RESUMO
Oral infections such as periodontitis and tooth decay are the most common diseases of humankind. Oleoresins from different copaifera species display antimicrobial and anti-inflammatory activities. Copaifera reticulata is the commonest tree of this genus and grows abundantly in several Brazilian states, such as Pará, Amazonas, and Ceará. The present study has evaluated the chemical composition and antimicrobial potential of the Copaifera reticulata oleoresin (CRO) against the causative agents of tooth decay and periodontitis and has assessed the CRO cytotoxic potential. Cutting edge analytical techniques (GC-MS and LC-MS) aided the chemical characterization of CRO. Antimicrobial assays included determination of the Minimum Inhibitory Concentration (MIC), determination of the Minimum Bactericidal Concentration (MBC), determination of the Minimum Inhibitory Concentration of Biofilm (MICB50), Time Kill Assay, and Checkerboard Dilution. Conduction of XTT assays on human lung fibroblasts (GM07492-A cells) helped to examine the CRO cytotoxic potential. Chromatographic analyses revealed that the major constituents of CRO were ß-bisabolene, trans-α-bergamotene, ß-selinene, α-selinene, and the terpene acids ent-agathic-15-methyl ester, ent-copalic acid, and ent-polyalthic acid. MIC and MBC results ranged from 6.25 to 200 µg/mL against the tested bacteria. The time-kill assay conducted with CRO at concentrations between 50 and 100 µg/mL showed bactericidal activity against Fusobacterium nucleatum (ATCC 25586) and Streptococcus mitis (ATCC 49456) after 4 h, Prevotella nigrescens (ATCC 33563) after 6 h, Porphyromonas gingivalis (ATCC 33277) and Lactobacillus casei (clinical isolate) after 12 h, and Streptococcus salivarius (ATCC 25975) and Streptococcus mutans (ATCC 25175) after 18 h. The fractional inhibitory concentration indexes (FICIs) revealed antagonistic interaction for Lactobacillus casei (clinical isolate), indifferent effect for Porphyromonas gingivalis (ATCC 33277), Fusobacterium nucleatum (ATCC 25586), Prevotella nigrescens (ATCC 33563), and Streptococcus salivarius (ATCC 25975), and additive effect for Streptococcus mutans (ATCC 25175) and Streptococcus mitis (ATCC 49456). Treatment of GM07492-A cells with CRO demonstrated that concentrations up to 39 µg/mL significantly reduced cell viability as compared to the negative control, being IC50 equal to 51.85 ± 5.4 µg/mL. These results indicated that CRO plays an important part in the search for novel sources of agents that can act against oral pathogens.
Assuntos
Antibacterianos/farmacologia , Fabaceae/química , Extratos Vegetais/farmacologia , Porphyromonas gingivalis/efeitos dos fármacos , Prevotella nigrescens/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/isolamento & purificação , Compostos Bicíclicos com Pontes/isolamento & purificação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cárie Dentária/microbiologia , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Humanos , Lacticaseibacillus casei/efeitos dos fármacos , Lacticaseibacillus casei/crescimento & desenvolvimento , Lacticaseibacillus casei/isolamento & purificação , Testes de Sensibilidade Microbiana , Sesquiterpenos Monocíclicos , Periodontite/microbiologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Porphyromonas gingivalis/crescimento & desenvolvimento , Porphyromonas gingivalis/isolamento & purificação , Prevotella nigrescens/crescimento & desenvolvimento , Prevotella nigrescens/isolamento & purificação , Sesquiterpenos/isolamento & purificação , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/isolamento & purificação , Streptococcus salivarius/efeitos dos fármacos , Streptococcus salivarius/crescimento & desenvolvimento , Streptococcus salivarius/isolamento & purificação , Terpenos/isolamento & purificaçãoRESUMO
Rheumatoid arthritis (RA) and periodontitis (PD) are chronic inflammatory disorders that cause bone loss. PD tends to be more prevalent and severe in RA patients. Previous experimental studies demonstrated that RA triggers alveolar bone loss similarly to PD. The aim of this study was to investigate if arthritis-induced alveolar bone loss is associated with modification in the oral microbiota. Checkerboard DNA-DNA hybridization was employed to analyze forty oral bacterial species in 3 groups of C57BL/6 mice: control (n = 12; without any challenge); Y4 (n = 8; received oral inoculation of Aggregatibacter Actinomycetemcomitans strain FDC Y4) and AIA group (n = 12; chronic antigen-induced arthritis). The results showed that AIA and Y4 group exhibited similar patterns of bone loss. The AIA group exhibited higher counts of most bacterial species analyzed with predominance of Gram-negative species similarly to infection-induced PD. Prevotella nigrescens and Treponema denticola were detected only in the Y4 group whereas Campylobacter showae, Streptococcus mitis and Streptococcus oralis were only found in the AIA group. Counts of Parvimonas micra, Selenomonas Noxia and Veillonella parvula were greater in the AIA group whereas Actinomyces viscosus and Neisseira mucosa were in large proportion in Y4 group. In conclusion, AIA is associated with changes in the composition of the oral microbiota, which might account for the alveolar bone loss observed in AIA mice.
