The objective of this study was to investigate the bacterial composition present in root canals of teeth associated with acute apical abscess by molecular identification (16S rRNA) of cultivable bacteria. Two hundred and twenty strains isolated by culture from 20 root canals were subjected to DNA extraction and amplification of the 16S rRNA gene (PCR), followed by sequencing. The resulting nucleotide sequences were compared to the GenBank database from the National Center of Biotechnology Information through BLAST. Strains not identified by sequencing were submitted to clonal analysis. The association of microbiological findings with clinical features and the association between microbial species were also investigated. Fifty-nine different cultivable bacteria were identified by 16S rRNA gene sequencing, belonging to 6 phyla, with an average number of 6 species per root canal. Molecular approaches allowed identification of 99% of isolates. The most frequently identified bacteria were Prevotella spp., Pseudoramibacter alactolyticus, Parvimonas micra, Dialister invisus, Filifactor alocis, and Peptostreptococcus stomatis. Positive association was found between Prevotella buccae and Pseudoramibacter alactolyticus and between Parvimonas micra and Prevotella nigrescens (both p<0.05). It was concluded that the microbiota of infected root canals associated with acute apical abscess is diverse and heterogeneous, composed mainly of anaerobic Gram-negative bacteria, with the great majority belonging to the phyla Firmicutes and Bacteroidetes.
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.
The aim of this study was to compare the biological activity of lipopolysaccharides (LPS) purified from Fusobacterium nucleatum and Porphyromonas gingivalis strains, both isolated from primary endodontic infection (PEI) in the levels of IL-1β and TNF-α released by macrophage cells. Moreover, LPS was purified from F. nucleatum and P. gingivalis American Type Collection (ATCC) and its biological activity was compared to respectively clinical isolates strains. F. nucleatum and P. gingivalis strains clinically isolated from PEI had their identification confirmed by sequencing the 16S rRNA gene. LPS from F. nucleatum and P. gingivalis and their respective ATCC strains were extracted by using Tri-reagent method. Macrophages (Raw 264.7) were stimulated with LPS at 100 ng/mL for 4, 8 and 12 h. Secretion of IL-1 β and TNF-α was also determined. Paired t-test, repeated measures ANOVA and one-way ANOVA were employed. All LPS induced significant production of IL-1β and TNF-α, with the former being secreted at higher levels than the latter in all time-points. F. nucleatum induced a higher expression of both cytokines compared to P. gingivalis (p<0.05). No differences were observed between clinical and ATCC strains, as both presented the same potential to induce pro-inflammatory response. It was concluded that F. nucleatum and P. gingivalis LPS presented different patterns of activation against macrophages as seen by the IL-1β and TNF-α production, which may contribute to the immunopathogenesis of apical periodontitis. Moreover, clinical and ATCC strains grown under the same in vitro environment conditions presented similar biological activity.
Objective: This study sought to investigate the prevalence of eight oral Treponemas (Treponema denticola, T. amylovorum, T. maltophilum, T. medium, T. pectinovorum, T. socranskii, T. vicentii and T. lecithinolyticum) in teeth with endodontic treatment failure and periapical lesion.Methods: Samples were taken from 40 root canals presenting endodontic failure and periapical lesion. DNA extraction was performed and Nested-PCR technique was used for the detection of Treponema species using specific primers.Results: Treponemas was detected in 56.5% of the samples analyzed (22/39). Individual root canals yielded a maximum of 6 target Treponema species. T. denticola (30.8%) and T. maltophilum (30.8%) were the most frequently detected species followed by T. medium (20.5%), T. socranskii (20.5%), T. pectinovorum (17.9%) and T. vicentii (17.9%). Positive association was verified between T. denticola and T. maltophilum such as T. medium (P<.05). T. lecithinolyticum was positively associated with intraradicular post (P<.05).Conclusion: The present study revealed that a wide variety of Treponema species plays a role in persistent/secondary infection turning the root canal microbiota even more complex than previously described by endodontic literature. (Eur J Dent 2013;7:61-68)
Introdução: Durante o tratamento endodôntico, devido às complexidades anatômicas dos canais radiculares, a ação mecânica dos instrumentos não é suficiente para a completa desinfecção dos condutos. Dessa forma, se faz necessário o uso de soluções irrigadoras que possam potencializar a desinfecção do sistema de canais radiculares. Objetivo: Realizar uma revisão integrativa da literatura para comparar as propriedades antimicrobianas da clorexidina com o hipoclorito de sódio. Metodologia: A busca na literatura foi realizada no período de setembro de 2019 a agosto de 2021, nas seguintes bases de dados: PUBMED/MEDLINE, LILACS e SCIELO. Utilizando os descritores: clorexidina (chlorhexidine), hipoclorito de sódio (sodium hypochlorite), irrigante do canal radicular (root canal irrigant) e limpeza (cleaning). Utilizou-se como critérios de busca, trabalhos experimentais laboratoriais in vitro, publicados entre os anos de 2017 e 2021. Resultados: Foram encontrados 165 artigos, dos quais 15 foram selecionados ao final do processo. 8 trabalhos não encontraram diferença estatisticamente significativa entre a clorexidina e o hipoclorito, 5 artigos apresentaram resultados superiores do NaOCl e em 2 a clorexidina foi superior. Conclusões: Após análise da literatura, observamos semelhança entre a ação antimicrobiana do hipoclorito de sódio e da clorexidina, e podemos concluir que ambas apresentam boa ação antimicrobiana, justificando seu uso clinicamente.
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