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1.
J Appl Oral Sci ; 24(2): 181-5, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-27119767

RESUMEN

Objective The aim of this study was to evaluate the association of Porphyromonas endodontalis, Filifactor alocis and Dialister pneumosintes with the occurrence of periodontitis. Material and Methods Thirty subjects with chronic periodontitis (ChP) and 10 with periodontal health (PH) were included in the study. Nine subgingival biofilm samples were collected as follows: i) PH group - from the mesial/buccal aspect of each tooth in two randomly chosen contralateral quadrants; ii) ChP group - from three sites in each of the following probing depth (PD) categories: shallow (≤3 mm), moderate (4-6 mm) and deep (≥7 mm). Checkerboard DNA-DNA hybridization was used to analyze the samples. Results We found the three species evaluated in a higher percentage of sites and at higher levels in the group with ChP than in the PH group (p<0.05, Mann-Whitney test). We also observed these differences when the samples from sites with PD≤4 mm or ≥5 mm of subjects with ChP were compared with those from subjects with PH (p<0.05, Mann-Whitney test). In addition, the prevalence and levels of D. pneumosintes, and especially of F. alocis were very low in healthy subjects (0.12x105 and 0.01x105, respectively). Conclusion F. alocis and D. pneumosintes might be associated with the etiology of ChP, and their role in the onset and progression of this infection should be further investigated. The role of P. endodontalis was less evident, since this species was found in relatively high levels and prevalence in the PH group.


Asunto(s)
Periodontitis Crónica/microbiología , Peptostreptococcus/patogenicidad , Porphyromonas endodontalis/patogenicidad , Veillonellaceae/patogenicidad , Adulto , Biopelículas , Brasil , Estudios de Casos y Controles , Recuento de Colonia Microbiana , Sondas de ADN , ADN Bacteriano , Placa Dental/microbiología , Femenino , Encía/microbiología , Humanos , Masculino , Persona de Mediana Edad , Peptostreptococcus/aislamiento & purificación , Porphyromonas endodontalis/aislamiento & purificación , Estadísticas no Paramétricas , Veillonellaceae/aislamiento & purificación
2.
J. appl. oral sci ; J. appl. oral sci;24(2): 181-185, Mar.-Apr. 2016. tab, graf
Artículo en Inglés | LILACS | ID: lil-779905

RESUMEN

ABSTRACT Objective The aim of this study was to evaluate the association of Porphyromonas endodontalis, Filifactor alocis and Dialister pneumosintes with the occurrence of periodontitis. Material and Methods Thirty subjects with chronic periodontitis (ChP) and 10 with periodontal health (PH) were included in the study. Nine subgingival biofilm samples were collected as follows: i) PH group - from the mesial/buccal aspect of each tooth in two randomly chosen contralateral quadrants; ii) ChP group - from three sites in each of the following probing depth (PD) categories: shallow (≤3 mm), moderate (4-6 mm) and deep (≥7 mm). Checkerboard DNA-DNA hybridization was used to analyze the samples. Results We found the three species evaluated in a higher percentage of sites and at higher levels in the group with ChP than in the PH group (p<0.05, Mann-Whitney test). We also observed these differences when the samples from sites with PD≤4 mm or ≥5 mm of subjects with ChP were compared with those from subjects with PH (p<0.05, Mann-Whitney test). In addition, the prevalence and levels of D. pneumosintes, and especially of F. alocis were very low in healthy subjects (0.12x105 and 0.01x105, respectively). Conclusion F. alocis and D. pneumosintes might be associated with the etiology of ChP, and their role in the onset and progression of this infection should be further investigated. The role of P. endodontalis was less evident, since this species was found in relatively high levels and prevalence in the PH group.


Asunto(s)
Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Peptostreptococcus/patogenicidad , Porphyromonas endodontalis/patogenicidad , Veillonellaceae/patogenicidad , Periodontitis Crónica/microbiología , Peptostreptococcus/aislamiento & purificación , Brasil , ADN Bacteriano , Recuento de Colonia Microbiana , Sondas de ADN , Estudios de Casos y Controles , Estadísticas no Paramétricas , Biopelículas , Porphyromonas endodontalis/aislamiento & purificación , Placa Dental/microbiología , Veillonellaceae/aislamiento & purificación , Encía/microbiología
3.
J Endod ; 41(6): 817-23, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25771141