Assuntos
Perda do Osso Alveolar/microbiologia , Processo Alveolar/microbiologia , Artrite Experimental/microbiologia , Maxila/microbiologia , Microbiota/genética , Periodontite/microbiologia , Aggregatibacter actinomycetemcomitans/classificação , Aggregatibacter actinomycetemcomitans/genética , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Perda do Osso Alveolar/patologia , Processo Alveolar/patologia , Animais , Artrite Experimental/patologia , Campylobacter/classificação , Campylobacter/genética , Campylobacter/isolamento & purificação , DNA Bacteriano/genética , Humanos , Masculino , Maxila/patologia , Camundongos , Camundongos Endogâmicos C57BL , Boca/microbiologia , Boca/patologia , Periodontite/patologia , Prevotella nigrescens/classificação , Prevotella nigrescens/genética , Prevotella nigrescens/isolamento & purificação , Streptococcus mitis/classificação , Streptococcus mitis/genética , Streptococcus mitis/isolamento & purificação , Streptococcus oralis/classificação , Streptococcus oralis/genética , Streptococcus oralis/isolamento & purificação , Treponema denticola/classificação , Treponema denticola/genética , Treponema denticola/isolamento & purificaçãoRESUMO
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.
Assuntos
Lipopolissacarídeos/análise , Ativação de Macrófagos/efeitos dos fármacos , Periodontite Periapical/microbiologia , Periodontite Periapical/terapia , Irrigantes do Canal Radicular/uso terapêutico , Preparo de Canal Radicular/métodos , Clorexidina/uso terapêutico , Humanos , Interleucina-1beta/análise , Ativação de Macrófagos/imunologia , Periodontite Periapical/imunologia , Porphyromonas endodontalis/isolamento & purificação , Porphyromonas gingivalis/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , Hipoclorito de Sódio/uso terapêutico , Fator de Necrose Tumoral alfa/análiseRESUMO
BACKGROUND: Suppuration (SUP) on probing may be an indication of active periodontal breakdown. The aim of the present study is to analyze which subgingival species are associated with SUP in patients with chronic (CP) and aggressive (AgP) periodontitis. METHODS: A total of 156 patients with CP and 66 with AgP were submitted to full-mouth periodontal examination and subgingival biofilm sampling (14 sites/patient). The counts of 44 bacterial species were determined by checkerboard. Comparisons between groups and sites were analyzed by the Mann-Whitney and Wilcoxon tests, respectively. Associations between frequency of SUP and bacterial species were analyzed by the Spearman correlation coefficient. RESULTS: The prevalence of SUP in patients with CP was 24.4%, and in patients with AgP it was 30.3%, and the percentage of SUP sites in the groups was 5.72% ± 1.06% and 6.96% ± 1.70%, respectively (P >0.05). SUP sites from patients with CP had significantly higher counts of Veillonella parvula, Dialister pneumosintes, Tannerella forsythia, and Prevotella nigrescens than SUP sites from patients with AgP (P <0.005). Significant positive correlations between high frequency of SUP and high levels of Actinomyces spp, Streptococcus spp., members of the orange complex, Acinetobacter baumannii, and Pseudomonas aeruginosa were observed in patients with CP (P <0.05). In patients with AgP, Actinomyces oris, Propionibacterium acnes, P. aeruginosa, Staphylococcus aureus, and Streptococcus sanguinis were positively associated with SUP, whereas Prevotella intermedia presented a negative association with SUP (P <0.05). CONCLUSIONS: SUP sites from patients with CP harbored significantly higher counts of several periodontal species than SUP sites from patients with AgP. Actinomyces spp., Streptococcus spp., members of the orange complex, T. forsythia, and certain non-oral pathogens were associated with a high number of sites with SUP.
