Analysis of Chemical Structure and Antibiofilm Properties of Exopolysaccharides from Lactiplantibacillus plantarum EIR/IF-1 Postbiotics.

Previous studies have indicated that the exopolysaccharides of lactic acid bacteria exhibit antibiofilm activity against non-oral bacteria by preventing their initial adhesion to surfaces and by downregulating the expression of genes responsible for their biofilm formation. The aims of this study were to (1) characterize the exopolysaccharides (EPSs) of Lactobacillus plantarum EIR/IF-1 postbiotics, (2) test their antibiofilm effect on dual biofilms, and (3) evaluate their bacterial auto-aggregation, co-aggregation, and hydrocarbon-binding inhibitory activity. The EPSs were characterized by FTIR, HPLC, and thermogravimetric analysis. Bacterial auto- and co-aggregation were tested by Kolenbrander's method and hydrocarbon binding was tested by Rosenberg's method. Dual biofilms were formed by culturing Fusobacterium nucleatum ATCC 25586 with one of the following bacteria: Prevotella denticola ATCC 33185, P. denticola AHN 33266, Porphyromonas gingivalis ATCC 33277, P. gingivalis AHN 24155, and Filifactor alocis ATCC 35896. The EPSs contained fractions with different molecular weights (51 and 841 kDa) and monosaccharides of glucose, galactose, and fructose. The EPSs showed antibiofilm activity in all the biofilm models tested. The EPSs may have inhibited bacterial aggregation and binding to hydrocarbons by reducing bacterial hydrophobicity. In conclusion, the EPSs of L. plantarum EIR/IF-1, which consists of two major fractions, exhibited antibiofilm activity against oral bacteria, which can be explained by the inhibitory effect of EPSs on the auto-aggregation and co-aggregation of bacteria and their binding to hydrocarbons.

Effects of sugar-free polyol chewing gums on gingival inflammation: a systematic review.

OBJECTIVES: A systematic review of published data was conducted with the aim of assessing the effects of sugar-free polyol chewing gums on gingival inflammation. MATERIALS AND METHODS: Electronic and hand searches were performed to find clinical studies concerning the effects of sugar-free chewing gums on gingival scores. Prospective randomized controlled clinical trials published between 1971 and 2021 were included in the review. RESULTS: The initial search identified 46 erythritol, 102 xylitol, 23 sorbitol, and nine maltitol chewing gum articles. After applying inclusion and exclusion criteria, seven xylitol chewing gum studies, one sorbitol, and one maltitol chewing gum study with either high or fair quality were reviewed. In five out of the seven xylitol studies, xylitol gum decreased gingival scores. In two studies, xylitol decreased gingival scores compared to a polyol gum, and in three studies compared to no gum/gum base. As for sorbitol and maltitol, only sorbitol gum chewing showed a small decrease in gingival scores compared to the controls. CONCLUSIONS: Habitual xylitol gum chewing may reduce gingival inflammation. The low number of studies and their heterogeneity provide clear indications that the effects of sugar-free polyol chewing gums on gingival inflammation need further, well-controlled studies. CLINICAL RELEVANCE: Sugar-free chewing gums, especially xylitol gum, may function as adjuncts to toothbrushing for reducing gingival inflammation, but the evidence so far is inconclusive.

Effects of xylitol chewing gum and candies on the accumulation of dental plaque: a systematic review.

OBJECTIVES: A systematic review of published data was conducted with the aim of assessing the effects of xylitol consumption on the amount of dental plaque. MATERIALS AND METHODS: Electronic and hand searches were performed to find clinical studies concerning the effects of xylitol chewing gum or candies on dental plaque. Prospective randomized controlled clinical trials published between 1971 and 2020 conducted in healthy subjects were included in the review. RESULTS: The initial search identified 424 xylitol articles. After applying inclusion and exclusion criteria, altogether 14 articles (16 studies) were reviewed. The review identified 12 of the total of 14 xylitol chewing gum studies as having fair or high quality. In 13 of the 14 chewing gum studies, xylitol gum decreased plaque accumulation. In six studies, xylitol gum chewing decreased plaque compared to sorbitol gum, and in three studies compared to gum base/no gum. In three fair-quality studies conducted with xylitol candies, plaque accumulation did not change. CONCLUSIONS: Habitual xylitol gum chewing appears to show plaque-reducing effects that differ from those of sorbitol gum. This suggests specific effects for xylitol on plaque accumulation. Xylitol candies appear not to decrease plaque. The heterogeneity of the studies warrants further research. Clinical relevance Habitual xylitol gum chewing is likely to decrease plaque.