RESUMEN

INTRODUCTION: This study investigated the presence of target bacterial species and the levels of endotoxins in teeth with apical periodontitis. Levels of inflammatory mediators (interleukin [IL]-1ß and tumor necrosis factor [TNF]-α) were determined after macrophage stimulation with endodontic content after different phases of endodontic therapy using different irrigants. METHODS: Thirty primarily infected root canals were randomly assigned into 3 groups according to the irrigant used for root canal preparation (n = 10 per group): GI: 2.5% sodium hypochlorite, GII: 2% chlorhexidine gel, and GIII (control group): saline solution. Root canal samples were taken by using paper points before (s1) and after root canal instrumentation (s2), subsequently to 17% EDTA (s3), after 30 days of intracanal medication (Ca[OH]2 + saline solution) (s4), and before root canal obturation (s5). Polymerase chain reaction (16S recombinant DNA) and limulus amebocyte lysate assay were used for bacterial and endotoxin detection, respectively. Macrophages were stimulated with the root canal contents for IL-1ß/TNF-α measurement using enzyme-linked immunosorbent assay. RESULTS: Porphyromonas gingivalis (17/30), Porphyromonas endodontalis (15/30), and Prevotella nigrescens (11/30) were the most prevalent bacterial species. At s1, endotoxins were detected in 100% of the root canals (median = 32.43 EU/mL). In parallel, substantial amounts of IL-1ß and TNF-α were produced by endodontic content-stimulated macrophages. At s2, a significant reduction in endotoxin levels was observed in all groups, with GI presenting the greatest reduction (P < .05). After a root canal rinse with EDTA (s3), intracanal medication (s4), and before root canal obturation (s5), endotoxin levels reduced without differences between groups (P < .05). IL-1ß and TNF-α release decreased proportionally to the levels of residual endotoxin (P < .05). CONCLUSIONS: Regardless of the use of sodium hypochlorite or CHX, the greatest endotoxin reduction occurs after chemomechanical preparation. Increasing steps of root canal therapy associated with intracanal medication enhances endotoxin reduction, leading to a progressively lower activation of proinflammatory cells such as macrophages.


Asunto(s)
Lipopolisacáridos/análisis , Activación de Macrófagos/efectos de los fármacos , Periodontitis Periapical/microbiología , Periodontitis Periapical/terapia , Irrigantes del Conducto Radicular/uso terapéutico , Preparación del Conducto Radicular/métodos , Clorhexidina/uso terapéutico , Humanos , Interleucina-1beta/análisis , Activación de Macrófagos/inmunología , Periodontitis Periapical/inmunología , Porphyromonas endodontalis/aislamiento & purificación , Porphyromonas gingivalis/aislamiento & purificación , Prevotella nigrescens/aislamiento & purificación , Hipoclorito de Sodio/uso terapéutico , Factor de Necrosis Tumoral alfa/análisis
4.
J Clin Periodontol ; 41(11): 1027-36, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-25139407

RESUMEN

AIM: To determine microbial profiles that discriminate periodontal health from different forms of periodontal diseases. METHODS: Subgingival biofilm was obtained from patients with periodontal health (27), gingivitis (11), chronic periodontitis (35) and aggressive periodontitis (24), and analysed for the presence of >250 species/phylotypes using HOMIM. Microbial differences among groups were examined by Mann-Whitney U-test. Regression analyses were performed to determine microbial risk indicators of disease. RESULTS: Putative and potential new periodontal pathogens were more prevalent in subjects with periodontal diseases than periodontal health. Detection of Porphyromonas endodontalis/Porphyromonas spp. (OR 9.5 [1.2-73.1]) and Tannerella forsythia (OR 38.2 [3.2-450.6]), and absence of Neisseria polysaccharea (OR 0.004 [0-0.15]) and Prevotella denticola (OR 0.014 [0-0.49], p < 0.05) were risk indicators of periodontal disease. Presence of Aggregatibacter actinomycetemcomitans (OR 29.4 [3.4-176.5]), Cardiobacterium hominis (OR 14.9 [2.3-98.7]), Peptostreptococcaceae sp. (OR 35.9 [2.7-483.9]), P. alactolyticus (OR 31.3 [2.1-477.2]), and absence of Fretibacterium spp. (OR 0.024 [0.002-0.357]), Fusobacterium naviforme/Fusobacterium nucleatum ss vincentii (OR 0.015 [0.001-0.223]), Granulicatella adiacens/Granulicatella elegans (OR 0.013 [0.001-0.233], p < 0.05) were associated with aggressive periodontitis. CONCLUSION: There were specific microbial signatures of the subgingival biofilm that were able to distinguish between microbiomes of periodontal health and diseases. Such profiles may be used to establish risk of disease.


