Subscribe to RSS

DOI: 10.1055/s-0044-1792165
Effects of Alternative Milk on Streptococcus Mutans Biofilm Formation and Enamel Demineralization in Human Primary Teeth

Abstract
Objectives This study was conducted to investigate the effects of alternative milk on Streptococcus mutans biofilm formation and their ability to demineralize enamel in primary teeth.
Materials and Methods First, to evaluate the effects of cow milk, lactose-free cow milk, goat milk, unsweetened pistachio milk, and sweetened pistachio milk on S. mutans biofilm formation, biofilm assay was conducted. The optical density (OD) was measured to determine S. mutans biofilm. Second, to assess the enamel demineralization, enamel slabs were prepared from 50 primary incisor teeth and divided into three test groups, along with positive and negative control groups. Enamel slabs were immersed in each type of milk three times a day for 5 days. The percentage of surface hardness loss (%SHL) for enamel demineralization was measured. One enamel slab was randomly selected from each group to visualize the enamel opacity in demineralization area by using a light microscope. Another slab was randomly selected from each group to stain with fluorescence dye and to observe the biofilm structure by using a confocal microscope.
Results The OD ± SD (standard deviation) measurements for S. mutans biofilm formation in cow milk, lactose-free cow milk, goat milk, unsweetened pistachio milk, and sweetened pistachio milk were 0.082 (± 0.002), 0.086 (± 0.004), 0.083 (± 0.007), 0.0952 (± 0.010), and 0.342 (± 0.072), respectively. The sweetened pistachio milk exhibited significantly more biofilm formation than the other milk (p < 0.05). Since there was no significant difference in biofilm formation among cow milk, lactose-free cow milk, goat milk, and unsweetened pistachio milk, we tested the enamel demineralization only with cow milk, unsweetened pistachio milk, and sweetened pistachio milk. The %SHL (± SD) for cow milk, unsweetened pistachio milk, and sweetened pistachio milk were 20.01 (± 2.618), 22.088 (± 3.4), and 35.49 (± 2.069), respectively. The %SHL on the enamel in sweetened pistachio milk was higher (p < 0.001) than other tested milk. White spot lesion was directly visualized on slabs in sweetened pistachio milk under light microscope. Biofilm formed in sweetened pistachio milk demonstrated a mushroom-like structure, whereas cow milk and unsweetened pistachio milk had a flat biofilm structure.
Conclusion Sweetened pistachio milk increases the risk for primary incisor teeth caries formation greater than cow milk and unsweetened pistachio milk regarding S. mutans biofilm formation was higher in quantity and ability to demineralization in primary teeth.
Ethical Consideration
This study was approved by the Ethics Approval Review Board, Faculty of Dentistry and the Faculty of Pharmacy, Mahidol University (MU-DT/PY-IRB 2020/015.2205).
Publication History
Article published online:
09 January 2025
© 2025. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India
-
References
- 1 Fejerskov O. Changing paradigms in concepts on dental caries: consequences for oral health care. Caries Res 2004; 38 (03) 182-191
- 2 Featherstone JD. Dental caries: a dynamic disease process. Aust Dent J 2008; 53 (03) 286-291
- 3 Twetman S. Prevention of dental caries as a non-communicable disease. Eur J Oral Sci 2018; 126 (Suppl. 01) 19-25
- 4 Philip N, Suneja B, Walsh LJ. Ecological approaches to dental caries prevention: paradigm shift or shibboleth?. Caries Res 2018; 52 (1–2): 153-165
- 5 Tanner AC, Kressirer CA, Faller LL. Understanding caries from the oral microbiome perspective. J Calif Dent Assoc 2016; 44 (07) 437-446
- 6 Tanzer JM, Livingston J, Thompson AM. The microbiology of primary dental caries in humans. J Dent Educ 2001; 65 (10) 1028-1037
- 7 Tantikalchan S, Mitrakul K. Association between Bifidobacterium and Scardovia Wiggsiae and caries-related factors in severe early childhood caries and caries-free Thai children: a quantitative real-time PCR analysis and a questionnaire cross-sectional study. Eur Arch Paediatr Dent 2022; 23 (03) 437-447
- 8 Mitrakul K, Akarapipatkul B, Thammachat P. Quantitative analysis of Streptococcus mutans, Streptococcus sobrinus and Streptococcus sanguinis and their association with early childhood caries. J Clin Diagn Res 2020; 14 (02) ZC23-ZC28
- 9 Mitrakul K, Chanvitan S, Jeamset A, Vongsawan K. Quantitative analysis of S. mutans, Lactobacillus and Bifidobacterium found in initial and mature plaques in Thai children with early childhood caries. Eur Arch Paediatr Dent 2017; 18 (04) 251-261
- 10 Kanasi E, Johansson I, Lu SC. et al. Microbial risk markers for childhood caries in pediatricians' offices. J Dent Res 2010; 89 (04) 378-383
