Effects of Consumption of Probiotic Yogurt on Salivary Streptococcus mutans, IL-1β and TNF-α Levels in Adults with Initial Stages of Dental Caries: a Randomized, Double‑Blind, Placebo‑Controlled Clinical Trial

doi:10.21203/rs.3.rs-4206212/v1

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Effects of Consumption of Probiotic Yogurt on Salivary Streptococcus mutans, IL-1β and TNF-α Levels in Adults with Initial Stages of Dental Caries: a Randomized, Double‑Blind, Placebo‑Controlled Clinical Trial

https://doi.org/10.21203/rs.3.rs-4206212/v1

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Background

Dental caries is one of the most common human diseases. The aim of the present study was to determine the effect of consumption of probiotic yogurt containing Bifidobacterium lactis Bb12 on salivary Streptococcus mutans (S. mutans), interleukin-1β (IL-1β) and tumor necrosis factor alpha (TNF-α) levels in adults with initial stages of dental caries.

Methods and Material:

Sixty six 18–30 years old students with initial stages of dental caries were selected for this 2 weeks double blind randomized placebo controlled clinical trial and were randomly assigned into two groups: intervention (300g/d probiotic yogurt), and control (300 g/d conventional yogurt) groups. Unstimulated fasting saliva sample (2–3 ml) was collected pre and post-intervention for S. mutans bacteria counting and measurement of IL-1β and TNF-α cytokines levels. All Data were analyzed using SPSS.

Results

Salivary S. mutans and IL-1β changes were statistically signifcant in intervention group compared to their baseline and compared to the control group post intervention.

Conclusion

Consumption of probiotic yogurt containing B. lactis Bb12 may be beneficial for preventing of the progression of dental caries.

Trial registration:

This study was registered in Iran clinical trials: IRCT2014062518240N1.

Probiotic yogurt

Bifidobacterium lactis

Tooth decay

Streptococcus mutans

Inflammatory mediators

Dental caries is the most prevalent chronic disease of the oral cavity with multi-factorial etiology [1]. It is strongly associated with the presence of cariogenic microorganisms, fermentable carbohydrates in diet, and susceptible teeth surface, and duration of exposure to cariogenic substances [2]. Several studies have shown an association between dental caries and Streptococcus mutans as the main cause of tooth decay [3, 4].The suggested mechanism for effective role of this micro-organism in development of dental caries is the ability of producing an insoluble substance in water called “branched-glucan” or “mutant” that facilitates S. mutans establishment in dental biofilm. This may subsequently lead to an acidogenic environment of oral cavity with low pH (less than 5.5) and rapid metabolism of carbohydrates [4].

Numerous studies also have evaluated the relation between development of carious lesions and the response of immune competent cells. Cytokines are products of the predominant cell types within periapical lesions including neutrophils, macrophages, T- and B-lymphocytes, mast cells, osteoclasts, osteoblasts, and fibroblasts [5, 6]. Elevated levels of the cytokines interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β) were found in saliva of patients with tooth decay and were associated with the progression of periodontal diseases [7]. Although the role of cytokines in the pathogenesis of dental caries is not clear, components of S. mutans were found to stimulate production of pro inflammatory cytokines [8–10]. In Hsukioja and et al study a positive association was observed between S. mutans and elevated levels of salivary IL-1β [11]. While in other studies such as Cogulu and et al study, results denied such a relationship [12].

Increasing interest is being shown in the use of alternative therapies to treat various inflammatory diseases, particularly through the use of probiotics [13–16]. Probiotic are alive non-digestible microbial food, when administrated in adequate amounts has a health benefit to the host [17]. The effect of probiotics on dental caries and its related risk factors have been evaluated in several studies, but most of these studies focused on reducing S.mutans aiming to dental caries prevention with varying degrees of success [18–24]. To date a few studies have been conducted about the effect of probiotic on the inflammatory markers in dental caries. Shyu PT et al in an experimental study showed that expression of IL-1β and TNF-α by lipopolysaccharide-treated macrophages was significantly down-regulated in cells with probiotic supplement [25]. Slawik S et al indicated that a daily consumption of probiotic milk drink reduces the plaque induced gingival inflammation [26].