Asunto(s)
Periodontitis Agresiva/microbiología , Biopelículas , Periodontitis Crónica/microbiología , Gingivitis/microbiología , Periodoncio/microbiología , Adulto , Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacteroides/aislamiento & purificación , Cardiobacterium/clasificación , Carnobacteriaceae/aislamiento & purificación , Femenino , Fusobacterium/clasificación , Fusobacterium nucleatum/aislamiento & purificación , Humanos , Masculino , Microbiota , Neisseria/clasificación , Peptostreptococcus/clasificación , Pérdida de la Inserción Periodontal/microbiología , Índice Periodontal , Bolsa Periodontal/microbiología , Porphyromonas/clasificación , Porphyromonas/aislamiento & purificación , Porphyromonas endodontalis/aislamiento & purificación , Prevotella/clasificación , Adulto Joven
5.
J Endod ; 40(6): 778-83, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24862703

RESUMEN

INTRODUCTION: Revascularization outcome depends on microbial elimination because apical repair will not happen in the presence of infected tissues. This study evaluated the microbial composition of traumatized immature teeth and assessed their reduction during different stages of the revascularization procedures performed with 2 intracanal medicaments. METHODS: Fifteen patients (7-17 years old) with immature teeth were submitted to the revascularization procedures; they were divided into 2 groups according to the intracanal medicament used: TAP group (n = 7), medicated with a triple antibiotic paste, and CHP group (n = 8), dressed with calcium hydroxide + 2% chlorhexidine gel. Samples were taken before any treatment (S1), after irrigation with 6% NaOCl (S2), after irrigation with 2% chlorhexidine (S3), after intracanal dressing (S4), and after 17% EDTA irrigation (S5). Cultivable bacteria recovered from the 5 stages were counted and identified by means of polymerase chain reaction assay (16S rRNA). RESULTS: Both groups had colony-forming unit counts significantly reduced after S2 (P < .05); however, no significant difference was found between the irrigants (S2 and S3, P = .99). No difference in bacteria counts was found between the intracanal medicaments used (P = .95). The most prevalent bacteria detected were Actinomyces naeslundii (66.67%), followed by Porphyromonas endodontalis, Parvimonas micra, and Fusobacterium nucleatum, which were detected in 33.34% of the root canals. An average of 2.13 species per canal was found, and no statistical correlation was observed between bacterial species and clinical/radiographic features. CONCLUSIONS: The microbial profile of infected immature teeth is similar to that of primarily infected permanent teeth. The greatest bacterial reduction was promoted by the irrigation solutions. The revascularization protocols that used the tested intracanal medicaments were efficient in reducing viable bacteria in necrotic immature teeth.


Asunto(s)
Antibacterianos/uso terapéutico , Antiinfecciosos Locales/uso terapéutico , Apexificación/métodos , Hidróxido de Calcio/uso terapéutico , Clorhexidina/uso terapéutico , Cavidad Pulpar/microbiología , Irrigantes del Conducto Radicular/uso terapéutico , Traumatismos de los Dientes/microbiología , Actinomyces/efectos de los fármacos , Actinomyces/aislamiento & purificación , Adolescente , Carga Bacteriana/efectos de los fármacos , Niño , Ciprofloxacina/uso terapéutico , Cavidad Pulpar/efectos de los fármacos , Necrosis de la Pulpa Dental/microbiología , Necrosis de la Pulpa Dental/terapia , Ácido Edético/uso terapéutico , Fusobacterium nucleatum/efectos de los fármacos , Fusobacterium nucleatum/aislamiento & purificación , Geles , Humanos , Metronidazol/uso terapéutico , Viabilidad Microbiana/efectos de los fármacos , Minociclina/uso terapéutico , Peptostreptococcus/efectos de los fármacos , Peptostreptococcus/aislamiento & purificación , Porphyromonas endodontalis/efectos de los fármacos , Porphyromonas endodontalis/aislamiento & purificación , Hipoclorito de Sodio/uso terapéutico , Ápice del Diente/efectos de los fármacos , Ápice del Diente/microbiología
6.
J Endod ; 38(8): 1082-6, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22794210

RESUMEN

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.


