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.

• 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.

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

Data will shared in a supplementary file in the published paper