Nitrate Mouthwash for Oral Microbiome

Overseen By Leigha Rock

Age: 18 - 65

Sex: Any

Trial Phase: Academic

Sponsor: Dalhousie University

Must not be taking: Antibiotics, Probiotics, Antimicrobials

Disqualifiers: Uncontrolled diseases, Oral conditions, others

Approved in 3 Jurisdictions This treatment is already approved in other countries

Compensation: Varies

Number of Visits: Unknown

Duration: 2 weeks

30 Participants Needed

Trial Summary

Will I have to stop taking my current medications?

The trial requires that you stop taking prescription medications, antibiotics, probiotics, or antimicrobial mouthrinses to participate.

What data supports the effectiveness of the treatment Nitrate Mouthwash for Oral Microbiome?

Research shows that a nitrate-rich diet, like beetroot juice, can reduce gingival inflammation and has antibacterial effects against harmful oral bacteria. Additionally, nitrate-reducing bacteria in the mouth can help improve oral health and may also benefit overall health by increasing nitric oxide levels, which can help with conditions like high blood pressure. ^{1 2 3 4 5}

Is nitrate mouthwash, or similar treatments like beetroot juice, safe for humans?

Research on nitrate-rich diets, including beetroot juice, suggests they are generally safe for humans and may even have beneficial effects on oral and gut health by altering microbiota and reducing inflammation. ^{4 5 6 7 8}

How does the nitrate mouthwash treatment differ from other treatments for oral microbiome health?

The nitrate mouthwash treatment is unique because it uses beetroot juice, which is rich in dietary nitrates, to promote beneficial changes in the oral microbiome by reducing harmful bacteria and inflammation. Unlike traditional mouthwashes that may kill all bacteria, this treatment supports the growth of nitrate-reducing bacteria, which can have additional health benefits like lowering blood pressure. ^{4 5 9 10 11}

What is the purpose of this trial?

The study is aimed to determine if changes to the oral microbiome occur after the use of a nitrate rich mouth rinse. Participants with be asked to use either a controlled (water) mouthrinse once daily for 2 weeks while maintaining normal oral hygiene practices, or use the nitrate rich mouthrinse once daily for 2 weeks. Primary Outcome measures: -changes in the composition of the oral microbiome Secondary Outcome measures * Blood Pressure * Salivary Nitric Oxide Levels

Research Team

Leigha Rock

Principal Investigator

Dalhousie University

Eligibility Criteria

This trial is for individuals interested in the effects of a nitrate-rich mouth rinse on their oral microbiome. Participants should maintain normal oral hygiene during the study and use either water or the provided mouth rinse daily for two weeks.

Inclusion Criteria

I do not have any uncontrolled illnesses.

Participants with mild chronic conditions such as asthma and type 1 diabetes mellitus

Exclusion Criteria

I am under 18 years old.

I am currently taking antibiotics, probiotics, or using antimicrobial mouthrinses.

I have current mouth or gum issues, including cold sores or cavities.

Timeline

Screening

Participants are screened for eligibility to participate in the trial

1-2 weeks

Treatment

Participants use either a nitrate-rich or water mouth rinse once daily for 2 weeks

2 weeks

Daily self-administration

Follow-up

Participants are monitored for changes in the oral microbiome, blood pressure, and salivary nitric oxide levels

2 weeks

Treatment Details

Interventions

• Beet Root

Trial Overview The study tests if using a beetroot-based, nitrate-rich mouth rinse once daily changes the makeup of bacteria in your mouth compared to rinsing with water. It also looks at blood pressure and saliva's nitric oxide levels as secondary outcomes.

Participant Groups

Group I: Nitrate Mouth Rinse Experimental Treatment 1 Intervention

Nitrate suspension with approximately 50-100mg nitrates formulated with water swished in mouth for 30 seconds and then expectorated once daily for 14 consecutive days.

Group II: Water Mouth Rinse Placebo Group 1 Intervention

Water mouth rinse swished in mouth for 30 seconds and then expectorated once daily for 14 consecutive days.

Beet Root is already approved in United States, European Union, Canada for the following indications:

Approved in United States as Beetroot Juice for:

• Dietary Supplement
• Blood Pressure Support

Approved in European Union as Beet Root Supplement for:

• Food Supplement
• Cardiovascular Health

Approved in Canada as Dietary Nitrate for:

• Natural Health Product
• Blood Pressure Regulation

Who Is Running the Clinical Trial?

