RESUMO
The present study aims to evaluate the longitudinal effects of induced experimental infections in gnotoxenic animals on the expression of inflammatory chemokines and their receptors in periradicular tissues. The null hypothesis tested was that Enterococcus faecalis and Fusobacterium nucleatum had no effect on CCR5, CCL5, CXCL10, CCL2/MCP-1, CXCR2 and CCR1 expression. Two groups of five animals (n = 5) aged between 8 and 12 weeks were used in this study. The animals were anaesthetized, and coronary access was performed in the first molar on the right and left sides. Microorganisms were inoculated into the left molar, and the right molar was sealed without contamination to function as a control. Animals were sacrificed 7 and 14 days after infection, and periapical tissues were collected. The cytokine mRNA expression levels were assessed using real-time PCR. The chemokine mRNA expression levels demonstrated that the experimental infection was capable of inducing increased chemokine expression on day 7 compared to that on day 14, except for CCR5 and CCL5, which showed no changes. The gnotoxenic animal model proved to be effective and allowed evaluation of the immune response against a known infection. Additionally, this study demonstrates that gene expression of chemokines and their receptors against the experimental infection preferentially prevailed during the initial phase of induction of the periradicular alteration (i.e., on day 7 post-infection).
Assuntos
Quimiocinas/análise , Cavidade Pulpar/imunologia , Doenças da Polpa Dentária/imunologia , Infecções por Fusobacterium/imunologia , Vida Livre de Germes , Infecções por Bactérias Gram-Positivas/imunologia , Receptores de Quimiocinas/análise , Animais , Quimiocinas/genética , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/microbiologia , Expressão Gênica , Camundongos , Doenças Periapicais/imunologia , Doenças Periapicais/microbiologia , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Quimiocinas/genética , Valores de Referência , Fatores de TempoRESUMO
Evidence shows the polymicrobial etiology of endodontic infections, in which bacteria and their products are the main agents for the development, progression, and dissemination of apical periodontitis. Microbial factors in necrotic root canals (e.g., endotoxin) may spread into apical tissue, evoking and supporting a chronic inflammatory load. Thus, apical periodontitis is the result of the complex interplay between microbial factors and host defense against invasion of periradicular tissues. This review of the literature aims to discuss the complex network between endodontic infectious content and host immune response in apical periodontitis. A better understanding of the relationship of microbial factors with clinical symptomatology is important to establish appropriate therapeutic procedures for a more predictable outcome of endodontic treatment.
Assuntos
Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/complicações , Doenças da Polpa Dentária/microbiologia , Periodontite Periapical/microbiologia , Infecções Bacterianas/complicações , Infecções Bacterianas/microbiologia , Citocinas/análise , Citocinas/fisiologia , Cavidade Pulpar/patologia , Doenças da Polpa Dentária/patologia , Endotoxinas/fisiologia , Humanos , Lipopolissacarídeos/fisiologia , Metaloproteinases da Matriz/análise , Metaloproteinases da Matriz/fisiologia , Periodontite Periapical/patologiaRESUMO
ABSTRACT Removal of bacterial biofilm from the root canal system is essential for the management of endodontic disease. Here we evaluated the antibacterial effect of N-acetylcysteine (NAC), a potent antioxidant and mucolytic agent, against mature multispecies endodontic biofilms consisting of Actinomyces naeslundii, Lactobacillus salivarius, Streptococcus mutans and Enterococcus faecalis on sterile human dentin blocks. The biofilms were exposed to NAC (25, 50 and 100 mg/mL), saturated calcium hydroxide or 2% chlorhexidine solution for 7 days, then examined by scanning electron microscopy. The biofilm viability was measured by viable cell counts and ATP-bioluminescence assay. NAC showed greater efficacy in biofilm cell removal and killing than the other root canal medicaments. Furthermore, 100 mg/mL NAC disrupted the mature multispecies endodontic biofilms completely. These results demonstrate the potential use of NAC in root canal treatment.
