Carbonated Beverage Consumption in pH and Bacterial Proliferation (pHSB)

Effects of Carbonated Beverage Consumption on Oral pH and Bacterial Proliferation in Adolescents: A Randomized Crossover Clinical Trial.

The objective of this research is to evaluate the oral modifications caused by different types of coke drinks (regular coke and diet coke). The salivary and the dental biofilm pH will be determined in the first minutes after their consumption. Additionally, the bacterial proliferation of dental biofilm will be evaluated.

Sugary soft drinks modify oral pH and favor bacterial proliferation and are associated with the development of caries. Information on the effects of consuming carbonated drinks without sucrose is limited. In this crossover clinical trial, salivary pH and dental biofilm pH will be determined. These will be registered at 0, 5, 10, 15, 30, 45, and 60 min after the participants ingested 355 ml of natural water, soft drink with sucrose, soft drink with aspartame/acesulfame K or carbonated water on different days (1 week between each other). In addition, dental biofilm cultures will be conducted at 0 and 120 minutes after intake of each beverage to determine Streptococcus mutans biofilm formation. The patients will be invited to participate and informed of the potential risks. Those who signed informed consent and have eligibility requirements will be randomized in a double-blind manner. The data collection will be carried out in records forms, including verifying the patient's previous conditions, identification data (ID, age, gender,) and possible adverse events. If any adverse effect could exist, the research team will be notified for the implementation of possible changes. A HANNA HI 221 potentiometer (HANNA Instruments Inc. Woonsocket-RI-USA, Romania) will be used to determine salivary pH and dental biofilm pH. The electrode will be calibrated using buffer solutions of pH 4.0 and 7.0 for correct records. The electrode will be washed with distilled water before and after each sample. The data will be collected by 2 verifiers, guaranteeing that the information obtained is the same as that indicated on the potentiometer; a stopwatch will indicate the exact time for obtaining the pH values. Samples of dental biofilm will be taken, and Streptococcus mutans biofilm formation will be evaluated at 0 and 120 minutes after taking each beverage. The samples will be cultivated in suitable conditions, identified and compared with ATCC. The samples obtained will be analyzed in the same place of collection to avoid possible contamination. Sample size with an alpha=0.05 and a beta=0.8 include 22, considering 20% losses. Variables will be described with frequencies and percentages or medians and interquartile range (IQR) according to the variable type. Salivary pH and dental biofilm at different times will be compared using ANOVA analysis with adjustment for multiple comparisons using Bonferroni correction. Changes in the bacterial proliferation of the dental biofilm at 0 and 120 min will be compared using the Wilcoxon test and intergroup changes will be compared using the Kruskal-Wallis test. The statistical program SPSS v. 22 will be used and statistical significance will be considered with a p ≤ 0.05

355 ml of water 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

355 ml of carbonated water 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

355 ml of drink of diet coke 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

355 ml of drink of regular coke 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

355 ml of regular coke, diet coke or mineral water 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

Inclusion Criteria: Habitual consumption of soft drinks DMFT (Decayed, Missing, and Filled Teeth) index of at least 3 Agree to participate in the study and sign informed consent Parents sign informed consent Any nutritional condition Exclusion Criteria: Undergoing orthodontic treatment Received a topical application of fluoride during the last 3 months Having a motor disability that interfered with tooth brushing Consuming drugs or being carriers of diseases that cause xerostomia Being under antibiotic therapy during the study period Having active periodontal infections.

The data will be shared with researchers interested in data analysis, request to email: americaml@hotmail.com, review of the request by MSc., PhD. America Liliana Miranda Lora

Gonzalez-Aragon Pineda AE, Garcia Perez A, Garcia-Godoy F. Salivary parameters and oral health status amongst adolescents in Mexico. BMC Oral Health. 2020 Jul 6;20(1):190. doi: 10.1186/s12903-020-01182-8.

Machiulskiene V, Campus G, Carvalho JC, Dige I, Ekstrand KR, Jablonski-Momeni A, Maltz M, Manton DJ, Martignon S, Martinez-Mier EA, Pitts NB, Schulte AG, Splieth CH, Tenuta LMA, Ferreira Zandona A, Nyvad B. Terminology of Dental Caries and Dental Caries Management: Consensus Report of a Workshop Organized by ORCA and Cariology Research Group of IADR. Caries Res. 2020;54(1):7-14. doi: 10.1159/000503309. Epub 2019 Oct 7.

Vieira AR, Modesto A, Marazita ML. Caries: review of human genetics research. Caries Res. 2014;48(5):491-506. doi: 10.1159/000358333. Epub 2014 May 21.

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.

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.

Gonzalez-Aragon Pineda AE, Borges-Yanez SA, Irigoyen-Camacho ME, Lussi A. Relationship between erosive tooth wear and beverage consumption among a group of schoolchildren in Mexico City. Clin Oral Investig. 2019 Feb;23(2):715-723. doi: 10.1007/s00784-018-2489-8. Epub 2018 May 13.

Lemos JA, Palmer SR, Zeng L, Wen ZT, Kajfasz JK, Freires IA, Abranches J, Brady LJ. The Biology of Streptococcus mutans. Microbiol Spectr. 2019 Jan;7(1):10.1128/microbiolspec.GPP3-0051-2018. doi: 10.1128/microbiolspec.GPP3-0051-2018.