Asunto(s)
Enfermedades de la Pulpa Dental/microbiología , Endotoxinas/análisis , Infecciones por Bacterias Gramnegativas/microbiología , Lipopolisacáridos/análisis , Enfermedades Asintomáticas , ADN Bacteriano/análisis , Cavidad Pulpar/microbiología , Enfermedades de la Pulpa Dental/terapia , Necrosis de la Pulpa Dental/microbiología , Necrosis de la Pulpa Dental/terapia , Infecciones por Fusobacterium/microbiología , Fusobacterium nucleatum/aislamiento & purificación , Infecciones por Bacterias Gramnegativas/terapia , Humanos , Prueba de Limulus , Dimensión del Dolor , Periodontitis Periapical/microbiología , Periodontitis Periapical/terapia , Reacción en Cadena de la Polimerasa , Porphyromonas endodontalis/aislamiento & purificación , Prevotella nigrescens/aislamiento & purificación , Recurrencia , Retratamiento , Supuración , Treponema/clasificación , Treponema denticola/aislamiento & purificación , Infecciones por Treponema/microbiología
7.
J Endod ; 38(2): 158-62, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-22244629

RESUMEN

INTRODUCTION: Acute endodontic infections harbor heterogeneous microbial communities in both the root canal (RC) system and apical tissues. Data comparing the microbial structure and diversity in endodontic infections in related ecosystems, such as RC with necrotic pulp and acute apical abscess (AAA), are scarce in the literature. The aim of this study was to examine the presence of selected endodontic pathogens in paired samples from necrotic RC and AAA using polymerase chain reaction (PCR) followed by the construction of cluster profiles. METHODS: Paired samples of RC and AAA exudates were collected from 20 subjects and analyzed by PCR for the presence of selected strict and facultative anaerobic strains. The frequency of species was compared between the RC and the AAA samples. A stringent neighboring clustering algorithm was applied to investigate the existence of similar high-order groups of samples. A dendrogram was constructed to show the arrangement of the sample groups produced by the hierarchical clustering. RESULTS: All samples harbored bacterial DNA. Porphyromonas endodontalis, Prevotella nigrescens, Filifactor alocis, and Tannerela forsythia were frequently detected in both RC and AAA samples. The selected anaerobic species were distributed in diverse small bacteria consortia. The samples of RC and AAA that presented at least one of the targeted microorganisms were grouped in small clusters. CONCLUSIONS: Anaerobic species were frequently detected in acute endodontic infections and heterogeneous microbial communities with low clustering behavior were observed in paired samples of RC and AAA.


Asunto(s)
Bacterias Anaerobias/clasificación , Necrosis de la Pulpa Dental/microbiología , Consorcios Microbianos , Absceso Periapical/microbiología , Algoritmos , Infecciones por Bacteroidaceae/microbiología , Bacteroides/aislamiento & purificación , Infecciones por Bacteroides/microbiología , Enterococcus faecalis/aislamiento & purificación , Fusobacterium/aislamiento & purificación , Infecciones por Fusobacterium/microbiología , Fusobacterium nucleatum/aislamiento & purificación , Infecciones por Bacterias Grampositivas/microbiología , Humanos , Reacción en Cadena de la Polimerasa , Porphyromonas endodontalis/aislamiento & purificación , Porphyromonas gingivalis/aislamiento & purificación , Prevotella/clasificación , Prevotella intermedia/aislamiento & purificación , Prevotella nigrescens/aislamiento & purificación
8.
J Periodontol ; 82(12): 1767-75, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21513472

RESUMEN

BACKGROUND: Microbial agents in root canal systems can induce periodontal inflammation. The aims of this study are to detect anaerobic microorganisms in endodontic-periodontal lesions, determine the genetic diversity among them, and assess the simultaneous colonization of the pulp and periodontal microenvironments by a single clone. METHODS: Twenty-seven teeth of patients with endodontic-periodontal lesions were selected. Samples were spread on an agar-blood medium, the detection of each species was performed using a polymerase chain reaction, and the determination of the simultaneous presence of the same species in the microenvironments by one or more clones was determined using arbitrarily primed PCR. RESULTS: Prevotella intermedia (Pi) was the most prevalent species of the colonies in periodontal pockets, whereas Porphyromonas gingivalis (Pg) and Pi were the more prevalent in root canals. Isolates of Pi and Pg were simultaneously identified in root canals and periodontal pockets. Eighteen percent of teeth exhibited the simultaneous colonization by Pg, Tannerella forsythia (previously T. forsythensis), and Porphyromonas endodontalis in the pulp and periodontal microenvironments. The presence of these species was noted even in niches from which no colonies were isolated. Seventeen different genotypes were found in periodontal and pulp sites, with the majority of sites colonized by one or two different genotypes. A high degree of genotype similarity was found for samples of Pg isolated from only one site as well as for those isolated from both microenvironments. CONCLUSION: Different clones of Pi and Pg with a high intraspecific genotype similarity were found to colonize the same anatomic sites in endodontic-periodontal infections.


