.

Antimicrobial and anti-inflammatory drug-delivery systems at endodontic reparative material: Synthesis and characterization.

OBJECTIVE: The aim of this study was to synthesize and characterize an experimental endodontic paste. METHODS: An experimental endodontic paste (EX) was characterized by its particle size, zeta potential, drug content and morphology. The powder of EX is composed of amoxicillin microspheres, calcium tungstate and α-tricalcium phosphate, mixed with an indomethacin nanocapsules suspension. Ultracal® (Ultradent), an iodoform-based paste (GP) and the EX were evaluated by its physical properties (flow, film thickness and radiopacity). The cytocompatibility was performed by MTT and SRB-colorimetric assays; the cell-migration was tested with scratch assay and cell-ability to remineralization with ALP and Alizarin Red S, with fibroblastic cell line. The antibacterial activity was assessed by the formation of inhibition zones and against planktonic bacteria. RESULTS: The EX and UL flow achieved ISO6876 standard, and GP was lower than 17mm. All pastes achieved the film thickness required. Radiopacity was equivalent to 1.81±0.25mmAl for EX, which did not differ from GP group 1.39±0.33mmAl (p>0.05). The UL presented 3.04±0.33mmAl. The values for SRB showed better citocompatibility in comparison with MTT for all materials. The ALP activity and formation of mineralized nodules demonstrated the remineralization potential for UL and EX. Cell migration showed continuous wound closure until complete cell healing, however, the EX accelerated the process (p<0.05). The EX showed the greatest inhibition zone (p<0.05) and was the only group with antibacterial activity against planktonic bacteria. SIGNIFICANCE: The synthesized endodontic paste demonstrated reliable physical and biological properties and could be a promising material for periapical tissue repair.

Photobiomodulation therapy improves multilineage differentiation of dental pulp stem cells in three-dimensional culture model.

Photobiomodulation therapy (PBM) has shown positive effects on stem cell differentiation in monolayer cell culture model, but little is known about its effect on three-dimensional (3-D) agarose gel culture. This study evaluated the PBM effect of human dental pulp stem cells (hDPSCs) differentiation and phosphatase alkaline activity (ALP) using an agarose 3-D model under different nutritional conditions. hDPSCs were characterized and seeded on a 0.3% agarose gel layer with different media (osteogenic, adipogenic, chondrogenic) and were assigned into four groups: control 10% fetal bovine serum (FBS), control 5% FBS, PBM 10% FBS, and PBM 5% FBS. Irradiation was performed with continuous-wave InGaAlP laser, 660 nm, 100 mW, 3,3 J / cm2, spot size 0.3 cm2, 10 s of exposure time, and 1 J of energy per point with 6-h interval between sessions. All groups were evaluated at 7 and 14 days. ALP assay was performed to analyze the deposition of mineralized tissue. At 7 days, PBM 5% FBS group presented better stimulation in osteogenic and adipogenic differentiation compared with control. After 14 days, hDPSCs cultured in 3-D exhibited osteogenic, adipogenic, and chondrogenic differentiation; furthermore, compared to control, PBM significantly stimulated all differentiation processes (p < 0.05). It can be concluded that hDPSCs cultured in 3-D agarose associated to PBM could be a promising tool for tissue engineering applications.

Cell Viability and Tissue Reaction of NeoMTA Plus: An In Vitro and In Vivo Study.