Regulation of hBD-2, hBD-3, hCAP18/LL37, and Proinflammatory Cytokine Secretion by Human Milk Oligosaccharides in an Organotypic Oral Mucosal Model.

Human milk oligosaccharides (HMOs), the third largest solid fraction in human milk, can modulate inflammation through Toll-like receptor signaling, but little is known about their immunomodulatory potential in the oral cavity. In this study, we determined whether the HMOs 2'-fucosyllactose (2'-FL) and 3-fucosyllactose (3-FL) regulate human-beta defensin (hBD)-2 and -3, cathelicidin (hCAP18/LL-37), and cytokine responses in human gingival cells using a three-dimensional oral mucosal culture model. The model was incubated with 0.1% or 1% 2'-FL and 3-FL, alone and in combination, for 5 or 24 h, and hBD-2, hBD-3, and hCAP18/LL-37 were analyzed by immunohistochemistry. The expression profiles of interleukin (IL)-1, IL-1RA, IL-8, and monocyte chemoattractant protein (MCP)-1 were determined by LUMINEX immunoassay. The combination of 1% 2'-FL and 1% 3-FL, and 1% 3-FL alone, for 24 h upregulated hBD-2 protein expression significantly (p < 0.001 and p = 0.016, respectively). No changes in the other antimicrobial peptides or proinflammatory cytokines were observed. Thus, 3-FL, alone and in combination with 2'-FL, stimulates oral mucosal secretion of hBD-2, without effecting a proinflammatory response when studied in an oral mucosal culture model.

Effects of xylitol and erythritol consumption on mutans streptococci and the oral microbiota: a systematic review.

OBJECTIVE: A systematic review of published data was conducted with the aim of assessing effects of xylitol and erythritol consumption on levels of mutans streptococci (MS) and the oral microbiota. MATERIALS AND METHODS: Electronic and hand searches were performed to find clinical microbiological studies concerning the consumption of xylitol and erythritol chewing gum or candies, and published between 2000 and 2019. Prospective randomized controlled clinical trials conducted in healthy subjects were included in the review. RESULTS: The initial search identified 561 xylitol and 83 erythritol studies. After applying inclusion and exclusion criteria, 21 xylitol studies and one erythritol study were reviewed. The review identified nine xylitol studies with a fair or high quality, four conducted in children and five in adults, all demonstrating a decrease in MS levels in association with habitual consumption of xylitol. The three microbiota studies employing multispecies probe approaches revealed no effects for xylitol on the microbiota. The only erythritol study fulfilling the inclusion criteria showed no consistent effects on MS levels. CONCLUSIONS: Xylitol consumption is likely to decrease MS counts but it may not change the overall microbiota. Xylitol shows thus properties of an oral prebiotic. More studies are needed to demonstrate the effects of erythritol on MS.

Xylitol and erythritol inhibit real-time biofilm formation of Streptococcus mutans.

