Mette Kirstine Keller

Short-term effect of chewing gums containing probiotic Lactobacillus reuteri on the levels of inflammatory mediators in gingival crevicular fluid

Objective. To investigate the effect of a chewing gum containing probiotic bacteria on gingival inflammation and the levels of selected inflammatory mediators in gingival crevicular fluid (GCF). Material and Methods. Forty-two healthy adults with moderate levels of gingival inflammation entered a double-blind placebo-controlled study design. The subjects were randomly assigned to one of three parallel arms: Group A/P was given one active and one placebo gum daily, Group A/A received two active chewing gums, and Group P/P two placebo gums. The chewing gums contained two strains of Lactobacillus reuteri: ATCC 55730 and ATCC PTA 5289 (1×108 CFU/gum, respectively). The subjects were instructed to chew the gums for 10 min over the course of 2 weeks. Bleeding on probing (BOP) and GCF sampling were conducted at baseline and after 1, 2 and 4 weeks. The levels of IL-1β, TNF-α, IL-6, IL-8 and IL-10 were determined using luminex technology and multiplex immunoassay kits. Results. BOP improved and GCF volume decreased in all groups during the chewing period, but the results were statistically significant (p<0.05) only in Groups A/P and A/A. The levels of TNF-α and IL-8 decreased significantly (p<0.05) in Group A/A compared with baseline after 1 and 2 weeks, respectively. A non-significant decreasing tendency was also observed concerning IL-1β during the chewing period. The levels of IL-6 and IL-10 were unaffected in all groups after 1 and 2 weeks. Conclusions. The reduction of pro-inflammatory cytokines in GCF may be proof of principle for the probiotic approach combating inflammation in the oral cavity.

Probiotics for Caries Prevention and Control

Modulation of the microbiota for restoring and maintaining health is a growing issue in medical science. A search for relevant clinical trials on the use of probiotic bacteria as a potential and clinically applicable anti-caries measure was performed. According to predetermined criteria, papers were selected and key data on study design, sample size, intervention, duration, and results were extracted. Two animal and 19 human studies were retrieved. Most studies were short-term and restricted to microbiological endpoints, and only 3 human studies reported a caries endpoint. A high degree of heterogeneity among the included investigations hampered the analysis. Significant reductions of mutans streptococci in saliva or plaque following daily intake of probiotic lactobacilli or bifidobacteria were reported in 12 out of 19 papers, whereas 3 reported an increase of lactobacilli. Three caries trials in preschool children and the elderly demonstrated prevented fractions of between 21% and 75% following regular intakes of milk supplemented with L. rhamnosus. No adverse effects or potential risks were reported. The currently available literature does not exclude the possibility that probiotic bacteria can interfere with the oral biofilm, but any clinical recommendation would be premature. Large-scale clinical studies with orally derived specific anti-caries candidates are still lacking.

Co-aggregation and growth inhibition of probiotic lactobacilli and clinical isolates of mutans streptococci: An in vitro study

Abstract 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.

Effect of chewing gums containing the probiotic bacterium Lactobacillus reuteri on oral malodour

Abstract Objective. To evaluate the effect of chewing gums containing probiotic bacteria on oral malodour. The null hypothesis was that no difference would be displayed compared with placebo gums. Materials and methods. Twenty-five healthy young adults with self-reported malodorous morning breath completed this randomized double-blind placebo-controlled cross-over trial. The design included run-in and wash-out periods interspersed by two intervention periods of 14 days each. The subjects were instructed to chew one gum in the morning and one in the evening containing either two strains of probiotic lactobacilli (L. reuteri DSM 17938 and L. reuteri ATCC PTA 5289) or placebo. The outcome measures were (i) organoleptic scores (0–5) by a certified test panel, (ii) concentration of volatile sulphur compounds (VSC) measured with a Halimeter and (iii) concentration of VSC after a cysteine rinse. Registrations were made at baseline and after each intervention period. Differences between the groups were assessed by non-parametric paired statistics and chi-square test. Results. The median organoleptic score was similar (score 2) in both groups at baseline. After 14 days of treatment, the organoleptic scores were significantly lower in the probiotic group compared with the placebo group (p < 0.05). Assessments of the VSC levels displayed no significant differences between the groups, either before or after rinsing with L-cysteine. No adverse effects were registered. Conclusion. The results demonstrated that probiotic chewing gums may have some beneficial effect on oral malodour assessed by organoleptic scores. The results indicate that the probiotic gum may affect bacteria that produce malodourous compounds other than VSCs.

