Optimized protocol for collecting root canal biofilms for in vitro studies.

Area of Science:

• Microbiology
• Endodontics
• Dental Materials

Background:

• Microbial persistence in the root canal system (RCS) is a primary cause of endodontic retreatment failure.
• Bacterial biofilms within the RCS exhibit increased pathogenicity, demanding novel disinfection approaches.

Purpose of the Study:

• To standardize and evaluate a novel in vitro protocol for collecting root canal system (RCS) biofilms.
• To compare the efficacy of the proposed protocol against the absorbent paper points method for biofilm removal.

Main Methods:

• Bovine incisors were prepared and inoculated with Enterococcus faecalis to form biofilms over 21 days.
• Two biofilm collection methods were tested: a proposed protocol and absorbent paper points.
• Colony-forming units (CFUs) and scanning electron microscopy (SEM) were used for analysis.

Main Results:

• The proposed biofilm collection protocol yielded significantly higher CFU counts (p < 0.0001) compared to absorbent paper points.
• The new protocol demonstrated a high efficiency rate of 95.85% ± 1.15% for biofilm removal.
• SEM analysis visually confirmed superior biofilm removal with the proposed method.

Conclusions:

• The novel in vitro protocol is highly effective for collecting RCS biofilms.
• This method offers a significant improvement over traditional absorbent paper points for biofilm assessment.
• The findings support the development of enhanced disinfection strategies in endodontic retreatment.