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ORIGINAL ARTICLE
Year : 2021  |  Volume : 39  |  Issue : 3  |  Page : 275-278
 

Role of probiotics and synbiotics on inhibiting Streptococcus mutans level in saliva of children: A randomized controlled trial


1 Department of Pediatric and Preventive Dentistry, Vinayaka Mission's Sankarachariyar Dental College, Vinayaka Mission's Research Foundation (Deemed to be University), Salem, Tamil Nadu, India
2 Department of Pediatric and Preventive Dentistry, KSR Institute of Dental Science and Research, Tiruchengode, Tamil Nadu, India
3 Department of Public Health Dentistry, Vinayaka Mission's Sankarachariyar Dental College, Vinayaka Mission's Research Foundation (Deemed to be University), Salem, Tamil Nadu, India
4 Department of Orthodontics and Dentofacial Orthopaedics, Vinayaka Mission's Sankarachariyar Dental College, Vinayaka Mission's Research Foundation (Deemed to be University), Salem, Tamil Nadu, India

Date of Submission26-Jul-2021
Date of Decision22-Sep-2021
Date of Acceptance22-Sep-2021
Date of Web Publication22-Nov-2021

Correspondence Address:
Dr. Vinola Duraisamy
Department of Pediatric and Preventive Dentistry, Vinayaka Mission's Sankarachariyar Dental College, Vinayaka Mission's Research Foundation (Deemed to be University), Salem, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jisppd.jisppd_270_21

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   Abstract 


Background: Probiotics and synbiotics through its direct and indirect interactions interferes with biofilm formation, competes with oral microorganisms thereby creating better oral health. Aim: This study aims to assess the effectiveness of Probiotics and Synbiotics on inhibiting Streptococcus mutans level in saliva of children after 15 days of daily intake of probiotic and synbiotic curd. Methodology: A double-blind randomized controlled trial was planned. Salivary samples at baseline were collected from forty children in the age group of 6–12 years who were divided into two groups of 20 each to receive probiotic and synbiotic curd respectively for 15 days. Salivary samples were collected after intervention and S. mutans levels were estimated. Statistical Analysis: The collected data were analyzed using Statistical Package for Social Sciences (SPSS) version 22. Intra- and inter-group comparison of mean S. mutans levels for both the groups were done using the paired and unpaired t-test respectively. The statistical significance was set at P ≤ 0.05. Results: A statistically significant reduction in salivary S. mutans counts was observed in both the groups after 15 days (P < 0.001). A significantly higher inhibition of S. mutans growth was present in the probiotic compared to synbiotic group (P < 0.001). Conclusion: Probiotics and Synbiotics were effective in inhibiting salivary S. mutans level of children. However, inhibition of S. mutans growth was found to be better in children who consumed Probiotic curd than the Synbiotic curd.


Keywords: Dental caries, probiotics, Streptococcus mutans, synbiotics


How to cite this article:
Duraisamy V, Geethapriya P R, Bharath C, Niveditha R S, John J B. Role of probiotics and synbiotics on inhibiting Streptococcus mutans level in saliva of children: A randomized controlled trial. J Indian Soc Pedod Prev Dent 2021;39:275-8

How to cite this URL:
Duraisamy V, Geethapriya P R, Bharath C, Niveditha R S, John J B. Role of probiotics and synbiotics on inhibiting Streptococcus mutans level in saliva of children: A randomized controlled trial. J Indian Soc Pedod Prev Dent [serial online] 2021 [cited 2021 Dec 5];39:275-8. Available from: https://www.jisppd.com/text.asp?2021/39/3/275/330706





   Introduction Top


Dental caries can be defined as a localized chemical loss of tooth structure carried by the metabolic activity of dental biofilm that covers the tooth surface.[1] Various factors such as bacterial colonization, saliva, fluoride exposure, consumption of dietary sugar influence the demineralization process and contribute to the development of dental caries.[2] In this process, Streptococcus mutans is still recognized as a major contributor to active dental caries due to its ability to produce acids at faster rates than that of other microbiota and is commonly deposited in the hard tissues of the oral cavity.[3] This results in bacterial adhesion, aggregation, and co-aggregation to colonize the oral cavity. Caries management strategies have traversed through various paths and recently it shows a paradigm shift toward preventive approach. Therapeutic antibiotics have relatively broad spectrum anti-microbial activity and they destroy both commensals as well as potentially destructive bacteria thereby creating an imbalance within the microflora. Alternate methods to selectively inhibit the oral pathogens can be achieved by the use of probiotics and synbiotics.[4]