BACKGROUND: Regular consumption of xylitol decreases the number of cariogenic streptococci in dental plaque. In vitro biofilm models to study the mechanism of xylitol action have been set-up, but the obtained results are contradictory. Biofilm growth is a dynamic process with time-specific characteristics that may remain undetected in conventional end-point biofilm tests. In this study we used an impedance spectroscopy instrument, xCELLigence Real Time Cell Analyzer (RTCA), that allows label-free, non-invasive real-time monitoring of biofilm formation, to explore effects of xylitol on biofilm formation by Streptococcus mutans. Based on the obtained information of biofilm dynamics, we assessed the number of viable bacteria, the polysaccharide content, and the expression levels of selected genes involved in glucan-mediated biofilm formation in different biofilm stages. Xylitol inhibition was compared with that of erythritol; another polyol suggested to have a positive impact on oral health. RESULTS: Our results showed that real-time monitoring provided new information of polyol-induced changes in S. mutans biofilm formation dynamics. The inhibitory effect of polyols was more pronounced in the early stages of biofilm formation but affected also the measured total amount of formed biofilm. Effects seen in the real-time biofilm assay were only partially explained by changes in CFU values and polysaccharide amounts in the biofilms. Both xylitol and erythritol inhibited real-time biofilm formation by all the nine tested S. mutans strains. Sensitivity of the strains to inhibition varied: some were more sensitive to xylitol and some to erythritol. Xylitol also modified the expression levels of gbpB, gtfB, gtfC and gtfD genes that are important in polysaccharide-mediated adherence of S. mutans. CONCLUSION: The erythritol- and xylitol- induced inhibition of biofilm formation was only partly explained by decrease in the number of viable S. mutans cells or the amount of polysaccharides in the biofilm matrix, suggesting that in addition to reduced proliferation also the matrix composition and thereby the surface attachment quality of biofilm matrix may be altered by the polyols.

Air Abrasion With Bioactive Glass Eradicates Streptococcus mutans Biofilm From a Sandblasted and Acid-Etched Titanium Surface.

Streptococcus mutans is able to form a high-affinity biofilm on material surfaces. S mutans has also been detected around infected implants. Bioactive glasses (BAGs) have been shown to possess antibacterial effects against S mutans and other microorganisms. This in vitro study was performed to investigate the influence of BAG air abrasion on S mutans biofilm on sandblasted and acid-etched titanium surfaces. Sandblasted and acid-etched commercially pure titanium discs were used as substrates for bacteria (n = 107). The discs were immersed in an S mutans solution and incubated for 21 hours to form an S mutans biofilm. Twenty colonized discs were subjected to air abrasion with Bioglass 45S5 (45S5 BAG), experimental zinc oxide containing BAG (Zn4 BAG), and inert glass. After the abrasion, the discs were incubated for 5 hours in an anaerobic chamber followed by an assessment of viable S mutans cells. Surface morphology was evaluation using scanning electron microscopy (n = 12). The thrombogenicity of the glass particle-abraded discs (n = 75) was evaluated spectrophotometrically using whole-blood clotting measurement at predetermined time points. Air abrasion with 45S5 and Zn4 BAG eradicated S mutans biofilm. Significantly fewer viable S mutans cells were found on discs abraded with the 45S5 or Zn4 BAGs compared with the inert glass (P < .001). No significant differences were found in thrombogenicity since blood clotting was achieved for all substrates at 40 minutes. Air abrasion with BAG particles is effective in the eradication of S mutans biofilm from sandblasted and acid-etched titanium surfaces. Zn4 and 45S5 BAGs had similar biofilm-eradicating effects, but Zn4 BAG could be more tissue friendly. In addition, the steady release of zinc ions from Zn4 may enhance bone regeneration around the titanium implant and may thus have the potential to be used in the treatment of peri-implantitis. The use of either BAGs did not enhance the speed of blood coagulation.

TiO2 Coating and UV Photofunctionalization Enhance Blood Coagulation on Zirconia Surfaces.