Assuntos
Humanos , Acetilcisteína/farmacologia , Streptococcus mutans/efeitos dos fármacos , Actinomyces/efeitos dos fármacos , Enterococcus faecalis/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Doenças da Polpa Dentária/microbiologia , Ligilactobacillus salivarius/efeitos dos fármacos , Antibacterianos/farmacologia , Streptococcus mutans/fisiologia , Actinomyces/fisiologia , Hidróxido de Cálcio/farmacologia , Clorexidina/farmacologia , Enterococcus faecalis/fisiologia , Cavidade Pulpar/microbiologia , Viabilidade Microbiana/efeitos dos fármacos , Ligilactobacillus salivarius/fisiologiaRESUMO
Removal of bacterial biofilm from the root canal system is essential for the management of endodontic disease. Here we evaluated the antibacterial effect of N-acetylcysteine (NAC), a potent antioxidant and mucolytic agent, against mature multispecies endodontic biofilms consisting of Actinomyces naeslundii, Lactobacillus salivarius, Streptococcus mutans and Enterococcus faecalis on sterile human dentin blocks. The biofilms were exposed to NAC (25, 50 and 100mg/mL), saturated calcium hydroxide or 2% chlorhexidine solution for 7 days, then examined by scanning electron microscopy. The biofilm viability was measured by viable cell counts and ATP-bioluminescence assay. NAC showed greater efficacy in biofilm cell removal and killing than the other root canal medicaments. Furthermore, 100mg/mL NAC disrupted the mature multispecies endodontic biofilms completely. These results demonstrate the potential use of NAC in root canal treatment.
Assuntos
Acetilcisteína/farmacologia , Actinomyces/efeitos dos fármacos , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Doenças da Polpa Dentária/microbiologia , Enterococcus faecalis/efeitos dos fármacos , Ligilactobacillus salivarius/efeitos dos fármacos , Streptococcus mutans/efeitos dos fármacos , Actinomyces/fisiologia , Hidróxido de Cálcio/farmacologia , Clorexidina/farmacologia , Cavidade Pulpar/microbiologia , Enterococcus faecalis/fisiologia , Humanos , Ligilactobacillus salivarius/fisiologia , Viabilidade Microbiana/efeitos dos fármacos , Streptococcus mutans/fisiologiaRESUMO
Abstract The present study aims to evaluate the longitudinal effects of induced experimental infections in gnotoxenic animals on the expression of inflammatory chemokines and their receptors in periradicular tissues. The null hypothesis tested was that Enterococcus faecalis and Fusobacterium nucleatum had no effect on CCR5, CCL5, CXCL10, CCL2/MCP-1, CXCR2 and CCR1 expression. Two groups of five animals (n = 5) aged between 8 and 12 weeks were used in this study. The animals were anaesthetized, and coronary access was performed in the first molar on the right and left sides. Microorganisms were inoculated into the left molar, and the right molar was sealed without contamination to function as a control. Animals were sacrificed 7 and 14 days after infection, and periapical tissues were collected. The cytokine mRNA expression levels were assessed using real-time PCR. The chemokine mRNA expression levels demonstrated that the experimental infection was capable of inducing increased chemokine expression on day 7 compared to that on day 14, except for CCR5 and CCL5, which showed no changes. The gnotoxenic animal model proved to be effective and allowed evaluation of the immune response against a known infection. Additionally, this study demonstrates that gene expression of chemokines and their receptors against the experimental infection preferentially prevailed during the initial phase of induction of the periradicular alteration (i.e., on day 7 post-infection).
Assuntos
Animais , Camundongos , Infecções por Bactérias Gram-Positivas/imunologia , Quimiocinas/análise , Receptores de Quimiocinas/análise , Cavidade Pulpar/imunologia , Doenças da Polpa Dentária/imunologia , Infecções por Fusobacterium/imunologia , Vida Livre de Germes , Doenças Periapicais/imunologia , Doenças Periapicais/microbiologia , Valores de Referência , Fatores de Tempo , Expressão Gênica , Quimiocinas/genética , Receptores de Quimiocinas/genética , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/microbiologia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Abstract: Evidence shows the polymicrobial etiology of endodontic infections, in which bacteria and their products are the main agents for the development, progression, and dissemination of apical periodontitis. Microbial factors in necrotic root canals (e.g., endotoxin) may spread into apical tissue, evoking and supporting a chronic inflammatory load. Thus, apical periodontitis is the result of the complex interplay between microbial factors and host defense against invasion of periradicular tissues. This review of the literature aims to discuss the complex network between endodontic infectious content and host immune response in apical periodontitis. A better understanding of the relationship of microbial factors with clinical symptomatology is important to establish appropriate therapeutic procedures for a more predictable outcome of endodontic treatment.