Bifidobacterium is one of the most commonly used probiotic bacterial in products [27]. A body of evidence suggests that bifidobacteria have benefits on general health such as reduced susceptibility to infections, reduction of allergies and lactose intolerance, as well as lowered blood pressure and serum cholesterol values [28], but little attention is paid to its possible effects on oral health and risk factors of caries [19]. So the aim of the present study was to determine the effect of short term consumption of probiotic yogurt containing bifidobacterium lactis on salivary S. mutans, IL1β and TNF-α level in adults with initial stages of dental caries.

Subjects

This randomized, parallel, placebo controlled intervention study conformed to the ethical guidelines of the 1975 Declaration of Helsinki and was approved by the Ethics Committee of Ahvaz Jundishapur University of medical Sciences (Ethical Code: AJUMS. REC.28. 2014 and registration code of Iran clinical trials: IRCT2014062518240N1). In this double blind (investigator and data analyzer) randomized clinical trial 66 students (18–30 years old) were selected from Ahvaz Jundishapur University of Medical Sciences. The sample size was determined based on the primary information obtained from the study by Nikawa et al [29] for S. mutans variable. For α value equal to 0.05 and a power of 80% (β = 0.2), \({z}_{1}-\frac{\alpha }{2}=1.96\) and \({\text{z}}_{\text{1}}-\beta =0.84\) and the proportion of reduction of S. mutans in the intervention group equal to 40% and the proportion of reduction of S. mutans in the control group equal to 10%, and the difference in proportions equals 0.3 the sample size was calculated 30 subjects per group. Considering the loss of 10%, sixty-six subjects selected for the study. Anthropometric measurements, demographic and oral health information were completed. In addition and dental examination in order to detect the DMFT (D= decay, M=missing, FT= filled teeth) (with inclusion criteria of 1–3 showing initial stages of dental caries) was performed [30]. The clinical examination of 66 children for dental caries was conducted by an examiner who was calibrated by a previously calibrated epidemiologist. Cohen’s kappa statistic was used to determine inter-examiner agreement. There was a high reproducibility (inter- and intra-examiner kappas > 0.8). Individuals were excluded if they having with greater than 3 decayed teeth or active caries, using mouth wash, brushing teeth more than twice daily, smokers, and people who have consumed any antioxidant supplements, anti-inflammatory drugs, antibiotics and probiotic products routinely during the last month. All subjects were asked to maintain their normal diet, brush their teeth 2 times a day and do not use xylitol gums and probiotic products during the intervention period. From all candidates participating in this study a consent form was obtained. Prior to collection of saliva, patients were kept seat for five minute relaxed and silent. After a thorough rinse with water and an initial swallow, un-stimulated saliva samples (2-3ml) were collected and were immediately transported in sterile containers to the laboratory of microbiology do bacteria counting and the cytokines experiments. Then subjects were randomly allocated to "A1" intervention (probiotic yogurt) and control "A2" (conventional yogurt) groups consisting of 33 subjects in each by another investigator (Fig. 1) using a random- number table. Participants and investigator were not informed about which group was allocated to "A1" and which one was allocated to A2. The subjects in the intervention group received 300g/d probiotic yogurt (containing a minimum count of 10⁶CFU/ml of B. lactis Bb12), and control group received 300 g/d conventional yogurt daily for 2 weeks. Both probiotic and conventional yogurts was produced in similar shape with the same trading brand containing 3% fat. The yogurts were distributed once before beginning of the intervention and once again one week after starting the intervention in order to monitor and ensure of the consumption of yogurts and in addition in order to remind to use of yogurts we phoned to the subjects. At the end of the study period, again saliva samples were collected from each participant. Figure 1 demonstrates the flowchart of participants. The CONSORT checklist was used for the present randomized trial [31].

Microorganisms and bacteria cultivation

Sample collection and preparation of dilution

At first dilution of 10^− 1 was prepared from salivary samples. For this purpose, 1 cc of each saliva sample was poured in a sterile tube and then 9 cc NaCl (0/9%) serum was added to the tube and shaken well.