Assuntos
Periodontite Agressiva/microbiologia , Bactérias/classificação , Periodontite Crônica/microbiologia , Acinetobacter baumannii/isolamento & purificação , Actinomyces/classificação , Adulto , Carga Bacteriana , Bacteroides/isolamento & purificação , Biofilmes , Placa Dentária/microbiologia , Feminino , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/isolamento & purificação , Humanos , Masculino , Prevotella intermedia/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , Propionibacterium acnes/isolamento & purificação , Pseudomonas aeruginosa/isolamento & purificação , Fumar , Staphylococcus aureus/isolamento & purificação , Streptococcus/classificação , Streptococcus sanguis/isolamento & purificação , Supuração/microbiologia , Veillonella/isolamento & purificaçãoRESUMO
This study investigated the presence of the black-pigmented bacteria Prevotella nigrescens and Prevotella intermedia, the non-black-pigmented bacteria Actinomyces spp and particularly the cariogenic pathogen Streptococcus mutans in the dental biofilms of patients with or without black extrinsic tooth stains, using the multiplex polymerase chain reaction (PCR) technique. Analysis of the dental biofilms of patients with (n=26) or without (n=26) black tooth stains was performed using duplex PCR for the 16S ribosomal RNA gene (P. nigrescens, P. intermedia, Actinomyces spp) and glucosyltransferase-I gene for S. mutans. P. nigrescens and S. mutans were the most frequent bacteria detected in both groups. The least frequently detected were P. intermedia and Actinomyces spp. The similar bacterial composition of dental biofilms of black tooth stains and healthy tooth surfaces indicates that black tooth stains are not free of cariogenic bacteria.
O objetivo deste estudo foi investigar a presença das bactérias pigmentadoras de negro Prevotella nigrescens e Prevotella intermedia, da não pigmentadora de negro Actinomyces spp e particularmente a bactéria cariogênica Streptococcus mutans, no biofilme dentário de pacientes com ou sem manchas dentárias extrínsecas negras, utilizando a técnica multiplex PCR (reação em cadeia da polimerase). Análises do biofilme dentário de pacientes com manchas (n=26) e sem manchas (n=26) foram realizadas utilizando a multiplex PCR para o gene 16S RNA ribosomal (P. nigrescens, P. intermedia, Actinomyces spp) e o gene glucosiltransferase-I para S. mutans. P. nigrescens e S. mutans foram as bactérias mais frequentemente detectadas em ambos os grupos. As menos frequentemente detectadas foram P. intermedia e Actinomyces spp. A similaridade entre a composição bacteriana dos biofilmes dentários das manchas dentárias extrínsecas negras e das superfícies dentárias sem manchas indicam que as manchas dentárias extrínsecas negras não estão livres de bactérias cariogênicas.
Assuntos
Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Actinomyces/isolamento & purificação , Placa Dentária/microbiologia , Prevotella intermedia/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , Streptococcus mutans/isolamento & purificação , Descoloração de Dente/microbiologia , Actinomyces/genética , Biofilmes , Reação em Cadeia da Polimerase Multiplex , Prevotella intermedia/genética , Prevotella nigrescens/genética , /análise , Streptococcus mutans/genéticaRESUMO
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.