Probiotics as a term were first used by Lilly and Stillwell (1965) to describe the 'substances secreted by one microorganism that stimulate the growth of another. The WHO defined probiotics as “live microorganisms which when administered in adequate amounts achieve a health benefit on the host.”[4] Probiotics through its direct and indirect interactions interferes with biofilm formation by competing with oral microorganisms by inhibiting the colonization of S. mutans on the tooth surface and prevent further development of dental caries to create better oral health.[5] Probiotic products are commercially available in form of milk, Yougurt, cheese, ice cream, lozenges, and chewing gums.

Gibson and Roberfroid (1995) introduced synbiotics as a nondigestable food ingredient that beneficially affects the host by selectively stimulating the growth and or activity of limited number of bacteria to improve host health.[6] A synbiotic is the combination of a microorganism shown to have some beneficial effect when consumed (i.e., a probiotic) and a compound that specifically favours growth (i.e., a prebiotic), having a synergistic effect when paired together.[7] The possible role of probiotics and synbiotics on inhibiting S. mutans level in saliva of children is less explored, no studies have been conducted to determine the effect of probiotics and synbiotics in inhibiting S. mutans levels in saliva of children. Hence, the present study was aimed to assess and compare the effectiveness of probiotic curd and synbiotic curd on inhibiting S. mutans level in saliva among 6–12 years old institutionalized school children. The null hypothesis set was, there was no difference between the effectiveness of probiotic curd and synbiotic curd in reduction of S. mutans level in saliva of children aged 6–12 years.


   Methodology Top


A randomized controlled study was planned, and the study carried after obtaining approval from the Institutional Research and Ethical Committee. The sample size estimation was done based on the previously published literature by Srivastava et al.[8] in the year 2016. A total of 150 children were screened. Based on the selection criteria, a purposive sample of 40 children aged 6–12 years were selected from an institutionalized home following uniform dietary pattern. The study protocol is described in [Figure 1].
Figure 1: Flow diagram of study protocol

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Inclusion criteria

  • Children/parents who gave informed consent to participate in the study
  • Children without any systemic health condition
  • Children with no active carious lesion
  • Children who were physically and mentally healthy.


Exclusion criteria

  • Children under antibiotic coverage or on any other drugs during the last 4 weeks before the commencement of study
  • Children with a history of allergy to milk products.


Informed oral and written consent was obtained from the institutional home authorities and the parents or their caretakers after explaining the protocol of the study in detail. Children were randomly divided into two groups based on the type of intervention, with 20 children in each group. Randomization was done using computer generated table of random numbers by an intern who was not involved in the study. The secondary investigator did the allocation concealment of the intervention using Sequentially Numbered Opaque Sealed Envelopes.

Group A: Children received 100 ml of Probiotic curd (Nilgiris Dahi Curd) once daily for 15 consecutive days.

Group B: Children received 100 ml of Synbiotic curd once daily for 15 consecutive days.

A commercially available probiotic curd (Nilgiris Dahi Curd) containing active Probiotic culture (0.1%) in 200 g of curd was given for Group A children. The children of Group B received probiotic curd (Nilgiris Dahi Curd) freshly mixed with commercially available Oats (Quaker brand) that was grounded into fine oats powder and heat-treated/cooked for 5 min at 85°C, prior to its addition.[9] All the study participants were supervised every day by an independent school teacher who was not involved in the study at 5.30 pm while consuming their respective products. The intervention was provided for 15 consecutive days at a particular specific time in the evening. The double blinding procedure was followed in which the study participants and the primary investigator were blinded of the intervention provided. The study participants were instructed to follow their routine oral hygiene practices and dietary patterns during the study period.