For microbiological cultivation, 20µl of each saliva sample was spread on Salivarius Mitis agar (Hi-Media, India) for counting total S.mutans. This culture media is specific to S.mutans bacteria. The bacteria cultivation was done using streak-plate method. Then all plates were placed in an anaerobic condition at 37°C for 3 days. Plates were examined after 3 days incubation at 37 ° C. If the colony had not grown, plates were returned to the anaerobic enclosure to be incubated for another week. After preparing the lam from the colonies on the culture medium, Gram staining was carried out. For identification of s. mutans Gram-positive colonies (cocci) were selected and catalase test was performed for them. Finally, gram- positive coccies and catalase- negative were incubated under biochemical tests for 48 hours at 37 ° C. Colonies that were Monitol- positive, Vogues-Proskauer (VP)-positive, Argeninie- negative, dextran -positive, urea- negative, Blie esculin- positive, were considered as s. mutants colonies. Finally, Streptococcus mutans colonies were counted and multiplied for each participant in the dilution coefficient and the number of colonies per ml of saliva was determined (CFU/ml).

Measurement of IL-6 and TNF-α

Salivary cytokines (IL-6 and TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA) method using E-Bio Science kit (Germany).

Statistical Analyses

All Data were analyzed using SPSS (ver.19, Chicago, IL, USA). Salivary concentrations of micro-organisms were calculated as log10 (colony-forming-units + 1) per ml. The data for bacterial counting were expressed as mean ± standard deviation (Mean ± SD). In order to determine whether data are normally distributed a visual examination of data and a goodness of fit test (Kolmorogov-Smirnoff) was conducted. A histogram was also used to confirm the normal distribution of the data. Independent sample T-test was used to compare the mean differences of variables between two groups at the baseline. Analysis of covariance (ANCOVA) was used to identify any differences between two groups at the end of the study, adjusting for baseline values. Paired sample T-test was used to compare the mean differences within groups. P < 0.05 was considered significant.

Sixty six subjects that randomly allocated to control group (n = 33), and intervention groups (n = 33) received interventions for two weeks and all of them completed the study (Fig. 1). Forty two of subjects were females (62%) and 24 were males (38%). In this study there were not any reports from subjects regarding any adverse effects or symptoms with two types of intervention provided during the study. The mean ± standard deviation for age, weight and DMFT was 22.1 ± 3.1 years, 67.85 ± 3.7 kg and 2.47 ± 0.38, respectively. All subjects had normal body mass index (BMI: 22.86 ± 2.47 kg/m2). There were no significant differences between groups in age, gender, weight, BMI and DMFT at the beginning of the study (P ≥ 0.05). Demographic and anthropometric data are shown in Table 1.

Table 1

Demographic and anthropometric characteristics of the subjects
Variable	Intervention group (n = 33)	Control group (n = 33)	p-value
Age(yr)	21.6 ± 2.5	22.4 ± 3.8	0.77
Male (n)	13	11	0.8
Female (n)	20	22	0.79
Weight (kg)	68.2 ± 4.4	66.3 ± 4.1	0.74
BMI (kg/m²)	22.77 ± 2.37	22.54 ± 2.25	0.87
DMFT	2.2 ± 0.7	2.2 ± 0.9	0.91
Notes: Abbreviations: BMI, Body Mass Index; DMFT, Decayed Missed Filled Teeth The results are described as mean ± Standard Deviation (SD) There were no significant differences between groups in age, gender, weight, BMI and DMFT at the beginning of the study (P ≥ 0.05).

There were no significant differences (P = 0.130) between two groups in the mean number of S. mutans at baseline. Within group analysis showed that the mean number of salivary S. mutans was significantly decreased in the intervention group post intervention (P = 0.001). While the reduction in S. mutans (P = 0.594) in control group post intervention. The results of analysis of covariance (ANCOVA) adjusted for baseline values showed significant differences between two groups in the mean number of S. mutans post intervention. At the end of the study the mean number of S. mutans was significantly lower in the intervention group compared with the control (3.28 ± 3.26 and 3.64 ± 3.43 CFU/ml; P = 0.001) (Fig. 2).

