Evaluation of the effects of probiotic and postbiotic toothpastes for children on cariogenic dental biofilms: an in vitro study

Background: This study aimed to evaluate the effects of commercially available probiotic- and postbiotic-containing toothpastes formulated for children on multispecies cariogenic biofilms under in vitro conditions. Methods: Multispecies biofilms composed of Streptococcus mutans ((American Type Culture Collection) ATCC 25175), Lactobacillus casei (ATCC 4646), and Scardovia wiggsiae ((German Collection of Microorganisms and Cell Cultures) DSM 22547) were formed on hydroxyapatite (HAP) discs. Seven experimental groups were evaluated, including a fluoride-containing positive control, a negative control, and commercially available probiotic-and postbiotic-containing children’s toothpastes. The biofilms were exposed to toothpaste slurries. Biofilm biomass and live/dead cell ratios were quantitatively evaluated using confocal laser scanning microscopy (CLSM) with subsequent analysis through COMSTAT software. Statistical comparisons were performed using the Kruskal-Wallis test followed by Dunn’s post hoc test (p < 0.05). Results: All toothpaste formulations significantly reduced live/dead bacterial biomass ratios compared with the negative control, while the fluoride-containing toothpaste (positive control) showed the strongest antibiofilm effect. Conclusions: Probiotic- and postbiotic-containing toothpastes modulated multispecies dental biofilm viability in vitro and may serve as complementary agents to fluoride-based formulations in pediatric oral care.

Toothpaste; Antibacterial; Biofilm; Probiotic; Postbiotic

[1] Pitts NB, Zero DT, Marsh PD, Ekstrand K, Weintraub JA, Ramos-Gomez F, et al. Dental caries. Nature Reviews Disease Primers. 2017; 3: 17030.

[2] Wang X, Liu M, Yu C, Li J, Zhou X. Biofilm formation: mechanistic insights and therapeutic targets. Molecular Biomedicine. 2023; 4: 49.

[3] Featherstone JD. Dental caries: a dynamic disease process. Australian Dental Journal. 2008; 53: 286–291.

[4] Al-Shahrani MA. Microbiology of dental caries: a literature review. Annals of Medical and Health Sciences Research. 2019; 9: 655–659.

[5] Spatafora G, Li Y, He X, Cowan A, Tanner AC. The evolving microbiome of dental caries. Microorganisms. 2024; 12: 121.

[6] Avila M, Ojcius DM, Yilmaz O. The oral microbiota: living with a permanent guest. DNA and Cell Biology. 2009; 28: 405–411.

[7] Lu Y, Lin Y, Li M, He J. Roles of Streptococcus mutans–Candida albicans interaction in early childhood caries: a literature review. Frontiers in Cellular and Infection Microbiology. 2023; 13: 1151532.

[8] Mulla SA, Mistry L, Kamath S, Deshpande S, Agarwal S, Kondkari SA, et al. Scardovia wiggsiae: a key cariopathogen. Indian Journal of Microbiology Research. 2025; 12: 34–39.

[9] Carlsson J, Grahnén H, Jonsson G. Lactobacilli and streptococci in the mouth of children. Caries Research. 1975; 9: 333–339.

[10] Marsh PD. Contemporary perspective on plaque control. British Dental Journal. 2012; 212: 601–606.

[11] Marsh PD, Zaura E. Dental biofilm: ecological interactions in health and disease. Journal of Clinical Periodontology. 2017; 44: S12–S22.

[12] Lippert F. An introduction to toothpaste—its purpose, history and ingredients. Monographs in Oral Science. 2013; 23: 1–14.

[13] Marinho VC, Higgins JP, Sheiham A, Logan S. Fluoride toothpastes for preventing dental caries in children and adolescents. Cochrane Database of Systematic Reviews. 2003; 2003: CD002278.

[14] Luo SC, Wei SM, Luo XT, Yang QQ, Wong KH, Cheung PCK, et al. How probiotics, prebiotics, synbiotics, and postbiotics prevent dental caries: an oral microbiota perspective. npj Biofilms and Microbiomes. 2024; 10: 14.

[15] Gruner D, Paris S, Schwendicke F. Probiotics for managing caries and periodontitis: systematic review and meta-analysis. Journal of Dentistry. 2016; 48: 16–25.

[16] Scott E, De Paepe K, Van De Wiele T. Postbiotics and their health modulatory biomolecules. Biomolecules. 2022; 12: 1640.

[17] Cai JN, Kim D. Biofilm ecology associated with dental caries: understanding of microbial interactions in oral communities leads to development of therapeutic strategies targeting cariogenic biofilms. Advances in Applied Microbiology. 2023; 122: 27–75.

[18] Golestannejad Z, Khozeimeh F, Abtahi R, Zarei Z, Sadeghalbanaei L, Sadeghian R. Inhibitory effects of ethanolic, methanolic, and hydroalcoholic extracts of olive (Olea europaea) leaf on growth, acid production, and adhesion of Streptococcus mutans. Dental Research Journal. 2020; 17: 179–185.

