Effect of supplementary sodium hypochlorite agitation techniques on an ex vivo oral multispecies biofilm during passive disinfection of simulated immature roots

Int Endod J. 2024 Jul;57(7):966-980. doi: 10.1111/iej.14053. Epub 2024 Feb 22.

Abstract

Aim: To compare the effect of different sodium hypochlorite (NaOCl) agitation techniques on an ex vivo oral multispecies biofilm during passive disinfection of simulated immature roots.

Methodology: Extracted human teeth were prepared to simulate immature roots. They were infected with a dental plaque-derived multispecies biofilm and cultured for 14 days. The roots were randomly designated into four groups: (1) negative control (PBS), (2) 1.5% NaOCl (CNI), (3) CNI + Ultrasonic activation (UA), (4) CNI + EasyClean agitation (ECA), (5) CNI + XP-endo finisher agitation (XPF), and (6) positive control (6% NaOCl). Biofilm samples were collected from the root canals and used to determine the number of viable cells (colony-forming units), scanning electron microscopy, and 16S rRNA gene sequencing. The mean colony-forming units per mL (CFU/mL) were analysed using One-way anova. 16S rRNA sequencing data were analysed for alpha (observed OTUs, Shannon index, and Chao1) and beta diversity (Bray-Curtis dissimilarities). The LEfSe analysis was used to determine the effect of treatment procedures on the abundance of root canal microbiota. The significance was set at .05.

Results: PBS and CNI samples had significantly higher CFU/mL counts than UA, ECA, XPF, and 6% NaOCl samples (p < .05). The pre-treatment, PBS, and CNI groups had significantly greater alpha diversity than the UA, ECA, XPF, and 6% NaOCl groups (p < .05). NaOCl agitation groups and the 6% NaOCl group achieved a more pronounced reduction in bacteria from the genera Fusobacterium, Actinomyces, Porphyromonas, and Capnocytophaga.

Conclusions: The effectiveness of passive disinfection protocols was enhanced by NaOCl agitation techniques, suggesting that this supplementary method can improve the outcome of revitalization procedures.

Keywords: biofilm; disinfection; endodontic infection; revitalization procedure.

MeSH terms

  • Biofilms* / drug effects
  • Dental Pulp Cavity / microbiology
  • Disinfection* / methods
  • Humans
  • In Vitro Techniques
  • Microscopy, Electron, Scanning
  • RNA, Ribosomal, 16S
  • Root Canal Irrigants / pharmacology
  • Sodium Hypochlorite* / pharmacology
  • Tooth Root / drug effects
  • Tooth Root / microbiology

Substances

  • Sodium Hypochlorite
  • Root Canal Irrigants
  • RNA, Ribosomal, 16S