Table 2 shows the mean levels of IL-1β and TNF-α at baseline and after intervention in the intervention and control groups. There were no significant differences in the mean level of TNF-α (P = 0.070) between the two groups at baseline. While significant difference in the mean level of IL-1β was observed between the groups at baseline (P = 0.001). The results showed that the mean level of IL-1β was decreased significantly after 2 weeks in the intervention group (P < 0.001), but there was no significant alteration in the control group (P = 0.380). The mean level of TNF-α had a non-significant reduction in intervention group (P = 0.060). While a significant increases in the mean level of TNF-α was observed in control group (P = 0.020). The results of analysis of covariance (ANCOVA) showed statistically significant differences between two studied groups in the mean changes of IL-1β levels post intervention, adjusted for baseline value (P < 0.001). While, no significant difference in the mean (P = 0.110) and mean change of TNF-α levels (P = 0.060) was observed between the two groups at the end of study.

Table 2

Mean ± SD for salivary IL-1 and TNF- levels within and between groups
βα
Variable	Measurement Period	Intervention group (n = 33)	Control group (n = 33)	Pvalue
IL-1β (Pg/ml)	Baseline After intervention Mean Change	81.25 ± 8.02 40.1 ± 6.4 41.15 ± 3.02	53.59 ± 13.44 45.49 ± 10.21 8.1 ± 3.02	0.001^a 0.062^b 0.001^b
Pvalue^c		0.001	0.06
TNF-α (Pg/ml)	Baseline After intervention Mean Change	7.6 ± 0.7 7.2 ± 1.3 0.4 ± 0.06	7.33 ± 0.9 8.18 ± 1.22 0.81 ± 0.2	0.07^a 0.11^b 0.06^b
Pvalue^c		0.06	0.02
Notes: Abbreviations: IL-1β, interleukin-1β; TNF-α, Tumor necrotic factor-α The results are described as mean ± Standard Deviation (SD) ^a Difference between groups in baseline, p value is reported based on Independent T-test . ^b Difference between groups after 2 weeks intervention, p value is reported based on analysis of covariance (ANCOVA) ^c Within group difference, p value is reported based on Paired t-test P < 0.05 was considered statistically significant.

This study is the first clinical trial that conducted to assess the effect of probiotic yogurt containing bifidobacterium lactis on salivary S.mutans and salivary IL-1β and TNF-α level in adults with initial stages of dental caries. The results showed that consumption of probiotic yogurt containing B. lactis Bb12 significantly reduced the number of S. mutans and the mean levels of IL-1β in saliva.

Probiotics may prevent the formation of dental plaque directly through adhering to tooth surface or indirectly through neutralizing free electrons [32]. Different types of probiotic bacteria have several effects on the growth of cariogenic bacteria. Bifidobacterium is one of the most commonly used probiotic bacteria in food products [33] that the inhibitory effect of foods or dairy products containing this bacterium, alone [19, 21] or in combination with other probiotic bacteria, has shown on growth and proliferation of S. mutans in saliva [34, 35]. It has shown that Bifidobacterium can reduce dental caries among people who do not have active caries [36]. Similarly the results of present article showed that consumption of probiotic yogurt containing B. lactis Bb12 significantly reduced the number of S. mutans in saliva. This finding was similar to the findings of previous clinical studies [19, 34, 35, 37–40]. It was found that Bifidobacteria could survive in saliva and bind to Fusobacterium nucleatum-covered hydroxy apatite, stressing the importance of other oral bacteria in the potential action of probiotic strains [41]. In contrast with the results of our study, Taipale T et al in a randomized clinical trial showed that consumption of twice a day carriers containing B. lactis- Bb12 probiotic bacteria for a long time (2 years) and from early childhood could not significantly reduce the number of S. mutans in the tooth surface and oral mucosa of volunteers [42]. Apart from differences in study design and participants characteristic, the diversity between findings of our study and Taipale T et al may be due to differences in the type of supplementation (tablets). The bacteria in the tablets are in a lyophilized form (freeze-drying to increase the shelf life of bacteria). Therefore time and enriched culture medium is essential for bacteria to become as an active and vegetative form. Physiologically, bacteria in tablets may not have the maximum growth, proliferation and producing activity of secondary metabolites. Therefore, the capability of probiotic bacteria in the inhibition of S. mutans growth, in the dried and encapsulated form is significantly different with vegetative forms [43]. In addition, a double blind study conducted by Nozari A et al reported that short-term (2 weeks) consumption of probiotic yogurt containing B. lactis Bb12 could not reduce the levels of salivary S. mutans in 6 to 12 year-old healthy children [44]. Different study design and characteristics of subjects may be factors involved in results of present and above mentioned study.

