THE EFFECT OF DENTIFRICE CONTAINING ENZYME ON ORAL MICROORGANISMS

THE EFFECT OF DENTIFRICE CONTAINING ENZYME ON ORAL MICROORGANISMS Soeherwin Mangundjaja * and Alexander Agung ** *Oral Biology Department of Faculty of Dentistry Universitas Indonesia and ** Research & Development Department of PT Enzym Bioteknologi Internusa ABSTRACT Saliva as a main mechanism of defense in its role against tooth caries process and periodontal disease. Several enzyme system and anti-bacterial in the saliva are still unclear statements.. The purpose of this paper is to obtain a picture of the effect of enzyme dentifrice on various oral diseases such as : Recurrent Aphthous Stomatitis, Halitosis and the use of enzyme dentifrice on caries activity of mutans streptococci in plaque, the growth of salivary mutans streptococci in orthodontic patients and on microbial contamination of toothbrushes. Data were obtained from those of various oral disease and the use of enzyme dentifrice on oral microorganisms showed that enzyme has the inhibiting effect toward oral microorganisms. It is a known fact that the enzyme systems in the saliva has the inhibiting effect toward the oral flora, but so far this inhibiting effect role toward oral disease is not popular among the dental profession or in the public. It can be concluded that the most aspect of oral and dental health is maintaining equilibrium of eco system in oral cavity, and it will involve antibacterial system in saliva as well as from out side the body. Key words: Enzyme - Oral Microorganisms INTRODUCTION There are various kinds of microorganisms which are normal flora in oral cavity, because oral cavity is a gate between external environment and internal environment of our body, so microorganisms may enter our body through it and then grow in our body. The existence of microorganisms which are normal flora in our cavity may give advantageous or disadvantageous effect to our body, there are three kinds of interaction between the host and microorganisms if e mutualism, commensalisms and parasitism relation. That interaction is dynamic interaction with three factors influencing each other, it e microorganisms, host and environment.microorganisms may cause infection or disorder if equilibrium among those three factors changes to the advantage of microorganisms. Several clinical manifestations of activity of bacteria which are flora in dental plaque and saliva are dental caries and periodontal diseases, besides oral cavity diseases I among others Recurrent Aphthous Stomatitis, due to imbalance condition in oral cavity and plaque. Dental caries is common dental health problem throughout the world although in some industrial countries its prevalence decreases ( Van Palenstein Helderman et ai, 1996). The opposite is true in Indonesia its prevalence tends to increase ( Effendi.1, 1984). The result of survey in special regions of Jakarta Indonesia shows d.ej.t index of

6.04 which means that every one child suffers the average of 6 dental caries (Ismu SS, 1984). The tendency of increasing prevalence of dental caries among others is due to increasing in take of carbohydrate from 7.7 kg annually in 1974 to 12.5 Kg annually in 1979 in Indonesia (Effendi 11984), however the current carbohydrate intake rate is unknown. By the increasing of sugar consumption, cariogenic activity among Indonesian population will also increase. this is a challenge for dentists to reduce dental caries and periodontal diseases prevalence in Indonesia. One of etiological factor of dental caries is specific role of microorganism i,e Streptococcus mutans.. One of factors of the host is factor of saliva, because besides containing anti bacterial component, it also contains organic component which may facilitate the growth and propagation of bacteria in oral cavity. So, it can be said that saliva has the function of protector for oral cavity bacteria, therefore dynamic equilibrium among host, microorganisms and environment can be maintained. Due to high prevalence of infectious diseases in Indonesia requirement for antibiotics is also high, and in turn it caus'e increasing resistance of bacteria against various types of antibiotics. Many preventive efforts against oral cavity infectious diseases have been carried out, among others by using antiseptic mouthwash, antibiotic and traditional medicine. However, up to now the prevalence of oral cavity infectious diseases still high, among others dental caries, periodontal diseases, and Stomatitis. Currently not many types of medicine containing enzyme which can accelerate recovery process of oral cavity infectious disease, can be found in society. However, now we can find tooth paste containing enzyme and fluoride but without detergent. It is known as tooth paste with typical quality of preventing various oral cavity infectious diseases. However, whether the use qf that toothpaste containing enzyme is medically accountable, shall need further careful and profound study, to know accurately the actual effect of that enzyme. What which will be presented here is toothpaste containing Amiloglucosidase (AG) and Glucoseoxidase (GO) enzymes, which are used to maintain dental and oral health. This study is intended to know how far the effectiveness of that enzyme against various diseases in society among others against dental caries, periodontal disease, halitosis and Stomatitis Aphthosa Recurrent. LITERATURE STUDY Among various microorganisms in oral cavity the cariogenics are Stretococcus mutans, Actinomyces viscosus and Lactobacillus acidophilus. The triggering factor of dental caries is Streptococcus mutans, because it can synthesizes glucan with a. bond ( 1-3) which is insoluble in water, from sucrose and it is also capable of producing lactic acid (Mayes, 1983) through homofermantation (Lehninger, 1975). Streptococcus mutans can form colony in hard surface and is more aciduric than other Streptococcus spp, Streptococcus mutans can grow in oral cavity if there is hard and solid surface such as teeth, prostheses appliance and -fix orthodontic appliance. 2