This in vitro study was designed to evaluate the effect of sol-gel derived TiO2 coating on blood coagulation, blood protein adsorption, and platelet response on zirconia surfaces. Square-shaped zirconia (n=96) (10x10x2 mm) was cut, ground, sintered, and finally cleansed ultrasonically in each of acetone and ethanol for 5 minutes. Three experimental groups (n=32) were fabricated: (a) zirconia coated with sol-gel derived TiO2, (b) zirconia coated with sol-gel derived TiO2 and treated with ultraviolet (UV) irradiation for 1 hour, and (c) non-coated zirconia as control. The coatings were prepared from tetraisopropyl orthotitanate solution by dip-coating. The thrombogenicity of the specimens was evaluated using a whole blood kinetic clotting time method where the extent of blood clotting was evaluated at 10, 20, 30, 40, 50, and 60 minutes (n=4/time point, total n=24/group). Scanning electron microscope images were taken to observe platelet morphologies after 1-hour incubation with platelet-rich plasma (PRP) (n=5/group). Surface characteristics were visualized using atomic force microscopy (n=1/group). Adsorption of plasma proteins and fibronectin on each surface was studied by gel electrophoresis (n=2/group). Significant differences were observed in blood coagulation between the test groups at 20-, 30-, 40-, and 50-minute time points (p<0.005). UV treated TiO2 coated specimens showed fastest blood coagulation followed by TiO2 coated and non-coated specimens. Furthermore, platelets appeared at a higher activation state on coated specimens. Gel electrophoresis revealed no difference in protein adsorption among the experimental groups. In summary, TiO2 coatings promoted blood coagulation, and it was further enhanced by UV treatment, which has the potential to hasten the wound healing process in vivo.

Ultraviolet photofunctionalization of nanostructured titanium surfaces enhances thrombogenicity and platelet response.

The purpose of this study was to evaluate blood and platelet response to nanostructured TiO2 coatings and to investigate the effect of Ultraviolet (UV) light treatment on blood clotting ability, platelet activation and protein adhesion. Ti-6Al-4V titanium alloy plates (n = 138) were divided into three groups; a sol-gel derived MetAliveTM coating (MA); hydrothermal coating (HT); and a non-coated group (NC). Sixty nine titanium substrates were further treated with UV light for 1 h. The thrombogenicity of the titanium substrates was assessed using fresh human blood with a whole blood kinetic clotting time method. The platelet adhesion test was conducted to evaluate the morphology and adhesion behavior of the platelets on the titanium substrates. Human diluted plasma and bovine fibronectin were used to evaluate protein adsorption. Total clotting time for the UV treated HT, MA and NC titanium substrates was almost 40 min compared to 60 min for non-UV substrates, the total clotting time for the UV treated groups were significantly lower than that of the non UV NC group (p < 0.05). UV light treatment had significantly enhanced coagulation rates. The HT and MA substrates presented more platelet aggregation, spreading and pseudopod formation in comparison with the NC substrates. UV treatment did not affect the platelet activation and protein adsorption. This in vitro study concluded that nanostructured titanium dioxide implant surfaces obtained by sol-gel and hydrothermal coating methods increased coagulation rates and enhanced platelet response when compared with non-coated surfaces. UV light treatment clearly improved thrombogenicity of all examined Ti-6Al-4V surfaces.

Antimicrobial efficacy and mechanical properties of BAC-modified hard and soft denture liners.

This study investigated the antimicrobial efficacy and mechanical strength of hard and soft denture liners modified with benzalkonium chloride (BAC). The specimens (1 mm thickness, 8 mm diameter) were prepared by mixing 0.5, 1, 2 and 5 wt% BAC with soft (Sofreliner Medium, Tokuyama) and hard (Rebase II, Tokuyama) denture liners (n = 5/group). BAC was not added to the controls. Candida albicans ATCC 28366 (A 550 = 0.5) and Streptococcus mutans Ingbritt suspensions (A 550 = 0.35) were pipetted onto the specimens, and incubated for 4 h. The viable cells were collected, and determined by plate-culturing (CFU). The tests were repeated after the specimens were soaked in distilled water for 7 days. The mechanical strengths were evaluated by tear and 4-point flexural strength tests for soft and hard liners, respectively. The data were analyzed with ANOVA and Tukey's HSD tests at p = 0.05. C. albicans viability was lost in all groups of BAC-modified soft liners (p < 0.001), and S. mutans viability was reduced (p < 0.01), except of soaked BAC 0.5 wt% group (p > 0.05). For the hard liner, BAC 5 wt% killed the C. albicans and S. mutans cells both before and after soaked in water (p < 0.001). BAC 2 wt% showed comparable tear strength with the soft liner control (p > 0.05). BAC did not reduce the flexural strength of the hard liner (p > 0.05), except of BAC 5 wt% group (p < 0.01). BAC can be a promising agent reducing the C. albicans and S. mutans viability on the soft and hard denture liner surfaces.

