Dental Caries, Microbiota, Oral Bacterial Infection
Conditions
Brief summary
caries-free (CF) individuals caries-active (CA) individuals (DMFT ≥ 6) will be recruited. Subjects will be instructed to brush their teeth twice daily for 3 min using Colgate® Sensitive Pro-Relief® toothpaste (containing 8% arginine and 1450 ppm NaF) for 2 weeks. Supra- and subgingival plaque, saliva, and in situ plaque samples will be collected before and after the treatment for laboratory analyses.
Detailed description
21 caries-free (CF) individuals with no clinical evidence of caries experience \[decayed, missing and filled teeth (DMFT) = 0\] and 21 caries-active (CA) individuals (DMFT ≥ 6) will be recruited in the clinical trial. The exclusion criteria are: smoker or former smoker, presence of any systemic disease that could alter the production or composition of saliva, treatment with antibiotics, steroids or any medication known to cause dry mouth in the last 3 months, and presence of dental prostheses or orthodontic devices. Subjects will be instructed to brush their teeth twice daily for 3 min using Colgate® Sensitive Pro-Relief® toothpaste (containing 8% arginine and 1450 ppm NaF) for 2 weeks. Volunteers are asked to refrain from brushing and flossing their teeth, eating, and drinking anything other than water for 12 h prior to sample collection visits at the end of both phases. Supra- and subgingival plaque (15 subjects in each group), saliva (21 subjects in each group), and in situ plaque samples (3 subjects in each group) will be collected before and after the treatment. Scraped plaque and saliva samples will be immediately transferred to and dispersed in sterile micro-centrifuge tubes containing 1×phosphate-buffered saline (PBS), and stored at -80 °C until needed for analysis. The in situ plaque samples will be transferred into 4% paraformaldehyde, kept under 4 °C for 16 h, and then stored in 50% (v/v) ethanol at -20 °C.
Interventions
OTHERColgate® Sensitive Pro-Relief® toothpaste
Subjects will be instructed to brush their teeth twice daily for 3 min using Colgate® Sensitive Pro-Relief® toothpaste for 2 weeks.
Sponsors
Sichuan University
Study design
Allocation
NON_RANDOMIZED
Intervention model
PARALLEL
Primary purpose
TREATMENT
Masking
NONE
Eligibility
Sex/Gender
ALL
Healthy volunteers
Yes
Inclusion criteria
* caries-free individuals with no clinical evidence of caries experience \[decayed, missing and filled teeth (DMFT) = 0\] and caries-active individuals (DMFT ≥ 6) were recruited
Exclusion criteria
* smoker or former smoker, presence of any systemic disease that could alter the production or composition of saliva, treatment with antibiotics, steroids or any medication known to cause dry mouth in the last 3 months, and presence of dental prostheses or orthodontic devices.
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | baseline , 2 week | Saliva and dental plaque samples will be collected, and the abundance of specific oral microbes such as Streptococcus mutans, Streptococcus sanguinis will be quantified by microbial 16S rRNA sequencing. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline ,2 week | Saliva and dental plaque samples will be collected and microbial enzyme activity will be quantified in the lab. Measures include relative enzyme activity of arginine deiminase, urease and lactase dehydrogenase as determined by standard laboratory enzymatic assay protocols. |
Participant flow
Recruitment details
The first recruitment took place on Oct. 2013, in the Department of Operative Dentistry and Endodontics, West China Hospital of Stomatology. 15 CF and 15 CA individuals were recruited by the end of Nov. 2013. The second recruitment took place on May in the same location. 2014. 6 CF and 6 CA individuals were recruited by the end of Jun. 2014.
Participants by arm
| Arm | Count |
|---|---|
| Caries-free Subjects Subjects will use Colgate® Sensitive Pro-Relief® toothpaste for 2 weeks,.
Colgate® Sensitive Pro-Relief® toothpaste: Subjects will be instructed to brush their teeth twice daily for 3 min using Colgate® Sensitive Pro-Relief® toothpaste for 2 weeks. | 21 |
| Caries-active Subjects Subjects will use Colgate® Sensitive Pro-Relief® toothpaste for 2 weeks,.
