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Probiotic Lactobacilli in the context of dental caries as a biofilm-mediated disease
Umeå University, Faculty of Medicine, Department of Odontology.
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Background: The World Health Organization defines probiotics as ‘live microorganisms which, when administered in adequate amounts, confer a health benefit to the host’. Traditionally, probiotic microorganisms have been used to prevent or treat gastrointestinal tract diseases. In the last 15 years, there has been increasing interest of a possible probiotic impact on the oral microbiota and dental caries. Dental caries is a multifactorial disease, and the causative factor in the oral microbiota includes a shift from a balanced microflora to a microflora that includes more aciduric species such as mutans streptococci (MS), non-mutans streptococci, and Actinomyces. MS is considered an opportunistic pathogen although several other bacteria also contribute to the disease. Early acquisition of MS is associated with early development of caries; therefore a desirable complement to other prophylactic measures would be a MS colonization inhibitor.

Objective: To better understand how selected strains of probiotic lactobacilli interact with MS in vitro and in vivo and to study the impact of probiotic lactobacilli on caries development during childhood.

Material and methods: The in vitro properties of probiotic lactobacilli were studied with regard to (i) acid production from sugars and sugar alcohols, (ii) growth inhibition capacity on clinical isolates and reference strains of MS as well as Candida albicans and (iii) the capacity to co-aggregate with MS. A randomized controlled trial (RCT) tested the short-term effect of intervention with two Lactobacillus reuteri strains on MS, which was evaluated after treatment with chlorhexidine. The re-growth patterns of MS and 19 other selected strains were also evaluated. In the second clinical study  we investigated the long-term effect on MS prevalence and dental caries after an intervention with Lactobacillus paracasei ssp. paracasei F19 (LF19) between 4 and 13 months of age.

Results: The results from the in vitro testing showed that strains of probiotic lactobacilli differed in their fermentation patterns, inhibition capacity and their capacity to co-aggregate, which should be kept in mind in the translation to clinical research. The clinical study on short-term effects of two L. reuteri strains on MS and other oral strains showed no effect on re-growth patterns after intervention. The clinical study on long-term effects of LF19 showed no effect on the prevalence of MS. Furthermore, the clinical follow-up at 9 years of age showed no differences in either decayed, missing, and filled surface (dmfs) or DMFS between the probiotic and placebo groups. Evaluation of saliva samples showed no signs of oral colonization with LF19 in the study group.

Conclusion: The in vitro testing showed potentials of the selected probiotic Lactobacillus strains for interference with MS and C. albicans. The results from the clinical studies showed no such effect on MS or dental caries. Evidence regarding the effectiveness of specific probiotic applications in the prevention of dental caries is limited and does not allow for conclusions concerning the use of probiotic bacteria as a preventive measure.

Place, publisher, year, edition, pages
Umeå: Umeå universitet , 2013. , 57 p.
Series
Umeå University odontological dissertations, ISSN 0345-7532 ; 125
Keyword [en]
caries prevention, co-aggregation, dental caries, growth inhibition, mutans streptococci, probiotic lactobacilli, re-colonization
National Category
Dentistry
Research subject
Odontology
Identifiers
URN: urn:nbn:se:umu:diva-67843ISBN: 978-91-7459-574-1 (print)OAI: oai:DiVA.org:umu-67843DiVA: diva2:615181
Public defence
2013-05-03, Sal B, Tandläkarhögskolan, Norrlands universitetssjukhus, Umeå, 13:00 (Swedish)
Opponent
Supervisors
Available from: 2013-04-12 Created: 2013-04-04 Last updated: 2013-04-12Bibliographically approved
List of papers
1. Sugar fermentation in probiotic bacteria: an in vitro study
Open this publication in new window or tab >>Sugar fermentation in probiotic bacteria: an in vitro study
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2008 (English)In: Oral Microbiology and Immunology, ISSN 0902-0055, E-ISSN 1399-302X, Vol. 23, no 6, 482-485 p.Article in journal (Refereed) Published
Abstract [en]

INTRODUCTION: Food supplemented with probiotic bacteria is a rapidly growing sector of the market. The aim of the present study was to evaluate and compare the acid production of selected probiotic strains available in commercial products.