Salivary sample collection

Before the start of the study, all the study participants were given instructions regarding their maintenance of oral hygiene. They were asked to brush with their regular toothpaste during the study period. The saliva samples were collected in the morning hours by the primary investigator few minutes after tooth brushing. Children were made to sit in an upright position in a well illuminated room and 3 ml of unstimulated saliva was collected from each participant. The saliva samples were collected at baseline and after the study period of 15 days (i.e., on the 16th day) to evaluate study the S. mutans level. The collected saliva samples were then transported to the laboratory immediately in an icebox and were subjected to microbiological analysis. One milliliter of saliva sample was inoculated into Mitis Salivarius Bacitracin Agar[10] using pour plate method and were inoculated at 37°C for 48 h. The colonies were then counted using manual colony counter and colony forming unit was estimated as number of colony forming units per ml of saliva.

Statistical analysis

The data obtained were statistically analyzed using the statistical package for the social sciences (IBM SPSS Inc., Chicago, II, USA, version 22.0). Descriptive and analytical statistical tests were performed. Intra and intergroup comparison of the means of S. mutans levels for both the groups were done using the paired and unpaired t-test, respectively. The statistical significance was set at P < 0.05.


   Results Top


A total of 40 children (mean age 10.20 ± 0.14) participated in the study. A significant reduction in the mean salivary S. mutans levels at post intervention was observed in both the probiotics group and synbiotic group (P < 0.001) [Table 1]. Intergroup comparison of mean S. mutans counts of postintervention showed a statistically significant reduction in the probiotic group (P < 0.001) [Table 2].
Table 1: Comparison of Streptococcus mutans levels of children in probiotic curd and synbiotic curd group

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Table 2: Intergroup comparison of Streptococcus mutans levels

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   Discussion Top


Dental caries is a multifactorial disease of bacterial origin that requires a multimodal approach for prevention and management. Curd being semi-solid has an extended beneficial effect compared to milk and milk products as they are retained for a longer period of time in the oral cavity.[10] Probiotic curd acts by neutralizing the acidic conditions in the mouth by interfering with cariogenic bacteria.[11] Hence, the present study was conducted using probiotic curd and synbiotic curd in inhibiting S. mutans level in saliva among 6–12 years old institutionalized school children. The present study results revealed that there was a significant reduction in the mean salivary S. mutans levels after 15 days of intervention in both the probiotics group and synbiotics group which was similar to the study done by Chinnappa et al.[12] However, the inhibition of S. mutans growth was significantly higher in the probiotic group. Probiotic therapy could be considered as an active means of inhibiting oral biofilm development and reducing the cascade of harmful immune-inflammatory reactions.[13] The consumption of dairy products containing probiotics could be considered as an alternative to improve the oral health in children which is in line with studies done by Näse et al.[14] and Ashwin et al.[15] Reduction in S. mutans count is found in an array of probiotics. However, intergroup comparison of mean S. mutans level post intervention showed a statistically significant reduction in probiotic group compared with synbiotic group. Hedayati-Hajikand et al.[16] had concluded in their randomized control trial that probiotic chewing tablets had an additive effect in reducing the development of early child hood caries when administered along with fluoride toothpaste. In the present study probiotics was better than synbiotics in inhibiting S. mutans level in saliva of children. Hence, probiotic curd having anti-cariogenic properties can be used for caries prevention as the pathogenic bacteria get inhibited due to their greater adherence to the tissues. Probiotic curd when administered can prevent dental caries with minimal adverse effects.[17] A significantly effective reduction in the salivary S. mutans levels was noted when yogurt milk was supplemented with sodium fluoride mouth rinse.[18] Studies have proved that children consuming milk containing probiotic, had significantly fewer dental caries and lower salivary S. mutans than controls.[19],[20] These promising results suggest a potentially beneficial application of probiotics in the prevention of dental caries. A larger sample size with longer follow-up period is required to study the long term effects of probiotics and synbiotics in reducing S. mutans level in saliva.


