Abstract
Objective
To evaluate total absorbance, planktonic growth, biofilm formation, viability, metabolic activity, and pH of Streptococcus mutans UA159 cultures when different dilutions of Stevia rebaudiana Bertoni were applied and to determine the minimum inhibitory concentration (MIC) and the minimum biofilm inhibitory concentration (MBIC) of Stevia on S. mutans.
Materials and methods
The effects of different dilutions of Stevia (0–400 mg/ml) on S. mutans total growth, planktonic growth, biofilm formation, viability, metabolic activity, and pH during a 72-h growth period were evaluated in this in vitro study. A stock solution was prepared by mixing 10 ml of tryptic soy broth (TSB) supplemented with 1% sucrose (TSBS) and 4 g of Stevia.
Results
S. mutans total growth and biofilm formation decreased with reduced concentrations of Stevia. Furthermore, the MIC was 25 mg/ml and the MBIC was 6.25 mg/ml. Complete eradication of S. mutans was not observed with any of the Stevia concentrations. Planktonic growth of S. mutans was not repressed by high concentrations of Stevia and most of the Stevia concentrations generated an increased pH.
Conclusion
Because Stevia reduces biofilm and acid production, Stevia can be considered a non-cariogenic sweetener.
Clinical relevance
This study confirms the anticariogenic effect of Stevia, like it has been previously reported, but more studies on the most effective concentration are needed, and in the present study, the minimum inhibitory concentration (MIC) and the minimum biofilm inhibitory concentration (MBIC) was determined in the presence of sucrose. Additionally, this is the first study to evaluate the effect of Stevia on S. mutans metabolic activity.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ojeada Garcés J, Oviedo García E, Salas L (2013) Streptococcus mutans and dental caries. Rev CES Odont 26(1):44–56 http://www.scielo.org.co/pdf/ceso/v26n1/v26n1a05.pdf
Selwitz R, Ismail A, Pitts N (2007) Dental caries. Lancet 369:51–59. https://doi.org/10.1016/S0140-6736(07)60031-2
Gupta P, Gupta N, Singh H (2014) Prevalence of dental caries in relation to body mass index, daily sugar intake and oral hygiene status in 12-year-old school children in Mathura City: a pilot study. Int J Pediatr 2014 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3945027/
Do T, Devine D, Marsh P (2013) Oral biofilms: molecular analysis, challenges and future prospects in dental diagnostics. Clin Cosmet Investig Dent 5:11–19 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3652372/
Gamboa F, Chaves M (2012) Antimicrobial potential of extracts from Stevia Rebaudiana leaves against bacteria of importance in dental caries. Acta Odontol Latinoam 25(2):171–175 http://www.scielo.org.ar/pdf/aol/v25n2/v25n2a03.pdf
Jiang S, Gao X, Jin L, Lo E (2016) Salivary microbiome diversity in caries free and caries affected children. Int J Mol Sci 17(12) https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5187778/pdf/ijms-17-01978.pdf
Takahashi N, Nyvad B (2011) The role of bacteria in the caries process: ecological perspectives. J Dent Res 90(3):294–303 https://journals.sagepub.com/doi/abs/10.1177/0022034510379602?rfr_dat=cr_pub%3Dpubmed&url_ver=Z39.882003&rfr_id=ori%3Arid%3Acrossref.org&journalCode=jdrb
Pedro Núñez D, García Bacallao L (2010) Biochemistry of dental caries. Revista Habanera de Ciencias Médicas 9(2):156–166 http://scielo.sld.cu/pdf/rhcm/v9n2/rhcm04210.pdf
Kidd E (2011) The implications of the new paradigm of dental caries. J Dent 39(Suppl 2):S3–S8 https://www.sciencedirect.com/science/article/pii/S0300571211002739?via%3Dihub
Huang R, Li M, Gregory R (2011) Bacterial interactions in dental biofilm. Virulence 2(5):435–444 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3322631/pdf/viru0205_0435.pdf
