RESUMO
AIM: To establish an SV40 T-Ag-transfected cell line of human pulp-derived cells in order to compare the cytotoxicity, genotoxicity and to investigate the activities of immunological biomarkers of several endodontic sealers. METHODOLOGY: Primary human pulp cells and transfected cells were cultured. Cell morphology and proliferation were analysed, and the expression of cell-specific gene transcripts and proteins was detected by RT-PCR and immunohistochemistry. Transfection of human pulp-derived cells resulted in an immortalized cell line retaining phenotypic characteristics from the primarily cells tested. The SV40 T-Ag-transfected cells were cultured and stimulated by sealers (Apexit Plus, Real Seal, AH Plus, and EndoREZ) to evaluate the cytotoxicity and genotoxicity by MTT and MTN assays, respectively. Immunological inflammatory biomarkers (IL6, IL8 and TNF-α) were determined by ELISA assay. The differences between median values were statistically analysed using Kruskal-Wallis and Dunn's tests at 5% significance level. RESULTS: The cytotoxicity assay revealed that multimethacrylate (Real Seal) was the most cytotoxic sealer (P < 0.05) and exhibited the highest inflammatory potential against the SV40 T-Ag-transfected cells (P < 0.05). All root canal sealers tested were able to stimulate the immortalized pulp cells to produce IL-6, IL-8 and TNF-α, with differences in relation to the control group (P < 0.05). Higher levels of IL-6, IL-8 and TNF-α were found in cell supernatant after stimulation with multimethacrylate (Real Seal) compared to all other sealers tested (P < 0.05). No differences were found comparing epoxy resin-based sealer (AHPlus), single-methacrylate sealer (EndoREZ) and calcium hydroxide-based sealer (Apexit Plus), regardless of the cytokine investigated (all P > 0.05). CONCLUSIONS: A SV40 T-Ag-transfected cell line of human pulp-derived cells was established. The methacrylate resin-based sealer (Real Seal) exhibited the greatest cytoxicity and inflammatory potential against immortalized pulp cells compared to an epoxy resin-based sealer (AH Plus), a methacrylate-based sealer (EndoRez) and a calcium hydroxide-based sealer (Apexit).
Assuntos
Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Materiais Restauradores do Canal Radicular/toxicidade , Biomarcadores/análise , Células Cultivadas , Citocinas/análise , Humanos , Teste de Materiais , TransfecçãoRESUMO
The aim of this study was to evaluate the amount of peroxide passage from the pulp chamber to the external enamel surface during the internal bleaching technique. Fifty bovine teeth were sectioned transversally 5 mm below the cemento-enamel junction (CEJ), and the remaining part of the root was sealed with a 2-mm layer of glass ionomer cement. The external surface of the samples was coated with nail varnish, with the exception of standardized circular areas (6-mm diameter) located on the enamel, exposed dentin, or cementum surface of the tooth. The teeth were divided into three experimental groups according to exposed areas close to the CEJ and into two control groups (n=10/group), as follows: GE, enamel exposure area; GC, cementum exposed area; GD, dentin exposed area; Negative control, no presence of internal bleaching agent and uncoated surface; and Positive control, pulp chamber filled with bleaching agent and external surface totally coated with nail varnish. The pulp chamber was filled with 35% hydrogen peroxide (Opalescence Endo, Ultradent). Each sample was placed inside of individual flasks with 1000 µL of acetate buffer solution, 2 M (pH 4.5). After seven days, the buffer solution was transferred to a glass tube, in which 100 µL of leuco-crystal violet and 50 µL of horseradish peroxidase were added, producing a blue solution. The optical density of the blue solution was determined by spectrophotometer and converted into microgram equivalents of hydrogen peroxide. Data were submitted to Kruskal-Wallis and Dunn-Bonferroni tests (α=0.05). All experimental groups presented passage of peroxide to the external surface that was statistically different from that observed in the control groups. It was verified that the passage of peroxide was higher in GD than in GE (p<0.01). The GC group presented a significantly lower peroxide passage than did GD and GE (p<0.01). It can be concluded that the hydrogen peroxide placed into the pulp chamber passed through the dental hard tissues, reaching the external surface and the periodontal tissue. The cementum surface was less permeable than were the dentin and enamel surfaces.
