Probiotic administration may favour caries prevention, as recent research has shown. This in vitro study aimed to investigate the growth of Lactobacillus rhamnosus GG (LGG) in experimental biofilms exposed to various carbohydrates, and also to assess its cariogenic potential. Multispecies experimental oral biofilms with or without LGG were grown with a sole-carbohydrate source (fructose/glucose/lactose/sorbitol/sucrose). The viable cells of LGG and structure of the biofilms were examined after 64.5 h of incubation, and pH values of spent media were measured at 16.5, 40.5, and 64.5 h. Fermentation profiles of LGG in biofilm media were assessed with study carbohydrate as the sole energy source. Our results showed that LGG reached higher viable cell numbers with glucose and sucrose in 64.5-h multispecies experimental oral biofilms compared to other carbohydrates. When LGG was incorporated in biofilms, no distinct pH changes at any time points were observed under any of the carbohydrates used; the pH values of spent media at each time point were lower when lactose was used, compared to other carbohydrates. The fermentation profiles of LGG in biofilm media were similar to its growth in MRS (no obvious growth with lactose or sucrose). In conclusion, LGG in our in vitro multispecies experimental oral biofilms was capable of surviving and growing well in each carbohydrate source. LGG might not have harmful effects on dental hard tissues. Another finding from our study was that the lowest pH values were observed in the presence of lactose, and the thickest biofilms were in sucrose.

Keywords:
Carbohydrate, Cross-feeding, Fluorescent in situ hybridisation, Probiotics
1.
Aminabadi NA, Erfanparast L, Ebrahimi A, Oskouei SG: Effect of chlorhexidine pretreatment on the stability of salivary lactobacilli probiotic in six- to twelve-year-old children: a randomized controlled trial. Caries Res 2011;45:148-154.
2.
Ardita CS, Mercante JW, Kwon YM, Luo LP, Crawford ME, Powell DN, Jones RM, Neish AS: Epithelial adhesion mediated by pilin SpaC is required for Lactobacillus rhamnosus GG-induced cellular responses. Appl Environ Microbiol 2014;80:5068-5077.
3.
Binkley J, Arnaud MB, Inglis DO, Skrzypek MS, Shah P, Wymore F, Binkley G, Miyasato SR, Simison M, Sherlock G: The Candida Genome Database: the new homology information page highlights protein similarity and phylogeny. Nucleic Acids Res 2014;42:D711-D716.
4.
Boonyanit T, Sroisiri T, Doan Minh T: Fermentation of various sugars and sugar substitutes by oral microorganisms. Asian Pac J Trop Med 2011;1:S258-S260.
5.
Dahlén G, Fiehn N-E, Olsen I, Dahlgren U: Oral Microbiology and Immunology. Copenhagen, Munksgaard Danmark, 2012.
6.
Douillard FP, Ribbera A, Jarvinen HM, Kant R, Pietila TE, Randazzo C, Paulin L, Laine PK, Caggia C, von Ossowski I, Reunanen J, Satokari R, Salminen S, Palva A, de Vos WM: Comparative genomic and functional analysis of Lactobacillus casei and Lactobacillus rhamnosus strains marketed as probiotics. Appl Environ Microbiol 2013;79:1923-1933.
7.
Estrela S, Trisos CH, Brown SP: From metabolism to ecology: cross-feeding interactions shape the balance between polymicrobial conflict and mutualism. Am Nat 2012;180:566-576.
8.
Floch MH, Walker WA, Sanders ME, Nieuwdorp M, Kim AS, Brenner DA, Qamar AA, Miloh TA, Guarino A, Guslandi M, Dieleman LA, Ringel Y, Quigley EMM, Brandt LJ: Recommendations for probiotic use-2015 update proceedings and consensus opinion. J Clin Gastroenterol 2015;49:S69-S73.
9.
Forssten SD, Bjorklund M, Ouwehand AC: Streptococcus mutans, caries and simulation models. Nutrients 2010;2:290-298.
10.
Gruner D, Paris S, Schwendicke F: Probiotics for managing caries and periodontitis: systematic review and meta-analysis. J Dent 2016;48:16-25.
11.
Guggenheim B, Giertsen E, Schupbach P, Shapiro S: Validation of an in vitro biofilm model of supragingival plaque. J Dent Res 2001;80:363-370.
12.
Gupta P, Gupta N, Pawar AP, Birajdar SS, Natt AS, Singh HP: Role of sugar and sugar substitutes in dental caries: a review. ISRN Dent 2013;2013:519421.
13.
