Oral Changes With Caloric and no Caloric Sweeteners

• Sponsor: Hospital Infantil de Mexico Federico Gomez (Other)
• Overall Status: Not yet recruiting
• CT.gov ID: NCT05852145
• Collaborator: (none)
• Enrollment: 52
• Arms: 3
• Anticipated Duration (Months): 14

Study Details

Study Description

Brief Summary

The objective of this clinical trial is to compare the effect that the intake of beverages added with non-caloric sweeteners (stevioside) and caloric sweeteners (sucrose) on oral pH and dental biofilm microbiome in Mexican adolescents.

Participants will drink on different occasions a beverage without sweetener, a beverage added with stevioside or a beverage added with sucrose. The researchers will compare the changes that each one causes in salivary pH, dental biofilm pH, dental biofilm bacterial proliferation and dental biofilm microbiome.

Condition or Disease	Intervention/Treatment	Phase
pH; Microbiome; Dental Plaque	Drug: Stevioside	Phase 1/Phase 2

Detailed Description

A randomized crossover clinical trial will be conducted including 52 healthy adolescents participants (sample size with alpha=0.05 and beta=0.8 ). The intervention will consist of ingesting 355 mL of tea lemon flavor (pH 7.1) in the first meeting, in subsequent meetings the tea will be added with 1) 2.2 grams (gr.) of stevioside or 2) 7.5 gr. of sucrose. The washout period between interventions will be 1 week.

Participants will be informed of potential risks and those who sign the informed consent and complete the inclusion criteria will be randomized in a double-blind manner.

The data will be collected in a special format including previous conditions, identification (age and sex) and possible adverse effects. If any possible adverse effect occurs, it will be notified to the research team in order to determine the changes.

• To determine the pH, saliva and dental biofilm samples will be obtained and analyzed using a HANNA potentiometer calibrated with buffer solutions.

• To determine bacterial proliferation, samples of dental biofilm will be obtained, cultivated in appropriate media and conditions, compared with the American Type Culture Collection (ATCC) catalog and the number of colony-forming units will be compared.

• To analyze the main components of the dental biofilm, genetic sequencing will be performed and the operative taxonomic units will be compared. (11 of the 52 participants will be randomized for microbiome analysis).

The data will be collected at a time indicated by a stopwatch and carried out by two verifiers.

The statistical analysis will be done according to the type of variable, these will be described with mean and standard deviation or frequencies and percentages. The pH will be compared by ANOVA analysis and adjusted by Bonferroni correction. Bacterial proliferation will be analyzed by Kruskal Wallis test. Statistical Package for the Social Sciences (SPSS)

1. 22 program will be used and statistical significance will be considered with p ≤ 0.05. For the microbiome, the principal component analysis (PCA) of each operational taxonomic unit (OTU) will be done. Non-parametric multivariate analysis of variance (Adonis) and an analysis of similarities (ANOSIM) will be used. The p values for Adonis and for ANOSIM will be calculated, identifying those that have differences in bacterial communities between the groups using the Genius (V3) software.

Study Design

• Study Type: Interventional
• Anticipated Enrollment: 52 participants
• Allocation: Randomized
• Intervention Model: Crossover Assignment
• Intervention Model Description: A Randomized Crossover Clinical TrialA Randomized Crossover Clinical Trial
• Masking: Triple (Participant, Care Provider, Investigator)
• Masking Description: colors will be assigned for each sweetener
• Primary Purpose: Basic Science
• Official Title: Effect of the Consumption of Beverages Added With Stevia Rebaudiana on Oral pH and Dental Biofilm in Adolescents
• Anticipated Study Start Date: Oct 1, 2023
• Anticipated Primary Completion Date: Oct 1, 2024
• Anticipated Study Completion Date: Dec 1, 2024