[19] Pratten J, Smith AW, Wilson M. Response of single species biofilms and microcosm dental plaques to pulsing with chlorhexidine. Journal of Antimicrobial Chemotherapy. 1998; 42: 453–459.

[20] Foster JS, Kolenbrander PE. Development of a multispecies oral bacterial community in a saliva-conditioned flow cell. Applied and Environmental Microbiology. 2004; 70: 4340–4348.

[21] Zhou Y, Millhouse E, Shaw T, Lappin DF, Rajendran R, Bagg J, et al. Evaluating Streptococcus mutans strain dependent characteristics in a polymicrobial biofilm community. Frontiers in Microbiology. 2018; 9: 1498.

[22] Kara D, Luppens SB, Cate JM. Differences between single- and dual-species biofilms of Streptococcus mutans and Veillonella parvula in growth, acidogenicity and susceptibility to chlorhexidine. European Journal of Oral Sciences. 2006; 114: 58–63.

[23] Thurnheer T, Belibasakis GN. Streptococcus oralis maintains homeostasis in oral biofilms by antagonizing the cariogenic pathogen Streptococcus mutans. Molecular Oral Microbiology. 2018; 33: 234–239.

[24] Jasim Al-Adool R, Muwafaq Hamdoon S, Faiz Hamodat H. Scardovia wiggsiae as recent player in caries process: a review. Mustansiria Dental Journal. 2023; 19: 117–123.

[25] Luo TL, Vanek ME, Gonzalez-Cabezas C, Marrs CF, Foxman B, Rickard AH. In vitro model systems for exploring oral biofilms: from single-species populations to complex multi-species communities. Journal of Applied Microbiology. 2022; 132: 855–871.

[26] Kreth J, Hagerman E, Tam K, Merritt J, Wong DTW, Wu BM, et al. Quantitative analyses of Streptococcus mutans biofilms with quartz crystal microbalance, microjet impingement and confocal microscopy. Biofilms. 2004; 1: 277–284.

[27] Serbiak B, Fourre T, Geonnotti AR, Gambogi RJ. In vitro efficacy of essential oil mouthrinse versus dentifrices. Journal of Dentistry. 2018; 69: 49–54.

[28] Cho MY, Kang SM, Lee ES, Kim BI. Antimicrobial activity of Curcuma xanthorrhiza nanoemulsions on Streptococcus mutans biofilms. Biofouling. 2020; 36: 825–833.

[29] Banakar M, Pourhajibagher M, Etemad-Moghadam S, Mehran M, Yazdi MH, Haghgoo R, et al. Antimicrobial effects of postbiotic mediators derived from Lactobacillus rhamnosus GG and Lactobacillus reuteri on Streptococcus mutans. Frontiers in Bioscience. 2023; 28: 88.

[30] Jeong D, Kim DH, Song KY, Seo KH. Antimicrobial and anti-biofilm activities of Lactobacillus kefiranofaciens DD2 against oral pathogens. Journal of Oral Microbiology. 2018; 10: 1472985.

[31] Radaic A, Ye C, Parks B, Gao L, Kuraji R, Malone E, et al. Modulation of pathogenic oral biofilms towards health with nisin probiotic. Journal of Oral Microbiology. 2020; 12: 1809302.

[32] Allaker RP, Stephen AS. Use of probiotics and oral health. Current Oral Health Reports. 2017; 4: 309–318.

[33] Inchingolo AD, Malcangi G, Semjonova A, Inchingolo AM, Patano A, Coloccia G, et al. Oralbiotica/Oralbiotics: the impact of oral microbiota on dental health and demineralization: a systematic review. Children. 2022; 9: 1014.

[34] Wasfi R, Abd El-Rahman OA, Zafer MM, Ashour HM. Probiotic Lactobacillus sp. inhibit growth, biofilm formation and gene expression of caries—inducing Streptococcus mutans. Journal of Cellular and Molecular Medicine. 2018; 22: 1972–1983.

[35] Chen Z, Schlafer S, Göstemeyer G, Schwendicke F. Probiotic effects on multispecies biofilm composition, architecture, and caries activity in vitro. Microorganisms. 2020; 8: 1272.

[36] Che J, Shi J, Fang C, Zeng X, Wu Z, Du Q, et al. Elimination of pathogen biofilms via postbiotics from lactic acid bacteria: a promising method in food and biomedicine. Microorganisms. 2024; 12: 704.

[37] Jung JI, Baek SM, Nguyen TH, Kim JW, Kang CH, Kim S, et al. Effects of probiotic culture supernatant on cariogenic biofilm formation and RANKL-induced osteoclastogenesis in RAW 264.7 macrophages. Molecules. 2021; 26: 733.

[38] Lin X, Chen X, Tu Y, Wang S, Chen H. Effect of probiotic lactobacilli on the growth of Streptococcus mutans and multispecies biofilms isolated from children with active caries. Medical Science Monitor. 2017; 23: 4175–4181.