Adjunctive Steps for Disinfection of Teeth With Primary Endodontic Infections

Bacterial Infections Clinical Trial

Official title:

Analysis of Metabolically Active Bacteria After Adjunctive Steps for Disinfection of Teeth With Primary Endodontic Infections: RNA- and DNA-based Molecular Study

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT03537664
• Other study ID #: FAPESP 2016/15473-0
• Status: Completed
• Phase: N/A
• Start date: August 4, 2017
• Est. completion date: May 29, 2020

Study information

• Verified date: September 2020
• Source: University of Sao Paulo
• Contact: n/a
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial Summary

This study aims to evaluate an endodontic treatment protocol using adjunctive steps for disinfection of teeth with primary endodontic infections.

Description:

The success of endodontic treatment in teeth with necrotic pulp and apical periodontitis depends on the reduction of viable microorganisms of root canals by endodontic disinfection procedures. This study aims to evaluate by molecular methods the effect of different disinfection procedures on reduction, diversity and microbial metabolic activity. Patients with necrotic pulps and apical periodontitis in single-rooted teeth will be selected. Microbiological samples will be taken from root canals after access cavity (S1), after chemomechanical preparation with Reciproc System and 2.5% NaOCl (S2), after irrigant activation using the XP-Endo Finisher instrument (S3a) and ultrasonic activation (S3b); and after intracanal medication with calcium hydroxide for 14 days (S4), followed by 2nd-visit root canal preparation (S5). DNA and RNA will be extracted from root canal samples, and complementary DNA (cDNA) synthetized using reverse transcription reaction. The effect of treatment protocols on total microbiota levels will be determined by DNA-based quantitative polymerase chain reaction (qPCR) using universal primers for Bacteria domain. RNA of root canal samples will be used to determine the diversity of metabolically active microbiota by Reverse Transcriptase Reaction (RT), followed by amplification (PCR) and high throughput sequencing of the hypervariable region V4-V5 of 16S rRNA gene. The metabolic activity of the most prevalent species / taxa will be calculated by rRNA- and DNA-based molecular methods. Ratios between rRNA and DNA levels will be calculated to search for active bacteria (rRNA/DNA ≥ 1) in root canal samples. Data will be analyzed by statistical tests, with 5% significance level.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 45
• Est. completion date: May 29, 2020
• Est. primary completion date: May 29, 2020
• Accepts healthy volunteers: No
• Gender: All
• Age group: 18 Years to 65 Years

Eligibility

Inclusion Criteria:

• Teeth with necrotic pulps and asymptomatic apical periodontitis

Exclusion Criteria:

• patients who had received antibiotics during the previous 3 months or had any general disease,

• teeth that could not be properly isolated with rubber dam,

• non-restored teeth,

• periodontal pockets depths greater than 4 mm,

• previous endodontic treatment,

• open apex,

• crown/root fracture

• root resorption or calcifications.

Related Conditions & MeSH terms

• Bacterial Infections
• Communicable Diseases
• Infection
• Periapical Periodontitis
• Periodontitis

Intervention

Procedure:
• Root canal preparation
First endodontic treatment session includes the root canal preparation with Reciproc System and NaOCl 2.5%, followed by final irrigation protocol using activation techniques: XP Endo-Finisher and ultrasonic activation.
• Intracanal medication
The second endodontic treatment includes the intracanal medication with calcium hydroxide paste, followed by an 2nd-visit root canal preparation

Locations

Country	Name	City	State
Brazil	University of São Paulo	São Paulo

Sponsors (1)

Lead Sponsor	Collaborator
University of Sao Paulo

Country where clinical trial is conducted

Brazil, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Other	Endodontic Success	Absence of clinical signs/ symptoms and reduction/ absence of apical radiolucency in radiologic examination	1 year after endodontic treatment
Primary	Metabolic Activity of Persistent Bacteria	The metabolic activity (rRNA/DNA ratio) of total bacteria, Bacteroidaceae sp. 272, Cutibacterium acnes, Selenomonas spp. and Enterococcus faecalis in root canal samples taken before and after endodontic procedures.	After 14 day treatment period
Secondary	Bacterial Levels	Quantitative data of total bacteria, Bacteroidaceae sp. HOT-272, Cutibacterium acnes, Selenomonas spp. and Enterococcus faecalis determined by DNA-based qPCR (quantitative polymerase chain reaction).	After 14 day treatment period
Secondary	Bacterial Community Diversity	Composition of the active microbiome by Next Generation Sequencing (NGS) analysis of the root canal samples.	After 14 day treatment period