Early Biofilm Formation on UV Light Activated Nanoporous TiO2 Surfaces In Vivo.

PURPOSE: To explore early S. mutans biofilm formation on hydrothermally induced nanoporous TiO2 surfaces in vivo and to examine the effect of UV light activation on the biofilm development. MATERIALS AND METHODS: Ti-6Al-4V titanium alloy discs (n = 40) were divided into four groups with different surface treatments: noncoated titanium alloy (NC); UV treated noncoated titanium alloy (UVNC); hydrothermally induced TiO2 coating (HT); and UV treated titanium alloy with hydrothermally induced TiO2 coating (UVHT). In vivo plaque formation was studied in 10 healthy, nonsmoking adult volunteers. Titanium discs were randomly distributed among the maxillary first and second molars. UV treatment was administered for 60 min immediately before attaching the discs in subjects' molars. Plaque samples were collected 24h after the attachment of the specimens. Mutans streptococci (MS), non-mutans streptococci, and total facultative bacteria were cultured, and colonies were counted. RESULTS: The plaque samples of NC (NC + UVNC) surfaces showed over 2 times more often S. mutans when compared to TiO2 surfaces (HT + UVHT), with the number of colonized surfaces equal to 7 and 3, respectively. CONCLUSION: This in vivo study suggested that HT TiO2 surfaces, which we earlier showed to improve blood coagulation and encourage human gingival fibroblast attachment in vitro, do not enhance salivary microbial (mostly mutans streptococci) adhesion and initial biofilm formation when compared with noncoated titanium alloy. UV light treatment provided Ti-6Al-4V surfaces with antibacterial properties and showed a trend towards less biofilm formation when compared with non-UV treated titanium surfaces.

Probiotic intervention influences the salivary levels of Matrix Metalloproteinase (MMP)-9 and Tissue Inhibitor of metalloproteinases (TIMP)-1 in healthy adults.

OBJECTIVE: To study the effect of orally administered Bifidobacterium animalis subsp. lactis BB-12 and Lactobacillus rhamnosus GG on the salivary levels of Matrix Metalloproteinases (MMP)-8, MMP-9 and of Tissue Inhibitor of Metalloproteinases (TIMP)-1 in healthy adults. Furthermore, the correlations between MMP-8, MMP-9 and TIMP-1 and plaque and gingival indices, salivary mutans streptococci and lactobacilli counts, and stimulated saliva secretion rate were analysed. DESIGN: The salivary samples originated from a randomized controlled trial where healthy student volunteers consumed probiotic or placebo lozenges twice a day for four weeks. The saliva samples were collected and clinical parameters measured at the baseline and at the end of the original study. For this study, the salivary levels of MMP-8, MMP-9 and TIMP-1 were analysed with immunofluorometric assay (IFMA) and enzyme-linked immunosorbent assay (ELISA). RESULTS: In the probiotic group (n=29), salivary MMP-9 levels increased (p<0.01) and TIMP-1 levels decreased (p<0.01) significantly during the intervention. Furthermore, MMP-9/TIMP-1 ratio differed significantly from the baseline level (p<0.01). These changes were not observed in the control group (n=31). In the whole data, salivary MMP-9 and gingival index correlated (r=0.260, p<0.05 at baseline and r=0.354, p<0.01 at the end of the study). Intergroup differences or correlations with other clinical parameters were not found. Probiotic consumption did not affect the saliva flow rate. CONCLUSIONS: Increased MMP-9 and decreased TIMP-1 levels in saliva may indicate that probiotics have immunomodulatory effects in the oral cavity. Furthermore, increased salivary MMP-9 levels may be an indication of the defensive potential of matrix metalloproteinases.

Dipeptidyl peptidase IV and quorum sensing signaling in biofilm-related virulence of Prevotella aurantiaca.