Assuntos
Humanos , Periodontite Periapical/microbiologia , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/complicações , Doenças da Polpa Dentária/microbiologia , Periodontite Periapical/patologia , Infecções Bacterianas/complicações , Infecções Bacterianas/microbiologia , Lipopolissacarídeos/fisiologia , Citocinas/análise , Citocinas/fisiologia , Metaloproteinases da Matriz/análise , Metaloproteinases da Matriz/fisiologia , Cavidade Pulpar/patologia , Doenças da Polpa Dentária/patologia , Endotoxinas/fisiologiaRESUMO
Endodontic infections are considered to be caused by the presence of various microorganisms within the root canal system. Recognition of this microbiota contributes to the successful treatment of infected root canals. This study investigated the microorganisms associated with primary and secondary endodontic infections via culture methods, biochemical tests, and molecular approaches in an Iranian population. Microbial specimens were collected from 36 patients with primary endodontic infection and 14 patients with a history of root canal therapy. Advanced microbiological culture techniques were used to isolate microbiota; subsequently, biochemical tests and 16S ribosomal RNA gene sequencing were performed to identify the microorganisms. Within the total 218 cultivable isolates, Veillonella parvula (20.6%) was found to occur with the highest frequency in primary endodontic infection, followed by Porphyromonas gingivalis (14.1%), and Aggregatibacter actinomycetemcomitans (9.2%). Enterococcus faecalis (36.6%) was the most predominant microorganism in secondary endodontic infections, followed by Candida albicans, Propionibacterium acnes, and V. parvula with frequencies of 20%, 2%, and 2%, respectively. It was concluded that V. parvula and E. faecalis was most frequently found in primary and secondary endodontic infections, respectively.
Assuntos
Bactérias Anaeróbias/isolamento & purificação , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/microbiologia , Adulto , Bactérias Anaeróbias/genética , Infecções Bacterianas/epidemiologia , Infecções Bacterianas/microbiologia , Contagem de Colônia Microbiana , Doenças da Polpa Dentária/epidemiologia , Feminino , Humanos , Irã (Geográfico)/epidemiologia , Masculino , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase , Prevalência , Adulto JovemRESUMO
Objetivo: identificar el complejo rojo periodontal, formado por Porphyromonas gingivalis, Treponema denticola y Tannerella forsythia, en la infección endodóntica primaria de necrosis pulpar, con cámara abierta y cerrada, utilizando técnicas de reacción en cadena de la polimerasa. Materiales y métodos: se realizó la toma para reacción en cadena de la polimerasa en 27 dientes con necrosis pulpar, 13 con cámara pulpar abierta y 14 con cámara cerrada. Resultados: en las muestras de necrosis abierta se identificaron P. gingivalis en un 92 por ciento, T. denticola en un 76 por ciento, T. forsythia en un 76 por ciento y el complejo rojo en un 61 por ciento. Las tomas de necrosis cerrada mostraron P. gingivalis en un 78 por ciento y T. denticola en un 57 por ciento; no se identificaron T. forsythia ni el complejo rojo. El análisis estadístico evidenció diferencias significativas entre los dos grupos (P<0,05). Conclusión: el sinergismo de las tres bacterias que forman el complejo rojo agravaría la patogénesis de la infección endodóntica y permitiría relacionar la microbiología endodóntica con la microbiología de periodontitis crónica.
Assuntos
Humanos , Necrose da Polpa Dentária/microbiologia , Exposição da Polpa Dentária/microbiologia , Periodontite/microbiologia , Doenças da Polpa Dentária/microbiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Reação em Cadeia da Polimerase/métodos , Treponema denticola/isolamento & purificação , Porphyromonas gingivalis/isolamento & purificação , Tannerella forsythia/isolamento & purificação , Interpretação Estatística de DadosRESUMO
OBJECTIVE: The aim of this study was to investigate the prevalence of isolated Candida albicans from periodontal endodontic lesions in diabetic and normoglycemic patients, and the fungi's virulence in different atmospheric conditions. MATERIAL AND METHODS: A case-control study was conducted on 15 patients with type 2 diabetes mellitus (G1) and 15 non-diabetics (G2) with periodontal endodontic lesions. Samples of root canals and periodontal pockets were plated on CHROMagar for later identification by polymerase chain reaction (PCR) and virulence test. RESULTS: C. albicans was identified in 79.2% and 20.8% of the 60 samples collected from diabetic and normoglycemic patients, respectively. Of the 30 samples collected from periodontal pockets, 13 showed a positive culture for C. albicans, with 77% belonging to G1 and 23% to G2. Of the 11 positive samples from root canals, 82% were from G1 and 18% from G2. Production of proteinase presented a precipitation zone Pz<0.63 of 100% in G1 and 72% in G2, in redox and negative (Pz=1), under anaerobic conditions in both groups. Hydrophobicity of the strains from G1 indicated 16.4% with low, 19.3% with moderate, and 64.3% with high hydrophobicity in redox. In G2, 42.2% had low, 39.8% had moderate, 18% had high hydrophobicity in redox. In anaerobic conditions, G1 showed 15.2% with low, 12.8% with moderate, and 72% with high hydrophobicity; in G2, 33.6% had low, 28.8% had moderate, and 37.6% had high hydrophobicity. There was statistical difference in the number of positive cultures between G1 and G2 (p<0.05) with predominance in G1. There was statistical difference for all virulence factors, except hemolysis (p=0.001). CONCLUSIONS: Candida albicans was isolated more frequently and had higher virulence in diabetic patients.
