Maintaining Oral Health With Bio-products

Study Overview
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Brief Summary
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This study will follow a randomised, double-blind, cross-over design. Participants will use mouth rinses containing plant extracts and/or fluoride, as well as a mouth rinse containing only water (placebo). They will use oral appliances containing sterilized enamel and dentine slabs, which will be later collected for analyses. The cross-over design will allow participants to use all rinses, allowing for a comparison between the rinses within each individual.

Detailed Description
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This study will be made in three in situ parts. Participants will wear an oral appliance containing sterilized enamel and dentine slabs. During this experimental phase, they will use the designated mouth rinse. After this period, the investigators will remove the enamel and dentine slabs, and will also collect salivary pellicle and any bacterial biofilm forming on the participant's teeth. Participants will also donate saliva for proteomic analysis (protein profile) to verify if there are any differences in proteins in saliva to proteins in the pellicle. Throughout the whole study, all participants will use standardized oral hygiene products. The experimental mouth rinses contain plant extracts that could interact with salivary proteins and influence the salivary pellicle and oral biofilm, thus bringing positive results against tooth (enamel and dentine) demineralisation.

Official Title
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Maintaining Oral Health With Bio-products: a Randomized, Cross-over, in Situ Study

Conditions
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Dental Caries, Dental Erosion

Intervention / Treatment
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* Other: Mouth rinse 1
* Other: Mouth rinse 2
* Other: Mouth rinse 3
* Other: Mouth rinse 4
* Other: Mouth rinse 5
* Other: Mouth rinse 6
* Other: Mouth rinse 7
* Other: Mouth rinse 8
* Other: Mouth rinse 9


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Participants will be healthy young adults of both sexes, with normal salivary flow, with no cavitated caries lesions, no gingivitis or periodontal disease, no severe erosive tooth wear, and no other serious general or oral conditions. Participants will have signed an informed consent form. Exclusion Criteria: Presenting oral diseases or conditions, such as untreated cavitated caries lesions, gingivitis or periodontitis, severe erosive tooth wear, etc. Participants who are under orthodontic treatment (wearing braces), Contraindications to the mouth rinses under study, e.g. known hypersensitivity or allergy to the investigational product, Participants using drugs that cause hyposalivation, Women who are pregnant or breast feeding, Intention to become pregnant during the course of the study, Individuals suffering from clinically significant concomitant disease states (e.g., renal failure, hepatic dysfunction, cardiovascular disease, etc.), Inability to follow the procedures of the study, e.g. due to language problems, psychological disorders, dementia, etc. of the participant, Participation in another study with investigational drug within the 30 days preceding and during the present study, Enrolment of the investigator, his/her family members, employees and other dependent persons, Specific exclusions for the disease under study, Specific concomitant therapy washout requirements prior to and/or during study participation

Ages Eligible for Study
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Minimum Age: 18 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: Prevention
Allocation: Randomized
Intervention Model: Crossover Assignment
Interventional Model Description: Cross-over study, where all participants will use all treatments. The treatment order will be set at random.
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: Placebo mouth rinse<br>Placebo: Deionized water (serving as negative control) | Other: Mouth rinse 1<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Water;|
| Active Comparator: Elmex mouth rinse<br>Commercial mouth rinse used as gold standard in erosion studies: elmex® Erosion Protection solution (which contains 800 ppm Sn2+, as SnCl2, and 500 ppm F-, as NaF and AmF) | Other: Mouth rinse 2<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Elmex Erosion Protection mouth rinse;|
| Active Comparator: Fluoride mouth rinse<br>Fluoride solution similar to many other commercial mouth rinses containing sodium fluoride (NaF at 500 ppm F-) | Other: Mouth rinse 3<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Fluoride mouth rinse;|
| Experimental: Plant extract A<br>Plant Extract A | Other: Mouth rinse 4<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Plant extract A;|
| Experimental: Plant extract A with fluoride<br>Plant Extract A + Fluoride | Other: Mouth rinse 5<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Plant extract A and fluoride;|
| Experimental: Plant extract B<br>Plant extract B | Other: Mouth rinse 6<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Plant extract B;|
| Experimental: Plant extract B with fluoride<br>Plant Extract B + Fluoride | Other: Mouth rinse 7<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Plant extract B and fluoride;|
| Experimental: Plant extract C<br>Plant extract C | Other: Mouth rinse 8<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Plant extract C;|
| Experimental: Plant extract C with fluoride<br>Plant Extract C + Fluoride | Other: Mouth rinse 9<br>* Participants will use 10mL of the respective mouth rinse, swish in the mouth for 30 seconds and spit it out. This will be made twice a day, for a total of 5 days.<br>* Other names: Plant extract C and fluoride;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Enamel mineral loss | Hardness of enamel slabs (Knoop hardness) | End of the study arm, expected to be after 1 week of using the mouth rinse | 
| Dentine mineral loss | Profilometry of dentine specimens (step height in µm) | End of the study arm, expected to be after 1 week of using the mouth rinse | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Identification of which proteins are present in the participants' saliva | Proteomic analyses of the saliva (list of proteins present) | End of the study arm, expected to be after 1 week of using the mouth rinse | 
| Identification of which proteins are present in the participants' salivary pellicle | Proteomic analyses of the salivary pellicle (list of proteins present) | End of the study arm, expected to be after 1 week of using the mouth rinse | 
| Thickness of the salivary pellicle | Measured using TEM (mineralization measured in Delta Z) | End of the study arm, expected to be after 1 week of using the mouth rinse | 
| pH of the participants' biofilm | Measured in vivo after using the mouth rinses (pH) | End of the study arm in part 3, expected to be after 1 day of using the mouth rinse | 
| Identification of the bacteria in the biofilm | Measured after collecting the biofilm (list of bacterial species) | End of the study arm in part 3, expected to be after 1 day of using the mouth rinse | 
| How many bacteria are adhered to the biofilm | Measured after collecting the biofilm (bacteria quantity) | End of the study arm in part 3, expected to be after 1 day of using the mouth rinse |