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This study aims to evaluate an endodontic treatment protocol using adjunctive steps for disinfection of teeth with primary endodontic infections.
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.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| Total bacteria analysis after 1rst- and 2nd-visit procedures | Experimental | DNA levels and activity (RNA/DNA ratio) of total bacteria after the first-visit procedures (root canal preparation with Reciproc System and NaOCl 2.5%, followed by final irrigation protocol using activation techniques: XP Endo-Finisher and ultrasonic activation) and the second-visit protocol (intracanal medication with calcium hydroxide paste, followed by an 2nd-visit root canal preparation). Additionally, the composition of the active microbiome will be assessed by Next Generation Sequencing (NGS) analysis of the root canal samples, and the success rate (apical repair) of the endodontic treatment after 1 follow-up period will be assessed by an intraoral radiograph and cone beam computed tomography (CBCT) analyses. |
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| Bacterial species analysis after root canal preparation | Other | DNA levels and activity (RNA/DNA ratio) of Bacteroidaceae sp. 272 , Cutibacterium acnes, Selenomonas spp., and Enterococcus faecalis after root canal preparation.Additionally, the success rate (apical repair) of the endodontic treatment after 1 follow-up period will be assessed by an intraoral radiograph and cone beam computed tomography (CBCT) analyses. |
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| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| Root canal preparation | Procedure | 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. |
| Measure | Description | Time Frame |
|---|---|---|
| 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 |
| Measure | Description | Time Frame |
|---|---|---|
| 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 |
| Bacterial Community Diversity |
| Measure | Description | Time Frame |
|---|---|---|
| Endodontic Success | Absence of clinical signs/ symptoms and reduction/ absence of apical radiolucency in radiologic examination | 1 year after endodontic treatment |
Inclusion Criteria:
Exclusion Criteria:
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| Name | Affiliation | Role |
|---|---|---|
| Ericka T Pinheiro, PhD | University of Sao Paulo | Principal Investigator |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| University of São Paulo | São Paulo | 05508000 | Brazil |
Samples from 20 patients were used for the analysis of total bacteria in 1st and 2nd visits. Additional 25 patients were necessary for the analysis of specific bacterial species in post-instrumentation samples (n=45 patients).
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| ID | Title | Description |
|---|---|---|
| FG000 | Total Bacteria Analysis After 1rst- and 2nd-visit Procedures | DNA levels and activity (RNA/DNA ratio) of total bacteria after the first-visit procedures (root canal preparation with Reciproc System and NaOCl 2.5%, followed by final irrigation protocol using activation techniques: XP Endo-Finisher and ultrasonic activation) and the second-visit protocol (intracanal medication with calcium hydroxide paste, followed by an 2nd-visit root canal preparation). Additionally, the composition of the active microbiome will be assessed by Next Generation Sequencing (NGS) analysis of the root canal samples, and the success rate (apical repair) of the endodontic treatment after 1 follow-up period will be assessed by an intraoral radiograph and cone beam computed tomography (CBCT) analyses. |
| FG001 | Bacterial Species Analysis After Root Canal Preparation | DNA levels and activity (RNA/DNA ratio) of Bacteroidaceae sp. 272 , Cutibacterium acnes, Selenomonas spp., and Enterococcus faecalis after root canal preparation. Additionally, the success rate (apical repair) of the endodontic treatment after 1 follow-up period will be assessed by an intraoral radiograph and cone beam computed tomography (CBCT) analyses. |
| Title | Milestones | Reasons Not Completed | ||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Overall Study |
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| ID | Title | Description |
|---|---|---|
| BG000 | Root Canal Preparation and Medication | 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. The second endodontic treatment includes the intracanal medication with calcium hydroxide paste. 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. |
| Units | Counts |
|---|---|
| Participants |
|
| Title | Description | Population Description | Parameter Type | Dispersion Type | Unit of Measure | Calculate Percentage | Denominator Units Selected | Denominators | Classes |
|---|---|---|---|---|---|---|---|---|---|
| Age, Categorical | Count of Participants |
| Type | Title | Description | Population Description | Reporting Status | Anticipated Posting Date | Parameter Type | Dispersion Type | Unit of Measure | Calculate Percentage | Time Frame | Units Analyzed | Denominator Units Selected | Arm/Group Information | Denominators | Classes | Analyses | |||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 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. | There was a lost of RNA samples from 5 cases. | Posted | Mean | Standard Deviation | RNA/DNA ratio | After 14 day treatment period |
|
0
Adverse Events were not monitored/assessed because the endodontic treatment does not cause mortality or serious adverse events.