Asunto(s)
Bacterias Anaerobias/genética , Cavidad Pulpar/microbiología , Periodontitis Periapical/microbiología , Bolsa Periodontal/microbiología , Adulto , Bacterias Anaerobias/clasificación , Bacteroides/genética , Bacteroides/aislamiento & purificación , Biodiversidad , Distribución de Chi-Cuadrado , Células Clonales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Tipificación Molecular , Porphyromonas endodontalis/genética , Porphyromonas endodontalis/aislamiento & purificación , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/aislamiento & purificación , Prevotella intermedia/genética , Prevotella intermedia/aislamiento & purificación
9.
J Endod ; 36(10): 1617-21, 2010 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-20850664

RESUMEN

INTRODUCTION: Bacteria located in the apical root canal system potentially participate in the pathogenesis of apical periodontitis. Detection and identification of apical bacteria can be compromised because of limitations in conventional sampling and identification procedures. This study identified several bacterial taxa in the apical and middle/coronal segments of primarily infected root canal system by using pulverized root segments and a culture-independent molecular method. METHODS: Seventeen extracted teeth with attached apical periodontitis lesions were sectioned to obtain 2 root fragments (apical and middle/coronal segments). Root fragments were cryogenically ground, and DNA was extracted from samples. After multiple displacement amplification, DNA from samples was used as template in a reverse-capture checkerboard hybridization assay targeting 28 bacterial taxa. RESULTS: Bacterial DNA was detected in all samples. The most prevalent taxa in the apical root canal system were Olsenella uli (76.5%), Prevotella baroniae (71%), Porphyromonas endodontalis (65%), Fusobacterium nucleatum (53%), and Tannerella forsythia (47%). O. uli, P. endodontalis, and Propionibacterium acnes were as frequently detected in apical samples as they were in middle/coronal samples. P. baroniae, T. forsythia, and F. nucleatum were found more frequently in the apical part of the canal as compared with matched coronal segments. Streptococcus species were more prevalent in middle/coronal samples. The median and mean of shared bacterial taxa between matched apical and middle/coronal segments were 27% and 41%, respectively. CONCLUSIONS: Several candidate endodontic pathogens were very prevalent in the apical root canal system. The apical microbiota was usually complex and differed in species composition when compared with the microbiota of middle/coronal samples from the same tooth.


Asunto(s)
Cavidad Pulpar/microbiología , Periodontitis Periapical/microbiología , Ápice del Diente/microbiología , Técnicas de Tipificación Bacteriana , Bacteroides/aislamiento & purificación , Criopreservación , ADN Bacteriano/análisis , Fusobacterium nucleatum/aislamiento & purificación , Humanos , Lactobacillus/aislamiento & purificación , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa , Porphyromonas endodontalis/aislamiento & purificación , Prevotella/aislamiento & purificación , ARN Bacteriano/análisis , ARN Ribosómico 16S/genética
10.
J Dent Res ; 88(1): 61-5, 2009 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-19131319

RESUMEN

As the breadth of bacterial diversity in the oral cavity has been deciphered by molecular studies, several newly identified species/phylotypes have emerged as potential pathogens. We hypothesized that many of these species/phylotypes could also be involved with the etiology of endodontic abscesses. Abscess aspirates from 42 persons were analyzed for the presence of 81 species/phylotypes by means of a reverse-capture checkerboard hybridization assay. Associations between the most frequently detected taxa were calculated. The most prevalent taxa were Fusobacterium nucleatum, Parvimonas micra, and Porphyromonas endodontalis. Other frequently found taxa included Olsenella uli, streptococci, Eikenella corrodens, some as-yet-uncultivated phylotypes (Bacteroidetes clone X083 and Synergistes clone BA121), and newly named species (Prevotella baroniae and Dialister invisus). Several positive bacterial associations were disclosed. Findings not only strengthen the association of many cultivable species with abscesses, but also include some newly named species and uncultivated phylotypes in the set of candidate pathogens associated with this disease.