Hedberg M, Hasslof P, Sjostrom I, Twetman S, Stecksen-Blicks C: Sugar fermentation in probiotic bacteria - an in vitro study. Oral Microbiol Immunol 2008;23:482-485.
14.
Hibberd PL, Kleimola L, Fiorino AM, Botelho C, Haverkamp M, Andreyeva I, Poutsiaka D, Fraser C, Solano-Aguilar G, Snydman DR: No evidence of harms of probiotic Lactobacillus rhamnosus GG ATCC 53103 in healthy elderly - a phase I open label study to assess safety, tolerability and cytokine responses. PLoS One 2014;9:e113456.
15.
Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, Morelli L, Canani RB, Flint HJ, Salminen S, Calder PC, Sanders ME: The International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastro Hepat 2014;11:506-514.
16.
Jiang Q, Stamatova I, Kainulainen V, Korpela R, Meurman JH: Interactions between Lactobacillus rhamnosus GG and oral micro-organisms in an in vitro biofilm model. BMC Microbiol 2016;16:149.
17.
Jiang Q, Stamatova I, Kari K, Meurman JH: Inhibitory activity in vitro of probiotic lactobacilli against oral Candida under different fermentation conditions. Benef Microbes 2015;6:361-368.
18.
Jorgensen MR, Castiblanco G, Twetman S, Keller MK: Prevention of caries with probiotic bacteria during early childhood: promising but inconsistent findings. Am J Dent 2016;29:127-131.
19.
Laleman I, Detailleur V, Slot DE, Slomka V, Quirynen M, Teughels W: Probiotics reduce mutans streptococci counts in humans: a systematic review and meta-analysis. Clin Oral Investig 2014;18:1539-1552.
20.
Laleman I, Teughels W: Probiotics in the dental practice: a review. Quintessence Int 2015;46:255-264.
21.
Lemos JA, Abranches J, Koo H, Marquis RE, Burne RA: Protocols to study the physiology of oral biofilms. Methods Mol Biol 2010;666:87-102.
22.
Li J, Helmerhorst EJ, Leone CW, Troxler RF, Yaskell T, Haffajee AD, Socransky SS, Oppenheim FG: Identification of early microbial colonizers in human dental biofilm. J Appl Microbiol 2004;97:1311-1318.
23.
Marsh PD: Are dental diseases examples of ecological catastrophes? Microbiology 2003;149:279-294.
24.
Marttinen A, Haukioja A, Karjalainen S, Nylund L, Satokari R, Ohman C, Holgerson P, Twetman S, Soderling E: Short-term consumption of probiotic lactobacilli has no effect on acid production of supragingival plaque. Clin Oral Investig 2012;16:797-803.
25.
Matsumoto M, Tsuji M, Sasaki H, Fujita K, Nomura R, Nakano K, Shintani S, Ooshima T: Cariogenicity of the probiotic bacterium Lactobacillus salivarius in rats. Caries Res 2005;39:479-483.
26.
Mayanagi G, Igarashi K, Washio J, Takahashi N: pH response and tooth surface solubility at the tooth/bacteria interface. Caries Res 2017;51:160-166.
27.
Meurman JH, Antila H, Korhonen A, Salminen S: Effect of Lactobacillus rhamnosus strain GG (ATCC 53103) on the growth of Streptococcus sobrinus in vitro. Eur J Oral Sci 1995;103:253-258.
28.
Moye ZD, Zeng L, Burne RA: Fueling the caries process: carbohydrate metabolism and gene regulation by Streptococcus mutans. J Oral Microbiol 2014;6:24878.
29.
Näse L, Hatakka K, Savilahti E, Saxelin M, Pönkä A, Poussa T, Korpela R, Meurman JH: Effect of long-term consumption of a probiotic bacterium, Lactobacillus rhamnosus GG, in milk on dental caries and caries risk in children. Caries Res 2001;35:412-420.
30.
Pande S, Kaftan F, Lang S, Svatos A, Germerodt S, Kost C: Privatization of cooperative benefits stabilizes mutualistic cross-feeding interactions in spatially structured environments. ISME J 2016;10:1413-1423.
31.
Pham LC, Hoogenkamp MA, Exterkate RA, Terefework Z, de Soet JJ, ten Cate JM, Crielaard W, Zaura E: Effects of Lactobacillus rhamnosus GG on saliva-derived microcosms. Arch Oral Biol 2011;56:136-147.
32.