Colgate® Sensitive Pro-Relief® toothpaste: Subjects will be instructed to brush their teeth twice daily for 3 min using Colgate® Sensitive Pro-Relief® toothpaste for 2 weeks. | 21 |
| Total | 42 |
Baseline characteristics
| Characteristic | Caries-active Subjects | Caries-free Subjects | Total |
|---|---|---|---|
| Age, Categorical <=18 years | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical >=65 years | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical Between 18 and 65 years | 21 Participants | 21 Participants | 42 Participants |
| Age, Continuous | 23 years | 23 years | 23 years |
| Region of Enrollment China | 21 Participants | 21 Participants | 42 Participants |
| Sex: Female, Male Female | 17 Participants | 11 Participants | 28 Participants |
| Sex: Female, Male Male | 4 Participants | 10 Participants | 14 Participants |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk |
|---|---|---|
| deaths Total, all-cause mortality | 0 / 21 | 0 / 21 |
| other Total, other adverse events | 0 / 21 | 0 / 21 |
| serious Total, serious adverse events | 0 / 21 | 0 / 21 |
Outcome results
Primary
The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing
Saliva and dental plaque samples will be collected, and the abundance of specific oral microbes such as Streptococcus mutans, Streptococcus sanguinis will be quantified by microbial 16S rRNA sequencing.
Time frame: baseline , 2 week
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: sm in supragingival plaque | 0.00756 percentage of bacteria | Standard Deviation 0.00726 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: sm in subgingival plaque | 0.00629 percentage of bacteria | Standard Deviation 0.00835 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: sm in saliva | 0.00608 percentage of bacteria | Standard Deviation 0.00575 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: ss in supragingival plaque | 1.007766667 percentage of bacteria | Standard Deviation 1.168793223 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: ss in subgingival plaque | 2.064646667 percentage of bacteria | Standard Deviation 2.595662011 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: ss in saliva | 0.812706667 percentage of bacteria | Standard Deviation 1.040344461 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: sm in supragingival plaque | 0.003406667 percentage of bacteria | Standard Deviation 0.003320384 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: sm in subgingival plaque | 0.0033 percentage of bacteria | Standard Deviation 0.006143173 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: sm in saliva | 0.002493333 percentage of bacteria | Standard Deviation 0.004110347 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: ss in supragingival plaque | 1.655106667 percentage of bacteria | Standard Deviation 1.274066247 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: ss in subgingival plaque | 4.588546667 percentage of bacteria | Standard Deviation 4.92217214 |
| Caries-free Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: ss in saliva | 1.3109 percentage of bacteria | Standard Deviation 0.885269549 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: ss in subgingival plaque | 3.232233333 percentage of bacteria | Standard Deviation 3.336652867 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: sm in supragingival plaque | 0.14636 percentage of bacteria | Standard Deviation 0.23221 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: sm in supragingival plaque | 0.009526667 percentage of bacteria | Standard Deviation 0.011458775 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: sm in subgingival plaque | 0.11956 percentage of bacteria | Standard Deviation 0.20686 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: ss in supragingival plaque | 3.21486 percentage of bacteria | Standard Deviation 3.436273455 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: sm in saliva | 0.01314 percentage of bacteria | Standard Deviation 0.01272 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: sm in subgingival plaque | 0.00284 percentage of bacteria | Standard Deviation 0.005101792 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: ss in supragingival plaque | 0.521506667 percentage of bacteria | Standard Deviation 0.511640082 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: ss in saliva | 2.040286667 percentage of bacteria | Standard Deviation 3.369773515 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: ss in subgingival plaque | 1.539913333 percentage of bacteria | Standard Deviation 1.369851357 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | 2 week: sm in saliva | 0.003006667 percentage of bacteria | Standard Deviation 0.002304488 |
| Caries-active Subjects | The Change of Abundance of Specific Oral Microbes After Intervention as Assessed by 16S rRNA Sequencing | Baseline: ss in saliva | 0.18324 percentage of bacteria | Standard Deviation 0.199811864 |
Secondary
The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays
Saliva and dental plaque samples will be collected and microbial enzyme activity will be quantified in the lab. Measures include relative enzyme activity of arginine deiminase, urease and lactase dehydrogenase as determined by standard laboratory enzymatic assay protocols.