Asunto(s)
Bacterias/clasificación , Absceso Periapical/microbiología , Actinobacteria/clasificación , Actinobacteria/aislamiento & purificación , Enfermedad Aguda , Adulto , Bacterias/aislamiento & purificación , Bacterias Anaerobias/clasificación , Bacterias Anaerobias/aislamiento & purificación , Bacteroides/clasificación , Bacteroides/aislamiento & purificación , Bacteroidetes/clasificación , Bacteroidetes/aislamiento & purificación , ADN Bacteriano/análisis , Eikenella corrodens/aislamiento & purificación , Fusobacterium nucleatum/aislamiento & purificación , Bacilos Gramnegativos Anaerobios Rectos, Curvos y Espirales/clasificación , Bacilos Gramnegativos Anaerobios Rectos, Curvos y Espirales/aislamiento & purificación , Humanos , Hibridación de Ácido Nucleico , Peptostreptococcus/clasificación , Peptostreptococcus/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Porphyromonas endodontalis/aislamiento & purificación , Prevotella/clasificación , Prevotella/aislamiento & purificación , ARN Bacteriano/análisis , ARN Ribosómico 16S/análisis , Selenomonas/aislamiento & purificación , Streptococcus/clasificación , Streptococcus/aislamiento & purificación , Treponema denticola/aislamiento & purificación
11.
Artículo en Inglés | MEDLINE | ID: mdl-17234549

RESUMEN

Black-pigmented anaerobic rods such as Prevotella spp. and Porphyromonas spp. are involved in the etiology and perpetuation of endodontic infections. The aim of this study was to evaluate the prevalence of these species in chronic endodontic infections by using culture and polymerase chain reaction (PCR) techniques. Samples of 100 patients with root canals displaying chronic endodontic infections were obtained by sterilized paper points. Bacterial identification was performed by using culture and PCR techniques. By culture, in 33% of the samples, P. intermedia-P. nigrescens (75.8%), P. gingivalis (27.3%), and P. endodontalis (9.1%) were identified, and by PCR 60% of the samples harbored P. nigrescens (43.3%), P. gingivalis (43.3%), P. intermedia (31.7%), and P. endodontalis (23.3%). The presence of these black-pigmented anaerobic rods alone or in association in chronic endodontic infections seems to be frequent. PCR is a very sensitive technique for detecting DNA from bacterial cells. Culturing is only able to reveal living bacteria and is less sensitive for the identification of low numbers of bacterial cells.


Asunto(s)
Técnicas de Tipificación Bacteriana , Infecciones por Bacteroidaceae/microbiología , Enfermedades de la Pulpa Dental/microbiología , Enfermedad Crónica , Recuento de Colonia Microbiana , ADN Bacteriano/análisis , Humanos , Reacción en Cadena de la Polimerasa , Porphyromonas endodontalis/aislamiento & purificación , Porphyromonas gingivalis/aislamiento & purificación , Prevotella intermedia/aislamiento & purificación , Prevotella nigrescens/aislamiento & purificación
12.
J Endod ; 32(12): 1135-8, 2006 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-17174667

RESUMEN

Evidence is mounting that the prevalence of some putative endodontic pathogens can significantly vary according to the geographic location in which samples were taken. This study aimed to provide additional knowledge on this subject by comparing the frequencies of 10 selected candidate endodontic pathogens in samples of acute apical abscesses obtained from two distinct geographic locations; Portland, OR, and Rio de Janeiro, Brazil. DNA was extracted from aspirates and used as template in nested PCR assays using 16S rRNA gene taxon-specific oligonucleotide primers. Of the target species/phylotypes, Treponema denticola (73% of the cases), Porphyromonas endodontalis (70%), and Tannerella forsythia (57%) were the most prevalent taxa found in Brazilian samples. Dialister invisus (70% of the cases), P. endodontalis (63%) and Dialister pneumosintes (55%) were the most frequent taxa in the Portland, OR samples. Data analysis revealed that T. denticola and T. forsythia were significantly more detected in Brazilian samples than in the Portland, OR samples. Although D. invisus, Filifactor alocis, and Synergistes oral clone W090 were detected in many more samples from the Portland, OR patients, differences were not found to be statistically significant. These findings confirmed that some bacterial taxa can markedly differ in the frequencies they occur in samples from different locations. It remains to be clarified whether this observation translates into relevant therapeutic implications.


