RESUMO
OBJECTIVE: This study aimed to examine programmed death protein 1 (PD-1) and programmed death lig-and 1 (PD-L1) expression on leukocytes from chronic apical periodontitis, and to determine the levels of cytokines in the apical periodontitis lesions. METHODS: Leukocytes from healthy gingival tissue (n=16) and chronic apical periodontitis (n=10) were eval-uated using flow cytometry. The PD-1 and PDL-1 expressions were evaluated using flow cytometry. The cy-tokine levels were evaluated by enzyme-linked immunosorbent assay. Data were analyzed using one-way ANOVA. The statistical significance level was set at P<0.05. RESULTS: Results showed that the apical periodontitis lesions are more infiltrated by PD-1+ and PDL1+ lym-phocytes than the control samples. In addition, the PDL-1 expression was detected on macrophages in the apical periodontitis lesions, and was significantly higher compared to leukocytes from healthy gingival tis-sue. The IFN-γ, TGF-ß, IL-10, and TNF-α levels were significantly higher in the apical periodontitis lesions com-pared to control samples. CONCLUSION: The PD-1, PD-L1, and CTLA-4 molecules are evident in apical periodontitis, and can be an impor-tant immune checkpoint in chronic periapical periodontitis.
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Objectives To evaluate the sealing ability of three root-end filling materials (white MTA, CPM, and MBPc) using an Enterococcus faecalis leakage model. Material and Methods Seventy single-root extracted human teeth were instrumented and root-ends were resected to prepare 3 mm depth cavities. Root-end preparations were filled with white MTA, CPM, and MBPc cements. Enterococcus faecalis was coronally introduced and the apical portion was immersed in BHI culture medium with phenol red indicator. The bacterial leakage was monitored every 24 h for 4 weeks. The statistical analysis was performed using the Wilcoxon-Gehan test (p<0.05). Results All cements showed bacterial leakage after 24 hours, except for the negative control group. The MBPc showed significantly less bacterial leakage compared with the MTA group (p<0.05). No significant differences were found between the CPM and the other groups. Conclusions The epoxy resin-based cement MBPc had lower bacterial leakage compared with the calcium silicate-based cements MTA and CPM.
Assuntos
Compostos de Alumínio/química , Compostos de Cálcio/química , Cimentos Dentários/química , Infiltração Dentária/microbiologia , Óxidos/química , Materiais Restauradores do Canal Radicular/química , Silicatos/química , Combinação de Medicamentos , Enterococcus faecalis , Humanos , Teste de Materiais , Reprodutibilidade dos Testes , Obturação Retrógrada/métodos , Estatísticas não Paramétricas , Fatores de Tempo , Raiz Dentária/efeitos dos fármacos , Raiz Dentária/microbiologiaRESUMO
ABSTRACT Objectives To evaluate the sealing ability of three root-end filling materials (white MTA, CPM, and MBPc) using an Enterococcus faecalis leakage model. Material and Methods Seventy single-root extracted human teeth were instrumented and root-ends were resected to prepare 3 mm depth cavities. Root-end preparations were filled with white MTA, CPM, and MBPc cements. Enterococcus faecalis was coronally introduced and the apical portion was immersed in BHI culture medium with phenol red indicator. The bacterial leakage was monitored every 24 h for 4 weeks. The statistical analysis was performed using the Wilcoxon-Gehan test (p<0.05). Results All cements showed bacterial leakage after 24 hours, except for the negative control group. The MBPc showed significantly less bacterial leakage compared with the MTA group (p<0.05). No significant differences were found between the CPM and the other groups. Conclusions The epoxy resin-based cement MBPc had lower bacterial leakage compared with the calcium silicate-based cements MTA and CPM.