Assuntos
Candida albicans/isolamento & purificação , Candida albicans/patogenicidade , Doenças da Polpa Dentária/microbiologia , Diabetes Mellitus Tipo 2/microbiologia , Doenças Periodontais/microbiologia , Adulto , Idoso , Estudos de Casos e Controles , DNA Fúngico , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/diagnóstico por imagem , Doenças da Polpa Dentária/fisiopatologia , Diabetes Mellitus Tipo 2/fisiopatologia , Eletroforese , Feminino , Humanos , Interações Hidrofóbicas e Hidrofílicas , Masculino , Pessoa de Meia-Idade , Oxirredução , Peptídeo Hidrolases/análise , Doenças Periodontais/diagnóstico por imagem , Doenças Periodontais/fisiopatologia , Bolsa Periodontal/microbiologia , Fosfolipases/análise , Reação em Cadeia da Polimerase , Radiografia Dentária , Estatísticas não Paramétricas , VirulênciaRESUMO
Abstract Pulpal and periodontal tissues have similar microbiota that allows cross-contamination between the pulp and periodontal tissues. Objective The aim of this study was to investigate the prevalence of isolated Candida albicans from periodontal endodontic lesions in diabetic and normoglycemic patients, and the fungi's virulence in different atmospheric conditions. Material and Methods A case-control study was conducted on 15 patients with type 2 diabetes mellitus (G1) and 15 non-diabetics (G2) with periodontal endodontic lesions. Samples of root canals and periodontal pockets were plated on CHROMagar for later identification by polymerase chain reaction (PCR) and virulence test. Results C. albicans was identified in 79.2% and 20.8% of the 60 samples collected from diabetic and normoglycemic patients, respectively. Of the 30 samples collected from periodontal pockets, 13 showed a positive culture for C. albicans, with 77% belonging to G1 and 23% to G2. Of the 11 positive samples from root canals, 82% were from G1 and 18% from G2. Production of proteinase presented a precipitation zone Pz<0.63 of 100% in G1 and 72% in G2, in redox and negative (Pz=1), under anaerobic conditions in both groups. Hydrophobicity of the strains from G1 indicated 16.4% with low, 19.3% with moderate, and 64.3% with high hydrophobicity in redox. In G2, 42.2% had low, 39.8% had moderate, 18% had high hydrophobicity in redox. In anaerobic conditions, G1 showed 15.2% with low, 12.8% with moderate, and 72% with high hydrophobicity; in G2, 33.6% had low, 28.8% had moderate, and 37.6% had high hydrophobicity. There was statistical difference in the number of positive cultures between G1 and G2 (p<0.05) with predominance in G1. There was statistical difference for all virulence factors, except hemolysis (p=0.001). Conclusions Candida albicans was isolated more frequently and had higher virulence in diabetic patients.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Doenças Periodontais/microbiologia , Candida albicans/isolamento & purificação , Candida albicans/patogenicidade , Doenças da Polpa Dentária/microbiologia , Diabetes Mellitus Tipo 2/microbiologia , Oxirredução , Peptídeo Hidrolases/análise , Doenças Periodontais/fisiopatologia , Doenças Periodontais/diagnóstico por imagem , Bolsa Periodontal/microbiologia , Fosfolipases/análise , Virulência , DNA Fúngico , Radiografia Dentária , Estudos de Casos e Controles , Reação em Cadeia da Polimerase , Estatísticas não Paramétricas , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/fisiopatologia , Doenças da Polpa Dentária/diagnóstico por imagem , Diabetes Mellitus Tipo 2/fisiopatologia , Eletroforese , Interações Hidrofóbicas e HidrofílicasRESUMO
Abstract: Endodontic infections are considered to be caused by the presence of various microorganisms within the root canal system. Recognition of this microbiota contributes to the successful treatment of infected root canals. This study investigated the microorganisms associated with primary and secondary endodontic infections via culture methods, biochemical tests, and molecular approaches in an Iranian population. Microbial specimens were collected from 36 patients with primary endodontic infection and 14 patients with a history of root canal therapy. Advanced microbiological culture techniques were used to isolate microbiota; subsequently, biochemical tests and 16S ribosomal RNA gene sequencing were performed to identify the microorganisms. Within the total 218 cultivable isolates, Veillonella parvula (20.6%) was found to occur with the highest frequency in primary endodontic infection, followed by Porphyromonas gingivalis (14.1%), and Aggregatibacter actinomycetemcomitans (9.2%). Enterococcus faecalis (36.6%) was the most predominant microorganism in secondary endodontic infections, followed by Candida albicans, Propionibacterium acnes, and V. parvula with frequencies of 20%, 2%, and 2%, respectively. It was concluded that V. parvula and E. faecalis was most frequently found in primary and secondary endodontic infections, respectively.