Assuntos
Doenças da Polpa Dentária/microbiologia , Endotoxinas/análise , Infecções por Bactérias Gram-Negativas/microbiologia , Lipopolissacarídeos/análise , Doenças Assintomáticas , DNA Bacteriano/análise , Cavidade Pulpar/microbiologia , Doenças da Polpa Dentária/terapia , Necrose da Polpa Dentária/microbiologia , Necrose da Polpa Dentária/terapia , Infecções por Fusobacterium/microbiologia , Fusobacterium nucleatum/isolamento & purificação , Infecções por Bactérias Gram-Negativas/terapia , Humanos , Teste do Limulus , Medição da Dor , Periodontite Periapical/microbiologia , Periodontite Periapical/terapia , Reação em Cadeia da Polimerase , Porphyromonas endodontalis/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , Recidiva , Retratamento , Supuração , Treponema/classificação , Treponema denticola/isolamento & purificação , Infecções por Treponema/microbiologiaRESUMO
OBJECTIVE: To compare the microbiota of endodontic infections in necrotic pulp from HIV-negative and HIV-positive subjects. MATERIALS AND METHODS: Root canal samples from necrotic pulp were collected from 40 HIV- and 20 HIV+ subjects. Pulps were amplified using multiple displacement amplification (MDA). Then, checkerboard DNA-DNA hybridization was employed to assess the levels of 107 microbial taxa. The percentage of DNA probe count and the percentage of teeth colonized by each test species were investigated. Significant differences between groups regarding proportions of taxa and prevalence of the test species were sought using the Mann-Whitney test and the Chi-square analysis, respectively. RESULTS: The most prevalent taxa detected were Dialister pneumosintes, Stenotrophomonas maltophilia, Streptococcus sobrinus, Corynebacterium diphteriae, and Helicobacter pylori among HIV- subjects and D. pneumosintes, Prevotella tannerae, Porphyromonas gingivalis, Parvimonas micra, Prevotella nigrescens, and Corynebacterium diphtheriae among HIV+ individuals. D. pneumosintes, C. diphtheria, and C. albicans were the most abundant species in the HIV- group, whereas the predominant taxa in HIV+ samples were P. tannerae, D. pneumosintes and Olsenella uli. P. tannerae, O. uli, Veilonella dispar, Bacteroides fragilis, and Actinomyces meyeri were significantly more abundant in HIV+ samples. CONCLUSIONS: There were significant differences in the prevalence and proportions of specific microbial taxa between HIV- and HIV+ individuals. The root canal microbiota may represent a reservoir of important oral and medical pathogens, mainly in HIV+ individuals.
Assuntos
Bactérias/classificação , Necrose da Polpa Dentária/microbiologia , Soronegatividade para HIV , Soropositividade para HIV/microbiologia , Actinomyces/isolamento & purificação , Adolescente , Adulto , Bacteroides fragilis/isolamento & purificação , Candida albicans/isolamento & purificação , Criança , Corynebacterium diphtheriae/isolamento & purificação , Sondas de DNA , Cavidade Pulpar/microbiologia , Feminino , Bacilos Gram-Negativos Anaeróbios Retos, Helicoidais e Curvos/classificação , Helicobacter pylori , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico , Hibridização de Ácido Nucleico/métodos , Peptostreptococcus/isolamento & purificação , Porphyromonas gingivalis/isolamento & purificação , Prevotella/classificação , Prevotella nigrescens/isolamento & purificação , Stenotrophomonas maltophilia/isolamento & purificação , Streptococcus sobrinus/isolamento & purificação , Veillonella/isolamento & purificação , Adulto JovemRESUMO
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.