INTRODUCTION: The aim of this study was to evaluate the cell viability and tissue reaction of NeoMTA Plus (NMP; Avalon Biomed Inc, Houston, TX) compared with mineral trioxide aggregate (MTA; Angelus, Londrina, PR, Brazil) and Biodentine (BD; Septodont, Saint-Maur-de-Fossés, France). METHODS: Fibroblasts (3T3) were plated and exposed to 1% extract from the test material before and after setting. Cytotoxicity assessment was performed using the 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide and sulforhodamine B assays. In vivo evaluation consisted of polyethylene tube implantation of the materials in rat subcutaneous tissue. Histologic analysis occurred at 7, 30, and 90 days, scoring inflammatory events and collagen fiber formation. Analysis of variance and the Tukey and t tests were used for cytocompatibility assays, and the Kruskal-Wallis test followed by the Dunn test were used for biocompatibility assays (P ≤ .05). RESULTS: The materials in the cytotoxicity assays presented greater viability after setting (P ≤ .05). NMP and MTA presented higher viability than the control (Dulbecco modified Eagle medium) on the 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide assay before and after setting (P ≤ .05). The sulforhodamine B assay showed that MTA and BD presented less viability than NMP and the control, and NMP was similar to the control before setting. After setting, MTA and BD presented higher viability when compared with the control group (P ≤ .05), and NMP was similar to control. Inflammatory infiltrate reduction occurred throughout the test periods for all materials. At 7 days, neutrophils were present in BD (P ≤ .05), and granuloma and giant cells were present in BD and MTA. At 30 days, BD showed intense inflammatory infiltrates and a large number of macrophages when compared with NMP, MTA, and the control (P ≤ .05). At 90 days, BD presented a thick fiber layer compared with NMP (P ≤ .05). CONCLUSIONS: NMP showed similar biocompatible behavior to MTA and BD.

Cell migration, viability and tissue reaction of calcium hypochlorite based-solutions irrigants: An in vitro and in vivo study.

OBJECTIVE: This study aimed to analyze in vitro cytotoxicity to cultured 3T3 fibroblasts and in vivo inflammatory reaction in rats by calcium hypochlorite (Ca(OCl)2) solutions compared with sodium hypochlorite (NaOCl) solutions. DESIGN: Cultured 3T3 fibroblasts were exposed to different concentrations of (Ca(OCl)2) and NaOCl solutions, and a scratch assay was performed. The viability rate was analyzed with trypan blue assay. Both solutions of 1% and 2.5% concentrations were injected into the subcutaneous tissue of 18 male Wistar rats aged 18 weeks. The inflammatory tissue reaction was evaluated at 2h, 24h, and 14days after the injections. The samples were qualitatively analyzed using a light microscope. Statistical analysis was performed with ANOVA and Tukey post hoc tests for in vitro assays and Kruskal-Wallis and Dunn post hoc tests for in vivo assays (α=0.05). RESULTS: In the scratch assay, Ca(OCl)2 showed no significant difference compared with the control group (culture medium) at 24h (p<0.05). Solutions of 0.0075% and 0.005% NaOCl and Ca(OCl)2 concentrations presented similar results compared with those in the positive control group (hydrogen peroxide) (p>0.05) in the trypan blue assay. In the in vivo assay, 1% Ca(OCl)2 group showed a significant decrease in neutrophils at 2h and 24h (p=0.041) and 2h and 14days (p=0.017). There was no statistically significant difference for lymphocyte/plasmocyte and macrophage counts among the different concentration groups. CONCLUSIONS: Ca(OCl)2 showed favorable results of viability and induced a low-level inflammatory response. Ca(OCl)2 presented acceptable cytotoxicity and biocompatibility as an irrigant solution.

Cytotoxicity and Bioactivity of Calcium Silicate Cements Combined with Niobium Oxide in Different Cell Lines.

The aim of this study was to evaluate the cytotoxicity and bioactivity of calcium silicate-based cements combined with niobium oxide (Nb2O5) micro and nanoparticles, comparing the response in different cell lines. This evaluation used four cell lines: two primary cultures (human dental pulp cells - hDPCs and human dental follicle cells - hDFCs) and two immortalized cultures (human osteoblast-like cells - Saos-2 and mouse periodontal ligament cells - mPDL). The tested materials were: White Portland Cement (PC), mineral trioxide aggregate (MTA), white Portland cement combined with microparticles (PC/Nb2O5µ) or nanoparticles (PC/Nb2O5n) of niobium oxide (Nb2O5). Cytotoxicity was evaluated by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) and trypan blue exclusion assays and bioactivity by alkaline phosphatase (ALP) enzyme activity. Results were analyzed by ANOVA and Tukey test (a=0.05). PC/Nb2O5n presented similar or higher cell viability than PC/Nb2O5µ in all cell lines. Moreover, the materials presented similar or higher cell viability than MTA. Saos-2 exhibited high ALP activity, highlighting PC/Nb2O5µ material at 7 days of exposure. In conclusion, calcium silicate cements combined with micro and nanoparticles of Nb2O5 presented cytocompatibility and bioactivity, demonstrating the potential of Nb2O5 as an alternative radiopacifier agent for these cements. The different cell lines had similar response to cytotoxicity evaluation of calcium silicate cements. However, bioactivity was more accurately detected in human osteoblast-like cell line, Saos-2.

