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

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.

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.

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

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

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

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

Chlorhexidine and proanthocyanidin enhance the long-term bond strength of resin-based endodontic sealer.

The aim of this study was to evaluate the effects of proanthocyanidin (PA) and chlorhexidine (CHX) on the bond strength (BS), failure pattern, and resin-dentin interface morphology of the endodontic sealers EndoREZ and AH Plus after 24 h and 6 months of water storage. A total of 120 prepared bovine roots were divided into six groups: AH Plus, CHX+AH Plus, PA+AH Plus, EndoREZ, CHX+EndoREZ, and PA+EndoREZ. Dentin was treated for 1 or 5 min with 2% CHX or 15% PA, respectively. Roots were filled and stored in water for 24 h or 6 months (n = 10). Root slices were subjected to push-out test and scanning electron microscopy (SEM). Data were compared using two-way ANOVA and student's t-test (α = 5%). BS decreased over time for AH Plus and untreated EndoREZ (p < 0.05). At 24 h, AH Plus had higher BS than EndoREZ (p < 0.001), with no differences among treatments for both sealers (p > 0.05). At 6 months, EndoREZ had higher BS values for CHX and PA than control (p < 0.05). AH Plus had higher BS than EndoREZ (p < 0.001), while with CHX or PA, similar BS was observed in both sealers (p > 0.05). Cohesive and mixed failures were observed in all groups. SEM revealed sealer tags in the root dentin. In conclusion, BS decreased with time and AH Plus had higher BS than EndoREZ in untreated dentin; however, CHX or PA enhanced long-term BS of EndoREZ. Overall, dentin treatment affected failure pattern and resin-dentin interface morphology, particularly for EndoREZ.

Local delivery of strontium ranelate promotes regeneration of critical size bone defects filled with collagen sponge.

The effect of local delivery of strontium ranelate (SR) on bone regeneration of critical size bone defects filled with collagen sponge was evaluated. Bone defects of 5 mm diameter created in rat calvaria were filled with collagen sponge (C); collagen sponge with 5 mM Sr2+ SR (C5SR) or collagen sponge with 50 mM Sr2+ SR (C50SR). After 2, 4, and 6 weeks, bone volume (BV), bone surface (BS), trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.Sp) were evaluated by computed microtomography. At 6 weeks, histological analysis was performed. Intragroup comparisons were made by the Friedman test, while comparisons between groups were made by Kruskal-Wallis test (α = 5%). All groups showed increased BV, BS, Tb.Th, and Tb.N over time, but only C50SR promoted the reduction of Tb.Sp (p < 0.05). No significant differences between groups were detected at weeks 2 and 4. However, C50SR showed the highest values of BV, BS, and Tb.Th at 6 weeks (p < 0.05). Histological analysis revealed connective tissue in C and C5SR and immature bone tissue in C50SR. Local delivery of SR 50 mM Sr2+ associated with collagen sponge increased and accelerated bone regeneration in critical bone defects. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 333-341, 2018.

Effects of EDTA gel and chlorhexidine gel on root dentin permeability.

The aim of this study was to evaluate the effect of 24% ethylenediaminetetraacetic acid (EDTA) gel and 2% chlorhexidine gel (CHX) in dentin permeability and smear layer removal from root canals instrumented with NiTi rotary system using histochemical staining and scanning electron microscopy (SEM) analysis. Overall, 43 premolars were classified into two experimental groups, EDTA (n = 20) and CHX (n = 20), and a negative control (NC) (n = 3). All specimens were instrumented and the irrigant solutions were used after each file change. The EDTA group received a final rinse with 5-ml 1% NaOCl followed by a 5-ml 0.9% saline solution; the CHX group received a final rinse with 10-ml 0.9% saline solution; and the negative control group received a final rinse with only 0.9% saline solution. Fifteen teeth from each group were prepared for histochemical staining and evaluation of dentin permeability using the image-scanning software Axion Vision (v.4.8.2). Five remaining teeth were prepared for analysis using SEM for morphological analysis. The study found that 24% EDTA gel increased the permeability of dentin in all thirds evaluated and also demonstrated an increased cleaning ability, with dentinal walls free of smear layer and open dentinal tubules, as compared to 2% CHX gel. It was concluded that EDTA was efficient in cleaning the dentinal tubules and increased dentin permeability.