   Conclusion Top


Probiotics and Synbiotics were effective in inhibiting S. mutans level in saliva of children. Inhibition of S. mutans growth was significantly higher in children of the probiotic group than synbiotic group after 15 days intervention.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

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Selwitz RH, Ismail AI, Pitts NB. Dental caries. Lancet 2007;369:51-9.  Back to cited text no. 1
    
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Featherstone JD. The continuum of dental caries – Evidence for a dynamic disease process. J Dent Res 2004;83:C39-42.  Back to cited text no. 2
    
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Paddick JS, Brailsford SR, Kidd EA, Gilbert SC, Clark DT, Alam S, et al. Effect of the environment on genotypic diversity of Actinomyces naeslundii and Streptococcus oralis in the oral biofilm. Appl Environ Microbiol 2003;69:6475-80.  Back to cited text no. 3
    
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Poorni S, Srinivasan MR, Nivedhitha MS. Probiotic Streptococcus strains in caries prevention: A systematic review. J Conserv Dent 2019;22:123-8.  Back to cited text no. 4
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Srivastava S, Saha S, Kumari M, Mohd S. Effect of probiotic curd on salivary pH and Streptococcus mutans: A double blind parallel randomized controlled trial. J Clin Diagn Res 2016;10:C13-6.  Back to cited text no. 8
    
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Mahrous H, El-Kholy WM, Elsanhoty RM. Production of new synbiotic yoghurt with local probiotic isolate and oat and study its effect on mice. J Adv Dairy Res 2014;2:1-7.  Back to cited text no. 9
    
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Bhalla M, Ingle NA, Kaur N, Yadav P. Mutans streptococci estimation in saliva before and after consumption of probiotic curd among school children. J Int Soc Prev Community Dent 2015;5:31-4.  Back to cited text no. 10
    
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Ferrazzano GF, Cantile T, Quarto M, Ingenito A, Chianese L, Addeo F. Protective effect of yogurt extract on dental enamel demineralization in vitro. Aust Dent J 2008;53:314-9.  Back to cited text no. 11
    
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Chinnappa A, Konde H, Konde S, Raj S, Beena JP. Probiotics for future caries control: A short-term clinical study. Indian J Dent Res 2013;24:547-9.  Back to cited text no. 12
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Patil RU, Dastoor PP, Unde MP. Comparative evaluation of antimicrobial effectiveness of probiotic milk and fluoride mouthrinse on salivary Streptococcus mutans counts and plaque scores in children – An in vivo experimental study. J Indian Soc Pedod Prev Dent 2019;37:378-82.  Back to cited text no. 13
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Näse L, Hatakka K, Savilahti E, Saxelin M, Pönkä A, Poussa T, 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;35:412-20.  Back to cited text no. 14
    
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Ashwin D, Ke V, Taranath M, Ramagoni NK, Nara A, Sarpangala M. Effect of probiotic containing ice cream on salivary mutans streptococci (SMS) levels in children of 612 years of age: A randomized controlled double blind study with sixmonths follow up. J Clin Diagn Res 2015;9:ZC069.  Back to cited text no. 15
    
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Hedayati-Hajikand T, Lundberg U, Eldh C, Twetman S. Effect of probiotic chewing tablets on early childhood caries – A randomized controlled trial. BMC Oral Health 2015;15:112.  Back to cited text no. 16
    
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Ghasempour M, Sefdgar SA, Moghadamnia AA, Ghadimi R, Gharekhani S, Shirkhani L. Comparative study of kefir yogurtdrink and sodium fluoride mouth rinse on salivary mutans streptococci. J Contemp Dent Pract 2014;15:2147.  Back to cited text no. 17
    
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Srinivasan S, Nandlal B, Rao MV. Assessment of plaque regrowth with a probiotic toothpaste containing Lactobacillus paracasei: A spectrophotometric study. J Indian Soc Pedod Prev Dent 2017;35:307-11.  Back to cited text no. 18
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Kavitha M, Prathima GS, Kayalvizhi G, Sanguida A, Ezhumalai G, Ramesh V. Evaluation of Streptococcus mutans serotypes e, f, and k in saliva samples of 6-12-year-old school children before and after a short-term daily intake of the probiotic lozenge. J Indian Soc Pedod Prev Dent 2019;37:67-74.  Back to cited text no. 19
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Jindal G, Pandey RK, Singh RK, Pandey N. Can early exposure to probiotics in children prevent dental caries? A current perspective. J Oral Biol Craniofac Res 2012;2:110-5.  Back to cited text no. 20
    


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