Soeherwin Mangundjaja., Abdul Muthalib., Ariadna Djais Department of Oral Biology Faculty of Dentistry Universitas Indonesia

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1 THE EFFECT OF DENTIFRICE CONTAINING ENZYME ON SALIVARY MUTANS STREPTOCOCCAL LEVEL IN ORTHODONTIC PATIENTS Soeherwin Mangundjaja., Abdul Muthalib., Ariadna Djais Department of Oral Biology Faculty of Dentistry Universitas Indonesia Abstract Objectives:The study was carried out to investigate the effect of dentifrice containing enzyme in inhibiting the growth of the salivary mutans streptococci. Method:Ten respondents wearing fixed orthodontic appliances par1icipated as the subjects, conducting two times of treatment as follows: ten as treatment group before and after tooth brushing with dentifrice containing enzyme, whereas ten subjects as control group before and after tooth brushing with dentifrice containing non-enzyme. Saliva samples were collected before and after tooth brushing with dentifrice containing enzyme and with non-enzyme, a serial dilution was made, followed by inoculating on TYS20B medium (Schaeken MJM, var. der Hoeven JS and Franken. 1986). Data which were obtained from Colony Forming Units of salivary mutans streptococci grew on the TYS20B medium before and after tooth brushing were analyzed in a descriptive and "t" test. Results: showed that there was no significant difference in the average amount of Streptococcus mutans colonies between before and after using non-enzyme. However, a significant difference (p=0.000) was found respectively as a result of before and after tooth brushing with dentifrice containing enzyme. Conclusion: Tooth brushing with dentifrice containing enzyme can inhibit the growth of salivary mutans streptococci. It can be used in preventing caries risk in orthodontic patients. Key words: Enzyme: Mutans streptococcal level, orthodontic. INRODUCTION Streptococcus mutans harbored in the dental plaque is thought to be the main agent to caries prevalence. Acidogenicity and aciduricity are important biochemical characteristics for cariogenicity of microorganism. The mutans streptococci have both of these properties and are considered the most cariogenic group within the oral micro flora. 1,.2

2 For this reason. early prevention is needed to maintain the oral health by rinsing through using mouthwash and tooth brushing. It is the most widely used and socially accepted form of oral hygiene 3. Mouth rinsing and tooth brushing are the principal way for mechanical removal of plaque and to prevent Streptococcus mutans colonized in teeth 3.4,5. The purpose of tooth brushing with dentifrice with chemotherapeutic agent is to reduce the population of plaque and salivary mutans streptococci. Recently, many kinds of dentifrice have been found in the market, with very many kinds of brands and utilities. Dentifrice containing enzyme of Amyloglucosidase (Am) and Glucoseoxidase (GO) 1.5% and Sodium fluoride 0.24 % has abroad bacteriostatic agent and is generally more effective against gram-positive than against gram -negative bacteria. This is a preliminary report to investigate the effectiveness of enzyme dentifrice on the growth of salivary mutans streptococci on the patients wearing fixed orthodontic appliances The amount of salivary mutans streptococci taken from orthodontic patients, before and after tooth brushing with enzyme dentifrice and non-enzyme as control are compared. This research is expected to contribute significantly to the world of science that toothpaste containing enzyme can inhibit the growth of salivary mutans streptococci, therefore in a long term of using dentifrice containing enzyme, caries can be prevented. MATERIAL and METHODS Ten subjects participated in this study. All were in good general and oral health, no evidence of progressive periodontal diseases and no untreated caries. All wore fixed orthodontic appliances and none was taking medications during the study. All subjects participated in two kinds of experiments, each lasting 30 days, and the experiments as follows : ten as treatment group. before and after tooth brushing with dentifrice containing enzyme of Amyloglucosidase (Am) and Glucoseoxidase (GO) 1.5 % and Sodium fluoride 0.24 % and also being as control group before and after tooth brushing with non-enzyme. The analyzed unit was salivary of mutans streptococci. It grew in the selective medium of Trypticase Yeast Extract Cystine with Bacitracin (TYS20B). 7 Respondents were examined their oral hygiene. Their oral hygiene shall be on the average. before tooth brushing with enzyme dentifrice and non-enzyme dentifrice. 2

3 Respondents chew a paraffin sterile for a period 30 seconds, then saliva samples were collected. Respondents were requested in the first month, as control group brushed their teeth twice a day with non enzyme dentifrice. In the next one month they brushed their teeth with dentifrice containing enzyme as treatment group. Saliva sample, before and after tooth brushing with enzyme dentifrice and non-enzyme dentifrice was then taken of 1 ml for a serial dilution for One per ten ml from a serial dilution of 10-4 was taken and it streaked on the TY20SB medium, incubated on 37 0 Celsius degree, anaerobic condition for a period 3 X 24 hours. The number or total Colony Forming Units of mutans streptococci on TY20SB agar were examined and counted with the aid of dissecting microscope. Data were obtained from Colony Forming Units (CFU) of salivary mutans streptococci grew on TY20SB agar before and after tooth brushing with dentifrice containing Amyloglucosidase (Am) and Glucoseoxidase (GO) 1.5 % and Sodium fluoride 0.24 % and non-enzyme were analyzed in a descriptive and " t test. RESULTS The results of analyzed CFU amount of Streptococcus mutans grown on TY20SB media before and after tooth brushing with dentifrice containing Amyloglucosidase (Am) and Glucoseoxidase (GO) 1.5 % and Sodium fluoride 0.24 % and non-enzyme are shown in table 1. Table 1. Mean value and standard deviation (SD) of CFU of Streptococcus mutans on treatment with enzyme and non-enzyme dentifrice. Dentifrice Mean SD Before-Non-enzyme After- Non-enzyme Before-Enzyme After- Enzyme Table 1 shows that Colony Forming Units of Streptococcus mutans after tooth brushing with dentifrice containing Amyloglucosidase (Am) and Glucoseoxidase (GO) 1.5 % and Sodium fluoride 0.24 % (X=41.38±34.80) 3