Cytotoxicity and genotoxicity of calcium silicate-based cements on an osteoblast lineage.

Several calcium silicate-based biomaterials have been developed in recent years, in addition to Mineral Trioxide Aggregate (MTA). The aim of this study was to evaluate the cytotoxicity, genotoxicity and apoptosis/necrosis in human osteoblast cells (SAOS-2) of pure calcium silicate-based cements (CSC) and modified formulations: modified calcium silicate-based cements (CSCM) and three resin-based calcium silicate cements (CSCR1) (CSCR 2) (CSCR3). The following tests were performed after 24 hours of cement extract exposure: methyl-thiazolyl tetrazolium (MTT), apoptosis/necrosis assay and comet assay. The negative control (CT-) was performed with untreated cells, and the positive control (CT+) used hydrogen peroxide. The data for MTT and apoptosis were submitted to analysis of variance and Bonferroni's posttest (p < 0.05), and the data for the comet assay analysis, to the Kruskal-Wallis and Dunn tests (p < 0.05). The MTT test showed no significant difference among the materials in 2 mg/mL and 10 mg/mL concentrations. CSCR3 showed lower cell viability at 10 mg/mL. Only CSC showed lower cell viability at 50 mg/mL. CSCR1, CSCR2 and CSCR3 showed a higher percentage of initial apoptosis than the control in the apoptosis test, after 24 hours exposure. The same cements showed no genotoxicity in the concentration of 2 mg/mL, with the comet assay. CSC and CSCR2 were also not genotoxic at 10 mg/mL. All experimental materials showed viability with MTT. CSC and CSCR2 presented a better response to apoptosis and genotoxicity evaluation in the 10 mg/mL concentration, and demonstrated a considerable potential for use as reparative materials.

Centering and transportation: in vitro evaluation of continuous and reciprocating systems in curved root canals.

CONTEXT: One of the goals of endodontic therapy is the shaping and cleaning of the root canal system. In recent years, there has been multiple systems instrumentation, and changes in their dynamics are central to maintain the original shape of the canal after preparation. AIMS: The aim of this study was to evaluate centering and transportation in curved root canals after using ProTaper(®) and MTwo(®) in continuous rotation, Reciproc(®) in reciprocating motion, and a step-down manual instrumentation technique. SETTINGS AND DESIGN: Mesiobuccal roots of human extracted the first and second maxillary molars were selected and the canals (n = 60) were divided into four groups according to the preparation techniques: PT-ProTaper(®); MT-MTwo(®); RE-Reciproc(®); MI-manual instrumentation. SUBJECTS AND METHODS: The final apical diameter was standardized to a size 25. Centering and transportation were evaluated by cone-beam computed tomography and Adobe Photoshop 8.0 software. STATISTICAL ANALYSIS USED: The data were statistically analyzed by ANOVA and Tukey post hoc. RESULTS: Results of transportation showed no statistical differences (P > 0.05) between groups, and significantly, difference (P < 0.05) between ProTaper(®) and Reciproc(®) was found when evaluating centering ability in the apical third. CONCLUSIONS: We concluded that there were no differences in transportation between the evaluated systems for the preparation of curved root canals with an apical instrumentation diameter of #25. For centering ability, in the apical third, ProTaper(®) presented worst behavior when compared to Reciproc(®).

Biocompatibility and bioactivity of calcium silicate-based endodontic sealers in human dental pulp cells.

