Analysis of Active Bacteria Persisting after Chemomechanical Procedures: An RNA- and DNA-based Molecular Study

Nardello, Laura Cristina Leite; Vilela, Bruna G.; Fernandes, Flávia Gonçalves; Bruno, Fernanda Pinheiro; Carvalho, Alexandre Pinheiro Lima de; Cazares, Roberto Xavier Romero; Nascimento, Laís Cunha Prado do; Honório, Heitor Marques; Ando, Ellen Sayuri; Mayer, Márcia Pinto Alves; Gavini, Giulio; Pinheiro, Éricka Tavares

doi:10.1016/j.joen.2020.08.004

Cited by 8 publications

(4 citation statements)

References 30 publications

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“…This fact may explain the resistance of this species to antimicrobial procedures commonly used during endodontic treatment [35]. C. acnes can persist active in root canals after root canal preparation, being a possible source for the establishment of persistent infections [36]. Other bacterial species commonly found in endodontic infections (e.g., Capnocytophaga sp., F. nucleatum, O. uli, P. micra, P. intermedia and T. forsythia ) expressed high numbers of AMR homologs.…”

Section: Discussionmentioning

confidence: 99%

“…Root canal samples were collected from ten patients, five with primary infections and five with persistent/secondary endodontic infections (called secondary infections in this study). Sampling procedures were performed as previously described [12,36,39]. After removal of dental plaque and caries, the tooth and rubber dam were disinfected with 30% H 2 O 2 , 2.5% NaOCl and 5% sodium thiosulfate.…”

Section: Methodsmentioning

confidence: 99%

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Metatranscriptome of infected root canals in teeth with apical periodontitis

Pinheiro

Russo

Candeiro

et al. 2022

Preprint

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Metatranscriptomics has not yet been applied to the study of the endodontic microbiome. Therefore, this study investigated the microbial composition and patterns of gene expression in teeth with asymptomatic apical periodontitis. We performed next-generation, microbiome- wide RNA-sequencing in eight root canal samples, including five primary and three secondary endodontic infections. Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria represented the dominant phyla, whereas Fusobacteria, Spirochaetes and Synergistetes were among the non-dominant phyla. At species level, the microbiome was dominated by Gram-negative and Gram-positive anaerobes. In particular,Prevotella spp.andOlsenella uliwere more abundant in primary than in secondary infections. Transcripts encoding moonlighting proteins were abundant, including glycolytic proteins, translational elongation factors, chaperonin, and heat shock proteins. Fusobacterium was highly active in primary infections, showing upregulation of genes involved in protein synthesis, responses to oxidative stress and resistance to antibiotics, especially tetracycline. In addition, other members of the community expressed high numbers of antimicrobial resistance genes. Overall, metatranscriptomics revealed the expression of putative virulence factors and the activity of potential endodontic pathogens, represented mainly by obligate anaerobes. In particular, the functional role of Fusobacterium in primary infections was highlighted. Furthermore, antimicrobial resistance, especially to tetracycline, was a common feature in endodontic samples.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Metatranscriptome of infected root canals in teeth with apical periodontitis

Pinheiro

Russo

Candeiro

et al. 2022

Preprint

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“…O tratamento endodôntico convencional reduz expressivamente o número de microrganismos persistentes, no entanto (Pinheiro et al 2015;Nardello et al 2020). A TFD, desta forma, em associação com o tratamento endodôntico pode agir como procedimento coadjuvante na redução da microbiota dos canais radiculares.…”

Section: Discussionunclassified

Avaliação da redução microbiana do sistema de canais radiculares após a Terapia Fotodinâmica (TFD) - estudo in vivo