Assuntos
Bactérias Anaeróbias/classificação , Necrose da Polpa Dentária/microbiologia , Consórcios Microbianos , Abscesso Periapical/microbiologia , Algoritmos , Infecções por Bacteroidaceae/microbiologia , Bacteroides/isolamento & purificação , Infecções por Bacteroides/microbiologia , Enterococcus faecalis/isolamento & purificação , Fusobacterium/isolamento & purificação , Infecções por Fusobacterium/microbiologia , Fusobacterium nucleatum/isolamento & purificação , Infecções por Bactérias Gram-Positivas/microbiologia , Humanos , Reação em Cadeia da Polimerase , Porphyromonas endodontalis/isolamento & purificação , Porphyromonas gingivalis/isolamento & purificação , Prevotella/classificação , Prevotella intermedia/isolamento & purificação , Prevotella nigrescens/isolamento & purificaçãoRESUMO
This study investigated the presence of the black-pigmented bacteria Prevotella nigrescens and Prevotella intermedia, the non-black-pigmented bacteria Actinomyces spp and particularly the cariogenic pathogen Streptococcus mutans in the dental biofilms of patients with or without black extrinsic tooth stains, using the multiplex polymerase chain reaction (PCR) technique. Analysis of the dental biofilms of patients with (n=26) or without (n=26) black tooth stains was performed using duplex PCR for the 16S ribosomal RNA gene (P. nigrescens, P. intermedia, Actinomyces spp) and glucosyltransferase-I gene for S. mutans. P. nigrescens and S. mutans were the most frequent bacteria detected in both groups. The least frequently detected were P. intermedia and Actinomyces spp. The similar bacterial composition of dental biofilms of black tooth stains and healthy tooth surfaces indicates that black tooth stains are not free of cariogenic bacteria.
Assuntos
Actinomyces/isolamento & purificação , Placa Dentária/microbiologia , Prevotella intermedia/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , Streptococcus mutans/isolamento & purificação , Descoloração de Dente/microbiologia , Actinomyces/genética , Adolescente , Adulto , Biofilmes , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Multiplex , Prevotella intermedia/genética , Prevotella nigrescens/genética , RNA Ribossômico 16S/análise , Streptococcus mutans/genéticaRESUMO
OBJECTIVE: In this study of patients with chronic periodontitis (CP), the severity of the disease and the main periodontal pathogens identified in patients with chronic kidney disease (CKD) were compared with those detected in individuals without systemic disease. DESIGN: Nineteen patients with CP without evidence of systemic disease (control group), 25 patients with CP and CKD who were in the pre-dialysis stages (pre-dialysis group), and 22 patients with CP and CKD who were on renal replacement therapy (RRT group) were examined. The severity of CP was based on the investigation of probing depth (PD) and clinical attachment level (CAL). The definition and stage of CKD were based on the criteria proposed by the Kidney Disease Outcomes Quality Initiative of the National Kidney Foundation. Glomerular filtration rate (GFR) was estimated using the equation of Modification of Diet in Renal Disease and the identification of microorganisms in subgingival plaque was performed using polymerase chain reaction (PCR). RESULTS: Candida albicans, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were more common in patients who were on RRT and pre-dialysis than in control subjects. CP was more severe in patients with CKD. A strong association was observed between the frequency of C. albicans (P = 0.056), P.gingivalis (P = 0.008), T. denticola (P = 0.013) and CAL, when CKD patients were compared with the control group. CONCLUSION: CP is more severe and is associated with increased frequency of C. albicans, P. gingivalis, T. forsythia, and T. denticola in patients with CKD.
Assuntos
Periodontite Crônica/microbiologia , Falência Renal Crônica/complicações , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Bacteroides/isolamento & purificação , Candida albicans/isolamento & purificação , Periodontite Crônica/classificação , Periodontite Crônica/complicações , Placa Dentária/microbiologia , Complicações do Diabetes , Eikenella corrodens/isolamento & purificação , Feminino , Fusobacterium nucleatum/isolamento & purificação , Hemorragia Gengival/classificação , Hemorragia Gengival/microbiologia , Retração Gengival/classificação , Retração Gengival/microbiologia , Taxa de Filtração Glomerular/fisiologia , Humanos , Falência Renal Crônica/microbiologia , Masculino , Pessoa de Meia-Idade , Perda da Inserção Periodontal/classificação , Perda da Inserção Periodontal/microbiologia , Bolsa Periodontal/classificação , Bolsa Periodontal/microbiologia , Reação em Cadeia da Polimerase , Porphyromonas gingivalis/isolamento & purificação , Prevotella intermedia/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , Terapia de Substituição Renal , Treponema denticola/isolamento & purificaçãoRESUMO
INTRODUCTION: Prophylactic programs to prevent dental biofilm accumulation must be implemented to minimize the risk for periodontal diseases in orthodontic patients. Therefore, we assessed the possible periodontal and microbiologic changes resulting from the use of 2 methods of orthodontic archwire ligation: elastomeric rings and steel ligatures. METHODS: The following parameters were measured: plaque index, gingival bleeding index, probing depth, and biofilm samples from the maxillary second premolars and the mandibular lateral incisors were evaluated in 14 subjects without clinical signs of gingival inflammation before orthodontic appliance placement and after 6 months of treatment. Each orthodontic arch was fixed with elastomeric rings on 1 side of the midline, and steel ligatures were used on the opposite side. Polymerase chain reaction analysis was used to detect Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and P nigrescens. RESULTS: The elastomeric rings were associated with a higher score for plaque index and bleeding than steel ligatures, as well as many positive sites of T forsythia and P nigrescens (P <0.05). CONCLUSIONS: Elastomeric rings favored these 2 periodontopathogens and harmed gingival conditions.