Asunto(s)
Infecciones Bacterianas/epidemiología , Absceso Periapical/microbiología , Técnicas de Tipificación Bacteriana , Bacteroides/aislamiento & purificación , Brasil/epidemiología , ADN Bacteriano/análisis , Métodos Epidemiológicos , Bacilos Gramnegativos Anaerobios Rectos, Curvos y Espirales/aislamiento & purificación , Humanos , Oregon/epidemiología , Reacción en Cadena de la Polimerasa/métodos , Porphyromonas endodontalis/aislamiento & purificación , Prevalencia , ARN Ribosómico 16S , Treponema denticola/aislamiento & purificación
13.
Oral Microbiol Immunol ; 20(4): 211-5, 2005 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-15943764

RESUMEN

he aim of this study was to investigate the presence of four black-pigmented bacteria, Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia and Prevotella nigrescens, in endodontic infections by culture and polymerase chain reaction (PCR) analyses. Microbial samples were obtained from 50 teeth with untreated necrotic pulps (primary infection) and from 50 teeth with failing endodontic treatment (secondary infection). Microbiological strict anaerobic techniques were used for serial dilution, plating, incubation, and identification. For PCR detection, the samples were analyzed using species-specific primers of 16S rDNA and the downstream intergenic spacer region. Culture and PCR detected the test species in 13/100 and 50/100 of the study teeth, respectively. The organisms were cultured from 11/50 (22%) of primarily infected root canal samples and from 2/50 (4%) of secondary root canal samples. PCR detection identified the target species in 32/50 (64%) and 18/50 (36%) of primary and secondary infections, respectively. P. gingivalis was rarely isolated by culture methods (1%), but was the most frequently identified test species by PCR (38%). Similarly, P. endodontalis was not recovered by culture from any tooth studied, but was detected by PCR in 25% of the sampled teeth. PCR-based identification also showed higher detection rates of P. intermedia (33%) and P. nigrescens (22%) than culture (13%). In conclusion, P. gingivalis, P. endodontalis, P. intermedia, and P. nigrescens were identified more frequently in teeth with necrotic pulp than in teeth with failing endodontic treatment. Also, a higher frequency of black-pigmented species was detected by PCR than by culture.


Asunto(s)
Infecciones por Bacteroidaceae/microbiología , Necrosis de la Pulpa Dental/microbiología , Fracaso de la Restauración Dental , Porphyromonas/aislamiento & purificación , Prevotella/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Recuento de Colonia Microbiana , ADN Bacteriano/análisis , Femenino , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Porphyromonas/genética , Porphyromonas/patogenicidad , Porphyromonas endodontalis/genética , Porphyromonas endodontalis/aislamiento & purificación , Porphyromonas endodontalis/patogenicidad , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/aislamiento & purificación , Porphyromonas gingivalis/patogenicidad , Prevotella/genética , Prevotella/patogenicidad , Prevotella intermedia/genética , Prevotella intermedia/aislamiento & purificación , Prevotella intermedia/patogenicidad , Prevotella nigrescens/genética , Prevotella nigrescens/aislamiento & purificación , Prevotella nigrescens/patogenicidad
14.
Artículo en Inglés | MEDLINE | ID: mdl-15829891

RESUMEN

OBJECTIVE: This study intended to compare the prevalence of 7 putative endodontic pathogens in samples of primary endodontic infections taken from patients of 2 distant geographic locations. STUDY DESIGN: Samples from infected root canals associated with asymptomatic periradicular lesions or from pus aspirated from acute periradicular abscesses were collected from patients in Rio de Janeiro, Brazil, and Seoul, South Korea. South Korean samples were frozen and delivered to Brazil, where all steps in the molecular analysis were performed. DNA was extracted and a species-specific nested polymerase chain reaction assay was used to detect 7 target bacterial species. RESULTS: The most prevalent species detected in Brazilian samples were Porphyromonas endodontalis (79% of the cases) , Treponema denticola (79%), and Dialister pneumosintes (76%). The most prevalent species found in South Korean samples were Fusobacterium nucleatum (38% of the cases) , Tannerella forsythia (26%), and Treponema maltophilum (24%). Overall, P endodontalis, D pneumosintes, Filifactor alocis, T denticola, and T forsythia were significantly more detected in Brazilian samples than in South Korean samples ( P < .05). CONCLUSIONS: Findings indicated that the prevalence of some species in infections of endodontic origin may significantly differ from one geographic location to another.