Biofilm formation and dipeptidyl peptidase IV (DPPIV) enzyme activity contribute to the virulence of oral bacteria, and these virulence factors are partly regulated by quorum sensing signaling system. We recently demonstrated that estradiol regulates growth properties and DPPIV activity of Prevotella intermedia, Prevotella nigrescens, and Prevotella pallens. Here, we examined the DPPIV dependency of biofilm formation of Prevotella aurantiaca. Three strains (two clinical strains AHN 37505 and 37552 and the type strain CCUG 57723) were incubated in three estradiol concentrations (30, 90, and 120 nmol/L). Regulation of DPPIV activity, biofilm and fimbria formation, and coaggregation of bacterial strains were analyzed after incubation with four concentrations (10 nM, 100 nM, 1 µM, 10 µM) of dihydroxy-2,3-pentaedione (DPD), the universal precursor of autoinducer -2 (AI-2), and analogs (ethyl-DPD, butyl-DPD, and isobutyl-DPD) for 24 h. Estradiol enhanced the planktonic growth, coaggregation, and biofilm formation of P. aurantiaca strains. The whole cell extract of AHN 37505 had the highest DPPIV activity, followed by CCUG 57723 and AHN 37552. Inhibition of DPPIV activity with di-isopropylfluorophosphate suppressed the effect of estradiol on biofilm formation. At 100 nM and 10 µM concentrations of DPD, butyl DPD, and isobutyl DPD, biofilm formation of P. aurantiaca was significantly inhibited. Fimbriae formation was enhanced up to concentrations of 100 nM and 1 µM followed by a significant inhibition at higher concentrations of DPD and all analogs. A slight but significant inhibitory effect of DPD and analogs on DPPIV activity was observed. Our results indicate that DPPIV plays a key role in the estradiol-regulated biofilm formation of P. aurantiaca. Quorum sensing autoinducer DPD and C1-alkyl analogs could inhibit biofilm-related virulence of P. aurantiaca.

Effects of Xylitol and Sucrose Mint Products on Streptococcus mutans Colonization in a Dental Simulator Model.

Few laboratory methods exist for evaluating the cariogenicity of food ingredients. In this study, a dental simulator was used to determine the effects of commercial sucrose and xylitol mint products on the adherence and planktonic growth of Streptococcus mutans. Solutions (3% w/v) of sucrose, xylitol, sucrose mints, xylitol mints, xylitol with 0.02% peppermint oil (PO), and 0.02% PO alone were used to test the levels of planktonic and adhered S. mutans. A dental simulator with continuous artificial saliva flow, constant temperature, and mixing was used as a test environment and hydroxyapatite (HA) discs were implemented into the model to simulate the tooth surface. Bacterial content was quantified by qPCR. Compared with the artificial saliva alone, sucrose and sucrose mints increased the numbers of HA-attached S. mutans, whereas xylitol decreased them. Similarly, planktonic S. mutans quantities rose with sucrose and declined with xylitol and xylitol mints. Versus sucrose mints, xylitol mints significantly reduced the counts of HA-bound and planktonic S. mutans. Similar results were observed with the main ingredients of both types of mints separately. PO-supplemented artificial saliva did not influence the numbers of S. mutans that attached to HA or planktonic S. mutans compared with artificial saliva control. In our dental simulator model, xylitol reduced the counts of adhering and planktonic S.mutans. The mints behaved similarly as their pure, main ingredients-sucrose or xylitol, respectively. PO, which has been suggested to have antimicrobial properties, did not influence S. mutans colonization.

Xylitol Chewing Gums on the Market: Do They Prevent Caries?

PURPOSE: To measure the xylitol content in sugar-free chewing gums available on the market in Gulf Cooperation Council (GCC) countries in the Middle East, in order to identify those products that can provide the recommended daily dose of xylitol for caries prevention (6-7 g). Acid production from chewing gums was also measured in vitro and in vivo. MATERIALS AND METHODS: Twenty-one chewing gums containing xylitol were identified and collected from the GCC market (Kuwait, Bahrain, Qatar, Saudi Arabia, UAE and Oman). Xylitol was extracted and its concentration was analysed using a special enzymatic kit. The pH of extracts was measured during 30-min incubation with Streptococcus mutans. Changes in saliva and plaque pH were noted in four subjects after the consumption of highly concentrated xylitol gums. RESULTS: The xylitol content in grams was clearly mentioned only on one product's label. Twelve products stated the percentage of xylitol (3.5% to 35%). The rest did not specify the amount. The mean measured weight of one piece of gum was 1.67 ± 0.38 g. The mean measured xylitol content/piece was 0.33 ± 0.21 g. Xylitol content was < 0.3 g/ piece in 9 products, 0.3-0.5 g in 7 and > 0.5 g in 5 products. None of the highly concentrated xylitol gums showed a pH drop in vitro or in vivo. One chewing gum, containing xylitol and glucose, resulted in a low pH level (< 5.5) when tested in vitro. CONCLUSION: The majority of xylitol chewing gums sold on the GCC market do not provide the consumers with the recommended daily dose of xylitol for caries prevention. Clear, accurate labeling is recommended.