Assuntos
Humanos , Compostos de Alumínio/química , Compostos de Cálcio/química , Cimentos Dentários/química , Infiltração Dentária/microbiologia , Óxidos/química , Materiais Restauradores do Canal Radicular/química , Silicatos/química , Combinação de Medicamentos , Enterococcus faecalis , Teste de Materiais , Reprodutibilidade dos Testes , Obturação Retrógrada/métodos , Estatísticas não Paramétricas , Fatores de Tempo , Raiz Dentária/efeitos dos fármacos , Raiz Dentária/microbiologiaRESUMO
This study investigated the efficacy of calcium hydroxide and chlorhexidine gel for the elimination of intratubular Candida albicans (C. albicans). Human single-rooted teeth contaminated with C. albicans were treated with calcium hydroxide, 2% chlorhexidine gel, calcium hydroxide plus 2% chlorhexidine gel, or saline (0.9% sodium chloride) as a positive control. The samples obtained at depths of 0-100 and 100-200 µm from the root canal system were analyzed for C. albicans load by counting the number of colony forming units and for the percentage of viable C. albicans using fluorescence microscopy. First, the antimicrobial activity of calcium hydroxide and the 2% chlorhexidine gel was evaluated by counting the number of colony forming units. After 14 days of intracanal medication, there was a significant decrease in the number of C. albicans colony forming units at a depth of 0-100 µm with chlorhexidine treatment either with or without calcium hydroxide compared with the calcium hydroxide only treatment. However, there were no differences in the number of colony forming units at the 100-200 µm depth for any of the medications investigated. C. albicans viability was also evaluated by vital staining techniques and fluorescence microscopy analysis. Antifungal activity against C. albicans significantly increased at both depths in the chlorhexidine groups with and without calcium hydroxide compared with the groups treated with calcium hydroxide only. Treatments with only chlorhexidine or chlorhexidine in combination with calcium hydroxide were effective for elimination of C. albicans.
Assuntos
Antifúngicos/farmacologia , Hidróxido de Cálcio/farmacologia , Candida albicans/efeitos dos fármacos , Clorexidina/farmacologia , Cavidade Pulpar/microbiologia , Irrigantes do Canal Radicular/farmacologia , Antifúngicos/administração & dosagem , Hidróxido de Cálcio/administração & dosagem , Clorexidina/administração & dosagem , Contagem de Colônia Microbiana , Corantes , Combinação de Medicamentos , Humanos , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Irrigantes do Canal Radicular/administração & dosagem , Preparo de Canal Radicular/métodos , Camada de Esfregaço , Fatores de TempoRESUMO
O potencial antimicrobiano dos seguintes enxaguatórios bucais: Periogard®, Plax®, Oral-B®, Flogoral ® e Listerine® foram avaliados in vitro, frente às seguintes cepas: Pseudomonas aeroginosa, Escherichiacoli, Cândida albicans, Staphilococcus áureos, Staphilococcus epidermidi e Enterococcus faecallis, por meio do método de difusão em Agar Muller Hinton, o qual foi distribuído em alíquotas de 25 ml, em placas de Pétri estéreis. Cinco discos de papel filtro foram colocados em pontos equidistantes e foram embebidos com 20 μl dos enxaguatórios testados. As leituras dos halos foram realizadas após 48 horas. Os melhores resultados médios foram obtidos pelo Periogard® e pelo Plax®.
Assuntos
Antissépticos Bucais/normas , Técnicas In Vitro , Placa Dentária/prevenção & controleRESUMO
OBJECTIVES: The aim of this study was to analyze the periodontal parameters of patients with chronic renal failure. MATERIAL AND METHODS: The periodontal status of 16 Brazilian patients aged 29 to 53 (41.7 ± 7.2) years with chronic renal failure (CRF) and another matched group of 14 healthy controls with periodontitis was assessed clinically and microbiologically. Probing pocket depth (PPD), gingival recession (GR), dental plaque index (PLI), gingival index (GI), and dental calculus index (CI) were the clinical parameters recorded for the entire dentition (at least 19 teeth), while the anaerobic periodontopathogen colonization in four sites with the highest PPD was evaluated using the BANA test ("PerioScan"; Oral B). RESULTS: The results for the CRF group and control group, respectively were: PPD: 1.77 ± 0.32 and 2.65 ± 0.53; GR: 0.58 ± 0.56 and 0.51 ± 0.36; PLI: 1.64 ± 0.56 and 1.24 ± 0.67; GI: 0.64 ± 0.42 and 0.93 ± 0.50; CI: 1.17 ± 0.54 and 0.87 ± 0.52. Comparison between groups using the "t" test revealed a significantly increased PPD (p<0.001) in the control group. Comparison of the other clinicial parameters by the Mann-Whitney test showed differences only for PLI, which was significantly higher (p<0.05) in the CRF group. Spearman's test applied to each group showed a positive correlation among all clinical parameters, except for GR (p<0.05). None of the groups showed any correlation between GR and GI, while a significant negative correlation between GR and PPD was observed for the CRF group. The percentage of BANA-positive sites was 35.9% for the CRF group and 35.7% for the control group. The BANA test correlated positively with PPD only in the control group and with GR only in the CRF group. CONCLUSIONS: In spite of a higher PLI and dense anaerobic microbial population even in shallow PPD, patients with CRF exhibited better periodontal conditions than periodontitis patients, which is an evidence of altered response to local irritants.