Asunto(s)
Periodontitis Periapical/microbiología , Adolescente , Adulto , Anciano , Técnicas de Tipificación Bacteriana , Bacteroides/aislamiento & purificación , Brasil , Distribución de Chi-Cuadrado , ADN Bacteriano/análisis , Fusobacterium/aislamiento & purificación , Bacilos Gramnegativos Anaerobios Rectos, Curvos y Espirales/aislamiento & purificación , Humanos , Corea (Geográfico) , Persona de Mediana Edad , Epidemiología Molecular , Reacción en Cadena de la Polimerasa/métodos , Porphyromonas endodontalis/aislamiento & purificación , Treponema/aislamiento & purificación
15.
Oral Microbiol Immunol ; 19(2): 71-6, 2004 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-14871344

RESUMEN

OBJECTIVES: The aim of this study was to investigate the root canal microbiota of primary and secondary root-infected canals and the association of constituent species with specific endodontic signs and symptoms. METHODS: Microbial samples were taken from 60 root canals, 41 with necrotic pulp tissues (primary infection) and 19 with failed endodontic treatment (secondary infection). Strict anaerobic techniques were used for serial dilution, plating, incubation and identification. RESULTS: A total of 224 cultivable isolates were recovered belonging to 56 different bacterial species. Individual root canals yielded a maximum of 10 bacterial species. Of the bacterial isolates, 70% were either strict anaerobes or microphilic. The anaerobes most frequently isolated were: Peptostreptococcus micros (35%), Fusobacterium necrophorum (23.3%), Fusobacterium nucleatum (11.7%), Prevotella intermedia/nigrescens (16.7%), Porphyromonas gingivalis (6.7%) and Porphyromonas endodontalis (5%). The root canal microflora of untreated teeth with apical periodontitis was found to be mixed, comprising gram-negative and gram-positive and mostly anaerobic microorganisms and usually containing more than 3 species per canal. On the other hand, facultative anaerobic and gram-positive bacteria predominated in canals with failed endodontic treatment, which harbored 1-2 species per canal. Suggested relationships were found between anaerobes, especially gram-negatives, and the presence or history of pain, tenderness to percussion and swelling (P<0.05). In particular, associations were found between: a) pain (n=29) and P. micros (P<0.01), P. intermedia/nigrescens and Eubacterium spp. (both P<0.05); b) history of pain (n=31) and P. micros (P<0.01) Porphyromonas and Fusobacterium spp. (P<0.05); c) tenderness to percussion (n=29) and Porphyromonas spp. (P<0.01), Peptostreptococcus and Fusobacterium spp. (P<0.001); d) swelling (n=20) and Peptostreptococcus spp. (P<0.01), Porphyromonas and Enterococcus spp. (P<0.05); e) wet canals (n=33) and Porphyromonas and Fusobacterium spp. (P<0.05); f) purulent exudate (n=20) and Porphyromonas, Peptostreptococcus and Fusobacterium spp. (P<0.05); previous endodontic treatment and Enterococcus faecalis, Streptococcus spp., P. micros, F. necrophorum (P<0.05). CONCLUSIONS: Our findings indicate potential complex interactions of species resulting in characteristic clinical pictures which cannot be achieved by individual species alone. They also indicate that the microbiota of primary infected canals with apical periodontitis differs in number and in species from the secondary infected canals by using the culture technique.


Asunto(s)
Cavidad Pulpar/microbiología , Necrosis de la Pulpa Dental/microbiología , Periodontitis Periapical/microbiología , Bacterias Anaerobias/aislamiento & purificación , Técnicas Bacteriológicas , Edema/microbiología , Fusobacterium necrophorum/aislamiento & purificación , Fusobacterium nucleatum/aislamiento & purificación , Bacterias Gramnegativas/aislamiento & purificación , Humanos , Dolor/microbiología , Peptostreptococcus/aislamiento & purificación , Absceso Periapical/microbiología , Porphyromonas endodontalis/aislamiento & purificación , Porphyromonas gingivalis/aislamiento & purificación , Prevotella intermedia/aislamiento & purificación , Prevotella nigrescens/aislamiento & purificación , Tratamiento del Conducto Radicular , Diente no Vital/microbiología
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