Double-blind Clinical Trials of Probiotic Products in Oral Health.

Study Overview
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Brief Summary
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In order to evaluate the effects of supplementary probiotics and their metabolites on oral environment and oral microbiota, Taiwan FDA No. 88037803 will be used to evaluate the effects of oral probiotic candidate strains from previous research results and develop oral probiotic products.

Official Title
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Double-blind Clinical Trials of Probiotic Products in Oral Health.

Conditions
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Probiotics, Oral Health, Probiotic Metabolites

Intervention / Treatment
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* Dietary Supplement: Combined probiotics (1)
* Dietary Supplement: Combined probiotics (2)
* Dietary Supplement: Combined heat-killed probiotics (1)
* Dietary Supplement: Combined heat-killed probiotics (2)
* Dietary Supplement: Probiotic metabolites
* Dietary Supplement: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Adults between 20 to 40 years old. More than 10^5 Streptococcus mutans (CFU/mL) of saliva. Exclusion Criteria: Smoking. Subject has major illness.

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 40 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: Parallel Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Combined probiotics (1)<br>Combined Lactobacillus salivarius subsp. salicinius AP-32, Bifidobacterium animalis subsp. lactis CP-9 and Lactobacillus paracasei ET-66. | Dietary Supplement: Combined probiotics (1)<br>* Subjects used one oral combined probiotics (1) tablet each morning, noon, and evening for 4 weeks.<br>|
| Experimental: Combined probiotics (2)<br>Combined Lactobacillus salivarius subsp. salicinius AP-32, Lactobacillus plantarum LPL28 and Lactobacillus paracasei ET-66. | Dietary Supplement: Combined probiotics (2)<br>* Subjects used one oral combined probiotics (2) tablet each morning, noon, and evening for 4 weeks.<br>|
| Experimental: Combined heat-killed probiotics (1)<br>Combined heat-killed Lactobacillus salivarius subsp. salicinius AP-32, Bifidobacterium animalis subsp. lactis CP-9 and Lactobacillus paracasei ET-66. | Dietary Supplement: Combined heat-killed probiotics (1)<br>* Subjects used one oral combined heat-killed probiotics (1) tablet each morning, noon, and evening for 4 weeks.<br>|
| Experimental: Combined heat-killed probiotics (2)<br>Combined heat-killed Lactobacillus salivarius subsp. salicinius AP-32 and Lactobacillus paracasei ET-66. | Dietary Supplement: Combined heat-killed probiotics (2)<br>* Subjects used one oral combined heat-killed probiotics (2) tablet each morning, noon, and evening for 4 weeks.<br>|
| Experimental: Probiotic metabolites<br>Combined Lactobacillus salivarius subsp. salicinius AP-32, Lactobacillus plantarum LPL28 and Lactobacillus paracasei ET-66 metabolites. | Dietary Supplement: Probiotic metabolites<br>* Subjects used one oral probiotic metabolites tablet each morning, noon, and evening for 4 weeks.<br>|
| Placebo Comparator: Placebo<br>Does not contain probiotics, heat-killed probiotics and related metabolites. | Dietary Supplement: Placebo<br>* Subjects used one oral placebo tablet each morning, noon, and evening for 4 weeks.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The change from baseline colony-forming unit (CFU) of Streptococcus mutans at 4 weeks. | The plaque on the inside and outside of teeth was collected by cotton swab and putting in the microbial culture broth before and after taking the oral tablets. The microbial culture broth will be diluted and smeared on the mitis salivarius-bacitracin agar (MSBA) plate for counting the CFU of Streptococcus mutans. | 4 weeks. | 
| The change from baseline concentration of saliva immunoglobulin A (IgA) at 4 weeks. | The saliva was collected by the centrifuge tube before and after taking the oral tablets. IgA will be detected by enzyme-linked immunosorbent assay (ELISA). | 4 weeks. | 
| The change from baseline microbiota at 4 weeks. | The saliva was collected by the next-generation sequencing (NGS) saliva collection tube before and after taking the oral tablets. The changes in microbial flora will be detected by NGS. | 4 weeks. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The change from baseline CFU of total oral microbiota at 4 weeks. | The plaque on the inside and outside of teeth was collected by cotton swab and putting in the microbial culture broth before and after taking the oral tablets. The microbial culture broth will be diluted and smeared on the plate count agar (PCA) plate for counting the CFU of total oral microbiota. | 4 weeks. | 
| The change from baseline CFU of probiotics at 4 weeks. | The plaque on the inside and outside of teeth was collected by cotton swab and putting in the microbial culture broth before and after taking the oral tablets. The microbial culture broth will be diluted and smeared on the De Man, Rogosa and Sharpe agar (MRSA) plate for counting the CFU of probiotics. | 4 weeks. |