Chlorhexidine and proanthocyanidin enhance the long-term bond strength of resin-based endodontic sealer

Abstract The aim of this study was to evaluate the effects of proanthocyanidin (PA) and chlorhexidine (CHX) on the bond strength (BS), failure pattern, and resin-dentin interface morphology of the endodontic sealers EndoREZ and AH Plus after 24 h and 6 months of water storage. A total of 120 prepared bovine roots were divided into six groups: AH Plus, CHX+AH Plus, PA+AH Plus, EndoREZ, CHX+EndoREZ, and PA+EndoREZ. Dentin was treated for 1 or 5 min with 2% CHX or 15% PA, respectively. Roots were filled and stored in water for 24 h or 6 months (n = 10). Root slices were subjected to push-out test and scanning electron microscopy (SEM). Data were compared using two-way ANOVA and student's t-test (α = 5%). BS decreased over time for AH Plus and untreated EndoREZ (p < 0.05). At 24 h, AH Plus had higher BS than EndoREZ (p < 0.001), with no differences among treatments for both sealers (p > 0.05). At 6 months, EndoREZ had higher BS values for CHX and PA than control (p < 0.05). AH Plus had higher BS than EndoREZ (p < 0.001), while with CHX or PA, similar BS was observed in both sealers (p > 0.05). Cohesive and mixed failures were observed in all groups. SEM revealed sealer tags in the root dentin. In conclusion, BS decreased with time and AH Plus had higher BS than EndoREZ in untreated dentin; however, CHX or PA enhanced long-term BS of EndoREZ. Overall, dentin treatment affected failure pattern and resin-dentin interface morphology, particularly for EndoREZ.

Oxidative nanopatterning of titanium surfaces promotes production and extracellular accumulation of osteopontin.

The bone-biomaterial interface has been characterized by layers of afibrillar extracellular matrix (ECM) enriched in non collagenous proteins, including osteopontin (OPN), a multifunctional protein that in bone controls cell adhesion and ECM mineralization. Physical and chemical aspects of biomaterial surfaces have been demonstrated to affect cell-ECM-substrate interactions. The present paper described the ability of oxidative nanopatterning of titanium (Ti) surfaces to control extracellular OPN deposition in vitro. Ti discs were chemically treated by a mixture of H2SO4/H2O2 for either 30 min [Nano(30') Ti] or 4 h [Nano(4h) Ti]. Non-etched Ti discs were used as control. Primary osteogenic cells derived from newborn rat calvarial bone were plated on control and etched Ti and grown under osteogenic conditions up to 7 days. High resolution scanning electron microscopy revealed that treated Ti discs exhibited a nanoporous surface and that areas of larger nanopits were noticed only for Nano(4h) Ti. Large extracellular OPN accumulation were detectable only for Nano(4h) Ti, which was associated with OPN-positive cells with typical aspects of migrating cells. At day 3, quantitative results in terms of areas of OPN labeling were as follows: Nano(4h) Ti > Nano(30') Ti > Control Ti. In conclusion, chemically nanostructured Ti surfaces may support the enhancement of endogenous extracellular OPN deposition by osteogenic cells in vitro depending on the etching time, a finding that should be taken into consideration in strategies to biofunctionalize implant surfaces with molecules with cell adhesion capacity.

Effects of a novel calcium aluminate cement on the early events of the progression of osteogenic cell cultures.

The present study evaluated the progression of osteogenic cell cultures exposed to a novel calcium aluminate cement (CAC+) in comparison with the gold standard mineral trioxide aggregate (MTA). Cells were enzimatically isolated from newborn rat calvarial bone, plated on glass coverslips containing either CAC+ or a control MTA samples in the center, and grown under standard osteogenic conditions. Over the 10-day culture period, roundening of sample edges was clearly noticed only for MTA group. Although both cements supported osteogenic cell adhesion, spreading, and proliferation, CAC+-exposed cultures showed significantly higher values in terms of total cell number at days 3 and 7, and total protein content and alkaline phosphatase activity at day 10. The present in vitro results indicate that the exposure to CAC+ supports a higher differentiation of osteogenic cells compared with the ones exposed to MTA. Further experimental studies should consider CAC+ as a potential alternative to MTA when the repair of mineralized tissues is one of the desired outcomes in endodontic therapy.

Effects of a novel calcium aluminate cement on the early events of the progressionof osteogenic cell cultures

The present study evaluated the progression of osteogenic cell cultures exposed to a novel calcium aluminate cement (CAC+) in comparison with the gold standard mineral trioxide aggregate (MTA). Cells were enzimatically isolated from newborn rat calvarial bone, plated on glass coverslips containing either CAC+ or a control MTA samples in the center, and grown under standard osteogenic conditions. Over the 10-day culture period, roundening of sample edges was clearly noticed only for MTA group. Although both cements supported osteogenic cell adhesion, spreading, and proliferation, CAC+-exposed cultures showed significantly higher values in terms of total cell number at days 3 and 7, and total protein content and alkaline phosphatase activity at day 10. The present in vitro results indicate that the exposure to CAC+ supports a higher differentiation of osteogenic cells compared with the ones exposed to MTA. Further experimental studies should consider CAC+ as a potential alternative to MTA when the repair of mineralized tissues is one of the desired outcomes in endodontic therapy.