Pham LC, van Spanning RJM, Roling WFM, Prosperi AC, Terefework Z, ten Cate JM, Crielaard W, Zaura E: Effects of probiotic Lactobacillus salivarius W24 on the compositional stability of oral microbial communities. Arch Oral Biol 2009;54:132-137.
33.
Quevedo B, Giertsen E, Zijnge V, Luthi-Schaller H, Guggenheim B, Thurnheer T, Gmur R: Phylogenetic group- and species-specific oligonucleotide probes for single-cell detection of lactic acid bacteria in oral biofilms. BMC Microbiol 2011;11:14.
34.
Salaun F, Mietton B, Gaucheron F: Buffering capacity of dairy products. Int Dairy J 2005;15:95-109.
35.
Sato Y, Yamagishi J, Yamashita R, Shinozaki N, Ye B, Yamada T, Yamamoto M, Nagasaki M, Tsuboi A: Inter-individual differences in the oral bacteriome are greater than intra-day fluctuations in individuals. PLoS One 2015;10.
36.
Schneider CA, Rasband WS, Eliceiri KW: NIH Image to ImageJ: 25 years of image analysis. Nat Methods 2012;9:671-675.
37.
Schwendicke F, Dorfer C, Kneist S, Meyer-Lueckel H, Paris S: Cariogenic effects of probiotic Lactobacillus rhamnosus GG in a dental biofilm model. Caries Res 2014a;48:186-192.
38.
Schwendicke F, Horb K, Kneist S, Dorfer C, Paris S: Effects of heat-inactivated Bifidobacterium BB12 on cariogenicity of Streptococcus mutans in vitro. Arch Oral Biol 2014b;59:1384-1390.
39.
Selwitz RH, Ismail AI, Pitts NB: Dental caries. Lancet 2007;369:51-59.
40.
Simark-Mattsson C, Emilson CG, Håkansson EG, Jacobsson C, Roos K, Holm S: Lactobacillus-mediated interference of mutans streptococci in caries-free vs. caries-active subjects. Eur J Oral Sci 2007;115:308-314.
41.
Snydman DR: The safety of probiotics. Clin Infect Dis 2008;46(suppl 2):S104-S111; discussion S144-S151.
42.
Soderling EM, Marttinen AM, Haukioja AL: Probiotic lactobacilli interfere with Streptococcus mutans biofilm formation in vitro. Curr Microbiol 2011;62:618-622.
43.
Stamatova I, Meurman JH, Kari K, Tervahartiala T, Sorsa T, Baltadjieva M: Safety issues of Lactobacillus bulgaricus with respect to human gelatinases in vitro. FEMS Immunol Med Microbiol 2007;51:194-200.
44.
Takahashi N, Nyvad B: The role of bacteria in the caries process: ecological perspectives. J Dent Res 2011;90:294-303.
45.
Tanzer JM, Chassy BM, Krichevsky MI: Sucrose metabolism by Streptococcus mutans, SL-I. Biochim Biophys Acta 1971;261:379-387.
46.
Tehrani MH, Akhlaghi N, Talebian L, Emami J, Keyhani SE: Effects of probiotic drop containing Lactobacillus rhamnosus, Bifidobacterium infantis, and Lactobacillus reuteri on salivary Streptococcus mutans and Lactobacillus levels. Contemp Clin Dent 2016;7:469-474.
47.
Thurnheer T, Gmur R, Guggenheim B: Multiplex FISH analysis of a six-species bacterial biofilm. J Microbiol Methods 2004;56:37-47.
48.
Thurnheer T, van der Ploeg JR, Giertsen E, Guggenheim B: Effects of Streptococcus mutans gtfC deficiency on mixed oral biofilms in vitro. Caries Res 2006;40:163-171.
49.
Wolcott R, Costerton JW, Raoult D, Cutler SJ: The polymicrobial nature of biofilm infection. Clin Microbiol Infect 2013;19:107-112.
50.
Wu CC, Lin CT, Wu CY, Peng WS, Lee MJ, Tsai YC: Inhibitory effect of Lactobacillus salivarius on Streptococcus mutans biofilm formation. Mol Oral Microbiol 2015;30:16-26.
51.
Xiao C, Ran S, Huang Z, Liang J: Bacterial diversity and community structure of supragingival plaques in adults with dental health or caries revealed by 16S pyrosequencing. Front Microbiol 2016;7:1145.
52.
Yamada T, Takahashi-Abbe S, Abbe K: Effects of oxygen on pyruvate formate-lyase in situ and sugar metabolism of Streptococcus mutans and Streptococcus sanguis. Infect Immun 1985;47:129-134.
© 2018 S. Karger AG, Basel
2018
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.