UNLABELLED: Mineral Trioxide Aggregate (MTA) is a calcium silicate-based material. New sealers have been developed based on calcium silicate as MTA Fillapex and MTA Plus. OBJECTIVE: The aim of this study was to evaluate biocompatibility and bioactivity of these two calcium silicate-based sealers in culture of human dental pulp cells (hDPCs). MATERIAL AND METHODS: The cells were isolated from third molars extracted from a 16-year-old patient. Pulp tissue was sectioned into fragments with approximately 1 mm3 and kept in supplemented medium to obtain hDPCs adherent cultures. Cell characterization assays were performed to prove the osteogenic potential. The evaluated materials were: MTA Plus (MTAP); MTA Fillapex (MTAF) and FillCanal (FC). Biocompatibility was evaluated with MTT and Neutral Red (NR) assays, after hDPCs exposure for 24 h to different dilutions of each sealer extract (1:2, 1:3 and 1:4). Unexposed cells were the positive control (CT). Bioactivity was assessed by alkaline phosphatase (ALP) enzymatic assay in cells exposed for one and three days to sealer extracts (1:4 dilution). All data were analyzed by ANOVA and Tukey post-test (p≤0.05%). RESULTS: MTT and NR results showed suitable cell viability rates for MTAP at all dilutions (90-135%). Cells exposed to MTAF and FC (1:2 and 1:4 dilutions) showed significant low viability rate when compared to CT in MTT. The NR results demonstrated cell viability for all materials tested. In MTAP group, the cells ALP activity was similar to CT in one and three days of exposure to the material. MTAF and FC groups demonstrated a decrease in ALP activity when compared to CT at both periods of cell exposure. CONCLUSIONS: The hDPCs were suitable for the evaluation of new endodontic materialsin vitro. MTAP may be considered a promising material for endodontic treatments.

Radiopacity and cytotoxicity of Portland cement associated with niobium oxide micro and nanoparticles.

OBJECTIVE: Mineral Trioxide Aggregate (MTA) is composed of Portland Cement (PC) and bismuth oxide (BO). Replacing BO for niobium oxide (NbO) microparticles (Nbµ) or nanoparticles (Nbη) may improve radiopacity and bioactivity. The aim of this study was to evaluate the radiopacity and cytotoxicity of the materials: (1) PC; (2) White MTA; (3) PC+30% Nbµ; (4) PC+30% Nbη. MATERIAL AND METHODS: For the radiopacity test, specimens of the different materials were radiographed along an aluminum step-wedge. For cell culture assays, Saos-2 osteoblastic-cells (ATCC HTB-85) were used. Cell viability was evaluated through MTT assay, and bioactivity was assessed by alkaline phosphatase activity assay. RESULTS: The results demonstrated higher radiopacity for MTA, followed by Nbµ and Nbη, which had similar values. Cell culture analysis showed that PC and PC+NbO associations promoted greater cell viability than MTA. CONCLUSIONS: It was concluded that the combination of PC+NbO is a potential alternative for composition of MTA.

Influence of photobiomodulation therapy on root development of rat molars with open apex and pulp necrosis

Abstract This study aimed to evaluate the role of photobiomodulation (PBM) in apexification and apexogenesis of necrotic rat molars with an open apex. Rat molars were exposed to the oral environment for 3 weeks. Canals were rinsed with 2.5% NaOCl and 17% EDTA, filled with antibiotic paste and sealed. After 7 days, canals were rinsed and divided into six groups (n=6): mineral trioxide aggregate (MTA); blood clot (BC); human dental pulp stem cells (hDPSC); MTA+PBM; BC+PBM; and hDPSC+PBM. In hDPSC groups, a 1% agarose gel scaffold was used. Two groups were not exposed: healthy tooth+PBM (n = 6), healthy tooth (n = 3); and one was exposed throughout the experiment: necrotic tooth (n = 3). In PBM groups, irradiation was performed with aluminum gallium indium phosphide (InGaAlP) diode laser for 30 days within 24-h intervals. After that, the specimens were processed for histological and immunohistochemical analyses. Necrotic tooth showed greater neutrophil infiltrate (p < 0.05). Necrotic tooth, healthy tooth, and healthy tooth+PBM groups showed absence of a thin layer of fibrous condensation in the periapical area. All the other groups stimulated the formation of a thicker layer of fibers (p < 0.05). All groups formed more mineralized tissue than necrotic tooth (p < 0.05). PBM associated with MTA, BC, or hDPSC formed more mineralized tissue (p < 0.05). MTA+PBM induced apexification (p < 0.05). Rabbit polyclonal anti-bone sialoprotein (BSP) antibody confirmed the histological findings of mineralized tissue formation, and hDPSC groups exhibited higher percentage of BSP-positive cells. It can be concluded that PBM improved apexification and favored apexogenesis in necrotic rat molars with an open apex.