Time frame: baseline ,2 week
Population: Among the 21 participants in each group，we only measured the enzyme activity from randomly choosed 15 participants from each group.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: urease activity in subgingival plaque | 0.71840 enzyme activity (μmol/min/g) | Standard Deviation 0.0877 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: ADS activity in supragingival plaque | 1.0771 enzyme activity (μmol/min/g) | Standard Deviation 0.23754 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: ADS activity in saliva plaque | 0.75870 enzyme activity (μmol/min/g) | Standard Deviation 0.09211 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: ADS activity in subgingival plaque | 0.74650 enzyme activity (μmol/min/g) | Standard Deviation 0.10275 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: urease activity in saliva | 0.72900 enzyme activity (μmol/min/g) | Standard Deviation 0.6621 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: ADS activity in saliva | 0.77410 enzyme activity (μmol/min/g) | Standard Deviation 0.11136 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: ADS activity in subgingival plaque | 0.63150 enzyme activity (μmol/min/g) | Standard Deviation 0.08604 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: LDH activity in supragingival plaque | 8.859050 enzyme activity (μmol/min/g) | Standard Deviation 0.6648584 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: urease activity in subgingival plaque | 0.73330 enzyme activity (μmol/min/g) | Standard Deviation 0.06165 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: urease activity in supragingival plaque | 0.6795 enzyme activity (μmol/min/g) | Standard Deviation 0.11518 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: urease activity in saliva | 0.73240 enzyme activity (μmol/min/g) | Standard Deviation 0.05464 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: LDH activity in subgingival plaque | 7.267220 enzyme activity (μmol/min/g) | Standard Deviation 0.2555969 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: LDH activity in supragingival plaque | 11.347060 enzyme activity (μmol/min/g) | Standard Deviation 0.9240766 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: ADS activity in supragingival plaque | 0.8945 enzyme activity (μmol/min/g) | Standard Deviation 0.1331 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: LDH activity in subgingival plaque | 8.664124 enzyme activity (μmol/min/g) | Standard Deviation 0.8184378 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: LDH activity in saliva | 7.508787 enzyme activity (μmol/min/g) | Standard Deviation 0.8707994 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: LDH activity in saliva | 9.496731 enzyme activity (μmol/min/g) | Standard Deviation 0.1797688 |
| Caries-free Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: urease activity in supragingival plaque | 0.5871 enzyme activity (μmol/min/g) | Standard Deviation 0.0797 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: LDH activity in saliva | 15.268450 enzyme activity (μmol/min/g) | Standard Deviation 0.3687766 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: ADS activity in supragingival plaque | 0.3653 enzyme activity (μmol/min/g) | Standard Deviation 0.06108 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: ADS activity in subgingival plaque | 0.29110 enzyme activity (μmol/min/g) | Standard Deviation 0.04347 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: ADS activity in saliva plaque | 0.23470 enzyme activity (μmol/min/g) | Standard Deviation 0.03328 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: urease activity in supragingival plaque | 0.3207 enzyme activity (μmol/min/g) | Standard Deviation 0.06317 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: urease activity in subgingival plaque | 0.34630 enzyme activity (μmol/min/g) | Standard Deviation 0.04611 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: urease activity in saliva | 0.17920 enzyme activity (μmol/min/g) | Standard Deviation 0.03047 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: LDH activity in supragingival plaque | 20.965090 enzyme activity (μmol/min/g) | Standard Deviation 1.5499 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: LDH activity in subgingival plaque | 17.576110 enzyme activity (μmol/min/g) | Standard Deviation 0.8875 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | baseline: LDH activity in saliva | 16.053870 enzyme activity (μmol/min/g) | Standard Deviation 2.36149 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: ADS activity in supragingival plaque | 0.8418 enzyme activity (μmol/min/g) | Standard Deviation 0.13711 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: ADS activity in subgingival plaque | 0.73600 enzyme activity (μmol/min/g) | Standard Deviation 0.07218 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: ADS activity in saliva | 0.34850 enzyme activity (μmol/min/g) | Standard Deviation 0.04391 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: urease activity in supragingival plaque | 0.6508 enzyme activity (μmol/min/g) | Standard Deviation 0.10083 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: urease activity in subgingival plaque | 0.69680 enzyme activity (μmol/min/g) | Standard Deviation 0.08427 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: urease activity in saliva | 0.25260 enzyme activity (μmol/min/g) | Standard Deviation 0.0456 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: LDH activity in supragingival plaque | 14.096910 enzyme activity (μmol/min/g) | Standard Deviation 0.3565668 |
| Caries-active Subjects | The Change of Relative Activities of Microbial Enzymes After Intervention as Determined by Laboratory Enzymatic Assays | 2 week: LDH activity in subgingival plaque | 11.649030 enzyme activity (μmol/min/g) | Standard Deviation 1.13521 |