Bifidobacteria inhibit the growth of Porphyromonas gingivalis but not of Streptococcus mutans in an in vitro biofilm model.

There is growing interest in the use of probiotic bifidobacteria for enhancement of the therapy, and in the prevention, of oral microbial diseases. However, the results of clinical studies assessing the effects of bifidobacteria on the oral microbiota are controversial, and the mechanisms of actions of probiotics in the oral cavity remain largely unknown. In addition, very little is known about the role of commensal bifidobacteria in oral health. Our aim was to study the integration of the probiotic Bifidobacterium animalis subsp. lactis Bb12 and of oral Bifidobacterium dentium and Bifidobacterium longum isolates in supragingival and subgingival biofilm models and their effects on other bacteria in biofilms in vitro using two different in vitro biofilms and agar-overlay assays. All bifidobacteria integrated well into the subgingival biofilms composed of Porphyromonas gingivalis, Actinomyces naeslundii, and Fusobacterium nucleatum and decreased significantly only the number of P. gingivalis in the biofilms. The integration of bifidobacteria into the supragingival biofilms containing Streptococcus mutans and A. naeslundii was less efficient, and bifidobacteria did not affect the number of S. mutans in biofilms. Therefore, our results suggest that bifidobacteria may have a positive effect on subgingival biofilm and thereby potential in enhancing gingival health; however, their effect on supragingival biofilm may be limited.

Supplementation of xylitol-containing chewing gum with probiotics: a double blind, randomised pilot study focusing on saliva flow and saliva properties.

The aim of this study was to investigate the impact of daily chewing, for 12 weeks, of 2 different probiotic gums compared with placebo on saliva flow rate, saliva IgA levels and saliva pH. The intervention study included 54 adult volunteers with hyposalivation in a double-blind, randomised and placebo-controlled design with three parallel groups. Volunteers were randomly assigned to 3 different groups: subjects in group A (n = 19) were given placebo chewing gum, group B (n = 17) received Bifidobacterium animalis ssp. lactis Bb12 (ATCC 27536) and group C (n = 18) received Lactobacillus rhamnosus LGG (ATCC 53103), Bifidobacterium longum 46 (DSM 14583) and Bifidobacterium longum 2C (DSM 14579) gums, during 3 months. Two volunteers from group B left the study for personal reasons leaving 19, 15 and 18 volunteers, respectively, for analyses. Clinical examinations, personal interviews, sialometries and saliva sampling were conducted at baseline and after 1, 2, 3 and 4 months. No statistically significant differences were found between probiotic and placebo groups for any of the parameters analysed. No side effects of probiotic or placebo chewing gums were observed. Chewing gum, with and without probiotics, had a positive impact on salivary flow rate and saliva pH and IgA levels.

Bifidobacterium animalis subsp. lactis BB-12 in reducing the risk of infections in early childhood.