Dalhousie University

Lead Sponsor

Trials

177

Recruited

402,000+

Findings from Research

A study isolated 62 bacteria from the mouths of healthy donors, identifying 53 as nitrate-reducing, which could help improve oral health and increase systemic nitric oxide levels, potentially benefiting conditions like hypertension and diabetes.

The research found that certain Rothia species not only effectively reduce nitrate but also do so better at an acidic pH, suggesting that these bacteria could be used as probiotics to enhance oral nitrate-reduction capacity and improve overall health.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Isolation and Characterization of Nitrate-Reducing Bacteria as Potential Probiotics for Oral and Systemic Health. Rosier, BT., Moya-Gonzalvez, EM., Corell-Escuin, P., et al.[2023]

This study found significant biological variability in the levels of nitric oxide (NO) markers (nitrate and nitrite) in both plasma and saliva after consuming nitrate-rich beetroot juice, indicating that individual responses can vary widely.

The research established critical difference thresholds for NO markers, which are essential for determining whether changes in these markers after dietary nitrate supplementation are meaningful, helping to clarify the mixed evidence surrounding the effects of dietary nitrate on exercise performance.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Variability in nitrate-reducing oral bacteria and nitric oxide metabolites in biological fluids following dietary nitrate administration: An assessment of the critical difference. Liddle, L., Burleigh, MC., Monaghan, C., et al.[2019]

In newborn infants, the conversion of nitrate to nitrite in saliva is significantly lower than in adults, with no measurable bacterial activity in the first two weeks of life, which may limit the protective effects of nitrite in their gastrointestinal tract.

The presence of oral nitrate-reducing bacteria in infants does not translate to effective nitrite production, potentially contributing to their increased vulnerability to hypoxic stress and gastrointestinal issues.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Nitrate reductase activity of bacteria in saliva of term and preterm infants. Kanady, JA., Aruni, AW., Ninnis, JR., et al.[2021]

A nitrate-rich diet, specifically through a daily intake of 200 mg from a lettuce juice beverage, significantly reduced gingival inflammation in 37 periodontal patients over a 2-week period.

This dietary intervention led to notable changes in the subgingival microbiome, including an increase in beneficial bacteria like Rothia and Neisseria, while the control group showed no significant microbial changes.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Nitrate-rich diet alters the composition of the oral microbiota in periodontal recall patients. Jockel-Schneider, Y., Schlagenhauf, U., Stölzel, P., et al.[2022]

Consuming 50 ml of nitrate-rich beetroot juice daily for two weeks did not affect bacterial lactate production or the pH levels in saliva and oral biofilm, indicating no caries-preventive properties.

Despite the accumulation of nitrate and nitrite in saliva after beetroot juice consumption, the study found no significant antibacterial effects against cariogenic bacteria under the tested conditions.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Influence of Consumption of Nitrate-rich Beetroot Juice on Lactate Production in Saliva and Oral Biofilm - A Clinical Trial. Sterzenbach, T., Hannig, C., Hertel, S.[2023]

Beetroot (Beta vulgaris L.) contains various bioactive compounds, such as dietary fiber, betalains, and phenolics, which have been shown to positively influence gut microbiota and promote gastrointestinal health.

These compounds can enhance the growth of beneficial gut bacteria, increase the production of short-chain fatty acids, and exhibit antioxidant and anti-inflammatory properties, making beetroot a valuable addition to a healthy diet and functional food products.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

A review on bioactive compounds of beet (Beta vulgaris L. subsp. vulgaris) with special emphasis on their beneficial effects on gut microbiota and gastrointestinal health. de Oliveira, SPA., do Nascimento, HMA., Sampaio, KB., et al.[2021]

A 14-day study on healthy adults showed that consuming red beetroot juice led to temporary changes in gut bacteria, including an increase in beneficial Akkermansia muciniphila and a decrease in Bacteroides fragilis after just 3 days.

The presence of betacyanins from beetroot in stool was linked to positive changes in certain gut bacteria and an increase in butyric acid, suggesting that red beetroot may promote both gut health and overall well-being.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Effect of two-week red beetroot juice consumption on modulation of gut microbiota in healthy human volunteers - A pilot study. Wang, Y., Do, T., Marshall, LJ., et al.[2023]

Freeze-dried red beet root (FDBR) and freeze-dried red beet stem and leaves (FDBSL) significantly increased beneficial bacteria like Lactobacillus and Bifidobacterium while decreasing harmful bacteria during 48 hours of in vitro colonic fermentation.