Assuntos
Humanos , Masculino , Feminino , Adulto , Adulto Jovem , Bactérias Anaeróbias/isolamento & purificação , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/microbiologia , Bactérias Anaeróbias/genética , Infecções Bacterianas/microbiologia , Infecções Bacterianas/epidemiologia , Contagem de Colônia Microbiana , Reação em Cadeia da Polimerase , Prevalência , Técnicas de Amplificação de Ácido Nucleico , Doenças da Polpa Dentária/epidemiologia , Irã (Geográfico)/epidemiologia , Pessoa de Meia-IdadeRESUMO
The aim of this study was to explore the bacterial diversity of 10 root canals with acute apical abscess using clonal analysis. Samples were collected from 10 patients and submitted to bacterial DNA isolation, 16S rRNA gene amplification, cloning, and sequencing. A bacterial genomic library was constructed and bacterial diversity was estimated. The mean number of taxa per canal was 15, ranging from 11 to 21. A total of 689 clones were analyzed and 76 phylotypes identified, of which 47 (61.84%) were different species and 29 (38.15%) were taxa reported as yet-uncultivable or as yet-uncharacterized species. Prevotella spp., Fusobacterium nucleatum, Filifactor alocis, and Peptostreptococcus stomatis were the most frequently detected species, followed by Dialister invisus, Phocaeicola abscessus, the uncharacterized Lachnospiraceae oral clone, Porphyromonas spp., and Parvimonas micra. Eight phyla were detected and the most frequently identified taxa belonged to the phylum Firmicutes (43.5%), followed by Bacteroidetes (22.5%) and Proteobacteria (13.2%). No species was detected in all studied samples and some species were identified in only one case. It was concluded that acute primary endodontic infection is characterized by wide bacterial diversity and a high intersubject variability was observed. Anaerobic Gram-negative bacteria belonging to the phylum Firmicutes, followed by Bacteroidetes, were the most frequently detected microorganisms.
Assuntos
Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/microbiologia , Bactérias Anaeróbias Gram-Negativas/isolamento & purificação , Abscesso Periapical/microbiologia , Clonagem Molecular , DNA Bacteriano/isolamento & purificação , Genoma Bacteriano , Biblioteca Genômica , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Microbiota , Reação em Cadeia da Polimerase , RNA Ribossômico 16S , Análise de Sequência de RNARESUMO
Desde 1928, los antibióticos eran las primeras armas contra los microoganismos. En 1998 apareció una bacteria resistente que permitió el incremento de las enfermedades infecciosas. Cuanto más expuesta esté una población bacteriana a un antibiótico, las bacterias que sobrevivan se harán más resistentes, aún para defenderse de futuros antibióticos. Pero su empleo está justificado como método de profilaxis en pacientes médicamente comprometidos. En principio, debe indicarse medicación local para reducir la carga microbiana. El proifesional debe saber cuándo recetar un antibiótico y no olvidar que éste es un coadyuvante del tratamiento, ya que es el propio sistema inmune del paciente el que realiza la curación.
Assuntos
Humanos , Resistência Microbiana a Medicamentos/fisiologia , Infecção Focal Dentária/tratamento farmacológico , Infecções Bacterianas/tratamento farmacológico , Antibioticoprofilaxia , Doenças da Polpa Dentária/microbiologia , Doenças Periodontais/microbiologia , Staphylococcus aureusRESUMO
Abstract The aim of this study was to explore the bacterial diversity of 10 root canals with acute apical abscess using clonal analysis. Samples were collected from 10 patients and submitted to bacterial DNA isolation, 16S rRNA gene amplification, cloning, and sequencing. A bacterial genomic library was constructed and bacterial diversity was estimated. The mean number of taxa per canal was 15, ranging from 11 to 21. A total of 689 clones were analyzed and 76 phylotypes identified, of which 47 (61.84%) were different species and 29 (38.15%) were taxa reported as yet-uncultivable or as yet-uncharacterized species. Prevotella spp., Fusobacterium nucleatum, Filifactor alocis, and Peptostreptococcus stomatis were the most frequently detected species, followed by Dialister invisus, Phocaeicola abscessus, the uncharacterized Lachnospiraceae oral clone, Porphyromonas spp., and Parvimonas micra. Eight phyla were detected and the most frequently identified taxa belonged to the phylum Firmicutes (43.5%), followed by Bacteroidetes (22.5%) and Proteobacteria (13.2%). No species was detected in all studied samples and some species were identified in only one case. It was concluded that acute primary endodontic infection is characterized by wide bacterial diversity and a high intersubject variability was observed. Anaerobic Gram-negative bacteria belonging to the phylum Firmicutes, followed by Bacteroidetes, were the most frequently detected microorganisms.
