Effect of Mouthwash Containing Bio-active Enzyme on Salivary Mutans Streptococci S. MANGUNDJAJA, E.I. AUERKARI, and A. DJAIS, Universitas Indonesia, Jakarta, Indonesia Abstract Objectives:The aim of the present study was to investigate the effect of antimicrobial agent of containing Bio-active Enzyme of Lysozyme,Lactoferrin,GlucoseOxidase,Lactoperoxidase to inhibit the growth of the salivary mutans streptococci. Methods: Before enrolled in the study, respondents fill and signature the informed consent Ten (10) respondents participated as subjects on the clinical trial were two times treated, before and after mouth washing with Biotene and as subject control,before and after mouth washing with Na Cl 0.9 %. Saliva samples were collected before and after mouth rinsing with Na Cl 0.9 %.A serial dilution was made, followed by pouring on the TYS20B medium. Data, which were obtaining from colony forming units (CFU) of salivary mutans streptococci grew on the TYS20B (Shaeken et al. 1986) medium before and after the Na Cl 0.9% and data obtained were analyzed in a descriptive and t test. Results: showed that there is no significance in the average amount of Streptococcus mutans colonies between before and after mouth rinsing with NaCL 0.9%.However, a significant difference was found respectively as results before and after mouth rinsing with Biotene..Conclusion: We concluded that Mouthwash Containing Bio-active Enzyme has anti microbial activity against local strains of Streptococcus mutans isolated from human harbouring species. Therefore mouth rinsing with Biotene caries can be prevented Key words: Mouthwash,- Biotene-Active Enzyne - Salivary Mutans Streptococci. Introduction The Directorate of Health Ministry of Oral Health Republic Indonesia detected that 60 80 % of population was suffering from dental caries.dental caries on population present a tremendous challenge to the dental profession in population. Streptococcus mutans as the prime cause of dental caries m their first symptoms by forming dental plaques. The dental plaques will cause the process of dental caries and periodontal diseases. The control of dental caries in the community has been done wide various methods,i.e with tooth brushing administration of fluoride either in the tooth paste or drinking water of certain community and gargling with antiseptic.
One methods of controlling dental caries by inhibiting the growth of mutans of Streptococcus mutans in saliva with enzyme in Biotene mouth wash. If the content of enzyme in Biotene the mouth wash can hamper the growth of mutans of Streptococcus mutans in saliva, then we find a acceptable method for the people to prevent dental caries. From the earlier study we have isolated Streptococcus mutans from the subjects in the Island of Panggang,Island of Kelapa Island of Palembang,. The Streptococcus mutans have labeled as S. mutans1, S. mutans2, S. mutans3, S. mutans4, S.mutans5, S.Mutans6(Soeherwin Mangundjaja et al, 1993,1995,1995) 1,2,3. Streptococcus mutans have virulent factors in their relation with dental caries, i.e in their adherent with enamel by forming dental plaque and producing acid(roitt & Lehner,1980) 4.Therefore it can be said that Streptococcus mutans are the most cariogenic bacteria for human and their activity can be seen in their high percentage in the population of bacteria in the fissures and surfaces of the teeth. Streptococcus mutans can produce acid from sucrose faster and with ph lower than the Lactobacillus(Soerodjo,1984) 5. And also have the acidity to make an optimal ph needed for the beginning of the destruction of the teeth. (Soeherwin Mangundjaja et al 2000) 6 stated that CHX mouth wash has been considered as having potential for inhibiting salivary levels of mutans streptococci. Base on study described, it is proper that we should do research toward the Enzyme Biotene against the Streptococcus mutans in saliva in relation with dental caries. ` D. The purpose of research Determining the level of the colony forming units of Streptococcus mutans in saliva of the treatment group who mouth rinsing with Enzyme biotene and the group who without mouth rinsing with Enzyme Biotene. E. The benefit of the research The results of the research can increase the knowledge of the people if by mouth rinsing the enzyme Biotene routinely can decrease the number of Streptococcus mutans in the saliva.so it can be used to anticipate against the occurring of the process of dental caries. F. Methodology The research is done by using experimental laboratory method in human with epidemiological approach. The subject of the research are the Dental students of the Faculty of Dentistry University of Indonesia,10 students 2
participated conducting two times treatment as treated group and being also as control group. The material used in this study is a commercial Biotene Mouthwash containing enzyme Lysozyme,Lactoferrin,GlucoseOxidase,Lactoperoxidase. Respondents are examined their oral hygiene, their oral hygiene shall be on average. Respondents fill and signature the informed consent. Before and after mouth rinsing with NaCl 0.09% and Enzyme in Biotene mouth wash, Respondents chew sterile paraffin to obtain considerable amount of saliva and the saliva is collected. 30 sec,45 sec and 60 sec,one hour after mouth rinsing,saliva are collected Saliva samples, then make a serial dilution. 1 ml of saliva is diluted with 9 ml sterile saline to make a serial dilution and from the tube of 1000 fold dilution, is then take 0.1 ml of solution to inoculate in the selective medium of TYS20B (Schaken et al, 1986) 7 All samples are incubated in anaerobic jar at temperature of 37 0 Celsius degree for 3 X 24 hours.. The Colony Forming Units (CFU) of Streptococcus mutans which grew in the TYS20B medium are then counted and recorded. Data which were obtained from colony forming units of salivary mutans streptococci grew in TYS20B medium before and after treatment were analyzed in a descriptive and t test. Results of the Research Data which were obtained from the colony forming units (CFU) of Streptococcus mutans which grew in the TYS20B bacteria medium before and after mouth rinsing with Na Cl 0/09% and Biotene were analyzed in a descriptive and t test Table 1. Mean value and standard deviation (SD) of CFU of Mutans Streptococci on treatment with Na Cl 0.09% and Biotene for a period of 30 seconds n = 10 Mean Standard deviation Before rinsing with Na Cl 0.09% After rinsing with Na CL 0.09% Before rinsing with Biotene After rinsing with Biotene 480,30 500.10 102.0. 47.80 330.50 330.10 1500.76 92.10 Results shown CFU of mutans streptococci after rinsing with Biotene for a period of 30 seconds lower than before rinsing with Biotene whether with Na CL 0.09% 3
Table 2. t test on treatment with Na Cl and Biotene period 30 seconds Different of treatment t p Before After rinsing with Na Cl 0.09% -0.720 0.420 Before After rinsing with Biotene 2.020 0.016 Before rinsing with Na Cl 0.09% and Biotene -1.014 0.370 After rinsing with Na Cl 0.09% and Biotene 3.400 0.0370 Results shown that Biotene effective to inhibit the growth of salivary mutans streptococci after rinsing with Biotene for a period 30 seconds. Table 3 Mean value and standard deviation (SD) of CFU of mutans streptococci on treatment with Na Cl 0.09% Biotene for a period of 45 seconds N = 10 Mean Standard deviation Before rinsing with Aqua drinking water After rinsing with Aqua drinking water Before rinsing with Biotene After rinsing with Biotene 21.39 39.50 92.90 3.60 23.02 23.80 20.38 2.68 Results shown CFU of salivary mutans streptococci after rinsing with Biotene for a period of 45 seconds lower than before rinsing with Biotene whether with Na Cl 0.09% Table 4. t test on treatment with Na Cl 0.09% and Biotene for a period 45 seconds Different treatment t p Before-after rinsing with Na Cl 0.09% Before-After rinsing with Biotene Before rinsing with Na Cl 0.09% and Biotene 15.10 2.120 2.011 0.120 0.015 0.080 After rinsing with Na Cl 0.09% and Biotene 3.487 0.005 4
Results shown that Bioyene effective to inhibit the growth of salivary mutans streptococci after rinsing with Biotene for a period 45 seconds.. Table 5. Mean value and standard deviation (SD) of CFU of mutans streptococci on treatment with Na Cl 0.09% and Biotene for a period of 60 seconds N = 10 Mean Standard deviation Before rinsing with Na Cl 0.09% After rinsing with Na Cl 0.09% Before rinsing with Biotene After rinsing with Biotene 755.00 920.35 42.80 6.07 1156.56 1150.00 47.80 6.60 Results shown CFU of salivary mutans streptococci after rinsing with Biotene for a period of 60 seconds lower than before rinsing with Biotene whether with Na Cl 0.09%. Table 6. t test on treatment with Na Cl 0.09% and Biotene for a period 60 seconds Different of treatment t p Before-after rinsing with Na Cl 0.09% Before-After rinsing with Biotene Before rinsing with Na Cl 0.09% and Biotene -0.400 2.690 1.0302 0.650 0.970 0.030 Non-sidnificant After rinsing with Na Cl 0.09% and Biotene 2.452 0.045 Results shown that Biotene effective to inhibit the growth of salivary mutans streptococci after rinsing with Biotene for a period 60 seconds.. Discussion From the results shown that Mouthwash Containing Bio-active Enzyme on treatment for a period of 30,45 and 60 seconds CFU of salivary mutans streptococci reduced significantly. Mouthwash Containing Bio-active Enzyme on treatment for a period 60 minutes is the most effective to reduce the population level of mutans streptococci in saliva. Currently known that Streptococcus mutans are triggering factor of caries because their characteristics always are related to caries process.streptococcus mutans can synthesize insoluble glucan (1-3) bond from sucrose, besides that Streptococcus mutans are also more aciduric than other Streptococci group 8. In the past study it had been proven that an enzyme in enzyme toothpaste can inhibit the growth of the population on oral microorganisms and caries activity of mutans streptococci in plaque 9,10. 5
The results of the research can increase the knowledge of the people if by mouth rinsing the enzyme Biotene routinely can decrease the number of Streptococcus mutans in the saliva.so it can be used to anticipate against the occurring of the process of dental caries. Conclusion and Suggestion Mouthwash containing Bio-active enzyme Lysozyme, Lactoferin,Glucose Oxidase,Lactoperoxidase has antimicrobial activity against salivary mutans streptococci.. Nevertheless, we can conclude that containing Bio-active enzyme of Lysozyme, Lactoferrin, Glucose Oxidase, Lactoperoxidase can inhibit the growth of salivary mutans streptococci, so we can anticipate dental caries. References 1. Soeherwin Mangundjaja,1993. Streptococcus mutans in Children in the Island of Panggang.Indonesia. Paper read at Asian Pacific Dental Congress 1993 Kuala Lumpur, Malaysia 2. S.Mangundjaja.,A Muthalib.,A Djais,1993. Streptococcus mutans in Caries free and Caries active subjects in Panggang Island Indonesia Paper read at International Association for Dental Research General session and Exhibition 1995, Singapore 3. S.Mangundjaja., A Muthalib, 1995. Activity of Streptococcus mutans Isolated from Human harboring Species in Kelapa Island Indonesia. Paper read at FDI Conference 1995, Hongkong 4. Roitt I.M and Lehner T 1980. Immunology of Oral Diseases Black Well Scientific Public.Oxford ed 2;346 349 5. Soerodjo,1989.Respon Imun Humoral terhadap Streptococcus mutans Sehubungan dengan Penyakit Karies Gigi. Disertasi Doktor Universitas Airlangga 1989 Surabaya 6. S.Mangundjaja.,R.Khairun Nisa.,S.Lasaryna.,E Fauzia.,Mutya,2000 Pengaruh Obat Kumur Khlorheksidin terhadap Populasi Kuman Streptococcus mutans di dalam Air Liur. Paper read at Indonesian Society for Microbiology Congress 27 28 June 2000 Denpasar bali 7. Shaeken M.J.M.,Vander hoeven C.S., Franken H.M.C 1986. Comparative Recovery of Streptococcus mutans on Five Isolation Media Including a New Simple Selective Medium. J,dent Res,1986;65;906-908 8. Mayes P.A. Metabolism of Carbohydrate,in Harper Review of Biochemistry. (Martin DW.,Mayes P.A and Rodwell V.W eds) ed 19 th Maruzen asian ed;page 62-167;Singapore 1983 9. Soeherwin Mangundjaja and Alexander Agung. The effect of Dentifrice Containing Enzyme on Oral Microorganisms. Presented at The Periodontology Association of Taiwan January 2003 Taiwan Taipe. 6
10. Mangundjaja S.,Djais S.,Angky S., Elina., Heriandi S., and Alexander Agung. The Effect of Enzyme Dentifrice on Caries Activity of Mutans Streptococci in Plaque.Presented at The FDI Annual World Dental Congress 27 September 1 October 2001 Kuala Lumpur Mala ---000--- Presented at The American Association of Dental Research Annual Meeting 2 6 April 2008 Dallas Texas USA 7