The main habitat of Streptococcus mutans is dental surface, but it is not uniform in the entire surfaces, even it is' often localized in certain surfaces (Newburn, 1989). Streptococcus mutans usually makes its colony in dental pit and fissures, occlusal surface, a proximal area, gingival sulcus (Schachtele, 1983) or in dental caries lesion. The development of dental caries and periodontal; diseases is preceded by dental plaque formation by Streptococcus mutans. Therefore virulence of Mutans streptococci is indicated by its capability to form dental plaque. A. THE EFFECT OF ENZYME DENTIFRICE ON CARIES ACTIVITY OF MUTANS STREPTOCOCCI INPLAQUE. (Soeherwin Mangundjaja, Angky S, Djais A, Elina Heriandi and, Agung A, 2001) reported result of their research, which shows that tooth brushing with dentifrice containing enzymes Amiloglucosidase (AG) and Glucoseoxidase (GO) can reduce caries activity of mutans streptococci in dental plaque for a period of one month. Prior to tooth brushing with enzyme dentifrice, the acidity in plaque is very high, the change of dental plaque acidity varies from 0 % through 80 %. Statistically the change of acidity before and after tooth brushing with enzyme dentifrice is significant. The enzymes effect bacteria's metabolic system. B. THE EFFECT OF LACTOPEROXIDASE - THIOCYANATE - HYDROGEN PEROXIDE ON THE METABOLISM OF CARIOGENIC MIDROORGANISM IN VITRO AND IN THE ORAL As stated in literature that LPO system ( Lactoperoxidase - thiocyanate Hydrogen Peroxide System) can inhibit the metabolism of cariogenic microorganism in the oral cavity.lactoperoxidase enzyme consists of six iso-enzymes which catalyze peroxidase reaction. Lactoperoxidase can be inactive easily and according to Pickering et al 1962, most of Streptococci can make the enzyme inactive by using Hydrogen peroxidase or H202 from their metabolism. The diagram is as follows: Lactoperoxidase Spontaneous H 2 O 2 Compound II H 2 O 2 Compound I 1 e (donor from SCN) catalase Compound III Compound IV Figure 1. Inactivation of lactoperoxidase enzyme. (Hendrik Doom 1974. The effect of Lactoperoxidase - Thiocyanate Hydrogen peroxide on the metabolism of cariogenic microorganism in vitro and in the oral caviy) 3