Assuntos
Humanos , Abscesso Periapical/microbiologia , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/microbiologia , Bactérias Anaeróbias Gram-Negativas/isolamento & purificação , DNA Bacteriano/isolamento & purificação , RNA Ribossômico 16S , Biblioteca Genômica , Reação em Cadeia da Polimerase , Infecções por Bactérias Gram-Negativas/microbiologia , Genoma Bacteriano , Clonagem Molecular , Análise de Sequência de RNA , MicrobiotaRESUMO
INTRODUCTION: This clinical study was conducted to correlate the levels of endotoxins and bacterial counts found in primary endodontic infection with the volume of periapical bone destruction determined by cone-beam computed tomography (CBCT) analysis. Moreover, the levels of bacteria and endotoxins were correlated with the development of clinical features. METHODS: Twenty-four root canals with primary endodontic disease and apical periodontitis were selected. Clinical features such as pain on palpation, pain on percussion, and previous episode of pain were recorded. The volume (cubic millimeters) of periapical bone destruction was determined by CBCT analysis. Endotoxins and bacterial samplings were collected by using sterile/apyrogenic paper points. Endotoxins were quantified by using limulus amebocyte lysate assay (KQCL test), and bacterial count (colony-forming units [CFU]/mL) was determined by using anaerobic culture techniques. Data were analyzed by Pearson correlation and multiple logistic regression (P < .05). RESULTS: Endotoxins and bacteria were detected in 100% of the root canal samples (24 of 24), with median values of 10.92 endotoxin units (EU)/mL (1.75-128 EU/mL) and 7.5 × 10(5) CFU/mL (3.20 × 10(5)-8.16 × 10(6) CFU/mL), respectively. The median volume of bone destruction determined by CBCT analysis was 100 mm(3) (10-450 mm(3)). The multiple regression analysis revealed a positive correlation between higher levels of endotoxins present in root canal infection and larger volume of bone destruction (P < .05). Moreover, higher levels of endotoxins were also correlated with the presence of previous pain (P < .05). CONCLUSIONS: Our findings revealed that the levels of endotoxins found in root canal infection are related to the volume of periapical bone destruction determined by CBCT analysis. Moreover, the levels of endotoxin are related to the presence of previous pain.
Assuntos
Infecções Bacterianas/diagnóstico , Tomografia Computadorizada de Feixe Cônico , Doenças da Polpa Dentária/microbiologia , Endotoxinas/análise , Periodontite Periapical/diagnóstico por imagem , Periodontite Periapical/microbiologia , Tratamento do Canal Radicular/efeitos adversos , Adulto , Perda do Osso Alveolar/diagnóstico por imagem , Carga Bacteriana , Humanos , Pessoa de Meia-Idade , Periodontite Periapical/terapia , Adulto JovemRESUMO
Background Enterococcus faecalis is considered to be one of most prevalent species in the oral cavity, particularly in endodontic infections. The aim of the present study was to investigate the prevalence of E. faecalis in dental root canals, clonal diversity by restriction fragment length polymorphism (RFLP) and randomly amplified polymorphic DNA (RAPD-PCR) analysis, and the antibiotic susceptibility of E. faecalis isolates. Results Among the bacterial strains isolated from dental root canal specimens (n = 82), E. faecalis was determined to have the highest prevalence followed by Streptococcus viridians, Leuconostoc mesenteroides, Staphylococcus aureus, Streptococcus mitis, and Pediococcus pentosaceus. Cluster analysis of RAPD-PCR and RFLP patterns of the E. faecalis isolates discriminated five and six different genotypes, respectively. Among the tested strains, 43%, 52% and 5% were susceptible, intermediate resistant, and resistant to erythromycin, respectively. In addition, one strain (E-12) was intermediate resistant to linezolid, and one isolate (E-16) was resistant to tetracycline. Interestingly, many of the intermediate resistant/resistant strains were grouped in clusters 5 and 6, according RAPD and to RFLP, respectively. Conclusions E. faecalis demonstrated the highest prevalence in the tested dental root canal specimens collected from Saudi patients and were grouped into five to six different genotypes. Different levels of antimicrobial susceptibility were observed in the tested E. faecalis strains, which clearly indicated that although bacterial strains may be similar, point mutations can result in extreme susceptibility or resistance to various antibiotics. This phenomenon is a cause for concern for clinicians in the treatment of dental infections caused by E. faecalis.