Both FDBR and FDBSL enhanced the metabolic activity of human colonic microbiota, leading to lower pH levels and increased production of short-chain fatty acids, suggesting their potential as effective prebiotic ingredients for gut health.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Investigating the effects of conventional and unconventional edible parts of red beet (Beta vulgaris L.) on target bacterial groups and metabolic activity of human colonic microbiota to produce novel and sustainable prebiotic ingredients. de Oliveira, SPA., de Albuquerque, TMR., Massa, NML., et al.[2023]

The study found that using an antiseptic mouthwash in rats significantly reduced the number of beneficial nitrate-reducing bacteria in the mouth, which in turn decreased the levels of bioactive nitric oxide and nitrite in the body.

As a result of the reduced bacteria, the protective effects of dietary nitrate on gastric mucosal defense were lost, and the expected decrease in blood pressure from nitrate supplementation was also eliminated, highlighting the importance of oral bacteria in cardiovascular and gastrointestinal health.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Gastroprotective and blood pressure lowering effects of dietary nitrate are abolished by an antiseptic mouthwash. Petersson, J., Carlström, M., Schreiber, O., et al.[2016]

Chlorhexidine mouthwash significantly inhibits the presence of nitrate-reducing bacteria, specifically Veillonella dispar, which could impact the body's ability to convert dietary nitrate into beneficial nitrite and nitric oxide.

In contrast, essential oil and povidone-iodine mouthwashes showed little effect on salivary nitrate/nitrite levels and the activity of nitrate-reducing bacteria, suggesting they may be safer options for maintaining oral nitrate-reducing activity.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

The effects of essential oil, povidone-iodine, and chlorhexidine mouthwash on salivary nitrate/nitrite and nitrate-reducing bacteria. Mitsui, T., Harasawa, R.[2018]

A diet high in nitrate and amines significantly increases the formation of carcinogenic nitrosamines, as shown by increased urinary and salivary levels in 12 volunteers consuming such foods.

Using an antibacterial mouthwash containing chlorhexidine effectively reduces salivary nitrite levels and urinary excretion of certain nitrosamines, suggesting it may help inhibit the formation of these harmful compounds.

NCBI (Pubmed)

pubmed.ncbi.nlm.nih.gov

Formation of nitrosamines during consumption of nitrate- and amine-rich foods, and the influence of the use of mouthwashes. van Maanen, JM., Pachen, DM., Dallinga, JW., et al.[2022]

References

1. Switzerland pubmed.ncbi.nlm.nih.gov

Isolation and Characterization of Nitrate-Reducing Bacteria as Potential Probiotics for Oral and Systemic Health. [2023]

2. United States pubmed.ncbi.nlm.nih.gov

Variability in nitrate-reducing oral bacteria and nitric oxide metabolites in biological fluids following dietary nitrate administration: An assessment of the critical difference. [2019]

3. United States pubmed.ncbi.nlm.nih.gov

Nitrate reductase activity of bacteria in saliva of term and preterm infants. [2021]

4. United States pubmed.ncbi.nlm.nih.gov

Nitrate-rich diet alters the composition of the oral microbiota in periodontal recall patients. [2022]

5. Germany pubmed.ncbi.nlm.nih.gov

Influence of Consumption of Nitrate-rich Beetroot Juice on Lactate Production in Saliva and Oral Biofilm - A Clinical Trial. [2023]

6. United States pubmed.ncbi.nlm.nih.gov

A review on bioactive compounds of beet (Beta vulgaris L. subsp. vulgaris) with special emphasis on their beneficial effects on gut microbiota and gastrointestinal health. [2021]

7. England pubmed.ncbi.nlm.nih.gov

Effect of two-week red beetroot juice consumption on modulation of gut microbiota in healthy human volunteers - A pilot study. [2023]

8. Canada pubmed.ncbi.nlm.nih.gov

Investigating the effects of conventional and unconventional edible parts of red beet (Beta vulgaris L.) on target bacterial groups and metabolic activity of human colonic microbiota to produce novel and sustainable prebiotic ingredients. [2023]

9. United States pubmed.ncbi.nlm.nih.gov

Gastroprotective and blood pressure lowering effects of dietary nitrate are abolished by an antiseptic mouthwash. [2016]

10. Japan pubmed.ncbi.nlm.nih.gov

The effects of essential oil, povidone-iodine, and chlorhexidine mouthwash on salivary nitrate/nitrite and nitrate-reducing bacteria. [2018]

11. England pubmed.ncbi.nlm.nih.gov

Formation of nitrosamines during consumption of nitrate- and amine-rich foods, and the influence of the use of mouthwashes. [2022]