Martignon S, Gomez J, Tellez M, Ruiz J, Marin L, Rangel M (2013) Current cariology education in dental schools in Spanish speaking Latin American countries. J Dent Educ 77(10):1330–1337 http://www.jdentaled.org/content/jde/77/10/1330.full.pdf
Minsalud (2012) Cuarto Estudio Nacional de Salud Bucal (ENSAB IV). https://www.minsalud.gov.co/sites/rid/Lists/BibliotecaDigital/RIDE/VS/PP/ENSAB-IV-Metodologia.pdf
Brambilla E, Cagetti M, Lonescu A, Campus G, Lingstrom P (2014) 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 48(1):19–23 https://www.karger.com/Article/Abstract/351650
Thomas J, Glade M (2010) Stevia: it’s not just about calories. Open Obes J 2:101–109 https://pdfs.semanticscholar.org/fd0b/5df46a776069b89244000885d65f0277318b.pdf
Contreras S (2013) Anticariogenic properties and effects on periodontal structures of Stevia rebaudiana Bertoni. Narrative review. J Oral Res 2(3):158–166 https://dialnet.unirioja.es/servlet/articulo?codigo=4995308
Salvador Reyes R, Sotelo Herrera M, Paucar Menacho L (2014) Study of Stevia (Stevia rebaudiana Bertoni) as a natural sweetener and its use in benefit of the health. Scientia Agropecuaria 5:157–163 https://es.scribd.com/document/362512646/stevia-pdf
Fambrizio Ferrazzano G, Cantile T, Alcidi B, Coda M, Ingenito A, Zarrelli A, Di Fabio G, Pollio A (2016) Is Stevia rebaudiana Bertoni a non-cariogenic sweetener? A review. Molecules 21(1):E38 https://www.mdpi.com/1420-3049/21/1/38
Giacaman R, Campos P, Muñoz Sandoval C, Castro R (2013) Cariogenic potential of commercial sweeteners in an experimental biofilm caries model on enamel. Arch Oral Biol 58(9):1116–1122 https://www.sciencedirect.com/science/article/pii/S0003996913000782?via%3Dihub
Blauth de Slavutzky S (2010) Stevia and sucrose effect on plaque formation. J FÜR VERBRAUCHERSCHUTZ Leb 5(2):213–216 https://link.springer.com/article/10.1007%2Fs00003-010-0555-5
Grembecka M (2015) Natural sweeteners in a human diet. Rocz Panstw Zakl Hig 66(3):195–202 https://www.ncbi.nlm.nih.gov/pubmed/26400114
Pierce CG, Uppuluri P, Tristan AR, Wormley FL, Mowat E, Ramage G, Lopez Ribot JL (2008) A simple and reproducible 96-well plate based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc 3(9):1494–1500 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2741160/pdf/nihms127871.pdf
Huang R, Li M, Gregory RL (2012) Effect of nicotine on growth and tabolism of Streptococcus mutans. Eur J Oral Sci 120(4):319–325 https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1600-0722.2012.00971.x
Das S, Das AK, Murphy RA, Punwani IC, Nasution MP, Kinghorn AD (1992) Evaluation of the cariogenic potential of the intense natural sweeteners stevioside and rebaudioside A. Caries Res 26(5):363–366 https://www.karger.com/Article/Abstract/261469
Acknowledgments
Special thanks to Dr. Ru Zhang for all the support and guidance throughout the study; to the Oral Health Research Institute of Indiana University; to Luis Gonzalo Alvarez for his professional help in statistics; and to Dr. Emery Alvarez for making this possible.
Funding
This investigation was fully funded by the Research and Innovation Department, CES University. The funding source had no involvement in data collection, analysis interpretation, or in the decision to submit the article for publication.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Informed consent
Not required.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Escobar, E., Piedrahita, M. & Gregory, R.L. Growth and viability of Streptococcus mutans in sucrose with different concentrations of Stevia rebaudiana Bertoni. Clin Oral Invest 24, 3237–3242 (2020). https://doi.org/10.1007/s00784-020-03197-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00784-020-03197-5