Effect of Probiotic Bacteria on Oral Candida in Frail Elderly

The aim of this study was to investigate the effect of a daily intake of probiotic lactobacilli on the prevalence and counts of oral Candida in frail elderly patients living in nursing homes. The study had a double-blind randomized placebo-controlled design with 2 parallel arms. The study group consisted of 215 older adults (range, 60 to 102 y) who were enrolled after informed consent. After baseline examination and randomization, the subjects were given 1 lozenge containing 2 strains of the probiotic bacterium Lactobacillus reuteri (DSM 17938 and ATCC PTA 5289) or placebo twice daily (morning and evening). The intervention period was 12 wk, and saliva and plaque samples were collected at baseline and follow-up. The primary end point was prevalence of high Candida counts assessed from chairside tests. Secondary end points were levels of dental plaque and gingival inflammation. The groups were balanced at baseline. The attrition rate to follow-up was 19%. There was a statistically significant reduction in the prevalence of high Candida counts in the probiotic group but not in the placebo group, and the difference was statistically significant in both saliva and plaque (P < 0.05). No significant differences between the groups were noted concerning the levels of supragingival plaque or bleeding on probing. Thus, daily use of probiotic lozenges may reduce the prevalence of high oral Candida counts in frail elderly nursing homes residents (ClinicalTrials.gov NCT02391532).

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

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.

Acid production in dental plaque after exposure to probiotic bacteria

BackgroundThe increasing interest in probiotic lactobacilli in health maintenance has raised the question of potential risks. One possible side effect could be an increased acidogenicity in dental plaque. The aim of this study was to investigate the effect of probiotic lactobacilli on plaque lactic acid (LA) production in vitro and in vivo.MethodsIn the first part (A), suspensions of two lactobacilli strains (L. reuteri DSM 17938, L. plantarum 299v) were added to suspensions of supragingival dental plaque collected from healthy young adults (n=25). LA production after fermentation with either xylitol or fructose was analyzed. In the second part (B), subjects (n=18) were given lozenges with probiotic lactobacilli (L. reuteri DSM 17938 and ATCC PTA 5289) or placebo for two weeks in a double-blinded, randomized cross-over trial. The concentration of LA in supragingival plaque samples was determined at baseline and after 2 weeks. Salivary counts of mutans streptococci (MS) and lactobacilli were estimated with chair-side methods.ResultsPlaque suspensions with L. reuteri DSM 17938 produced significantly less LA compared with L. plantarum 299v or controls (p<0.05). Fructose gave higher LA concentrations than xylitol. In part B, there were no significant differences in LA production between baseline and follow up in any of the groups and no differences between test and placebo were displayed. The salivary MS counts were not significantly altered during the intervention but the lactobacilli counts increased significantly in the test group (p<0.05).ConclusionLactic acid production in suspensions of plaque and probiotic lactobacilli was strain-dependant and the present study provides no evidence of an increase in plaque acidity by the supply of selected probiotic lactobacilli when challenged by fructose or xylitol. The study protocol was approved by The Danish National Committee on Biomedical Research Ethics (protocol no H-2-2010-112).Trial registrationNCT01700712

Lactobacillus reuteri supplements do not affect salivary IgA or cytokine levels in healthy subjects : A randomized, double-blind placebo-controlled, cross-over trial