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Adulto Jovem , Infecções Bacterianas/microbiologia , Enterococcus faecalis/isolamento & purificação , Enterococcus faecalis/genética , Farmacorresistência Bacteriana , Doenças da Polpa Dentária/microbiologia , Variação Genética , Polimorfismo de Fragmento de Restrição , Testes de Sensibilidade Microbiana , Técnica de Amplificação ao Acaso de DNA Polimórfico , GenótipoRESUMO
INTRODUCTION: This systematic review and meta-regression analysis aimed to calculate a combined prevalence estimate and evaluate the prevalence of different Treponema species in primary and secondary endodontic infections, including symptomatic and asymptomatic cases. METHODS: The MEDLINE/PubMed, Embase, Scielo, Web of Knowledge, and Scopus databases were searched without starting date restriction up to and including March 2014. Only reports in English were included. The selected literature was reviewed by 2 authors and classified as suitable or not to be included in this review. Lists were compared, and, in case of disagreements, decisions were made after a discussion based on inclusion and exclusion criteria. A pooled prevalence of Treponema species in endodontic infections was estimated. Additionally, a meta-regression analysis was performed. RESULTS: Among the 265 articles identified in the initial search, only 51 were included in the final analysis. The studies were classified into 2 different groups according to the type of endodontic infection and whether it was an exclusively primary/secondary study (n = 36) or a primary/secondary comparison (n = 15). The pooled prevalence of Treponema species was 41.5% (95% confidence interval, 35.9-47.0). In the multivariate model of meta-regression analysis, primary endodontic infections (P < .001), acute apical abscess, symptomatic apical periodontitis (P < .001), and concomitant presence of 2 or more species (P = .028) explained the heterogeneity regarding the prevalence rates of Treponema species. CONCLUSIONS: Our findings suggest that Treponema species are important pathogens involved in endodontic infections, particularly in cases of primary and acute infections.
Assuntos
Doenças da Polpa Dentária/epidemiologia , Doenças da Polpa Dentária/microbiologia , Infecções por Treponema/epidemiologia , Humanos , Abscesso Periapical/epidemiologia , Abscesso Periapical/microbiologia , Periodontite Periapical/epidemiologia , Periodontite Periapical/microbiologia , PrevalênciaRESUMO
O objetivo principal deste estudo foi investigar a interação de 24 cepas de E. faecalis isoladas de infecções endodônticas primárias às proteínas de matriz dentinária, como também a moléculas de matriz presentes em lesões de endocardites. A análise desta interação foi feita através de técnica enzimática, com confirmação pela técnica de fluorescência. Além disto, foi realizada a confirmação do isolamento da espécie E. faecalis, através da técnica de PCR para o gene 16SrRNA e a análise da presença de genes de virulência da referida espécie microbiana para aderência às supostas proteínas de matriz incluindo às de ligação ao colágeno (ace, gelE, esp, agg e efaA). O maior padrão de interação das cepas ocorreu com a fibronectina (83,4%), seguido pelo fibrinogênio (62,5%) e colágeno humano tipo I (52%). Curiosamente, a aderência observada para o colágeno do tipo I, foi de pequena magnitude, quando comparado com a amostra padrão da ATCC 29212. As cepas ATCC 29212, A1, A43 e A68 interagiram com todas as proteínas de matriz utilizadas neste estudo. Um percentual expressivo das cepas testadas apresentou amplificação para efaA (86,9%) e para ace (73,9%). Paralelamente, todas as cepas apresentaram amplificação para gelE e foram negativas para os genes agg e esp. Adicionalmente, não houve correlação entre a detecção dos genes de virulência e a interação às proteínas de matriz, evidenciando que, mesmo com a detecção dos genes nas amostras, se faz necessário avaliar a expressão gênica por qPCR...