Antimicrobial Brazilian Propolis (EPP-AF) Containing Biocellulose Membranes as Promising Biomaterial for Skin Wound Healing.

Among remarkable discoveries concerning propolis, such as antifungal, antiviral, and antioxidant activities, its anti-inflammatory, and mainly its antibacterial, properties deserve special attention when skin wound healing is concerned. Based on this and knowing the distinctive performance of bacterial (BC) membranes on wound healing, in this work it is proposed to demonstrate the potent antimicrobial activity and wound healing properties of a novel propolis containing biocellulose membrane. The obtained propolis/BC membrane was able to adsorb propolis not only on the surface, but also in its interstices demonstrated by scanning electron microscopy, X-ray diffraction, Fourier transform infrared (FT-IR) spectroscopy, and thermogravidimetric assays. Additionally, the polyphenolic compounds determination and the prominent antibacterial activity in the membrane are demonstrated to be dose dependent, supporting the possibility of obtaining propolis/BC membranes at the desired concentrations, taking into consideration its application and its skin residence time. Finally, it could be suggested that propolis/BC membrane may favor tissue repair in less time and more effectively in contaminated wounds.

Cytotoxicity and Bioactivity of Calcium Silicate Cements Combined with Niobium Oxide in Different Cell Lines

Abstract The aim of this study was to evaluate the cytotoxicity and bioactivity of calcium silicate-based cements combined with niobium oxide (Nb2O5) micro and nanoparticles, comparing the response in different cell lines. This evaluation used four cell lines: two primary cultures (human dental pulp cells - hDPCs and human dental follicle cells - hDFCs) and two immortalized cultures (human osteoblast-like cells - Saos-2 and mouse periodontal ligament cells - mPDL). The tested materials were: White Portland Cement (PC), mineral trioxide aggregate (MTA), white Portland cement combined with microparticles (PC/Nb2O5µ) or nanoparticles (PC/Nb2O5n) of niobium oxide (Nb2O5). Cytotoxicity was evaluated by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) and trypan blue exclusion assays and bioactivity by alkaline phosphatase (ALP) enzyme activity. Results were analyzed by ANOVA and Tukey test (a=0.05). PC/Nb2O5n presented similar or higher cell viability than PC/Nb2O5µ in all cell lines. Moreover, the materials presented similar or higher cell viability than MTA. Saos-2 exhibited high ALP activity, highlighting PC/Nb2O5µ material at 7 days of exposure. In conclusion, calcium silicate cements combined with micro and nanoparticles of Nb2O5 presented cytocompatibility and bioactivity, demonstrating the potential of Nb2O5 as an alternative radiopacifier agent for these cements. The different cell lines had similar response to cytotoxicity evaluation of calcium silicate cements. However, bioactivity was more accurately detected in human osteoblast-like cell line, Saos-2.