Assuntos
Cemento Dentário/metabolismo , Esmalte Dentário/metabolismo , Dentina/metabolismo , Peróxido de Hidrogênio/uso terapêutico , Peróxidos/farmacocinética , Clareadores Dentários/uso terapêutico , Clareamento Dental/métodos , Animais , Bovinos , Permeabilidade do Esmalte Dentário/efeitos dos fármacos , Cavidade Pulpar/metabolismo , Permeabilidade da Dentina/efeitos dos fármacos , Corantes Fluorescentes , Violeta Genciana , Umidade , Peróxido de Hidrogênio/administração & dosagem , Temperatura , Clareadores Dentários/administração & dosagemRESUMO
AIM: The aim of this paper was to evaluate the antimicrobial activity of 2% chlorhexidine gel (CLX) associated with various intracanal medicaments against Candida albicans and Enterococcus faecalis inoculated in root canals. METHODS: Thirty six human single-rooted teeth were contaminated with C.albicans and E.faecalis. The canals were instrumented using 2% CLX gel and were divided into three groups according to the intracanal medicaments (ICM) used. Group 1: calcium hydroxide paste [Ca(OH)2], Group 2: 2% chlorhexidine gel (CLX) and Group 3: 2% CLX gel + Ca(OH)2. The root canal collections were performed after 21 days of contamination (control collection), after instrumentation (1st collection), after 14 days of intracanal medicament (2nd collection) and 7 days after medicament removal (3rd collection). The microbiological samples were plated in culture media and incubated for 48 hours. The results were submitted to Kruskal-Wallis test (P ≤ 0.05). RESULTS: It was verified that the instrumentation with CLX reduced the number of CFU/ml significantly when compared with the confirmation collection (control). However, the use of the ICM was only capable to eliminate completely the microorganisms in the root canals without difference statistics between them. CONCLUSION: Although the use of 2% chlorherixidine gel reduces the number of microorganisms significantly, only the ICM calcium hydroxide and calcium hydroxide associated with chlorhexidine are able to eliminate these microorganisms completely.
Assuntos
Candida albicans/efeitos dos fármacos , Clorexidina/análogos & derivados , Cavidade Pulpar/microbiologia , Enterococcus faecalis/efeitos dos fármacos , Carga Bacteriana , Hidróxido de Cálcio/farmacologia , Candida albicans/crescimento & desenvolvimento , Candida albicans/isolamento & purificação , Clorexidina/farmacologia , Instrumentos Odontológicos , Sinergismo Farmacológico , Enterococcus faecalis/crescimento & desenvolvimento , Enterococcus faecalis/isolamento & purificação , Contaminação de Equipamentos , Géis , Humanos , Técnicas In Vitro , Testes de Sensibilidade MicrobianaRESUMO
AIM: The aim of this study was to evaluate the cytotoxicity and genotoxicity of the new castor oil bean cement (COB) material in comparison to commonly used pulp capping materials. METHODOLOGY: Specimens of COB, calcium hydroxide (Hydro C), and mineral trioxide aggregate (white and gray MTA) were extracted in culture medium (91.6 mm(2) sample surface mL(-1)). Transfected human pulp cells (tHPCs) were exposed to dilutions of the extracts for 1 h, and the generation of reactive oxygen species (ROS) was determined by flow cytometry (FACS) using H(2)DCF-DA as a dye. Survival of tHPCs was measured photometrically using a crystal violet assay after a 24-h exposure period. Genotoxicity as indicated by the formation of micronuclei in V79 cells, and the modification of the normal cell cycle by extracts of the materials was analysed by FACS. RESULTS: Clear cytotoxic effects were detected only with extracts of Hydro C under the current experimental conditions. The two MTA preparations induced an insignificant reduction in the number of cells. In contrast, the extracts of COB slightly induced cell proliferation. Extracts of Hydro C caused a twofold increase in ROS production, whilst the other tested materials were ineffective. An increase in the number of micronuclei was not detected with any material tested; Hydro C slightly increased the number of cells in G1 and G2. CONCLUSIONS: The COB and the two MTA preparations did not negatively influence cell survival or ROS production and may thus be further considered for pulp capping studies.