The main objective of this study was to investigate the interaction of 24 E. faecalis strains isolated from primary endodontic infections with dentin matrix proteins, as well as to the matrix molecules present in endocarditis lesions. The analysis of this interaction was made by enzyme assay, with confirmation by the fluorescence technique. In addition, confirmation of the isolation of the species E. faecalis was performed by PCR for 16S rRNA gene and the analysis of the presence of virulence genes for matrix proteins including type I collagen (ace, gelE, esp , agg and efA). The strains interacted mostly with fibronectin (83.4%), followed by fibrinogen (62.5%) and human collagen type I (52%). Interestingly, the adhesion to type I collagen occurred in small magnitude, when compared with the E. faecalis type strain ATCC 29212. The strains ATCC 29212, A1, A43 and A68 interacted with all matrix proteins investigated in this study. A significant percentage of the tested strains showed amplification for efaA (86.9%) and ace (73.9%). In parallel, all strains showed amplification for gelE and were negative for the genes esp and agg. Additionally, there was no correlation between the detection of virulence genes and the interaction with matrix proteins, showing that even with the detection of genes in a particular strain, it is necessary to evaluate the gene expression by qPCR...
Assuntos
Humanos , Doenças da Polpa Dentária/microbiologia , Endodontia , Enterococcus faecalis , Fatores de Virulência , Brasil , Ensaio de Imunoadsorção EnzimáticaRESUMO
OBJECTIVES: To determine the phenotypic and molecular characteristics of Enterococcus faecalis recovered from primary endodontic infections in Brazilian patients. METHODS: Twenty isolates of E. faecalis recovered from 43 Brazilian patients with primary endodontic infections were identified by biochemical profiling (API20Strep) and 16S rDNA sequencing. Antimicrobial susceptibility was ascertained by agar dilution, using the recommended protocol of the Clinical and Laboratory Standards Institute (CLSI). PCR with validated primers was used to detect genes associated with antibiotic resistance and specific virulence factors. RESULTS: All isolates were deemed susceptible to penicillin G, erythromycin and vancomycin. However, nine isolates had a minimum inhibitory concentration of 4µg/mL to vancomycin (the resistance breakpoint). Fourteen isolates (70% of isolates) were also resistant to tetracycline with MICs of >64µg/mL. PCR products for tetracycline resistance genes were detected in test isolates, while erythromycin and vancomycin resistance genes were not evident. Gelatinase, aggregation substance and enteroccocal surface protein genes were detected in 20, 18 and 12 isolates, respectively. CONCLUSIONS: Endodontic E. faecalis isolates exhibit high level of resistance to tetracycline, an antibiotic that has use in local treatment of dental infections. This opens up a much-needed debate on the role and efficacy of this antibiotic for oral infections. Furthermore, these isolates were shown to possess genes that could contribute to pathogenicity in the pulp cavity.
Assuntos
Doenças da Polpa Dentária/microbiologia , Farmacorresistência Bacteriana , Enterococcus faecalis/fisiologia , Infecções por Bactérias Gram-Positivas/microbiologia , Antibacterianos/farmacologia , Proteínas de Bactérias/análise , Técnicas Bacteriológicas , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/genética , Enterococcus faecalis/patogenicidade , Eritromicina/farmacologia , Gelatinases/análise , Perfilação da Expressão Gênica , Proteínas de Membrana/análise , Testes de Sensibilidade Microbiana , Penicilina G/farmacologia , Fenótipo , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Resistência a Tetraciclina , Vancomicina/farmacologia , Virulência , Fatores de Virulência/análiseRESUMO
INTRODUCTION: This clinical study was conducted to compare the levels of endotoxins (lipopolysaccharides [LPSs]) found in primary and secondary endodontic infections with apical periodontitis by correlating LPS contents with clinical/radiographic findings. In addition, the presence of target gram-negative anaerobic bacteria was also investigated. METHODS: Samples were taken from 15 root canals with primary infections and 15 with secondary infections by using paper points. The limulus amebocyte lysate assay was used to quantify endotoxins, and the polymerase chain reaction technique (16S rDNA) was used for bacterial investigation. RESULTS: Endotoxins were detected in 100% of the root canal samples collected from primary (15/15) and secondary (15/15) infections with median values of 7.49 EU/mL and 3.96 EU/mL, respectively (P < .05). The median value of endotoxins found in the presence of clinical symptoms was significantly higher than in asymptomatic teeth with primary infections (P < .05). A positive correlation was found between endotoxin contents and a larger size of the radiolucent area (>3 mm) (P < .05). Prevotella nigrescens (10/15, 4/15), Fusobacterium nucleatum (5/15, 1/15), Treponema denticola (3/15, 1/15), and Treponema socranskii (5/15, 1/15) were detected in teeth with primary and secondary infections, respectively. P. endodontalis was present only in teeth with primary infections (5/15). CONCLUSIONS: Teeth with primary endodontic infections had higher contents of endotoxins and a more complex gram-negative bacterial community than teeth with secondary infections. Moreover, the levels of endotoxins were related to the severity of bone destruction in periapical tissues as well as the development of clinical features in teeth with primary infections.