- 11 Marsh PD, Head DA, Devine DA. Ecological approaches to oral biofilms: control without killing. Caries Res 2015; 49 (Suppl. 01) 46-54
- 12 Rirattanapong P, Vongsavan K, Suratit R. et al. Effect of various forms of calcium in dental products on human enamel microhardness in vitro. Southeast Asian J Trop Med Public Health 2012; 43 (04) 1053-1058
- 13 Lee J, Townsend JA, Thompson T. et al. Analysis of the cariogenic potential of various almond milk beverages using a streptococcus mutans biofilm model in vitro. Caries Res 2018; 52 (1–2): 51-57
- 14 Bowen WH, Koo H. Biology of Streptococcus mutans-derived glucosyltransferases: role in extracellular matrix formation of cariogenic biofilms. Caries Res 2011; 45 (01) 69-86
- 15 Huang Y, Thompson T, Wang Y. et al. Analysis of cariogenic potential of alternative milk beverages by in vitro Streptococcus mutans biofilm model and ex vivo caries model. Arch Oral Biol 2019; 105: 52-58
- 16 Ceballos LS, Morales ER, de la Torre Adarve G, Castro JD, Martínez LP, Sampelayo MRS. Composition of goat and cow milk produced under similar conditions and analyzed by identical methodology. Int J Food Sci Nutr 2009; 22 (04) 322-329
- 17 Song N, Chen Y, Luo J. et al. Negative regulation of αS1-casein (CSN1S1) improves β-casein content and reduces allergy potential in goat milk. J Dairy Sci 2020; 103 (10) 9561-9572
- 18 Dashper SG, Saion BN, Stacey MA. et al. Acidogenic potential of soy and bovine milk beverages. J Dent 2012; 40 (09) 736-741
- 19 Bergsson G, Arnfinnsson J, Steingrímsson O, Thormar H. Killing of Gram-positive cocci by fatty acids and monoglycerides. Acta Pathol Microbiol Scand Suppl 2001; 109 (10) 670-678
- 20 Wimolsantirungsri K, Asvanund Y, Mitrakul K, Srisatjaluk RL. Effect of bovine and plant based milks on Streptococcus mutans biofilm formation, biofilm pH Level and enamel demineralization in human primary teeth. Southeast Asian J Trop Med Public Health 2023; 54 (02) 71-86
- 21 Muñoz-Sandoval C, Muñoz-Cifuentes MJ, Giacaman RA, Ccahuana-Vasquez RA, Cury JA. Effect of bovine milk on Streptococcus mutans biofilm cariogenic properties and enamel and dentin demineralization. Pediatr Dent 2012; 34 (07) e197-e201
- 22 Giacaman RA, Muñoz-Sandoval C. Cariogenicity of different commercially available bovine milk types in a biofilm caries model. Pediatr Dent 2014; 36 (01) 1E-6E
- 23 Bergsson G, Arnfinnsson J, Steingrímsson O, Thormar H. In vitro killing of Candida albicans by fatty acids and monoglycerides. Antimicrob Agents Chemother 2001; 45 (11) 3209-3212
- 24 Mandalari G, Bisignano C, D'Arrigo M. et al. Antimicrobial potential of polyphenols extracted from almond skins. Lett Appl Microbiol 2010; a 51 (01) 83-89
- 25 Shen P, Walker GD, Yuan Y. et al. Effects of soy and bovine milk beverages on enamel mineral content in a randomized, double-blind in situ clinical study. J Dent 2019; 88: 103160
- 26 Xiao J, Hara AT, Kim D, Zero DT, Koo H, Hwang G. Biofilm three-dimensional architecture influences in situ pH distribution pattern on the human enamel surface. Int J Oral Sci 2017; 9 (02) 74-79
- 27 Paraschos S, Magiatis P, Mitakou S. et al. In vitro and in vivo activities of Chios mastic gum extracts and constituents against Helicobacter pylori . Antimicrob Agents Chemother 2007; 51 (02) 551-559
- 28 Magi G, Marini E, Brenciani A. et al. Chemical composition of Pistacia vera L. oleoresin and its antibacterial, anti-virulence and anti-biofilm activities against oral streptococci, including Streptococcus mutans . Arch Oral Biol 2018; 96: 208-215
- 29 Lemos JA, Palmer SR, Zeng L. et al The biology of Streptococcus mutans. Microbiol Spectr 2019; 7 (01)
- 30 Assaf D, Steinberg D, Shemesh M. Lactose triggers biofilm formation by Streptococcus mutans . Int Dairy J 2015; 42: 51-57
- 31 Ricomini Filho AP, de Assis ACM, Costa Oliveira BE, Cury JA. Cariogenic potential of human and bovine milk on enamel demineralization. Caries Res 2021; 55 (04) 260-267
- 32 Shimizu A, Yamamoto T, Nakashima S, Nikaido T, Sugawara T, Momoi Y. Measurement of surface hardness of primary carious lesions in extracted human enamel—measurement of Knoop hardness using Cariotester. Dent Mater J 2015; 34 (02) 252-256
- 33 Dikmen B. Icdas II criteria (international caries detection and assessment system). J Istanb Univ Fac Dent 2015; 49 (03) 63-72
- 34 Prabhakar AR, Kurthukoti AJ, Gupta P. Cariogenicity and acidogenicity of human milk, plain and sweetened bovine milk: an in vitro study. J Clin Pediatr Dent 2010; 34 (03) 239-247
- 35 Kim D, Barraza JP, Arthur RA. et al. Spatial mapping of polymicrobial communities reveals a precise biogeography associated with human dental caries. Proc Natl Acad Sci USA 2020; 117 (22) 12375-12386
- 36 Bowen WH, Pearson SK, Rosalen PL, Miguel JC, Shih AY. Assessing the cariogenic potential of some infant formulas, milk and sugar solutions. J Am Dent Assoc 1997; 128 (07) 865-871