Arms and Interventions

Arm	Intervention/Treatment
Placebo Comparator: Tea lemon flavor 355 ml de tea lemon flavor should be drunk. Salivary pH will be collected at 0,5,10,15,30,45 and 60 minutes later. Dental biofilm pH will be collected at 0,5,10,15,30, 45 and 60 minutes later. Streptococcus mutans dental biofilm formation ( Colony Forming Units) will be collected at 0 and 120 minutes later. Operative Taxonomic Units of dental biofilm will be analyzed at 0 and 120 minutes later.	Drug: Stevioside; 355 ml of tea lemon flavor, tea lemon flavor added with 2.2 gr. of stevioside or tea lemon flavor added with 7.5 gr. of sucrose should be drunk. Salivary pH will be determined at 0, 5, 10, 15, 30, 45 and 60 minutes later. Dental biofilm pH will be determined at 0, 5, 10, 15, 30, 45 and 60 minutes later. Streptococcus mutans dental biofilm formation ( Colony Forming Units) will be conducted at 0 and 120 minutes later. Operative Taxonomic Units of dental biofilm will be analyzed at 0 and 120 minutes later.; Other Names: Non caloric sweetener
Experimental: Stevioside 355 ml de tea lemon flavor added with 2.2 gr. of stevioside should be drunk. Salivary pH will be collected at 0,5,10,15,30,45 and 60 minutes later. Dental biofilm pH will be collected at 0,5,10,15,30, 45 and 60 minutes later. Streptococcus mutans dental biofilm formation ( Colony Forming Units) will be collected at 0 and 120 minutes later. Operative Taxonomic Units of dental biofilm will be analyzed at 0 and 120 minutes later.	Drug: Stevioside; 355 ml of tea lemon flavor, tea lemon flavor added with 2.2 gr. of stevioside or tea lemon flavor added with 7.5 gr. of sucrose should be drunk. Salivary pH will be determined at 0, 5, 10, 15, 30, 45 and 60 minutes later. Dental biofilm pH will be determined at 0, 5, 10, 15, 30, 45 and 60 minutes later. Streptococcus mutans dental biofilm formation ( Colony Forming Units) will be conducted at 0 and 120 minutes later. Operative Taxonomic Units of dental biofilm will be analyzed at 0 and 120 minutes later.; Other Names: Non caloric sweetener
Active Comparator: Sucrose 355 ml de tea lemon flavor added with 7.5 gr of sucrose should be drunk. Salivary pH will be collected at 0,5,10,15,30,45 and 60 minutes later. Dental biofilm pH will be collected at 0,5,10,15,30, 45 and 60 minutes later. Streptococcus mutans dental biofilm formation ( Colony Forming Units) will be collected at 0 and 120 minutes later. Operative Taxonomic Units of dental biofilm will be analyzed at 0 and 120 minutes later.	Drug: Stevioside; 355 ml of tea lemon flavor, tea lemon flavor added with 2.2 gr. of stevioside or tea lemon flavor added with 7.5 gr. of sucrose should be drunk. Salivary pH will be determined at 0, 5, 10, 15, 30, 45 and 60 minutes later. Dental biofilm pH will be determined at 0, 5, 10, 15, 30, 45 and 60 minutes later. Streptococcus mutans dental biofilm formation ( Colony Forming Units) will be conducted at 0 and 120 minutes later. Operative Taxonomic Units of dental biofilm will be analyzed at 0 and 120 minutes later.; Other Names: Non caloric sweetener