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| ID | Title | Description | Deaths (Affected) | Deaths (At Risk) | Serious Events (Affected) | Serious Events (At Risk) | Other Events (Affected) | Other Events (At Risk) |
|---|---|---|---|---|---|---|---|---|
| EG000 | Root Canal Preparation and Medication | 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. The second endodontic treatment includes the intracanal medication with calcium hydroxide paste. Root canal samples from 20 patients were used to the analysis of total bacteria, Bacteroidaceae sp. HOT-272 and Cutibacterium acnes), while root canal samples from 25 patients were used for the analysis of total bacteria, Selenomonas spp. and Enterococcus faecalis |
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| Title | Organization | Phone | Extension | |
|---|---|---|---|---|
| Dr. Ericka Pinheiro | University of Sao Paulo | 551126488293 | erickapinheiro@usp.br |
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| Type | Includes Protocol | Includes SAP | Includes ICF | Document Label | Document Date | Document Uploaded Date | Document File Name |
|---|---|---|---|---|---|---|---|
| Prot_SAP | Yes | Yes | No | Study Protocol and Statistical Analysis Plan | Jul 20, 2020 | Jul 24, 2020 | Prot_SAP_000.pdf |
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| ID | Term |
|---|---|
| D010485 | Periapical Periodontitis |
| D001424 | Bacterial Infections |
| ID | Term |
|---|---|
| D010483 | Periapical Diseases |
| D007571 | Jaw Diseases |
| D009057 | Stomatognathic Diseases |
| D010510 | Periodontal Diseases |
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| ID | Term |
|---|---|
| D018915 | Root Canal Preparation |
| ID | Term |
|---|---|
| D012390 | Root Canal Therapy |
| D004708 | Endodontics |
| D003813 | Dentistry |
| D019246 | Tooth Preparation |
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| Intracanal medication | Procedure | The second endodontic treatment includes the intracanal medication with calcium hydroxide paste, followed by an 2nd-visit root canal preparation |
|
Composition of the active microbiome by Next Generation Sequencing (NGS) analysis of the root canal samples. |
| After 14 day treatment period |
| Participants |
|
| Age, Continuous | Mean | Full Range | years |
|
| Sex: Female, Male | Count of Participants | Participants |
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| Race (NIH/OMB) | Count of Participants | Participants |
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| Region of Enrollment | Number | participants |
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| Number of participants with infected root canals | Count of Participants | Participants |
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| OG001 | Bacterial Species Analysis After Root Canal Preparation | RNA/DNA ratio of Bacteroidaceae sp. 272 , Cutibacterium acnes, Selenomonas spp., and Enterococcus faecalis after root canal preparation (active bacteria with rRNA/DNA ≥ 1). |
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| 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). | There was a lost of RNA samples from 5 cases | Posted | Median | Full Range | DNA copies (Bacterial levels) | After 14 day treatment period |
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| Secondary | Bacterial Community Diversity | Composition of the active microbiome by Next Generation Sequencing (NGS) analysis of the root canal samples. | Root canal samples from 10 patients were included in the sequencing analysis. | Posted | Number | relative abundance of phyla (percentage) | After 14 day treatment period |
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| Other Pre-specified | Endodontic Success | Absence of clinical signs/ symptoms and reduction/ absence of apical radiolucency in radiologic examination | A subset of 30 patients had both intra-oral radiograph and cone beam computed tomography (CBCT) scans for analysis of the treatment outcome. After 1 year, 8 patients were lost during the follow-up period and 22 patients were analysed. | Posted | Count of Participants | Participants | 1 year after endodontic treatment |
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| 0 |
| 0 |
| 0 |
| 0 |
| 0 |
| 0 |
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| D009059 |
| Mouth Diseases |
| D010518 | Periodontitis |
| D001423 | Bacterial Infections and Mycoses |
| D007239 | Infections |
| Total bacteria after root canal Preparation (S2) |
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| Total bacteria after adjunctive procedures (S3) |
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| Total bacteria after intracanal medication (S4) |
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| Total bacteria after 2nd-visit preparation (S5) |
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| Bacteroidaceae sp. 272 at S1 |
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| Bacteroidaceae sp. 272 at S2 |
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| Cutibacterium acnes at S1 |
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| Cutibacterium acnes at S2 |
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| Selenomonas spp. at S1 |
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| Selenomonas spp. at S2 |
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| Enterococcus faecalis at S1 |
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| Enterococcus faecalis at S2 |
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| Title | Measurements |
|---|---|
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| Proteobacteria |
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| Actinobacteria |
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| Spirochaetes |
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