4 is lower than before with brushing Amyloglucosidase (Am) and Glucoseoxidase (GO)1.5% and Sodium fluoride 0.24 % (X=190.75±133.80). The Colony Forming Unit of Streptococcus mutans after brushing with Amyloglucosidase (Am) and Glucoseoxidase (GO) 1.5 % and Sodium fluoride 0.24 % (X=41.38±34.80) is also lower than after brushing with nonenzyme (X=199.30± ). Student t test analysis was done to know the significance of the effectiveness of tooth brushing with dentifrice containing Amyloglucosidase (Am) and Glucoseoxidase (GO) 1.5 % and Sodium fluoride 0.24 %.the results can be seen in table 2. Table 2. "t" value on treatment with brushing with dentifrice containing enzyme and non-enzyme dentifrice. Difference of Treatment t p Before-after tooth brushing (non-enzyme) Before-after tooth brushing ( enzyme) Before tooth brushing (Non-enzyme - Enzyme) After tooth brushing (Non-enzyme-Enzyme) No significant difference was found between the Colony Forming Units of Streptococcus mutans before and after tooth brushing with nonenzyme (p = t = 0.197). As expected the amount of salivary of mutans streptococci showed a very significant difference between the Colony Forming Units before and after tooth brushing with enzyme dentifrice (p =3.450,t = 0.005). When Colony Forming Units of Streptococcus mutans before brushing with non-enzyme compared to before brushing with enzyme dentifrice (t = 0.452,p = 0.734) meaning that there is no significant difference after tooth brushing with non-enzyme compared to after brushing with enzyme dentifrice also showed significant difference ( p = 0.010, t = 0,734) DISCUSSION From the results shown That the.amount of Colony Forming Units Streptococcus mutans after tooth brushing with dentifrice containing Amyloglucosidase (AM) and Glucoseoxidase (GO) 1.5 % and Sodium fluoride 0.24 % found to be lower compared to the amount of Colony Forming Units of Streptococcus mutans after tooth brushing with non- 4

5 enzyme. The Colony Forming Units of Streptococcus mutans has been decreased % of the CFU before using enzyme dentifrice. The virulence of mutans streptococci is caused by its ability to produce glucose polymers (glucan) of which the material to form dental plaque, Streptococcus mutans is able to synthesize glucan from sucrose catalyzed by glucosyltransferase through anaerobic glycolysis to become lactate, propionate, and acetate acid. The product of acid which ph will decrease in a few minutes. This acidic environment gives selective growth and amplification for oral acid tolerant bacteria such as Streptococcus mutans. Due to this condition the use of enzyme dentifrice might reduce the population of mutans streptococci, this is in accordance with previous findings that enzyme dentifrice is effective in suppressing on caries activity of mutans streptococci in plaque 9. In the previous study, one case was found Pseudomonas aeruginosa, It is gram negative bacteria, anaerobe facultative, am! scattered in soil. Pseudomonas aeruginosa is an opportunistic bacteria. If the environments are favorable for this bacteria. It will be a pathogenic microorganism. 10 Fixed orthodontic appliance gave the opportunity to growth and amplification for anaerobic oral bacteria such as Pseudomonas aeruginosa, and also highly colonized with mutans streptococci. CONCLUSION Tooth brushing with dentifrice containing enzyme will decrease population of salivary mutans streptococci. It is expected that it can be used in preventing caries risk in orthodontic patients. ACKNOWLEDGMENT We would like to thank Enzim PT Indonesia Dental Industries which has supported the research from the first beginning. References 1. Ernilson CG. Lidquisl B, Krasse B. Colonization pattern of two serotype. of mutans streptococci implanted in human Journal Caries Res 1989;

6 2. Kohler B, Birkhed, Olsson S. Acid production by human strains of Streptococcus mutans and Streptococcus sobrinus. Journal Caries Res 1995;29: Newbrun E.Cariology 3 th ed. Chicago:Quentessence Publishing Co Inc.1989; Erickson L. Oral health promotion and prevention for older adult. Journal Dent Clinics of North America. 1997; 41 (4): Harris NO and Christen. Primary preventive dentistry 4 th ed. Stamford Connecticut: Appleton and Lange 1995; Hendrik Hoogemdoorn. The effect of lactoperoxidase thyocyanate hydrogen peroxide on the metabolism of cariogenic microorganisms in vitro and in the oral cavity. Netherland: Technische Hogenschool Delf Doctor Dessertation. 1974: Schaeken MJM. van der Hoeven JS, Franken HCM. Comparative recovery of Streptococcus mutans on five isolation media including a new simple selective medium. Journal Dent Res 1986; 65: Trahan, L. Xylitol A review of its action on mutans streptococci and dental plaque its significance. Inter Dental.J 1995; 48: Mangundjaja S, Djais A, Angky S, Elina, Heriandi S. The effect of Enzyme dentifrice on Caries Activity of Mutans streptococci in Plaque. Presented at The FDI Annual World Congress in 200I. Kuala Lumpur: Malaysia 10. Mangundjaja S., A Muthalib,. Djais A., Auerkari L. The effect of chlorhexidine mouthwash treatment on salivary mutans streptococci levels in Orthodontic Patients. Jurnal Kedokteran Gigi Universitas Indonesia Edisi Khlusus 2000;7: JURNAL KEDOKTERAN GIGI - USU Dentika Dental Journal Vol 6 No 1, 2001: