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Journal of Indian Society of Pedodontics and Preventive Dentistry Official publication of Indian Society of Pedodontics and Preventive Dentistry
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ORIGINAL ARTICLE
Year : 2016  |  Volume : 34  |  Issue : 2  |  Page : 120-123
 

The role of cacao extract in reduction of the number of mutans streptococci colonies in the saliva of 12-14 year-old-children


1 Department of Pediatric Dentistry, Hasanuddin University, Makassar, Indonesia
2 Department of Oral Biology, Hasanuddin University, Makassar, Indonesia

Date of Web Publication14-Apr-2016

Correspondence Address:
Dr. Fajriani
Department of Pediatric Dentistry, Hasanuddin University, JL Perintis Kemerdekaan Tamalanrea, Makassar
Indonesia
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0970-4388.180414

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   Abstract 

Background: Cacao bean husk ethanol extract (CBHEE) has polyphenol that acts as an antibacterial agent, specially anti-glucosyltransferase. Aim: This study is aimed to see the effectiveness of CBHEE to reduce the number of mutans streptococci colonies in the saliva of children (with young permanent teeth), when CBHE is used as mouth rinse. Materials and Methods: This study used cross-sectional study design with time-series experimental study and used simple random sampling on 30 subjects. The chosen subjects are those who have middle oral hygiene status (OHI-S). Each subject was given the same intervention; in the first step, saliva was collected from the subjects (prior to intervention), in the second step, the subjects were given 15 mL of CBHEE 0.1% mouth rinse to rinse their mouth for about 30 s. After intervention, their saliva was collected twice in 15 min and 30 min after intervention. Furthermore, the number of mutans streptococci colonies were measured in colony-forming units (CFU) and the data was statistically analyzed using analysis of variance (ANOVA) and paired t-test. The data was analyzed and proccessed using Statistical Package for the Social Sciences (SPSS) 22.0 for windows versions. Results: The data statistically showed, significantly, the reduction of Streptococcus mutans colonies before and after 30 min of intervention. Before intervention, there were 59.10 CFU/mL of mutans streptococci, and after 15 min of intervention it showed reduction of mutans streptococci to 25.73 CFU/mL and after 30 min of intervention, the counts of mutans streptococci showed a reduction to 9.40 CFU/mL. From the test results, statistical value of this research was P = 0.000 (P < 0.05), which means that the reduction of the mutans streptococci count was significant. Conclusion: Using CBHEE as mouth rinse for children has been proven highly effective in reducing mutans streptococci colony counts in the mouth.


Keywords: Cacao extract, mutans streptococci, 12-14 years


How to cite this article:
Fajriani, Mustamin AW, Asmawati. The role of cacao extract in reduction of the number of mutans streptococci colonies in the saliva of 12-14 year-old-children. J Indian Soc Pedod Prev Dent 2016;34:120-3

How to cite this URL:
Fajriani, Mustamin AW, Asmawati. The role of cacao extract in reduction of the number of mutans streptococci colonies in the saliva of 12-14 year-old-children. J Indian Soc Pedod Prev Dent [serial online] 2016 [cited 2021 Apr 18];34:120-3. Available from: https://www.jisppd.com/text.asp?2016/34/2/120/180414



   Introduction Top


Preventive action for dental caries can be realized by protecting oral hygiene; for example, the habit of brushing teeth everyday in the morning after breakfast and in the evening before sleep. But, in some cases, protecting oral hygiene is not enough. Some authors said that the addition of antimicrobial substance to oral health products is important as well. [1],[2],[3],[4]

Polyphenol is a natural phytochemical substance that is found in some plants such as fruits, vegetables, and beans such as coffee and cacao, as well as tea leaves and grapes. [5] Most polyphenols are catechin and epigallocatechin, known as antimicrobial substances that are effective against mutans streptococci. As mentioned before, cacao (Theobroma cacao L). has polyphenol substances. Cacao is rich in polyphenols that are always useful as additional material in a medicinal treatment. [1],[4]

The Indonesian cacao sector has experienced tremendous growth in the past 25 years. The main cocoa-producing area is the Sulawesi island, Indonesia, specially South Sulawesi, Indonesia that produces up to 70% of all Indonesian cacao products, the name of the beans is "Sulawesi bean." One of the biggest cacao factories in Asia is Barry Callebaut Comextra Indonesia (BCCI) located in Makassar, South Sulawesi, Indonesia. Beside producing cacao beans, this factory produces waste materials as well, that is cacao bean husk (shell). Generally, the society deems that only cacao bean has benefits and cacao bean husk is just waste material. But now there are studies about the advantage of chemical substances in cacao bean husk and it has been found that cacao bean husk has polyphenol substances such as antioxidants and antimicrobial agents. [2],[6],[7]

Additionally, in dentistry, there are some studies which have proven that Cacao bean husk ethanol extract (CBHEE) has significant benefit in oral health. A study by Ooshima et al. (2000) showed that CBHEE has antibacterial activity as well as anti-glucosyltransferase activity that effectively capable inhibits mutans streptococci growth as pathogenic bacterial that causes dental caries. [8],[9]

For that reason, the author thought that a research about the effectiveness of CBHEE cacao bean husk extract in reducing the number of mutans streptococci colony, when used as mouth rinse by children with young permanent teeth, is needed.


   Materials and Methods Top


This study used cross-sectional study design with time-series experimental study and used simple random sampling to get 30 subjects. The population consisted of students of 25 Junior High Schools of Makassar, Indonesia, who had attained the age of 12-14 years. The chosen subjects were those who had middle OHI-S status. This study was conducted in the Phytochemistry Laboratory of the Faculty of Pharmacy, Hasanuddin University, 25 Junior High Schools of Makassar, Indonesia, and Microbiology Laboratory of the Faculty of Medicine, Hasanuddin University.

Each subject was given the same intervention; the first step was the collection of saliva from subjects (prior to intervention), in the second step, subjects were given 15 mL of CBHEE solution 0.1% to rinse their mouth for about 30 s. After intervention, the saliva was collected twice in 15 min and 30 min after intervention. Furthermore, the number of mutans streptococci colonies were measured in colony-forming units (CFU) and the data was statistically analyzed using the analysis of variance (ANOVA) and paired t-test. The data was analyzed and proccessed using Statistical Package for the Social Sciences (SPSS) 22.0 for windows versions.


   Result Top


The data was analyzed and proccessed using SPSS 22.0 for windows versions. The result showed in the distribution table as follows:

[Table 1] showed sample characteristic distribution of this study that has 30 samples. According to the table above, the number of males is more than the number of females, that is 18 males (60%) and 12 females (40%). In accordance with the inclusion criteria that had been determined by the author, that oral hygiene status (OHI-S) must be in middle status, so the average OHI-S of samples gathered in this research was 1.45. The number of mutans streptococci colony (CFU/mL) prior to intervention (pretest) was 59.10 CFU/mL. And then, after intervention of the CBHEE solution 0.1% in posttest 1 (after 15 min) became 25.73 CFU/mL and in posttest 2 (after 30 min) had decreased up to 9.40 CFU/mL. It showed that there is significant reduction in the number of mutan streptococci colony from before intervention to 15 min and 30 min after intervention.
Table 1: The sample characteristic distribution


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[Table 2] and [Figure 1] showed the effectiveness of CBHEE solution 0.1% that was used as intervention material in this study. Table V.2 and Figure 2.8 showed reduction in the number of mutans streptococci colony in samples prior to intervention, to 15 min and 30 min after intervention. The reduction is statistically significant. Based on the result of the ANOVA test, the P value was 0.000 (P < 0.05; "significant"). In other words, this CBHEE solution 0.1% has significant effectivenes in reducing the number of mutans streptococci colony.
Figure 1: Reduction the number of mutans streptococci colony in sample 1 and 2 based on study time interval (pre-post 1-post 2)
Source: Author personal documentation


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Table 2: Difference effectiveness of cacao bean husk ethanol extract solution 0.1% based on time interval


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   Discussion Top


The research done by Osawa et al. (2000) has drawn attention because they reported that cacao bean husk has polyphenol substances that contain strong anti-glucosyltransferase activity. The main substance of polyphenols found in cacao bean husk is catechin and epicatechin. Beside that, there are free fatty acids as well such as oleic acid and linoleic acid that showed strong bactericidal activity against mutans streptococci. Glucosyltransferase is one of the mutans streptococci products that play a role in plaque-forming and facilitate enamel demineralization. According to Fukui et al. in 1980, both free fatty acids showed their act to inhibit mutans streptococci growth. Some derrivates of flavonoid are found in cacao bean husk as well. [11],[12],[13],[14]

In this research, the author used CBHEE solution 0.1% that has polyphenol substances, specially flavonoid derivates, in the form of catechin and epigallocatechin which have effectiveness as that can inhibit mutans streptococci activity in plaque-forming as the beginning of tooth decay forming. The author decided to use this material with this concentration (0.1%) because some studies showed that 0.1% of CBHEE has significant influence in reducing mutans streptococci in the saliva and the plaque of the teeth of childen, just like a study in India by Shrikanti et al. [10],[12],[15],[16]

From the obtained data, it can be seen that there are differences in the number of mutans streptococci colony from before intervention to after intervention (pretest-posttest 1-posttest 2). Based on [Table 2] we can see that the mouth rinse solution is effective in reducing the number of mutans streptococci.

The effectiveness of CBHEE solution 0.1% that was used as mouth rinse solution in the intervention of this study, has been proven showing the reduction of mutans streptococci colony. Beside that, according to Srikanth et al. CBHEE solution 0.1% is effective in reducing plaque accumulation in the teeth of children in India as well. Some authors, in addition, have said that there are possibilities of CBHEE in the prevention of oral ulceration and oral cell cancer forming. [9],[10],[17],[18]

Thus, we can see that intervention material used in this study is effective in reducing the number of mutans streptococci colony in the saliva of children. Additionally, it is needed to be known that some safe solvents in extracting cacao bean husk are aseton, ethanol, and methanol. [13],[19],[20],[21]

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
   References Top

1.
Badan Penelitian dan Pengembangan Kesehatan Departemen Kesehatan, Republik Indonesia. Riset kesehatan dasar 2007. p. 130-47.  Back to cited text no. 1
    
2.
Angela A. Pencegahan primer pada anak yang berisiko karies tinggi. Maj Ked Gigi (Dent J) 2005:38:130-1.  Back to cited text no. 2
    
3.
Ismu SS. Karies Gigi Pada Anak Dengan Berbagai Faktor Etiologi. Jakarta: EGC; 1992. p. 6.  Back to cited text no. 3
    
4.
Forssten SD, Björklund M, Ouwehand AC. Ouwehand. Streptococcus mutans, caries and simulation models. Nutrients 2010;2:290-8.  Back to cited text no. 4
    
5.
Keerthi M, Lakshmi Prasanna J, Santhosh Aruna M, Rama Rao N. Review of polyphenol as nature gift. WJPP 2014;3:445-55.  Back to cited text no. 5
    
6.
I Wayan Yasa. Indonesian Cocoa Beans: Current situation. Available from: http://www.icco.org/sites/www.roundtablecocoa.org/documents/8%20Mr.%20I.%20Wayan%20Yasa%20-%20Indonesia.pdf. [Last accessed on 2014 Sep 5].   Back to cited text no. 6
    
7.
Langkong J, Ishak E, Bilang M, Muhidong J. Pemetaan lemak dari biji kakao (Theobroma cacao L) di Sulawesi Selatan. [Last accessed on 2014 Sep 5].  Back to cited text no. 7
    
8.
Ooshima T, Osaka Y, Sasaki H, Osawa K, Yasuda H, Matsumura M, et al. Caries inhibitory activity of cacao bean husk extract in in-vitro and animal experiments. Arch Oral Biol 2000;45:639-45.  Back to cited text no. 8
    
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Petti S, Scully C. Polyphenols, oral health and disease: A review. J Dent 2009;37:413-23.  Back to cited text no. 9
    
10.
Institut Pertanian Bogor. Available from: http://repository.ipb.ac.id/bitstream/handle/123456789/53015/BAB%20II%20Tinjauan%20Pustaka.pdf?sequence=3. [Last accesed on 2013 Des 12].  Back to cited text no. 10
    
11.
Hii CL, Law CL, Suzannah S, Misnawi, Cloke M. Polyphenols in cocoa (Theobroma cacao L.). Asian J Food Agro-Industry 2009;2:702-4.  Back to cited text no. 11
    
12.
Srikanth RK, Shashikiran ND, Subha Reddy VV. Chocolate mouth rinse: Effect on plaque accumulation and mutansstreptococci counts when used by children. J Indian Soc Pedod Prevent Dent 2008;26:67-70.  Back to cited text no. 12
    
13.
Osawa K, Miyazaki K, Shimura S, Okuda J, Matsumoto M, Ooshima T. Identification of cariostatic substances in the cocoa bean husk: Their anti-glucosyltransferase and antibacterial activities. J Dent Res 2001;80:2000-4.  Back to cited text no. 13
    
14.
Jeon JG, Rosalen PL, Falsetta ML, Koo H. Natural products in caries research: Current (limited) knowledge, challenges and future perspective. Caries Res 2011;45:243-4.  Back to cited text no. 14
    
15.
Nugraha AW. Si Plak Dimana-mana "Streptococcus mutans". Yogyakarta: Fakultas Farmasi USD; 2008. p. 1-2.  Back to cited text no. 15
    
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Gazzani G, Daglia M, Papetti A. Food components with anticaries activity. Curr Opin Biotechnol 2012;23:153-9.  Back to cited text no. 16
    
17.
Ferrazzano GF, Amato I, Ingenito A, Zarrelli A, Pinto G, Pollio A. Plant polyphenols and their anti-cariogenic properties: A review. Molecules 2011;16:1486-507.  Back to cited text no. 17
    
18.
Kofink M, Papagiannopoulos M, Galensa R. (-)-Catechin in cocoa and chocolate: Occurence and analysis of an atypical flavan-3-ol enantiomer. Molecules 2007;12:1274-88.  Back to cited text no. 18
    
19.
Jan W. The Contents and Effects of Polyphenols in Chocolate. Dissertation. Germany: University of Gießen; 2004. p. 20-4.  Back to cited text no. 19
    
20.
Nsor-Atindana J, Zhong F, Mothibe KJ; Mohamed Lamine Bangoura dan Camel Lagnika. Quantification of Total Polyphenolic Content and Antimicrobial Activity of Cocoa (Theobroma cacao L.) Bean Shells. Pakistan J Nutr 2012;11:574-9.  Back to cited text no. 20
    
21.
Purnamasari DA, Munadziroh E, Yogiartono RM. Konsentrasi ekstrak biji kakao sebagai material alam dalam menghambat pertumbuhan Streptococcus mutans (Concentration of Cacao ceed extract as a natural material in prevent Streptococcus mutans growth). J PDGI 2009;59:17.  Back to cited text no. 21
    


    Figures

  [Figure 1]
 
 
    Tables

  [Table 1], [Table 2]



 

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