Resumo O objetivo deste estudo foi avaliar a citotoxicidade e bioatividade de cimentos à base de silicato de cálcio associados com óxido de nióbio (Nb2O5) micro e nanoparticulados, e comparar a resposta em diferentes linhagens celulares. Foram utilizadas quatro linhagens celulares: duas culturas primárias (células da polpa dentária humana - hDPCs e células do folículo dentário humano - hDFCs) e duas culturas imortalizadas (células osteoblásticas humanas - Saos-2 e células do ligamento periodontal de ratos - mPDL). Os materiais analisados foram: Cimento Portland branco (PC); Agregado trióxido mineral (MTA); PC associado com micropartículas (PC/Nb2O5µ) ou nanopartículas (PC/Nb2O5n) de óxido de nióbio (Nb2O5). A citotoxicidade foi avaliada pelos ensaios de brometo de metil-tiazolil-difeniltetrazólio (MTT) e azul de tripan, e a bioatividade pela atividade da enzima fosfatase alcalina (ALP). Os resultados foram analisados por ANOVA e teste de Tukey (a=0,05). O grupo do PC/Nb2O5n apresentou viabilidade celular semelhante ou maior do que o grupo do PC/Nb2O5μ em todas as linhagens celulares. Além disso, ambos os grupos apresentaram viabilidade celular semelhante ou maior do que o MTA. Saos-2 apresentaram maior atividade de ALP, com destaque para o material PC/Nb2O5μ aos 7 dias de exposição. Concluiu-se que cimentos de silicato de cálcio associados com Nb2O5 micro ou nanoparticulado apresentaram citocompatibilidade e bioatividade, demonstrando potencial do Nb2O5 como agente radiopacificador alternativo para estes cimentos. As linhagens celulares estudadas apresentaram resposta semelhante na avaliação da citotoxicidade de cimentos de silicato de cálcio. No entanto, a bioatividade é melhor detectada na linhagem de células osteoblásticas humanas, Saos-2.

Cytotoxicity and genotoxicity of calcium silicate-based cements on an osteoblast lineage

Abstract Several calcium silicate-based biomaterials have been developed in recent years, in addition to Mineral Trioxide Aggregate (MTA). The aim of this study was to evaluate the cytotoxicity, genotoxicity and apoptosis/necrosis in human osteoblast cells (SAOS-2) of pure calcium silicate-based cements (CSC) and modified formulations: modified calcium silicate-based cements (CSCM) and three resin-based calcium silicate cements (CSCR1) (CSCR 2) (CSCR3). The following tests were performed after 24 hours of cement extract exposure: methyl-thiazolyl tetrazolium (MTT), apoptosis/necrosis assay and comet assay. The negative control (CT-) was performed with untreated cells, and the positive control (CT+) used hydrogen peroxide. The data for MTT and apoptosis were submitted to analysis of variance and Bonferroni's posttest (p < 0.05), and the data for the comet assay analysis, to the Kruskal-Wallis and Dunn tests (p < 0.05). The MTT test showed no significant difference among the materials in 2 mg/mL and 10 mg/mL concentrations. CSCR3 showed lower cell viability at 10 mg/mL. Only CSC showed lower cell viability at 50 mg/mL. CSCR1, CSCR2 and CSCR3 showed a higher percentage of initial apoptosis than the control in the apoptosis test, after 24 hours exposure. The same cements showed no genotoxicity in the concentration of 2 mg/mL, with the comet assay. CSC and CSCR2 were also not genotoxic at 10 mg/mL. All experimental materials showed viability with MTT. CSC and CSCR2 presented a better response to apoptosis and genotoxicity evaluation in the 10 mg/mL concentration, and demonstrated a considerable potential for use as reparative materials.

Avaliação de três técnicas de instrumentação com diferentes diâmetros apicais na formação de smear layer / Evaluation of three instrumentation techniques with different apical diameters on smear layer formation

Objetivo: avaliar os efeitos de diferentes instrumentações na formação de smear layer no terço apical de 90 raízes mesiobucais de molares superiores por microscopia eletrônica de varredura (MEV). Materiais e método: três grupos foram formados baseados nas técnicas utilizadas: instrumentação manual, sistema rotatório K3 e sistema reciprocante NSK. Os grupos foram subdivididos em três, de acordo com o diâmetro apical: #30, #35 ou #40. Após o preparo, as raízes foram seccionadas no sentido transversal, separando os terços apicais do restante das raízes; esses terços foram divididos em duas metades e preparados para MEV. A formação de smear layer na superfície do canal radicular e os túbulos den-tinários foram avaliados por escores num aumento de 1.000×. Os dados foram analisados pelo teste Kruskal-Wallis complementado pelo teste de Dunn (p < 0.05). Resultados: o aumento no diâmetro do preparo apical não influenciou na formação de smear layer nas paredes dentinárias. Conclusão: apesar das técnicas e dos diâmetros apicais utilizados durante o preparo, nenhuma parede livre de smear layer foi observada.