Bechir F, Pacurar M, Tohati A, Bataga SM. Comparative Study of Salivary pH, Buffer Capacity, and Flow in Patients with and without Gastroesophageal Reflux Disease. Int J Environ Res Public Health. 2021 Dec 25;19(1):201. doi: 10.3390/ijerph19010201.

Peng X, Han Q, Zhou X, Chen Y, Huang X, Guo X, Peng R, Wang H, Peng X, Cheng L. Effect of pH-sensitive nanoparticles on inhibiting oral biofilms. Drug Deliv. 2022 Dec;29(1):561-573. doi: 10.1080/10717544.2022.2037788.

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.

Aiuchi H, Kitasako Y, Fukuda Y, Nakashima S, Burrow MF, Tagami J. Relationship between quantitative assessments of salivary buffering capacity and ion activity product for hydroxyapatite in relation to cariogenic potential. Aust Dent J. 2008 Jun;53(2):167-71. doi: 10.1111/j.1834-7819.2008.00027.x.

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.

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.

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.

Ahmadi-Motamayel F, Falsafi P, Goodarzi MT, Poorolajal J. Comparison of Salivary pH, Buffering Capacity and Alkaline Phosphatase in Smokers and Healthy Non-Smokers: Retrospective cohort study. Sultan Qaboos Univ Med J. 2016 Aug;16(3):e317-21. doi: 10.18295/squmj.2016.16.03.009. Epub 2016 Aug 19.

Gornowicz A, Tokajuk G, Bielawska A, Maciorkowska E, Jablonski R, Wojcicka A, Bielawski K. The assessment of sIgA, histatin-5, and lactoperoxidase levels in saliva of adolescents with dental caries. Med Sci Monit. 2014 Jun 29;20:1095-100. doi: 10.12659/MSM.890468.

Acquier AB, Pita AK, Busch L, Sanchez GA. Comparison of salivary levels of mucin and amylase and their relation with clinical parameters obtained from patients with aggressive and chronic periodontal disease. J Appl Oral Sci. 2015 May-Jun;23(3):288-94. doi: 10.1590/1678-775720140458.

Gabryel-Porowska H, Gornowicz A, Bielawska A, Wojcicka A, Maciorkowska E, Grabowska SZ, Bielawski K. Mucin levels in saliva of adolescents with dental caries. Med Sci Monit. 2014 Jan 18;20:72-7. doi: 10.12659/MSM.889718.

Hideaki W, Tatsuya H, Shogo M, Naruto Y, Hideaki T, Yoichi M, Yoshihiro O, Kazuo U, Hidenori T. Effect of 100 Hz electroacupuncture on salivary immunoglobulin A and the autonomic nervous system. Acupunct Med. 2015 Dec;33(6):451-6. doi: 10.1136/acupmed-2015-010784. Epub 2015 Oct 8.

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.

Moynihan PJ, Kelly SA. Effect on caries of restricting sugars intake: systematic review to inform WHO guidelines. J Dent Res. 2014 Jan;93(1):8-18. doi: 10.1177/0022034513508954. Epub 2013 Dec 9.

Olivier B, Serge AH, Catherine A, Jacques B, Murielle B, Marie-Chantal CL, Sybil C, Jean-Philippe G, Sabine H, Esther K, Perrine N, Fabienne R, Gerard S, Irene M. Review of the nutritional benefits and risks related to intense sweeteners. Arch Public Health. 2015 Oct 1;73:41. doi: 10.1186/s13690-015-0092-x. eCollection 2015. Erratum In: Arch Public Health. 2015;73:49.

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.

Jawale BA, Bendgude V, Mahuli AV, Dave B, Kulkarni H, Mittal S. Dental plaque pH variation with regular soft drink, diet soft drink and high energy drink: an in vivo study. J Contemp Dent Pract. 2012 Mar 1;13(2):201-4. doi: 10.5005/jp-journals-10024-1121.

Sanchez GA, Fernandez De Preliasco MV. Salivary pH changes during soft drinks consumption in children. Int J Paediatr Dent. 2003 Jul;13(4):251-7. doi: 10.1046/j.1365-263x.2003.00469.x.

Uma E, Theng KS, Yi LLH, Yun LH, Varghese E, Soe HHK. Comparison of Salivary pH Changes after Consumption of Two Sweetened Malaysian Local Drinks among Individuals with Low Caries Experience: A Pilot Study. Malays J Med Sci. 2018 Jul;25(4):100-111. doi: 10.21315/mjms2018.25.4.10. Epub 2018 Aug 30.

Roos EH, Donly KJ. In vivo dental plaque pH variation with regular and diet soft drinks. Pediatr Dent. 2002 Jul-Aug;24(4):350-3.

Llena-Puy C. The role of saliva in maintaining oral health and as an aid to diagnosis. Med Oral Patol Oral Cir Bucal. 2006 Aug;11(5):E449-55. English, Spanish.

Saeed S, Al-Tinawi M. Evaluation of acidity and total sugar content of children's popular beverages and their effect on plaque pH. J Indian Soc Pedod Prev Dent. 2010 Jul-Sep;28(3):189-92. doi: 10.4103/0970-4388.73783.