Results of a Pilot Trial Assessing the Effects of Proper Oral Hygiene and a Probiotic Dietary Supplement on Oral Health in Volunteers with Oral Malodor
Abstract
:1. Introduction
2. Materials and Methods
2.1. Participants and Inclusion/Exclusion Criteria
- Individuals that were 18–55 years old;
- Individuals with a persistent malodor (≥3 months);
- Individuals with organoleptic scores 2–5.
- Individuals who were less than 18 years of age or over 55 years of age at enrollment;
- Individuals who had minor persistent malodor scores (scores 0–1);
- Individuals who had dentures;
- Individuals who reported a professional dental cleaning within 1 month of randomization;
- Individuals who reported any gastrointestinal (Reflux esophagitis, gastritis, ulcer, hiatal hernia, Zencker diverticulum, achalasia) and ENT disorders (pharyngitis, tonsillitis, sinusitis, bronchiectasis);
- Female individuals who reported being pregnant or intending to become pregnant in the next two months;
- Individuals who reported the use of immunosuppressive therapy;
- Individuals who reported immune deficiencies.
2.2. Study Design
2.2.1. Office Visits Activities
2.2.2. At-Home Malodor Measurements
2.3. Probiotic and Placebo Lozenges
2.4. Outcomes
- Salivary inflammatory/anti-inflammatory cytokine levels;
- The DNA levels of the probiotic strains in the tongue coatings by Quantitative PCR (qPCR) analysis;
- Salivary pH.
2.5. Probiotic DNA Levels Analysis
- L. plantarum KABP051 forward 5′-AGGGTTGGACGAGACAA-3′;
- L. plantarum KABP051 reverse 5′-CCAAACTCATCGGACCTATTC-3′;
- L. brevis KABP052 forward 5′-TTAGCGTCGTTAGTTGTTATAGG-3′;
- L. brevis KABP052 reverse 5′-ACGTTCTTGGTCATCGTAATC-3′;
- P. acidilactici KABP053 forward 5′-AGCTACAGCTACCCAATCT-3′;
- P. acidilactici KABP053 reverse 5′-GTCGGTTGTGTCCATTAAGT-3′.
- Forward Primer BACT1369F CGGTGAATACGTTCYCGG;
- Reverse Primer PROK1492R GGWTACCTTGTTACGACTT.
2.6. Salivary Cytokines
2.7. Salivary pH
2.8. Statistical Analysis
2.8.1. Summary Statistics
2.8.2. Malodor, Tongue Coating, and Cytokine Concentration Analysis
2.8.3. PCR Data Analysis
3. Results
3.1. General Characteristics of Participants
3.2. Effect of Proper Dental Hygiene and Probiotic Use on Malodor
3.3. Effect of Proper Dental Hygiene and Probiotic Use on Tongue Coating
3.4. Effect of Proper Dental Hygiene and Probiotic Use on Salivary pH
3.5. Effect of Proper Dental Hygiene and Probiotic Use on the Probiotics’ DNA Levels in the Tongue Coating
3.6. Effect of Proper Dental Hygiene and Probiotic Use on Salivary Cytokines
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Analyte | Copies of DNA per μL of the Extracted Sample |
---|---|
L. plantarum | X = 10^((CT-42.994)/−2.7788) |
L. brevis | X = 10^((CT-40.775)/−1.7513) |
P. acidilactici | X = 10^((CT-48.583)/−3.91) |
16S | X = 10^((CT-38.939)/−2.4257) |
Placebo Cohort, n = 13 | Probiotic Cohort, n = 18 | Total Number of Participants (%) n = 31 | |
---|---|---|---|
Race/ethnicity, n (%) | |||
White-Non-Hispanic | 9 (69%) | 9 (50%) | 18 (58%) |
Asian | 4 (31%) | 4 (22%) | 8 (26%) |
White-Hispanic | 0 (0%) | 4 (22%) | 4 (13%) |
Black, Non-Hispanic | 0 (0%) | 1 (6%) | 1 (3%) |
Gender, n (%) | |||
Female | 7 (54%) | 11 (61%) | 16 (52%) |
Male | 6 (46%) | 7 (39%) | 13 (48%) |
Age | |||
Mean + SD | 28.1 + 3.5 | 27.3 + 4.9 | N/A * |
Placebo Cohort, n = 13 | Probiotic Cohort, n = 18 | |
---|---|---|
Visit 1 (baseline) | ||
Mean + SE | 0.75 + 0.09 | 0.72 + 0.12 |
Visit 2 (30 days post lozenge’s use) | ||
Mean + SE | 0.39 + 0.12 | 0.40 + 0.09 |
Visit 3 (30 days follow-up) | ||
Mean + SE | 0.42 + 0.13 | 0.59 + 0.08 |
Placebo Cohort, n = 13 | Probiotic Cohort, n = 18 | |
---|---|---|