METHODS: Six Lactobacillus strains (Lactobacillus plantarum 299v and 931; Lactobacillus rhamnosus GG and LB21; Lactobacillus paracasei subsp. paracasei F19, and Lactobacillus reuteri PTA 5289) were cultivated at 37 degrees C in an anaerobic atmosphere on Man, Rogosa, Shape (MRS) agar for 48 h or MRS broth for 16 h. After centrifugation, the cells were washed and resuspended in sterile phosphate-buffered saline and immediately subjected to a fermentation assay with 12 different carbohydrates (nine sugars and three sugar alcohols) in microtiter plates with a pH indicator. The plates were examined for color changes after 24, 48, and 72 h of incubation under aerobic and anaerobic conditions. Three scores were used: negative (pH > 6.8); weak (pH 5.2-6.8), and positive (pH < 5.2). The strains were characterized with the API 50 CH system to confirm their identity.

RESULTS: L. plantarum fermented all the sugars except for melibiose, raffinose, and xylitol. Both L. rhamnosus strains were generally less active although L. rhamnosus GG was slightly more active than strain LB21 in the 5% CO(2) setting. The latter strain exhibited negative reactions for sucrose, maltose, arabinose, and sorbitol under anaerobic conditions. The assays with L. paracasei and L. reuteri had negative or weak reactions for all tested sugars under both aerobic and anaerobic conditions.

CONCLUSION: The metabolic capacity to form acid from dietary sugars differed significantly between the various probiotic strains.

National Category
Dentistry
Identifiers
urn:nbn:se:umu:diva-26381 (URN)10.1111/j.1399-302X.2008.00457.x (DOI)18954354 (PubMedID)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
2. Growth inhibition of oral mutans streptococci and candida by commercial probiotic lactobacilli: an in vitro study
Open this publication in new window or tab >>Growth inhibition of oral mutans streptococci and candida by commercial probiotic lactobacilli: an in vitro study
2010 (English)In: BMC Oral Health, ISSN 1472-6831, E-ISSN 1472-6831, Vol. 10, 18- p.Article in journal (Refereed) Published
Abstract [en]

The selected probiotic strains showed a significant but somewhat varying ability to inhibit growth of oral mutans streptococci and Candida albicans in vitro.

National Category
Dentistry
Identifiers
urn:nbn:se:umu:diva-40681 (URN)10.1186/1472-6831-10-18 (DOI)000208596700018 ()20598145 (PubMedID)
Available from: 2011-03-05 Created: 2011-03-05 Last updated: 2017-12-11Bibliographically approved
3. Co-aggregation and growth inhibition of probiotic lactobacilli and clinical isolates of mutans streptococci: an in vitro study
Open this publication in new window or tab >>Co-aggregation and growth inhibition of probiotic lactobacilli and clinical isolates of mutans streptococci: an in vitro study
2011 (English)In: Acta Odontologica Scandinavica, ISSN 0001-6357, E-ISSN 1502-3850, Vol. 69, no 5, 263-268 p.Article in journal (Refereed) Published
Abstract [en]

Objective: Co-aggregation and growth inhibition abilities of probiotic bacteria may play a key role in their interference with the oral biofilm. The aim was to investigate the in vitro ability of selected commercial probiotic lactobacilli to co-aggregate and inhibit growth of oral mutans steptococci isolated from adults with contrasting levels of caries.

Materials and methods: Mutans streptococci (MS) strains were isolated from caries-free (n = 3) and caries-susceptible (n = 5) young adults and processed with eight commercial probiotic lactobacilli strains. One laboratory reference strain (S. mutans Ingbritt) was selected as control. Co-aggregation was determined spectrophotometrically and growth inhibition was assessed with the agar overlay technique.

Results: All probiotic lactobacilli showed an ability to co-aggregate with the isolated MS strains. Statistically significant differences (p < 0.05) were found between strains from different individuals when compared with the reference strain. The selected lactobacilli inhibited MS growth, but the ability varied between the strains and was clearly related to pH. No differences were observed between the different MS strains from caries-free and caries-susceptible individuals.

Conclusions: The selected lactobacilli displayed co-aggregation activity and inhibited growth of clinical mutans streptococci. The growth inhibition was strain-specific and dependent on pH and cell concentration. The findings indicate that the outcome of lactobacilli-derived probiotic therapy might vary between individuals and depend on the specific strain used.