Furthermore the potential role of the probiotics in modulating immune response has led to an interest in using of them as preventive and therapeutic intervention. Furrie E et al demonstrated that administration of a symbiotic containing bifidobacterium longum for a month, lead to decrease serum levels of IL-1𝛽 and TNF-𝛼 in patient with active ulcerative colitis [45]. Shyu PT et al showed that expression of IL-1𝛽 and TNF-𝛼 by lipopolysaccharide-treated macrophages was significantly down regulated in cells with probiotic supernatants compared to those exposed to MRS medium [25]. To date a few studies have been conducted about the effect of probiotic on the inflammatory markers in dental caries. This study is the first clinical trial that conducted to assess the effect of probiotic yogurt containing bifidobacterium lactis on salivary IL-1β and TNF-α level in adults with initial stages of dental caries. The results showed that the short-term (2weeks) consumption of probiotic yogurt containing B. lactis Bb12 caused significant reduction in the mean level of IL-1β in saliva of intervention group compared to the control group. The results of present study were consistent with previous studies [46–48]. In Contrast with the results of our study, in a randomized clinical trial, Asemi et al demonstrated that daily consumption of 200 (gr) probiotic yogurt containing lactobacillus acidophilus for 9 weeks had no effect on TNF-α level in pregnant women [49]. In addition, in the Ouwehand et al study over a period of 6 months from 55 institutionalized elderly subjects, who were enrolled in a double-blind placebo-controlled study a non-significant association was found between serum level of TNF-𝛼 and consumption of pill containing bifidobacterium [47].

Although the exact mechanism of the effects of probiotics in oral cavity is not fully understood, several mechanisms have been reported, i.e., secretion of various antimicrobial substances such as organic acids, hydrogen peroxide, and bacteriocins. These substances fight against S. mutans strains in the oral cavity to replace the naturally occurring cariogenic strains. In addition, they compete with pathogenic agents for nutrients, growth factors and adhesion sites on the oral mucosa. Probiotics can also alter the surrounding environment through modulating the pH and/ or the oxidation-reduction potential, which can adversely, affect the pathogenic microbes and facilitate their elimination [50].

As we mentioned, to date most of the studies focused on reducing S.mutans as the main cause of tooth decay, and a few studies have been conducted about the effect of probiotic on the inflammatory markers in dental caries, but according to the role of pathogenic bacteria and inflammation in the initiation and progression of dental caries and considering positive effects of probiotics on both pathogenic bacteria [19, 34, 35, 37, 38, 40, 51] and inflammatory factors [46–48], we can suggest that probiotics consumption may be beneficial in prevention of dental caries by decreasing both pathogenic bacteria and inflammatory factors through above mentioned mechanisms and moreover through stimulating the non-specific immunity and modulate the cellular and humeral immune response [19, 52, 53].

Present study had several strengths including collection of unstimulated saliva from each participant, because the analysis of unstimulated saliva is more sensitive than analysis of stimulated saliva [54]. Moreover, the using of randomized controlled trial design caused that many of the confounders were controlled. This study was adequately powered to detect mean difference in number of bacteria between the groups. However present study had some limitation including a small sample size, so our study findings cannot be generalized to other populations, and the short study duration. Also the effects of probiotic microorganisms after a period of no consumption of probiotic product were not considered. The study design, characteristics of the target group, the type of probiotic, culture medium and the concentration of probiotics, may be factors causing diversity in the results of present study with other previous studies. It is recommended that more clinical trial studies with longer duration and more sample size be conducted on susceptible groups that are at risk of dental caries to assess the effectiveness of alternative therapy with probiotic products in preventing the development of caries. In addition, in future studies on the effect of consumption of probiotic products, the Saliva buffering capacity factor also be considered. Furthermore, because of the complex structure of dental caries pathogenesis and progression, it is difficult to identify factors that predict the caries process. More clinical trials with considering different species of probiotic bacteria and additional cytokines may be required in future.