Assuntos
Benzoilarginina-2-Naftilamida , Falência Renal Crônica/complicações , Índice Periodontal , Diálise Renal , Adulto , Bacteroides/isolamento & purificação , Estudos de Casos e Controles , Periodontite Crônica/classificação , Periodontite Crônica/microbiologia , Cálculos Dentários/classificação , Placa Dentária/microbiologia , Índice de Placa Dentária , Feminino , Retração Gengival/classificação , Retração Gengival/microbiologia , Humanos , Falência Renal Crônica/terapia , Masculino , Pessoa de Meia-Idade , Índice de Higiene Oral , Bolsa Periodontal/classificação , Bolsa Periodontal/microbiologia , Periodontite/classificação , Periodontite/microbiologia , Porphyromonas gingivalis/isolamento & purificação , Treponema denticola/isolamento & purificaçãoRESUMO
OBJECTIVES: The aim of this study was to analyze the periodontal parameters of patients with chronic renal failure. MATERIAL AND METHODS: The periodontal status of 16 Brazilian patients aged 29 to 53 (41.7±7.2) years with chronic renal failure (CRF) and another matched group of 14 healthy controls with periodontitis was assessed clinically and microbiologically. Probing pocket depth (PPD), gingival recession (GR), dental plaque index (PLI), gingival index (GI), and dental calculus index (CI) were the clinical parameters recorded for the entire dentition (at least 19 teeth), while the anaerobic periodontopathogen colonization in four sites with the highest PPD was evaluated using the BANA test ("PerioScan"; Oral B). RESULTS: The results for the CRF group and control group, respectively were: PPD: 1.77±0.32 and 2.65±0.53; GR: 0.58±0.56 and 0.51±0.36; PLI: 1.64±0.56 and 1.24±0.67; GI: 0.64±0.42 and 0.93±0.50; CI: 1.17±0.54 and 0.87±0.52. Comparison between groups using the "t" test revealed a significantly increased PPD (p<0.001) in the control group. Comparison of the other clincial parameters by the Mann-Whitney test showed differences only for PLI, which was significantly higher (p<0.05) in the CRF group. Spearman's test applied to each group showed a positive correlation among all clinical parameters, except for GR (p<0.05). None of the groups showed any correlation between GR and GI, while a significant negative correlation between GR and PPD was observed for the CRF group. The percentage of BANA-positive sites was 35.9 percent for the CRF group and 35.7 percent for the control group. The BANA test correlated positively with PPD only in the control group and with GR only in the CRF group. CONCLUSIONS: In spite of a higher PLI and dense anaerobic microbial population even in shallow PPD, patients with CRF exhibited better periodontal conditions than periodontitis patients, which is an evidence of altered response to local irritants.
Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Falência Renal Crônica/complicações , Índice Periodontal , Diálise Renal , Bacteroides/isolamento & purificação , Estudos de Casos e Controles , Periodontite Crônica/classificação , Periodontite Crônica/microbiologia , Índice de Placa Dentária , Cálculos Dentários/classificação , Placa Dentária/microbiologia , Retração Gengival/classificação , Retração Gengival/microbiologia , Falência Renal Crônica/terapia , Índice de Higiene Oral , Bolsa Periodontal/classificação , Bolsa Periodontal/microbiologia , Periodontite/classificação , Periodontite/microbiologia , Porphyromonas gingivalis/isolamento & purificação , Treponema denticola/isolamento & purificaçãoRESUMO
Candida albicans is recognized by phagocytic cells through a set of recognition receptors patterns. Recently, we showed the importance of TLR2 in the regulation of neutrophil survival after C. albicans infection. In the present work, we analyzed the involvement of TLR4 in the recognition of C. albicans by neutrophils and macrophages. Our results show that the absence of functional TLR4 resulted in lower chemotaxis of neutrophils to the site of infection, lower levels of TNF-α, CXCL1 and nitric oxide, and dissemination and persistence of the pathogen in lymph nodes and spleen. In vitro, the phagocytic activity, nitric oxide production and myeloperoxidase activity, CXCL1, IL-1ß production by neutrophils from TLR4-defective mice were not changed. In contrast, macrophages from TLR4-defective mice demonstrated lower phagocytosis and lower levels of CXCL1, IL-1ß and TNF-α. Together, these data demonstrate that TLR4 signals are important for the recognition of C. albicans by macrophages and their absence allows persistence of the infection.