Place, publisher, year, edition, pages
Oslo: Taylor & Francis, 2011
Keyword
Dental caries, oral microorganisms, probiotics
National Category
Dentistry
Identifiers
urn:nbn:se:umu:diva-46614 (URN)10.3109/00016357.2011.554863 (DOI)21306197 (PubMedID)
Available from: 2011-09-07 Created: 2011-09-07 Last updated: 2017-12-08Bibliographically approved
4. Probiotic supplements (Lactobacillus reuteri DSM 17938 and ATCC PTA 5289) do not affect regrowth of mutans streptococci after full-mouth disinfection with chlorhexidine: a randomized controlled multicenter trial
Open this publication in new window or tab >>Probiotic supplements (Lactobacillus reuteri DSM 17938 and ATCC PTA 5289) do not affect regrowth of mutans streptococci after full-mouth disinfection with chlorhexidine: a randomized controlled multicenter trial
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2012 (English)In: Caries Research, ISSN 0008-6568, E-ISSN 1421-976X, Vol. 46, no 2, 140-146 p.Article in journal (Refereed) Published
Abstract [en]

The aim of this study was to investigate the effectiveness of tablets containing two probiotic Lactobacillus reuteri strains in inhibiting regrowth of salivary mutans streptococci (MS) after full-mouth disinfection (FMD) with chlorhexidine. The null hypothesis was that the levels of MS would not differ in comparison with a placebo protocol. The study population was comprised of 62 young adults (mean age 23 years) with moderate or high counts of salivary MS who volunteered after informed consent. The study was a double-blinded randomized controlled trial with two parallel groups. After a 3-day chlorhexidine regimen, the subjects were randomly assigned to a test group (n = 32) with probiotic lozenges (2/day) or a placebo group (n = 30). The intervention period was 6 weeks, and stimulated whole saliva was collected at baseline and after 1, 6, and 12 weeks. The samples were processed for MS by a chair-side test and DNA-DNA hybridization as an estimate of 19 bacterial strains associated with oral health and disease. There was no significant difference between the groups at inclusion, and FMD reduced the salivary MS levels significantly in both groups. The MS suppression lasted less than 6 weeks and there were no statistical differences in salivary MS regrowth between the test and control groups at any of the follow-ups. Likewise, there were no major differences in the regrowth patterns of the checkerboard panel between the two groups. We conclude that daily oral administration of L. reuteri did not seem to affect or delay the regrowth of salivary MS after FMD with chlorhexidine. Copyright (c) 2012 S. Karger AG, Basel

Place, publisher, year, edition, pages
S. Karger, 2012
Keyword
Biofilm, Caries, DNA hybridization, Mutans streptococci, Saliva
National Category
Dentistry
Identifiers
urn:nbn:se:umu:diva-55682 (URN)10.1159/000337098 (DOI)000303368300008 ()
Available from: 2012-05-28 Created: 2012-05-28 Last updated: 2017-12-07Bibliographically approved
5. Early intervention with probiotic Lactobacillus paracasei F19 has no long-term effect on caries experience
Open this publication in new window or tab >>Early intervention with probiotic Lactobacillus paracasei F19 has no long-term effect on caries experience
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2013 (English)In: Caries Research, ISSN 0008-6568, E-ISSN 1421-976X, Vol. 47, no 6, 559-565 p.Article in journal (Refereed) Published
Abstract [en]

The aim of the study was to evaluate possible long-term effects of a cereal diet supplemented with Lactobacillus paracasei F19 (LF19) during weaning on caries experience, mutans streptococci (MS) and lactobacilli (LBC) in a group of 9-year-old children. A secondary aim was to evaluate if the intervention resulted in the permanent integration of LF19 as part of the oral microbiota. The study followed up on a double-blind placebo-controlled randomised trial. Among 179 infants that were randomised to a daily diet that included cereals with or without LF19 from 4 to 13 months of age, 56 from the probiotic group and 62 from the placebo group participated in the follow-up at 9 years. Data were collected by oral clinical examination and questionnaires. MS and LBC levels were assessed with conventional cultivation; LF19 was detected by using randomly amplified polymerase chain reactions (RAPD-PCR). At the follow-up, neither decayed, missing and filled surfaces for primary teeth (dmfs) nor decayed, missing and filled surfaces for permanent teeth (DMFS) differed significantly between the probiotic and placebo groups (p > 0.05). MS and LBC levels were similar in both groups (p > 0.05). RAPD-PCR showed no evidence of oral colonisation with LF19 in the study group. It is concluded that an early intervention with LF19 did not affect the frequency of dental caries, MS or LBC. LF19 did not establish itself as a permanent facet of the oral microbiota in any of the subjects included in this study.

Place, publisher, year, edition, pages
Karger, 2013
Keyword
Dental caries, Lactobacilli, Mutans streptococci, Probiotic intervention

National Category
Dentistry
Research subject
Odontology
Identifiers
urn:nbn:se:umu:diva-67782 (URN)10.1159/000350524 (DOI)
Available from: 2013-04-04 Created: 2013-04-03 Last updated: 2017-12-06Bibliographically approved

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Citation style
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  • modern-language-association-8th-edition
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