In conclusion it is suggested that short-term consumption of probiotic yogurt containing B.lactis Bb12 may prevent the progression of dental caries through reducing the population of S. mutans (cariogenic bacteria) and the levels of some inflammatory cytokines in saliva in those who are at risk of active dental caries.

Ethics approval and consent to participate

This study was approved by the Ethics Committee of Ahvaz Jundishapur University of medical Sciences (Ethical Code: AJUMS. REC.28. 2014 and registration code of Iran clinical trials: IRCT2014062518240N1) and carried out based on the Declaration of Helsinki. Informed consent was obtained from all subjects involved in the study.

Consent for publication

Not applicable as no personal data were used in this article.

Availability of data and materials

The datasets collected and/or analyzed during the present study are not publicly accessible due to ethical concerns but corresponding author may provide datasets upon reasonable request.

Competing interests

The authors declare no competing interests.

Funding

This study was part of the M.Sc thesis of Mr. Essam Amerian and was financially supported by the Deputy of Research and Technology, Ahvaz Jundishapur University of Medical Sciences (AJUMS. REC. 28. 2014). The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript

Author contributions
AZJ and EA initially conceptualized and designed the study. LB and AE upgraded the design. EA and LB contribute sampling. MHH and LMN were responsible for statistical analysis. The manuscript was written by LMN and EA. AZJ performed English editing. All authors read and approved the final manuscript.

Acknowledgement

We thank the Faculty of Arvand International Division, Research Committee and Laboratory of Microbiology, School of Allied Medical Sciences of Ahvaz Jundishapur University of Medical Sciences (AJUAMS), and the staff and participants of the present study.