Assuntos
Fios Ortodônticos/microbiologia , Adolescente , Análise de Variância , Bacteroides/genética , Bacteroides/isolamento & purificação , Biofilmes , DNA Bacteriano/análise , Índice de Placa Dentária , Elastômeros , Feminino , Humanos , Masculino , Doenças Periodontais/microbiologia , Índice Periodontal , Prevotella nigrescens/genética , Prevotella nigrescens/isolamento & purificação , Aço Inoxidável , Estatísticas não Paramétricas , Dente/microbiologiaRESUMO
AIM: This study aimed at evaluating the prevalence of putative periodontal pathogens (Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella nigrescens, Treponema denticola) in saliva of children with mixed dentition at two different time points, correlating these findings with a clinical parameter of gingival health. DESIGN: Polymerase chain reaction (PCR) detection was used to determine the prevalence of these bacteria in saliva of 64 children in 2003 and 60 children in 2004. Gingival health was assessed by gingival index. RESULT: Sixty-two (96.9%) and 50 (83.3%) children presented low gingival inflammation, whereas 2 (3.1%) and 10 (16.7%) had moderate scores in 2003 and 2004, respectively. Majority of the children (81.3% in 2003 and 73.3% in 2004) had detectable levels of at least one of the bacteria. The prevalence found was of 4.7% and 1.7% for A. actinomycetemcomitans, 6.3% and 8.3% for P. gingivalis, 23.4% and 48.3% for P. nigrescens (P < 0.05), and 71.9% and 50% for T. denticola (P < 0.05) in 2003 and 2004, respectively. No significant relationship between gingival index and presence of these bacteria and combination of different species was found. CONCLUSION: A high percentage of children harboured at least one of the putative periodontal pathogens in saliva, but presented periodontally healthy conditions.
Assuntos
Dentição Mista , Gengivite/microbiologia , Bactérias Gram-Negativas/isolamento & purificação , Saliva/microbiologia , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Brasil , Criança , Contagem de Colônia Microbiana , Feminino , Seguimentos , Humanos , Estudos Longitudinais , Masculino , Índice Periodontal , Reação em Cadeia da Polimerase , Porphyromonas gingivalis/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , Treponema denticola/isolamento & purificaçãoRESUMO
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.
Assuntos
Técnicas de Tipagem Bacteriana , Infecções por Bacteroidaceae/microbiologia , Doenças da Polpa Dentária/microbiologia , Doença Crônica , Contagem de Colônia Microbiana , DNA Bacteriano/análise , Humanos , Reação em Cadeia da Polimerase , Porphyromonas endodontalis/isolamento & purificação , Porphyromonas gingivalis/isolamento & purificação , Prevotella intermedia/isolamento & purificação , Prevotella nigrescens/isolamento & purificaçãoRESUMO
BACKGROUND AND AIM: Specific microbial profiles that may distinguish between generalized aggressive-periodontitis (GAgP) and generalized chronic-periodontitis (GCP) have, to date, not been described. The purpose of the present study was to describe the subgingival microbial composition of Mexican subjects with GAgP and compare it with that of individuals with GCP and periodontal health (PH). MATERIAL AND METHODS: Seventy-seven subjects with GAgP (n=19), GCP (n=39) and PH (n=19) were selected. Clinical measurements included plaque accumulation, gingival erythema, bleeding on probing, suppuration, pocket depth and attachment level. Up to 28 subgingival plaque samples were obtained from each subject and analysed using the checkerboard DNA-DNA hybridization technique. RESULTS: GAgP and GCP subjects harboured significantly higher levels and/or proportion of Porphyromonas gingivalis, Tannerella forsythia (levels: p<0.001, proportion: p<0.01), Prevotella nigrescens (p<0.05 levels) and "red" complex species (p<0.001 proportion) than PH subjects. All GAgP subjects were carriers of P. gingivalis and P. nigrescens. No significant differences in any of the 40 microbial species tested were detected between GAgP and GCP subjects. CONCLUSIONS: Our results revealed that the microbial differences between GAgP and GCP subjects were only discrete and none of the bacterial species tested seemed to specifically differentiate the subgingival microbial profile of either periodontitis group.