Mean + SE | 6.83 + 0.07 | 6.76 + 0.08 |
Mean + SE | 6.75 + 0.06 | 6.81 + 0.06 |
Mean + SE | 6.73 +0.06 | 6.66 + 0.06 |
Cohorts | L. plantarum | L. brevis | P. acidilactici |
---|---|---|---|
Placebo, n = 13 | 5 | 2 | 9 |
Probiotics, n = 18 | 7 | 3 | 12 |
Placebo Cohort, n = 13 | Probiotic Cohort, n = 18 | p Value | |
---|---|---|---|
L. plantarum | |||
Baseline | |||
Mean + SE | 197,482 + 195,685 | 999 + 681 | 1 |
30 days intervention | |||
Mean + SE | 240 + 257 * | 10,681 + 7883 † | 0.047 |
30 days follow-up | |||
Mean + SE | 5081 + 3775 * | 412 + 393 † | 0.466 |
L. brevis | |||
Baseline | |||
Mean + SE | 7 + 4 | 737 + 709 | 1 |
30 days intervention | |||
Mean + SE | 14 + 9 * | 26,708 + 26,335 † | 0.304 |
30 days follow-up | |||
Mean + SE | 77,301 + 77,300 * | 125 + 90 † | 0.079 |
P. acidilactici | |||
Baseline | |||
Mean + SE | 839,724 + 795,780 | 141,452 + 125,536 | 1 |
30 days intervention | |||
Mean + SE | 2933 + 2917 * | 36,180 + 288,769 † | 0.584 |
30 days follow-up | |||
Mean + SE | 807,375 + 470,904 * | 21,348 + 20,908 † | 0.823 |
Cytokines | Placebo Cohort, n = 13 | Probiotic Cohort, n = 18 | p Values |
---|---|---|---|
IL-1β (pg/mL) | |||
Baseline | |||
Mean + SE, pg/mL | 298.35 + 93.37 | 371.69 + 115.81 | 0.564 |
30 days intervention | |||
Mean + SE, pg/mL | 248.17 + 52.84 * | 320.06 + 88.23 * | 0.554 |
30 days follow-up | |||
Mean + SE, pg/mL | 159.80 + 43.09 * | 271.75 + 79.69 * | 0.1044 |
IL-8 (pg/mL) | |||
Baseline | |||
Mean + SE, pg/mL | 1371.30 + 308.07 | 2295.15 + 386.26 | 0.081 |
30 days intervention | |||
Mean + SE, pg/mL | 1531.58 + 238.46 * | 2103.40 + 378.26 * | 0.671 |
30 days follow-up | |||
Mean + SE, pg/mL | 1253.64 + 287.87 * | 1602.88 + 421.08 * | 0.755 |
TNF-α (pg/mL) | |||
Baseline | |||
Mean + SE, pg/mL | 5.15 + 1.17 | 10.88 + 7.85 | 0.086 |
30 days intervention | |||
Mean + SE, pg/mL | 6.11 + 1.18 * | 11.66 + 2.87 * | 0.6884 |
30 days follow-up | |||
Mean + SE, pg/mL | 5.21 + 1.04 * | 7.33 + 1.93 * | 0.989 |
IL-6 (pg/mL) | |||
Baseline | |||
Mean + SE, pg/mL | 5.46 + 0.79 | 32.34 + 21.82 | 0.271 |
30 days intervention | |||
Mean + SE, pg/mL | 8.05 + 1.93 * | 39.96 + 27.94 * | 0.9207 |
30 days follow-up | |||
Mean + SE, pg/mL | 6.35 + 1.22 * | 9.07 + 2.59 * | 0.9726 |
IL-10 (pg/mL) | |||
Baseline | |||
Mean + SE, pg/mL | 1.61 + 0.56 | 2.76 + 0.73 | 0.224 |
30 days intervention | |||
Mean + SE, pg/mL | 1.40 + 0.19 * | 3.16 + 0.84 * | 0.772 |
30 days follow-up | |||
Mean + SE, pg/mL | 1.50 + 0.49 * | 2.02 + 0.62 * | 0.863 |
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Share and Cite
Enioutina, E.Y.; Keddington, R.J.; Hauck, K.G.; Chavez, A.; Clifford, J.J.; Cao, T.; Smith, B.; Job, K.M.; Balch, A. Results of a Pilot Trial Assessing the Effects of Proper Oral Hygiene and a Probiotic Dietary Supplement on Oral Health in Volunteers with Oral Malodor. Microorganisms 2024, 12, 1821. https://doi.org/10.3390/microorganisms12091821
Enioutina EY, Keddington RJ, Hauck KG, Chavez A, Clifford JJ, Cao T, Smith B, Job KM, Balch A. Results of a Pilot Trial Assessing the Effects of Proper Oral Hygiene and a Probiotic Dietary Supplement on Oral Health in Volunteers with Oral Malodor. Microorganisms. 2024; 12(9):1821. https://doi.org/10.3390/microorganisms12091821
Chicago/Turabian StyleEnioutina, Elena Y., R. James Keddington, Kurtis G. Hauck, Amarina Chavez, Jeffrey J. Clifford, Thy (April) Cao, Bryce Smith, Kathleen M. Job, and Alfred Balch. 2024. "Results of a Pilot Trial Assessing the Effects of Proper Oral Hygiene and a Probiotic Dietary Supplement on Oral Health in Volunteers with Oral Malodor" Microorganisms 12, no. 9: 1821. https://doi.org/10.3390/microorganisms12091821