Lenander-Lumikari M, Loimaranta V. Saliva and dental caries. Adv Dent Res. 2000;14:40–7.
Ozdemir D. Dental Caries: The Most Common Disease Worldwide and Preventive Strategies. Int J Biology. 2013;5(4):55–61.
Teanpaisan R, Dahlen G. Use of polymerase chain reaction techniques and sodium dodecyl sulfate-polyacrylamide gel electrophoresis for differentiation of oral Lactobacillus species. Oral Microbiol Immunol. 2006;21(2):79–83.
Hoorizad M, et al. Effect of probiotic yoghurt on the salivary PH of orthodontic patients- A clinical trial study. J Res Dent Sci. 2013;10(3):155–9.
Shapiro H, Lutaty A, Ariel A. Macrophages, meta-inflammation, and immuno-metabolism. ScientificWorldJournal. 2011;11:2509–29.
Chang SK et al. Cadherin-11 regulates fibroblast inflammation. Proceedings of the National Academy of Sciences, 2011. 108(20): pp. 8402–8407.
Gornowicz A, et al. Pro-inflammatory cytokines in saliva of adolescents with dental caries disease. Ann Agric Environ Med. 2012;19(4):711–6.
Russell MW, Mansson-Rahemtulla B. Interaction between surface protein antigens of Streptococcus mutans and human salivary components. Oral Microbiol Immunol. 1989;4(2):106–11.
BENABDELMOUMENE S, et al. Activation of Human Monocytes by Streptococcus mutans Serotype f Polysaccharide: Immunoglobulin G Fc Receptor Expression and Tumor Necrosis Factor and Interleukin-1 Production. Volume 59. INFECTION AND IMMUNITY; 1991. pp. 3261–6. 9.
Soell M et al. Binding of Streptococcus mutans SR protein to human monocytes: production of tumor necrosis factor, interleukin 1, and interleukin 6. INFECTION AND IMMUNITY, 1994. 62(5): pp. 1805–1812.
Haukioja A. Probiotics and Oral Health. Eur J Dentistry. 2010;4(3):348–55.
Cogulu D, et al. Associations of interleukin (IL)-1beta, IL-1 receptor antagonist, and IL-10 with dental caries. J Oral Sci. 2015;57(1):31–6.
Cummings JH, Macfarlane GT, Macfarlane S. Intestinal bacteria and ulcerative colitis. Curr Issues Intest Microbiol. 2003;4(1):9–20.
Macfarlane GT, Cummings JH. Probiotics, infection and immunity. Curr Opin Infect Dis. 2002;15(5):501–6.
Macfarlane GT, Cummings JH. Probiotics and prebiotics: can regulating the activities of intestinal bacteria benefit health? BMJ: Br Med J. 1999;318(7189):999–1003.
Steed H, Macfarlane GT, Macfarlane S. Prebiotics, synbiotics and inflammatory bowel disease. Mol Nutr Food Res. 2008;52(8):898–905.
Gibson GR, Roberfroid MB. Dietary modulation of the human colonic microbiota: introducing the concept of prebiotics. J Nutr. 1995;125(6):1401–12.
Becker MR, et al. Molecular analysis of bacterial species associated with childhood caries. J Clin Microbiol. 2002;40(3):1001–9.
Caglar E, et al. Short-term effect of ice-cream containing Bifidobacterium lactis Bb-12 on the number of salivary mutans streptococci and lactobacilli. Acta Odontol Scand. 2008;66(3):154–8.
Nase L, et al. 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(6):412–20.
Cildir SK, et al. Reduction of salivary mutans streptococci in orthodontic patients during daily consumption of yoghurt containing probiotic bacteria. Eur J Orthod. 2009;31(4):407–11.
Ahola AJ, et al. Short-term consumption of probiotic-containing cheese and its effect on dental caries risk factors. Arch Oral Biol. 2002;47(11):799–804.
Montalto M, et al. Probiotic treatment increases salivary counts of lactobacilli: a double-blind, randomized, controlled study. Digestion. 2004;69(1):53–6.
Chuang LC, et al. Probiotic Lactobacillus paracasei effect on cariogenic bacterial flora. Clin Oral Investig. 2011;15(4):471–6.
Cytotoxicity of Probiotics from Philippine Commercial Dairy Products on Cancer Cells and the Effect on Expression of cfos and cjun Early Apoptotic-Promoting Genes and Interleukin-1β and Tumor Necrosis Factor-α Proinflammatory Cytokine Genes. BioMed Research International, 2014. 2014: p. 9.
Slawik S, et al. Probiotics affect the clinical inflammatory parameters of experimental gingivitis in humans. Eur J Clin Nutr. 2011;65(7):857–63.
Akhlaghi N, Mortazavi S. The role of probiotics in oro-dental health. J Isfahan Dent School. 2011;7(2):187–99.
Reid G, et al. Potential Uses of Probiotics in Clinical Practice. Clin Microbiol Rev. 2003;16(4):658–72.
Nikawa H, et al. Lactobacillus reuteri in bovine milk fermented decreases the oral carriage of mutans streptococci. Int J Food Microbiol. 2004;95(2):219–23.
Zadik Y, Bechor R. Hidden occlusal caries: challenge for the dentist. N Y State Dent J. 2008;74(4):46–50.
Schulz KF, et al. CONSORT 2010 Statement: updated guidelines for reporting parallel group randomised trials. BMC Med. 2010;8(1):18.
Doron S, Gorbach SL. Probiotics: their role in the treatment and prevention of disease. Expert Rev Anti Infect Ther. 2006;4(2):261–75.