Leite

Saguchi

Oshiro

et al. 2021

RSD

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O tratamento endodôntico tem o objetivo de remover tecidos infectados e microrganismos do sistema de canal radicular (SCR). Devido à complexidade do SCR, os métodos convencionais de irrigação e tornam limitados, assim diferentes técnicas foram introduzidas a fim de potencializar a limpeza e desinfecção dos condutos radiculares. O objetivo deste trabalho foi comparar a redução microbiana intracanal promovida pelo tratamento endodôntico convencional e terapia fotodinâmica (TFD) com emissão de laser na entrada e no interior do sistema radicular. O presente estudo submeteu pacientes com necessidades de tratamento endodôntico de acordo com os critérios de inclusão. Distribuindo em grupos G1(10) seguindo o protocolo de preparo químico- cirúrgico (PQC), G2(10) com protocolo PQC+TFD com ponteiras na entrada do canal e G3(10) no protocolo PQC+TFD com ponteira no interior do canal. Foi realizado o teste Two-way ANOVA p>0,05 para comparação entre os grupos. Os resultados mostraram que houve redução microbiana, mas não houve diferença significativa. Observou-se também que houve maior homogeneidade dos dados nos grupos irradiados (G2 e G3). Conclui-se que o TDF pode ser utilizado como coadjuvante no tratamento endodôntico com o aumento do índice de sucesso do mesmo.

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“…On the other hand, remaining dead cells might trigger host inflammatory reactions, leading to persistence of apical periodontitis. Moreover, a study conducted to analyse the microbial cells activity in ribosomal RNA and compare it to the rDNA levels, suggested a low risk of false-positive detection when targeting rDNA (Nardello et al, 2020). Finally, although patients and radiographic examiners were blinded to the treatment protocol used, the treatment was visually distinguishable to the operators and, thus, might have resulted in an overall bias, which is a frequent limitation in dental randomized trials (Yi et al, 2020).…”

Section: Restoration Qualitymentioning

confidence: 99%

The impact of an enhanced infection control protocol on molar root canal treatment outcome – a randomized clinical trial

et al. 2021

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Aim To evaluate the effect of an enhanced infection control protocol on root canal treatment outcomes and on microbial load within root canals after chemomechanical preparation. Methodology A total of 144 molar teeth from 139 healthy patients receiving primary root canal treatment were block randomized to a standard protocol (StP) or an enhanced infection control protocol (EnP). Both treatment arms adhered to current best practice recommendations, while the EnP comprised additional steps that included replacing rubber dams, gloves, files, all instruments and surface barriers at the time of canal filling to reduce the chances of iatrogenic contamination. Patients and radiographic examiners were blinded to the protocol used. Intracanal microbial samples were taken at baseline (S1) and after completion of chemomechanical preparation (S2). Microbial 16S rDNA copy numbers were enumerated by quantitative polymerase chain reaction (qPCR). Cone beam computed tomography (CBCT) scans were taken before treatment and at one‐year follow‐up. The outcome was assessed clinically and radiographically using CBCT by logistic regression modelling. Results At one‐year follow‐up, 115 teeth were analysed (54 in StP and 61 in EnP). The percentage of favourable outcomes assessed by CBCT was 85.2% in the EnP and 66.7% in the StP. The odds of 12‐month success was three times higher in the EnP group compared with the StP group (OR=2.89; p=0.022, CI: 1.17 – 7.15). The median bacterial reads were reduced from 8.1×103 in S1 samples to 3.5×103 in the StP group and from 8.6×103 to 1.3×103 in the EnP group. The enhanced protocol significantly reduced bacterial counts in pre‐canal filling samples when compared to the standard protocol (p=0.009). Conclusions The implementation of a facile, enhanced infection control protocol in primary root canal treatment resulted in less detectable bacterial DNA before canal filling and significantly more successful outcomes at one year.

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Analysis of Active Bacteria Persisting after Chemomechanical Procedures: An RNA- and DNA-based Molecular Study

Cited by 8 publications

References 30 publications

Metatranscriptome of infected root canals in teeth with apical periodontitis

Metatranscriptome of infected root canals in teeth with apical periodontitis

Avaliação da redução microbiana do sistema de canais radiculares após a Terapia Fotodinâmica (TFD) - estudo in vivo

The impact of an enhanced infection control protocol on molar root canal treatment outcome – a randomized clinical trial

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