Outcome Measures

Primary Outcome Measures

1. Mean salivary pH with tea lemon flavor [0 minutes]

   Logarithm of hydrogen ion concentration

2. Mean salivary pH with tea lemon flavor [5 minutes]

   Logarithm of hydrogen ion concentration

3. Mean salivary pH with tea lemon flavor [10 minutes]

   Logarithm of hydrogen ion concentration

4. Mean salivary pH with tea lemon flavor [15 minutes]

   Logarithm of hydrogen ion concentration

5. Mean salivary pH with tea lemon flavor [30 minutes]

   Logarithm of hydrogen ion concentration

6. Mean salivary pH with tea lemon flavor [45 minutes]

   Logarithm of hydrogen ion concentration

7. Mean salivary pH with tea lemon flavor [60 minutes]

   Logarithm of hydrogen ion concentration

8. Mean salivary pH with tea lemon flavor added with 2.2 gr. of stevioside [0 minutes]

   Logarithm of hydrogen ion concentration

9. Mean salivary pH with tea lemon flavor added with 2.2 gr. of stevioside [5 minutes]

   Logarithm of hydrogen ion concentration

10. Mean salivary pH with tea lemon flavor added with 2.2 gr. of stevioside [10 minutes]

    Logarithm of hydrogen ion concentration

11. Mean salivary pH with tea lemon flavor added with 2.2 gr. of stevioside [15 minutes]

    Logarithm of hydrogen ion concentration

12. Mean salivary pH with tea lemon flavor added with 2.2 gr. of stevioside [30 minutes]

    Logarithm of hydrogen ion concentration

13. Mean salivary pH with tea lemon flavor added with 2.2 gr. of stevioside [45 minutes]

    Logarithm of hydrogen ion concentration

14. Mean salivary pH with tea lemon flavor added with 2.2 gr. of stevioside [60 minutes]

    Logarithm of hydrogen ion concentration

15. Mean salivary pH with tea lemon flavor added with 7.5 gr. sucrose [0 minutes]

    Logarithm of hydrogen ion concentration

16. Mean salivary pH with tea lemon flavor added with 7.5 gr. sucrose [5 minutes]

    Logarithm of hydrogen ion concentration

17. Mean salivary pH with tea lemon flavor added with 7.5 gr. sucrose [10 minutes]

    Logarithm of hydrogen ion concentration

18. Mean salivary pH with tea lemon flavor added with 7.5 gr. sucrose [15 minutes]

    Logarithm of hydrogen ion concentration

19. Mean salivary pH with tea lemon flavor added with 7.5 gr. sucrose [30 minutes]

    Logarithm of hydrogen ion concentration

20. Mean salivary pH with tea lemon flavor added with 7.5 gr. sucrose [45 minutes]

    Logarithm of hydrogen ion concentration

21. Mean salivary pH with tea lemon flavor added with 7.5 gr. sucrose [60 minutes]

    Logarithm of hydrogen ion concentration

Secondary Outcome Measures

1. Mean dental biofilm pH with tea lemon flavor [0 minutes]

   Logarithm of hydrogen ion concentration

2. Mean dental biofilm pH with tea lemon flavor [5 minutes]

   Logarithm of hydrogen ion concentration

3. Mean dental biofilm pH with tea lemon flavor [10 minutes]

   Logarithm of hydrogen ion concentration

4. Mean dental biofilm pH with tea lemon flavor [15 minutes]

   Logarithm of hydrogen ion concentration

5. Mean dental biofilm pH with tea lemon flavor [30 minutes]

   Logarithm of hydrogen ion concentration

6. Mean dental biofilm pH with tea lemon flavor [45 minutes]

   Logarithm of hydrogen ion concentration

7. Mean dental biofilm pH with tea lemon flavor [60 minutes]

   Logarithm of hydrogen ion concentration

8. Mean dental biofilm pH with tea lemon flavor added with 2.2 gr. of stevioside [0 minutes]

   Logarithm of hydrogen ion concentration

9. Mean dental biofilm pH with tea lemon flavor added with 2.2 gr. of stevioside [5 minutes]

   Logarithm of hydrogen ion concentration

10. Mean dental biofilm pH with tea lemon flavor added with 2.2 gr. of stevioside [10 minutes]

    Logarithm of hydrogen ion concentration

11. Mean dental biofilm pH with tea lemon flavor added with 2.2 gr. of stevioside [15 minutes]

    Logarithm of hydrogen ion concentration

12. Mean dental biofilm pH with tea lemon flavor added with 2.2 gr. of stevioside [30 minutes]

    Logarithm of hydrogen ion concentration

13. Mean dental biofilm pH with tea lemon flavor added with 2.2 gr. of stevioside [45 minutes]

    Logarithm of hydrogen ion concentration

14. Mean dental biofilm pH with tea lemon flavor added with 2.2 gr. of stevioside [60 minutes]

    Logarithm of hydrogen ion concentration

15. Mean dental biofilm pH with tea lemon flavor added with 7.5 gr. sucrose [0 minutes]

    Logarithm of hydrogen ion concentration

16. Mean dental biofilm pH with tea lemon flavor added with 7.5 gr. sucrose [5 minutes]

    Logarithm of hydrogen ion concentration

17. Mean dental biofilm pH with tea lemon flavor added with 7.5 gr. sucrose [10 minutes]

    Logarithm of hydrogen ion concentration

18. Mean dental biofilm pH with tea lemon flavor added with 7.5 gr. sucrose [15 minutes]

    Logarithm of hydrogen ion concentration

19. Mean dental biofilm pH with tea lemon flavor added with 7.5 gr. sucrose [30 minutes]

    Logarithm of hydrogen ion concentration

20. Mean dental biofilm pH with tea lemon flavor added with 7.5 gr. sucrose [45 minutes]

    Logarithm of hydrogen ion concentration

21. Mean dental biofilm pH with tea lemon flavor added with 7.5 gr. sucrose [60 minutes]

    Logarithm of hydrogen ion concentration

Other Outcome Measures

1. Mean Colony Forming Units Streptococcus mutans dental biofilm with tea lemon flavor [0 minutes]

   Number of viable colonies on a semisolid agar culture medium that are visible and separable

2. Mean Colony Forming Units Streptococcus mutans dental biofilm with tea lemon flavor [120 minutes]

   Number of viable colonies on a semisolid agar culture medium that are visible and separable