Akhlaghi N, Mortazavi S. The role of probiotics in oro-dental health. J Isfahan Dent School. 2011;7(2):187–99.
Jindal G, et al. A comparative evaluation of probiotics on salivary mutans streptococci counts in Indian children. Eur Arch Paediatr Dent. 2011;12(4):211–5.
Singh RP, Damle SG, Chawla A. Salivary mutans streptococci and lactobacilli modulations in young children on consumption of probiotic ice-cream containing Bifidobacterium lactis Bb12 and Lactobacillus acidophilus La5. Acta Odontol Scand. 2011;69(6):389–94.
Tahmourespour A, et al. The effect of lactobacillus fermentumATCC9338 as a probiotic on the adhesion of oral streptococci in vitro. Iran J Med Microbiol. 2008;2(1):45–51.
Caglar E, et al. Effect of yogurt with Bifidobacterium DN-173 010 on salivary mutans streptococci and lactobacilli in young adults. Acta Odontol Scand. 2005;63(6):317–20.
Moeiny P, et al. Effect of a probiotic yogurt produced in Iran on the salivary counts of Streptococcus mutans. J Res Dent Sci. 2013;10(2):73–82.
Salem RG, Abd-El-Aziz AM, Erfan DM. Assessment of the Effect of Probiotic Yoghurt and Different Probiotic Strains on Salivary Streptococcus Mutans in children: An In Vivo and an In Vitro Study. Egypt J Med Microbiol 2016 25 (2): p. 35–41.
Ghasempour M, et al. Comparative study of Kefir yogurt-drink and sodium fluoride mouth rinse on salivary mutans streptococci. J Contemp Dent Pract. 2014;15(2):214–7.
Haukioja A, et al. Oral adhesion and survival of probiotic and other lactobacilli and bifidobacteria in vitro. Oral Microbiol Immunol. 2006;21(5):326–32.
Taipale T, et al. Bifidobacterium animalis subsp. lactis BB-12 administration in early childhood: a randomized clinical trial of effects on oral colonization by mutans streptococci and the probiotic. Caries Res. 2012;46(1):69–77.
Vinderola CG, Reinheimer JA. Culture media for the enumeration of Bifidobacterium bifidum and Lactobacillus acidophilus in the presence of yoghurt bacteria. Int Dairy J. 1999;9(8):497–505.
Nozari A, et al. The Effect of Iranian Customary Used Probiotic Yogurt on the Children's Salivary Cariogenic Microflora. J Dent (Shiraz). 2015;16(2):81–6.
Furrie E, et al. Synbiotic therapy (Bifidobacterium longum/Synergy 1) initiates resolution of inflammation in patients with active ulcerative colitis: a randomised controlled pilot trial. Gut. 2005;54(2):242–9.
Steed H, et al. Clinical trial: the microbiological and immunological effects of synbiotic consumption - a randomized double-blind placebo-controlled study in active Crohn's disease. Aliment Pharmacol Ther. 2010;32(7):872–83.
Ouwehand AC, et al. Bifidobacterium microbiota and parameters of immune function in elderly subjects. FEMS Immunol Med Microbiol. 2008;53(1):18–25.
Hatakka K, et al. Effects of probiotic therapy on the activity and activation of mild rheumatoid arthritis–a pilot study. Scand J Rheumatol. 2003;32(4):211–5.
Asemi Z, et al. Effects of daily consumption of probiotic yoghurt on inflammatory factors in pregnant women: a randomized controlled trial. Pak J Biol Sci. 2011;14(8):476–82.
Cogulu D, et al. Potential effects of a multistrain probiotic-kefir on salivary Streptococcus mutans and Lactobacillus spp. J Dent Sci. 2010;5(3):144–9.
Salem RG, Abd-El-Aziz AM, Erfan DM. Assessment of the Effect of Probiotic Yoghurt and Different Probiotic Strains on Salivary Streptococcus Mutans in children: An In Vivo and an In Vitro Study. Egypt J Med Microbiol. 2016;25(2):35–41.
Erickson KL, Hubbard NE. Probiotic immunomodulation in health and disease. J Nutr. 2000;130(2S Suppl):S403–9.
Isolauri E, et al. Probiotics: effects on immunity. Am J Clin Nutr. 2001;73(2 Suppl):S444–50.
Kaufman E, Lamster IB. The diagnostic applications of saliva–a review. Crit Rev Oral Biol Med. 2002;13(2):197–212.

No competing interests reported.

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Effects of Consumption of Probiotic Yogurt on Salivary Streptococcus mutans, IL-1β and TNF-α Levels in Adults with Initial Stages of Dental Caries: a Randomized, Double‑Blind, Placebo‑Controlled Clinical Trial

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Abstract

Background

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Results

Conclusion

Trial registration:

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Introduction

Materials and Methods

Subjects

Microorganisms and bacteria cultivation

Measurement of IL-6 and TNF-α

Statistical Analyses

Results

Discussion

Conclusion

Declarations

References

Additional Declarations

Supplementary Files

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