Assuntos
Placa Dentária/microbiologia , Gengiva/microbiologia , Periodontite/classificação , Actinomyces/classificação , Actinomyces/isolamento & purificação , Adolescente , Adulto , Idoso , Bacteroides/classificação , Bacteroides/isolamento & purificação , Criança , Contagem de Colônia Microbiana , Placa Dentária/classificação , Eritema/microbiologia , Feminino , Doenças da Gengiva/microbiologia , Hemorragia Gengival/microbiologia , Humanos , Masculino , México , Pessoa de Meia-Idade , Perda da Inserção Periodontal/microbiologia , Periodontite/microbiologia , Porphyromonas gingivalis/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , SupuraçãoRESUMO
BACKGROUND: Actinobacillus actinomycetemcomitans is considered a major etiologic agent of aggressive periodontitis (AgP). Other periodontopathic bacteria such as Porphyromonas gingivalis are also suspected of participating in aggressive periodontitis although the evidence to support this is controversial. The aim of the present study was to determine the prevalence of eight periodontopathic bacteria in Chilean patients with AgP. METHODS: Subgingival plaque samples were collected from 36 aggressive, 30 localized, and six generalized periodontitis patients. Samples from 17 advanced chronic periodontitis (CP) patients were taken as controls. Samples collected from the four deepest periodontal pockets in each patient were pooled in prereduced transport fluid (RTF) and cultured. Periodontal bacteria were primarily identified by colony morphology under stereoscopic microscope and rapid biochemical tests. The identity of some bacterial isolates was confirmed by colony polymerase chain reaction (PCR). RESULTS: AgP showed a significatively higher prevalence of C. rectus than CP (P = 0.036). The only statistical difference found was for C. rectus. Patients with AgP showed a higher, but not statistically significant, prevalence of P. gingivalis, E. corrodens, P. micros, and Capnocytophaga sp. A similar prevalence in both groups of patients was observed for F. nucleatum and P. intermedia/nigrescens, and A. actinomycetemcomitans was less prevalent in AgP than CP patients. In localized AgP, P. intermedia/nigrescens, E. corrodens, F. nucleatum, and P. micros were the more prevalent pathogens in contrast to generalized AgP patients who harbored A. actinomycetemcomitans, P. gingivalis, and Capnocytophaga sp. as the most prevalent bacteria. CONCLUSIONS: C. rectus, P. gingivalis, E. corrodens, P. micros, and Capnocytophaga sp. were the most predominant periodontopathic bacteria of AgP in this Chilean population, but the only statistical difference found here between AgP and CP was for C. rectus, suggesting that the differences in clinical appearance may be caused by factors other than the microbiological composition of the subgingival plaque of these patients. In this study, the prevalence of A. actinomycetemcomitans was much lower than that of P. gingivalis.
