Comparison of Biofilm Characteristics Between an in Situ Biofilm Model of Supragingival Plaque and Natural Teeth

Study Overview
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Brief Summary
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The purpose of this study is to determine in situ the bacterial viability, thickness and bacterial diversity of a biofilm formed on different substrates in comparison to a naturally tooth-formed oral biofilm. In addition, it will be evaluated whether the restraint of oral hygiene measures may influence the development of the PL-Biofilm.

Detailed Description
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To calculate an a priori sample size, the following statistical criteria were established: an effect size of 0.35, an alpha error of 0.05 and a statistical power of 87%. Assuming these criteria and using the repeated measures ANOVA test, a sample size of 15 subjects was required. Bacterial viability and thickness will be analyse by Confocal Laser Scanning Microscope and Live/Dead BackLight staining solution. Bacterial diversity will be analyse by Pyrosequencing the 16S DNAr gene.

Official Title
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Comparison of Biofilm Characteristics Between an in Situ Biofilm Model of Supragingival Plaque and Natural Teeth

Conditions
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Biofilm, Dental Plaque

Intervention / Treatment
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* Other: Toothbrushing


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: systemically healthy good oral health status Exclusion Criteria: smoker or former smoker, presence of dental prostheses or orthodontic devices, antibiotic treatment or routine use of oral antiseptics in the previous 3 months, presence of any systemic disease that could alter the production or composition of saliva

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 45 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| No Intervention: No toothbrushing during experiment and Pyrosequencing<br>Volunteers will not brush their teeth during 48 hours. Dental plaque samples will be analyse by CLSM (confocal Laser scanning microscope) and 16S DNA pyrosequencing. | |
| No Intervention: No toothbrushing during experiment<br>Volunteers will not brush their teeth during 48 hours. Dental plaque samples will be analyse by CLSM (confocal Laser scanning microscope). | |
| Active Comparator: Toothbrushing during experiment<br>48hours-Dental plaque samples will be analyse by CLSM (confocal Laser scanning microscope). | Other: Toothbrushing<br>* Normal toothbrushing without toothpaste or antiseptic mouthwash.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Bacterial viability | It will be measured by analysis of images taken by Confocal laser Scanning Microscope | Bacterial viability measured in proportion of viable/no viable bacteria after 48 hours of exposition to oral cavity | 
| Bacterial diversity | It will be measured by 16S rDNA gene pyrosequencing and posterior bioinformatic analysis. | Bacterial diversity measured by microbial estimators of abundance and richness after 48 hours of exposition to oral cavity | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Biofilm thickness | It will be measured by analysing sectional images taken by Confocal Laser Scanning Microscope. | Oral biofilm thickness measured by analysis of sectional images after 48 hours of exposition to oral cavity | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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pyrosequencing, 16S DNA gene, Confocal Laser Scanning Microscope, bacterial viability, bacterial diversity