3. Mean Colony Forming Units Streptococcus mutans dental biofilm with tea lemon flavor added with 2.2 gr. stevioside [0 minutes]

   Number of viable colonies on a semisolid agar culture medium that are visible and separable

4. Mean Colony Forming Units Streptococcus mutans dental biofilm with tea lemon flavor added with 2.2 gr. stevioside [120 minutes]

   Number of viable colonies on a semisolid agar culture medium that are visible and separable

5. Mean Colony Forming Units Streptococcus mutans dental biofilm with tea lemon flavor added with 7.5 gr sucrose [0 minutes]

   Number of viable colonies on a semisolid agar culture medium that are visible and separable

6. Mean Colony Forming Units Streptococcus mutans dental biofilm with tea lemon flavor added with 7.5 gr sucrose [120 minutes]

   Number of viable colonies on a semisolid agar culture medium that are visible and separable

7. Principal component analysis of dental biofilm with tea lemon flavor [0 minutes]

   Operative Taxonomic Units

8. Principal component analysis of dental biofilm with tea lemon flavor [120 minutes]

   Operative Taxonomic Units

9. Principal component analysis of dental biofilm with tea lemon flavor added with 2.2 gr. of stevioside. [0 minutes]

   Operative Taxonomic Units

10. Principal component analysis of dental biofilm with tea lemon flavor added with 2.2 gr. of stevioside. [120 minutes]

    Operative Taxonomic Units

11. Principal component analysis of dental biofilm with tea lemon flavor added with 7.5 gr. of sucrose [0 minutes]

    Operative Taxonomic Units

12. Principal component analysis of dental biofilm with tea lemon flavor added with 7.5 gr. of sucrose [120 minutes]

    Operative Taxonomic Units

Eligibility Criteria

Criteria

• Ages Eligible for Study: 12 Years to 18 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• Habitual consumption of soft drinks

• Decayed, Missing, and Filled Teeth (DMF) index of at least 3

• Agree to participate in the study and sign informed consent

• Parents sign informed consent

• Any nutritional condition

Exclusion Criteria:

• Orthodontic treatment

• Topical application of fluoride during the last 3 months

• Having a motor disability that interfered with tooth brushing

• Xerostomia

• Antibiotic therapy during the study period

• Periodontal infections

Contacts and Locations

Locations

No locations specified.

Sponsors and Collaborators

• Hospital Infantil de Mexico Federico Gomez

Investigators

• Principal Investigator: Carolina Barajas, MsC, Children´s Hospital Federico Gómez. México

Study Documents (Full-Text)