Abstract Objectives: To evaluate the effect of daily ingestion of probiotic lactobacilli on the levels of secretory IgA (sIgA) and selected cytokines in whole saliva of healthy young adults. Materials and methods: The study group consisted of 47 healthy adults (18–32 years) who volunteered for a randomized, double-blind, placebo-controlled, cross-over trial after informed consent. During intervention, the subjects ingested two lozenges per day containing two strains of the probiotic bacterium Lactobacillus reuteri (DSM 17938 and ATCC PTA 5289) or placebo lozenges. The intervention and wash-out periods were 3 weeks. Saliva samples were collected at baseline, immediately after each intervention period and 3 weeks post-intervention. ELISA was used to measure sIgA and luminex technology was used to measure the interleukins (IL)-1β, IL-6, IL-8 and IL-10. For statistical analyses a mixed ANOVA model was employed to calculate changes in the salivary outcome variables. Results: Forty-one subjects completed the study and reported a good compliance. No significant differences in the concentrations of salivary sIgA or cytokines were recorded between the L. reuteri and placebo interventions or between baseline and 3 weeks post-intervention levels. No side- or adverse effects were reported. Conclusions: Supplementation with two strains of the probiotic L. reuteri did not affect sIgA or cytokine levels in whole saliva in healthy young adults. The results thereby indicate that daily oral supplementation with L. reuteri do not seem to modulate the salivary oral immune response in healthy young subjects (ClinicalTrials.gov NCT02017886).

Lactobacillus reuteri Influences Regrowth of Mutans Streptococci after Full-Mouth Disinfection: A Double-Blind, Randomised Controlled Trial

This study assessed whether the persistence of Lactobacillus reuteri DSM 17938 and ATCC PTA 5289 in saliva could delay the regrowth of mutans streptococci (MS) after a full-mouth disinfection with chlorhexidine (CHX). A randomised, double-blind, placebo-controlled study with a 6-week intervention period and 3- and 6-month follow-up was performed. 62 healthy subjects with moderate to high counts of MS were randomly assigned to a test group (n = 32) or a placebo group (n = 30). Before onset of the intervention, subjects received two sessions of professional cleaning, flossing, and application of CHX varnish and rinsed their mouth with a CHX solution between the sessions (2 days). Thereafter, the test group used probiotic lozenges (2/day) containing L. reuteri (DSM 17938 and ATCC PTA 5289; 1 × 108 CFU of each strain), and the placebo group used identical lozenges lacking the lactobacilli. Saliva samples were collected and cultured onto selective media, and isolates of L. reuteri as well as DNA directly extracted from saliva were tested by polymerase chain reaction (PCR) with specific primers. Presence of salivary MS was analysed with a chair-side test. L. reuteri was frequently detected by culture during the intervention period but in only 3 test group subjects at follow-ups. Regrowth of MS statistically significantly differed depending on the presence or absence of L. reuteri DSM 17938 detected by PCR. We conclude that cultivable L. reuteri strains may only sporadically be confirmed after termination of the intervention, but subjects with PCR-detected L. reuteri demonstrated slower regrowth of MS.

Effect of tablets containing probiotic bacteria (Lactobacillus reuteri) on early caries lesions in adolescents: a pilot study.

The objective of the study was to investigate the effect of tablets containing probiotic lactobacilli on early caries lesions in adolescents with quantitative light-induced fluorescence (QLF). 36 healthy adolescents of both sexes (12-17 years of age) were enrolled and randomly allocated to a placebo-controlled trial with two parallel groups. The test group received two tablets daily containing two strains of Lactobacillus reuteri (DSM 17938 and ATCC PTA 5289) for a period of three months, while the control group got identical placebo tablets without live bacteria. The primary outcome was QLF-readings (change in fluorescence, ΔF and lesion area, mm2) at baseline and after 3 months, conducted at two buccal sites of each individual, pre-selected with clearly visible clinical signs of enamel demineralisation (white spots). Significantly more premolars were allocated to the placebo group, while the test group had more incisors (P<0.05). There were no statistically significant differences in fluorescence values between the groups, neither at baseline, nor at the follow-up. There was however a significant decrease in fluorescence over time in the test group, but not in the placebo group (P<0.05). No alterations of the lesion area (ΔA) were found in any group. The inter-examiner intra-class correlation coefficient-value for QLF-readings was excellent. No side- or adverse effects were reported during the intervention period. This pilot study found a significant decrease over time in the test group. However, no statistically significant differences in fluorescence values between the groups were found. Hence, the null hypothesis could not be rejected.