Assuntos
Óleo de Rícino/toxicidade , Citotoxinas/toxicidade , Cimentos Dentários/toxicidade , Capeamento da Polpa Dentária , Polpa Dentária/efeitos dos fármacos , Mutagênicos/toxicidade , Compostos de Alumínio/toxicidade , Animais , Compostos de Cálcio/toxicidade , Hidróxido de Cálcio/toxicidade , Ciclo Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cricetinae , Combinação de Medicamentos , Fibroblastos/efeitos dos fármacos , Citometria de Fluxo , Fluoresceínas , Corantes Fluorescentes , Violeta Genciana , Cimentos de Ionômeros de Vidro/toxicidade , Humanos , Micronúcleos com Defeito Cromossômico/induzido quimicamente , Óxidos/toxicidade , Fotometria , Espécies Reativas de Oxigênio/análise , Silicatos/toxicidade , Fatores de TempoRESUMO
The purpose of this study was to evaluate in vitro the efficacy of root reinforcements by light-cured composite resin or zirconium fiber post in simulated immature non-vital teeth. Fifty-six bovine incisors teeth were used for this study. The crown of each tooth was removed in the medium third to obtain a standard length of 30 mm. The specimens were divided into four groups (n = 14): G1) the root canals were instrumented and enlarged to simulate immature non-vital teeth and were reinforced with a light-cured composite resin using a translucent curing post (Luminex system); (G2) the specimens were instrumented, enlarged and they received root reinforcement with zirconium fiber post; G3 (positive control): they received similar treatment to the G1 and G2 groups, but did not receive root reinforcement; G4 (negative control): the roots were not weakened and did not receive reinforcement. Every tooth was submitted to compressive force using an Instron testing machine with an angle of 45 degrees at a speed of 1 mm min(-1) until the fracture. The results showed a markedly increased resistance to fracture in the G1 and G2 (122.38 and 122.08 kg f, respectively). Among the results of G1 and G2 there was not any significant difference (P > 0.05) but they were significantly different from the control groups (P < 0.05). The conclusion is that the use of root reinforcements with zirconium fiber post or composite resin can increase significantly the structural resistance of the weakened teeth, decreasing the risk of the fracture.
Assuntos
Técnica para Retentor Intrarradicular , Fraturas dos Dentes/prevenção & controle , Raiz Dentária/fisiopatologia , Dente não Vital/fisiopatologia , Animais , Bovinos , Resinas Compostas , Força Compressiva , Análise do Estresse Dentário , Raiz Dentária/crescimento & desenvolvimento , ZircônioRESUMO
AIM: To evaluate in vitro the effectiveness of sodium hypochlorite (NaOCl), chlorhexidine (CHX) and five intracanal medicaments on microorganisms within root canals. METHODOLOGY: Ninety-six human single-rooted extracted teeth were used. After removing the crowns, canal preparation was completed and the external root surfaces were coated with epoxy resin. Following sterilization, the teeth were contaminated with Candida albicans and Enterococcus faecalis, and were incubated at 37 +/- 1 degrees C for 7 days. The teeth were divided according to the irrigant solution or intracanal medicament: group 1, sterile physiologic solution (SPS) and calcium hydroxide (Ca(OH)2) paste; group 2, SPS and camphorated paramonochlorophenol (CPMC); group 3, SPS and tricresol formalin; group 4, SPS and CaOH2 + CPMC paste; group 5, SPS and PMC furacin; group 6, 2.5% NaOCl without intracanal medication; group 7, 2.0% CHX without intracanal medication and group 8, SPS without intracanal medication (control group). Microbiological samples were collected with sterile paper points, and bacterial growth was determined. The data were submitted to the analysis of variance (anova, P = 0.05). RESULTS: For C. albicans, groups 3 and 8 were statistically less effective than groups 1, 2, 4 and 5 (Kruskal-Wallis (K-W) = 65.241; gl = 7; P = 0.001). For E. faecalis, groups 6 and 8 were statistically less effective than groups 1-4 and 7 (K-W = 61.048; gl = 7; P = 0.001). CONCLUSIONS: Ca(OH)2 + CPMC paste was the most effective intracanal medicament for the elimination of the two microorganisms; 2.0% CHX solution was more effective than 2.5% NaOCl against E. faecalis.