BACKGROUND: Specific probiotic bacteria have proven to be effective in the prevention and treatment of infectious diseases in early life in at-risk populations. The impact of administration of Bifidobacterium animalis subsp. lactis BB-12 (BB-12) on the risk of acute infectious diseases was studied in healthy children. METHODS: In this double-blind, placebo-controlled study, 109 1-mo-old infants were assigned randomly to a probiotic group receiving a BB-12-containing tablet (n = 55) or a placebo (n = 54). Test tablets were administered to the infants twice a day (daily dose of BB-12 10 billion colony-forming units) until the age of 2 y with a novel slow-release pacifier or a spoon. Breastfeeding habits, pacifier use, dietary habits, medications, and all signs and symptoms of acute infections were registered in diaries by parents and in questionnaires by trained professionals. RESULTS: The infants receiving BB-12 were reported to have experienced fewer respiratory tract infections (RTIs; 87 vs. 100%; risk ratio: 0.87; 95% confidence interval: 0.76, 1.00; P = 0.033) than the controls. No significant differences between the groups were observed in reported gastrointestinal symptoms, otitis media, or fever. The baseline characteristics of the two groups were similar, as was the duration of breastfeeding. CONCLUSION: Administration of BB-12 in early childhood may reduce RTIs.

Does estradiol have an impact on the dipeptidyl peptidase IV enzyme activity of the Prevotella intermedia group bacteria?

Initiation and development of pregnancy-associated gingivitis is seemingly related to the microbial shift towards specific gram-negative anaerobes in subgingival biofilms. It is known that Prevotella intermedia sensu lato is able to use estradiol as an alternative source of growth instead of vitamin K. The aim of the present study was to investigate the impact of estradiol on the bacterial dipeptidyl peptidase IV (DPPIV) enzyme activity in vitro as a virulent factor of the Prevotella intermedia group bacteria, namely P. intermedia, Prevotella nigrescens, Prevotella pallens, and Prevotella aurantiaca. In all experiments, 2 strains of each Prevotella species were used. Bacteria were incubated with the concentrations of 0, 30, 90, and 120 nmol/L of estradiol and were allowed to build biofilms at an air-solid interface. DPPIV activities of biofilms were measured kinetically during 20 min using a fluorometric assay. The enzyme activity was later related to the amount of protein produced by the same biofilm, reflecting the biofilm mass. Estradiol significantly increased DPPIV activities of the 8 Prevotella strains in a strain- and dose-dependent manner. In conclusion, our in vitro experiments indicate that estradiol regulates the DPPIV enzyme activity of P. intermedia, P. nigrescens, P. pallens, and P. aurantiaca strains differently. Our results may, at least partly, explain the role of estradiol to elicit a virulent state which contributes to the pathogenesis of pregnancy-related gingivitis.

High sucrose intake at 3 years of age is associated with increased salivary counts of mutans streptococci and lactobacilli, and with increased caries rate from 3 to 16 years of age.

OBJECTIVE: We aimed to study the long-term associations between sucrose intake (SI), selected representatives of the cariogenic oral flora, and the dental health of children from 3 to 16 years of age. METHODS: At 7 months of age 1,062 infants (540 intervention; 522 controls) were included in the prospective, randomised STRIP-project aimed at restricting the child's saturated fat and cholesterol intake to prevent atherosclerosis when they become adults. At 3 years of age, every fifth child was invited (n = 178) to an oral sub-study, and 148 (78 boys) children attended. A restudy was conducted on 135 children aged 6, 127 aged 9, 114 aged 12 and 88 aged 16. SI using 4-day food records, plate-cultured mutans streptococci (MS), salivary lactobacilli (LB) and yeasts using commercial kits (Orion Diagnostica, Espoo Finland), toothbrushing frequency using fluoridated toothpaste and dental health expressed as d 3 mft/D 3 MFT were regularly recorded. RESULTS: The SI of children whose intake was ≥ 10 E% (high SI) at 3 years remained high throughout the entire follow-up (p < 0.001, GLM for repeated measures) period, and they had higher salivary MS and LB counts (p = 0.024 and p = 0.068, respectively, GLM) than their counterparts whose SI was below 10 E% (low SI). No differences in toothbrushing habits were found between the high and low SI-groups. Caries-survival was strongly associated with low 6-year-counts of MS (p = 0.008, Cox regression analysis), and the d 3 mft/D 3 MFTscores of the high SI-group were higher than those of the low SI-group (p = 0.046, GLM). CONCLUSIONS: High SI at 3 years was associated with high MS-counts ( ≥ 10 5 cfu/ml) and with a high risk for caries.