O objetivo do presente estudo foi avaliar a progressão de cultura de células osteogênicas expostas a um novo cimento de aluminato de cálcio (CAC+) em comparação ao agregado de trióxido mineral (MTA). As células foram obtidas por digestão enzimática de calvária de ratos recém-nascidos, plaqueadas sobre lamínulas de vidro contendo em sua área central discos de CAC+ ou MTA e crescidas em condições osteogênicas por até 10 dias. Durante a cultura primária, observou-se o arredondamento das bordas das amostras de cimento apenas para MTA. Embora ambos os cimentos tenham permitido a adesão, o espraiamento e a proliferação celulares, as culturas crescidas em contato com CAC+ exibiram valores maiores de número total de células em 3 e 7 dias, e de conteúdo de proteína total e atividade de fosfatase alcalina em 10 dias. Os resultados indicam que a exposição ao CAC+ permite o desenvolvimento de uma proporção maior de células em estágios mais avançados da diferenciação osteoblástica, quando comparado ao MTA. Deve-se considerar em futuros estudos experimentais a utilização do CAC+ como um material alternativo ao MTA especialmente quando um dos objetivos do tratamento endodôntico é o de reparação dos tecidos mineralizados da região periapical.

Oxidative nanopatterning of titanium surfaces promotes production and extracellular accumulation of osteopontin

The bone-biomaterial interface has been characterized by layers of afibrillar extracellular matrix (ECM) enriched in non collagenous proteins, including osteopontin (OPN), a multifunctional protein that in bone controls cell adhesion and ECM mineralization. Physical and chemical aspects of biomaterial surfaces have been demonstrated to affect cell-ECM-substrate interactions. The present paper described the ability of oxidative nanopatterning of titanium (Ti) surfaces to control extracellular OPN deposition in vitro. Ti discs were chemically treated by a mixture of H2SO4/H2O2 for either 30 min [Nano(30') Ti] or 4 h [Nano(4h) Ti]. Non-etched Ti discs were used as control. Primary osteogenic cells derived from newborn rat calvarial bone were plated on control and etched Ti and grown under osteogenic conditions up to 7 days. High resolution scanning electron microscopy revealed that treated Ti discs exhibited a nanoporous surface and that areas of larger nanopits were noticed only for Nano(4h) Ti. Large extracellular OPN accumulation were detectable only for Nano(4h) Ti, which was associated with OPN-positive cells with typical aspects of migrating cells. At day 3, quantitative results in terms of areas of OPN labeling were as follows: Nano(4h) Ti > Nano(30') Ti > Control Ti. In conclusion, chemically nanostructured Ti surfaces may support the enhancement of endogenous extracellular OPN deposition by osteogenic cells in vitro depending on the etching time, a finding that should be taken into consideration in strategies to biofunctionalize implant surfaces with molecules with cell adhesion capacity.

A interface osso-implante é caracterizada pela presença de uma camada de matriz extracellular (MEC) afibrilar rica em proteínas não-colágenas, incluindo osteopontina (OPN), cujas funções no tecido ósseo estão relacionadas à adesão celular e ao controle do processo de mineralização da MEC (crescimento de cristais). Aspectos físicos e químicos das superfícies de biomateriais podem afetar as interações célula-MEC-substrato. O objetivo do presente estudo foi demonstrar a capacidade de aspectos nanotopográficos de superfície de titânio (Ti) de controlar a deposição extracelular de OPN in vitro. Discos de Ti foram tratados quimicamente por solução de H2SO4/H2O2 durante 30 min [Nano(30') Ti] ou 4 h [Nano(4h) Ti]. Superfícies de Ti não tratadas foram usadas como controle. Células osteogênicas primárias derivadas de calvárias de ratos recém-nascidos foram plaqueadas sobre os discos de Ti e cultivadas em condições osteogênicas por até 7 dias. Microscopia eletrônica de varredura de alta resolução revelou que os discos de Ti tratados quimicamente exibiam superfície nanoporosa, com áreas de nanoporos maiores para Nano(4h) Ti. Apenas para esse grupo detectavam-se acúmulos extensos de OPN extracelular, os quais se distribuíam em áreas adjacentes a células OPN-positivas, com aspectos morfológicos típicos de células em migração. Em conclusão, a nanoestruturação química de superfície de Ti pode favorecer o aumento da deposição extracelular de OPN endógena por células osteogênicas in vitro, dependendo do tempo de condicionamento utilizado, o que deve ser considerado no desenvolvimento de estratégias para funcionalizar superfícies de implantes com moléculas com reconhecido efeito no processo de adesão celular.