• Informed Consent Form - May 5, 2023

More Information

Publications

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• Behjati S, Tarpey PS. What is next generation sequencing? Arch Dis Child Educ Pract Ed. 2013 Dec;98(6):236-8. doi: 10.1136/archdischild-2013-304340. Epub 2013 Aug 28.
• Brambilla E, Cagetti MG, Ionescu A, Campus G, Lingstrom P. An in vitro and in vivo comparison of the effect of Stevia rebaudiana extracts on different caries-related variables: a randomized controlled trial pilot study. Caries Res. 2014;48(1):19-23. doi: 10.1159/000351650. Epub 2013 Nov 6.
• Carvalho RN, Letieri ADS, Vieira TI, Santos TMPD, Lopes RT, Neves AA, Pomarico L. Accuracy of visual and image-based ICDAS criteria compared with a micro-CT gold standard for caries detection on occlusal surfaces. Braz Oral Res. 2018 Jul 10;32:e60. doi: 10.1590/1807-3107bor-2018.vol32.0060.
• Edelstein BL. The dental caries pandemic and disparities problem. BMC Oral Health. 2006 Jun 15;6 Suppl 1(Suppl 1):S2. doi: 10.1186/1472-6831-6-S1-S2.
• Escobar E, Piedrahita M, Gregory RL. Growth and viability of Streptococcus mutans in sucrose with different concentrations of Stevia rebaudiana Bertoni. Clin Oral Investig. 2020 Sep;24(9):3237-3242. doi: 10.1007/s00784-020-03197-5. Epub 2020 Mar 18.
• Foglio-Bonda PL, Brilli K, Pattarino F, Foglio-Bonda A. Salivary flow rate and pH in patients with oral pathologies. Eur Rev Med Pharmacol Sci. 2017 Jan;21(2):369-374.
• Garcia-Cortes JO, Mejia-Cruz JA, Medina-Cerda E, Orozco-De la Torre G, Medina-Solis CC, Marquez-Rodriguez S, Navarrete-Hernandez Jde J, Islas-Granillo H. [Experience, prevalence, severity, treatment needs for dental caries and care index in Mexican adolescents and young adults]. Rev Invest Clin. 2014 Nov-Dec;66(6):505-11. Spanish.
• He J, Li Y, Cao Y, Xue J, Zhou X. The oral microbiome diversity and its relation to human diseases. Folia Microbiol (Praha). 2015 Jan;60(1):69-80. doi: 10.1007/s12223-014-0342-2. Epub 2014 Aug 23.
• Humphrey SP, Williamson RT. A review of saliva: normal composition, flow, and function. J Prosthet Dent. 2001 Feb;85(2):162-9. doi: 10.1067/mpr.2001.113778.
• Ilie O, van Turnhout AG, van Loosdrecht MC, Picioreanu C. Numerical modelling of tooth enamel subsurface lesion formation induced by dental plaque. Caries Res. 2014;48(1):73-89. doi: 10.1159/000354123. Epub 2013 Nov 14.
• Irigoyen ME, Mejia-Gonzalez A, Zepeda-Zepeda MA, Betancourt-Linares A, Lezana-Fernandez MA, Alvarez-Lucas CH. Dental caries in Mexican schoolchildren: a comparison of 1988-1989 and 1998-2001 surveys. Med Oral Patol Oral Cir Bucal. 2012 Sep 1;17(5):e825-32. doi: 10.4317/medoral.18008.
• Kaur A, Kwatra KS, Kamboj P. Evaluation of non-microbial salivary caries activity parameters and salivary biochemical indicators in predicting dental caries. J Indian Soc Pedod Prev Dent. 2012 Jul-Sep;30(3):212-7. doi: 10.4103/0970-4388.105013.
• Koo H, Falsetta ML, Klein MI. The exopolysaccharide matrix: a virulence determinant of cariogenic biofilm. J Dent Res. 2013 Dec;92(12):1065-73. doi: 10.1177/0022034513504218. Epub 2013 Sep 17.
• Krzysciak W, Pluskwa KK, Piatkowski J, Krzysciak P, Jurczak A, Koscielniak D, Skalniak A. The usefulness of biotyping in the determination of selected pathogenicity determinants in Streptococcus mutans. BMC Microbiol. 2014 Aug 5;14:194. doi: 10.1186/1471-2180-14-194.
• Mathur MR, Tsakos G, Millett C, Arora M, Watt R. Socioeconomic inequalities in dental caries and their determinants in adolescents in New Delhi, India. BMJ Open. 2014 Dec 12;4(12):e006391. doi: 10.1136/bmjopen-2014-006391.
• Moradi G, Mohamadi Bolbanabad A, Moinafshar A, Adabi H, Sharafi M, Zareie B. Evaluation of Oral Health Status Based on the Decayed, Missing and Filled Teeth (DMFT) Index. Iran J Public Health. 2019 Nov;48(11):2050-2057.
• Pandey P, Reddy NV, Rao VA, Saxena A, Chaudhary CP. Estimation of salivary flow rate, pH, buffer capacity, calcium, total protein content and total antioxidant capacity in relation to dental caries severity, age and gender. Contemp Clin Dent. 2015 Mar;6(Suppl 1):S65-71. doi: 10.4103/0976-237X.152943.
• Skrivele S, Care R, Berzina S, Kneist S, de Moura-Sieber V, de Moura R, Borutta A, Maslak E, Tserekhava T, Shakovets N, Wagner M. Caries and its risk factors in young children in five different countries. Stomatologija. 2013;15(2):39-46.
• Struzycka I. The oral microbiome in dental caries. Pol J Microbiol. 2014;63(2):127-35.

• Responsible Party: Guadalupe Barajas, Principal Investigator, Hospital Infantil de Mexico Federico Gomez
• ClinicalTrials.gov Identifier: NCT05852145
• Other Study ID Numbers: HIM 2022-049 FF
• First Posted: May 10, 2023
• Last Update Posted: May 10, 2023
• Last Verified: Apr 1, 2023

• Individual Participant Data (IPD) Sharing Statement: Yes
• Plan to Share IPD: Yes
• Studies a U.S. FDA-regulated Drug Product: Yes
• Studies a U.S. FDA-regulated Device Product: No
• Product Manufactured in and Exported from the U.S.: Yes
• Keywords provided by Guadalupe Barajas, Principal Investigator, Hospital Infantil de Mexico Federico Gomez: artificial sweetener; stevioside; pH; caries; sucrose; dental biofilm
• Additional relevant MeSH terms: Dental Plaque

Study Results

No Results Posted as of May 10, 2023