Assuntos
Periodontite/microbiologia , Doença Aguda , Adolescente , Adulto , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Aggregatibacter actinomycetemcomitans/patogenicidade , Campylobacter rectus/isolamento & purificação , Campylobacter rectus/patogenicidade , Capnocytophaga/isolamento & purificação , Capnocytophaga/patogenicidade , Distribuição de Qui-Quadrado , Doença Crônica , Contagem de Colônia Microbiana , Estudos Transversais , Placa Dentária/microbiologia , Eikenella corrodens/isolamento & purificação , Eikenella corrodens/patogenicidade , Feminino , Fusobacterium nucleatum/isolamento & purificação , Fusobacterium nucleatum/patogenicidade , Humanos , Masculino , Pessoa de Meia-Idade , Peptostreptococcus/isolamento & purificação , Peptostreptococcus/patogenicidade , Porphyromonas gingivalis/isolamento & purificação , Porphyromonas gingivalis/patogenicidade , Prevotella intermedia/isolamento & purificação , Prevotella intermedia/patogenicidade , Prevotella nigrescens/isolamento & purificação , Prevotella nigrescens/patogenicidadeRESUMO
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.
Assuntos
Infecções por Bacteroidaceae/microbiologia , Necrose da Polpa Dentária/microbiologia , Falha de Restauração Dentária , Porphyromonas/isolamento & purificação , Prevotella/isolamento & purificação , Técnicas de Tipagem Bacteriana , Contagem de Colônia Microbiana , DNA Bacteriano/análise , Feminino , Humanos , Masculino , Reação em Cadeia da Polimerase , Porphyromonas/genética , Porphyromonas/patogenicidade , Porphyromonas endodontalis/genética , Porphyromonas endodontalis/isolamento & purificação , Porphyromonas endodontalis/patogenicidade , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/isolamento & purificação , Porphyromonas gingivalis/patogenicidade , Prevotella/genética , Prevotella/patogenicidade , Prevotella intermedia/genética , Prevotella intermedia/isolamento & purificação , Prevotella intermedia/patogenicidade , Prevotella nigrescens/genética , Prevotella nigrescens/isolamento & purificação , Prevotella nigrescens/patogenicidadeRESUMO
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.
Assuntos
Cavidade Pulpar/microbiologia , Necrose da Polpa Dentária/microbiologia , Periodontite Periapical/microbiologia , Bactérias Anaeróbias/isolamento & purificação , Técnicas Bacteriológicas , Edema/microbiologia , Fusobacterium necrophorum/isolamento & purificação , Fusobacterium nucleatum/isolamento & purificação , Bactérias Gram-Negativas/isolamento & purificação , Humanos , Dor/microbiologia , Peptostreptococcus/isolamento & purificação , Abscesso Periapical/microbiologia , Porphyromonas endodontalis/isolamento & purificação , Porphyromonas gingivalis/isolamento & purificação , Prevotella intermedia/isolamento & purificação , Prevotella nigrescens/isolamento & purificação , Tratamento do Canal Radicular , Dente não Vital/microbiologiaRESUMO
OBJECTIVE: To compare periodontal conditions in children with and without cleft. DESIGN: Clinical examinations and microbiological analysis of 57 selected children, including 30 with unilateral complete cleft lip and palate (experimental group) and 27 without clefts (control group). SETTING: Hospital of Rehabilitation of Craniofacial Anomalies (HRCA) in Bauru, Sao Paulo, Brazil. PATIENTS, PARTICIPANTS: All children examined were healthy and between the ages of 5 and 6 years. RESULTS: The mean plaque index (PI) in the experimental group was higher (1.82 +/- 0.3) than in the control group (1.63 +/- 0.38), although this difference was not statistically significant. The mean gingival index (GI) in the experimental group (1.82 +/- 0.38) was found to be significantly higher (p <.05) than that of the control group (0.79 +/- 0.33). The cleft area in the experimental group, with a mean PI of 2.04 +/- 0.58 and mean GI of 1.11 +/- 0.26, compared with the posterior area, with a mean PI of 1.74 +/- 0.37 and mean GI of 1.04 +/- 0.26, showed a statistically significant difference only in the PI. Most of the children in both experimental and control groups presented a moderate PI degree (73.33% and 81.48%, respectively) and a high prevalence of mild gingivitis (53.33% and 70.37%, respectively). Analysis of the organisms showed that Prevotella nigrescens was detected in 16.67% of the experimental group and 11.11% of the control, whereas Porphyromonas gingivalis and Treponema denticola were not detected. CONCLUSION: Children with clefts showed greater gingival inflammation, despite the same amount of plaque and prevalence of microorganisms.