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Candida albicans/imunologia , Candidíase/imunologia , Macrófagos/imunologia , Receptor 4 Toll-Like/imunologia , Animais , Movimento Celular , Quimiocina CXCL1/metabolismo , Quimiotaxia , Modelos Animais de Doenças , Interleucina-1beta/metabolismo , Linfonodos/microbiologia , Macrófagos/microbiologia , Masculino , Camundongos , Camundongos Endogâmicos C3H , Camundongos Knockout , Neutrófilos/imunologia , Neutrófilos/microbiologia , Óxido Nítrico/metabolismo , Baço/microbiologia , Receptor 4 Toll-Like/deficiência , Fator de Necrose Tumoral alfa/metabolismoRESUMO
OBJECTIVE: In this study, presence of dentin infection in root canals, obturated with 4 techniques submitted to the bacterial leakage test, was evaluated using histologic methods. STUDY DESIGN: The canals of palatal roots of 160 molars were instrumented and divided into different groups, according to the obturation technique used (lateral condensation, MicroSeal system, Touch 'n Heat + Ultrafil, and Tagger's hybrid technique) and extent of the remaining obturation material (5 mm and 10 mm). Ten additional roots were used as control samples. The roots were sterilized in ethylene oxide and mounted on a device for evaluation of bacterial leakage using the bacteria Enterococcus faecalis for 120 days. After the leakage test, roots were microscopically analyzed for the presence of dentin infection in the root canals and dentinal tubules. RESULTS: A total of 154 specimens were analyzed using both methodologies in the experimental groups; 50 root canals (32.4%) showed bacterial leakage at the end of the experimental period, and 118 (76.6%) showed the presence of bacteria in the root canals using the histologic criteria. The lateral condensation technique allowed lower penetration of bacteria in the root canals and dentinal tubules, followed by Touch 'n Heat + Ultrafil, MicroSeal, and Tagger's hybrid technique, which allowed significantly greater penetration of bacteria. Root canals with 10 mm of remaining obturation material presented similar bacterial penetration as root canals with 5 mm. CONCLUSIONS: Even when an adequate seal of the apical foramen was shown by the absence of turbidity in the bacterial leakage test, E. faecalis dentin infection was present in a high percentage of the root canals after 120 days of root filling exposure to the bacteria. Tagger's hybrid technique presented greater quantity of bacteria in histologic sections than root canals obturated with the other techniques.
Assuntos
Infiltração Dentária/microbiologia , Cavidade Pulpar/microbiologia , Obturação do Canal Radicular/métodos , Colagem Dentária/métodos , Desinfetantes de Equipamento Odontológico/uso terapêutico , Cavidade Pulpar/patologia , Dentina/microbiologia , Dentina/patologia , Enterococcus faecalis/crescimento & desenvolvimento , Óxido de Etileno/uso terapêutico , Infecções por Bactérias Gram-Positivas/microbiologia , Guta-Percha/uso terapêutico , Humanos , Materiais Restauradores do Canal Radicular/uso terapêutico , Irrigantes do Canal Radicular/uso terapêutico , Hipoclorito de Sódio/uso terapêutico , Esterilização/métodos , Fatores de Tempo , Ápice Dentário/microbiologia , Ápice Dentário/patologiaRESUMO
OBJECTIVE: Aggregatibacter actinomycetemcomitans is an oral Gram-negative bacterium that contributes to periodontitis progression. Isolated antigens from A. actinomycetemcomitans could be activating innate immune cells through Toll-like receptors (TLRs). In this study, we evaluated the role of TLR4 in the control of A. actinomycetemcomitans infection. MATERIAL AND METHODS: We examined the mechanisms that modulate the outcome of A. actinomycetemcomitans-induced periodontal disease in TLR4(-/-) mice. The production of cytokines was evaluated by ELISA. The bacterial load was determined by counting the number of colony-forming units per gram of tissue. RESULTS: The results showed that TLR4-deficient mice developed less severe periodontitis after A. actinomycetemcomitans infection, characterized by significantly lower bone loss and inflammatory cell migration to periodontal tissues. However, the absence of TLR4 facilitated the A. actinomycetemcomitans dissemination. Myeloperoxidase activity was diminished in the periodontal tissue of TLR4(-/-) mice. We observed a significant reduction in the production of tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-1beta in the periodontal tissue of TLR4(-/-) mice. CONCLUSION: The results of this study highlighted the role of TLR4 in controlling A. actinomycetemcomitans infection.