Lactoperoxidase reacts with one molecule of H 2 O 2, to yield Compound(C) I. An electron from SCN- reduces CI to be CII. SCN ion can act as electron donor. It protects lactoperoxidase against this form of oxidation. CII is compound to be maintained because it can stimulate lactoperoxidase synthesis. CII reacts with H 2 O 2, to yield CIII, which spontaneously turns to inactive CIV. CII also can spontaneously turn to CI again. In order that CII can be maintained, two mechanisms are needed: a. CIII shall be turned to CII by catalase. b. CI needs an electron from SCN - to be come CII. Therefore SCN-is very important for lactoperoxidase system. Lactoperoxidase is an important oxidative enzyme of saliva. Lactoperoxidase acts as bacteriostatic when combined with H20 and SCN-. The combination creates lactoperoxidase system (LPO system). Thiocyanate (SCN-) is sufficient in human saliva to active LPO system.hydrogen - peroxide (H 2 0 2 ) is produced by oral microorganism, but its level in saliva is insufficient to active the system. The optimal level of H 2 0 2 in saliva to active the system should be 30 to 300!J.M. That is the reason why additional external H 2 0 2 is needed, for example from dentifrice. So, LPO system is applied in dentifrice by adding two specific enzymes to increase H202 in saliva. The enzymes are Amiloglucosidase (AG) and Glucoseoxidase (GO). B. THE EFFECT OF DENTIFRICE CONTAINING ENZYME ON THE GROWTH OF SALIVARY MUTANS STREPTOCOCCI IN ORTHODONTIC PATIENTS (Soeherwin Mangundjaja, Abdul Muthalib and Ariadna Djais, 2001) reported that the result of research indicated that Colony Forming Units of salivary mutans streptococci after tooth brushing with dentifrice containing enzyme are lower than CFU of Streptococcus mutans bacteria after tooth brushing without it. Previous research found out one case of Pseudomonas aeruginosa bacteria in patient with fixed orthodontic appliance ( Soeherwin Mangundjaja, Abdul Muthalib, Ariadna Djais and Elza Auerkari, 2000). Those bacteria are opportunistic bacteria which will be pathogen if the condition is favorable for them. It is hope that dentifrice containing enzyme may prevent caries and growth of pathogenic bacteria in fixed orthodontic appliance user patients. D.EFFECT OF ENZYME DENTIFRICE ON MICROBIAL CONTAMINATION OF TOOTHBRUSHES { Soeherwin Mangundjaja, Heriandi S, Titi P, Sarworini and Agung A, 2002) reported that the results of initial research indicated that bacterial load of contaminant bacteria on tooth brush after tooth brushing for 3 weeks using dentifrice containing enzyme is still very high, or about 105 through 106 CFU. As mentioned above that tooth brushing with dentifrice containing enzyme still shows high bacterial load on tooth brush I however it shows decreasing CFU population of Mutans streptococci after tooth brushing for three weeks.. It can be conclude that the presence of contaminant bacteria on tooth brush is due to in proper care and keeping of tooth brush. 4

E.THE EFFECT OF DENTIFRICE CONTAINING ENZYME ON HEALING PROCESS OF RECURRENT APHTOUS STOMATITIS Recurrent stomatitis is often called cancerous wound and is the most common lesion in oral cavity, its prevalence is around 20 % and it is commonly found in middle class and upper society (Sonis et ai, 1984). Base on the latest 40 years of study, etiological factor of Recurrent Aphthous Stomatitis is not due to herpes simplex virus (HSV) as it was assumed before. The latest concept of Recurrent Aphthous Stomatitis is clinical syndrome with various possibilities of etiological factors, the identified main etiological factors among other are hereditary factor, allergy, hematology deficiency, and immunology disorder ( Sonis et ai, 1984, Lynch, 1994), while ( Miller et ai, 1991) found that 1303 children from 530 families showed increasing sensitivity to Recurrent Aphthous Stomatitis of their parents were Recurrent Aphthous Stomatitis positive. Result of study of Oral Medicine Department, Faculty of Dentistry, University of Indonesia 1994 in Cipto Mangunkusumo Hospital shows that Recurrent Aphthous Stomatitis makes 26.6 % of the existing oral disease, whereas its etiology has not been known exactly. Donatsky & Bendixen I 1972) stated that Streptococcus sanguis has a play role for triggering of occurrence Recurrent Aphthous Stomatitis, while ( Altman, 1998). 'stated that treatment with stable isotope in water may act as immunological modulator., and (Sri Wendari.H and Soeherwin Mangundjaja, 1999).stated in initial research has proven that natural stability of isotopes in water has the ability to cure Recurrent Aphthous Stomatitis and the ability to stimulate in the immune response of the cell. From empirical study,data were collected based on patients suffering Recurrent Aphthous Stomatitis, enzyme dentifrice seems effective for treating Recurrent Aphthous Stomatitis, however still required scientific evidence. According to dentifrice containing enzyme is used in treatment of Recurrent Aphthous Stomatitis for certain patients, it assumed that it can relieve pain, accelerate healing process of Recurrent Aphthous Stomatitis lesion, and reduce population of Streptococcus sanguis in oral cavity. Therefore temporarily it can be assumed that the enzyme can inhibit the growth of one of bacterial etiological factors of Recurrent Aphthous Stomatitis. And the activity of the enzyme is assumed to be related to immunology response. F.THE EFFECT OF DENTIFRICE CONTAINING ENZYME ON HALITOSIS Halitosis (bad breath, fetor ex ore, fetor oris) is not a disease, instead it is symptom of a disease ( Burket W Lister in RR Darwita et al, 2001). Halitosis is also symptom of various diseases, among others various dental and oral diseases, systemic disease, upper respiratory tract disease ( i,e, tonsillitis, sinusitis, tumor, neurological disorder), digestive tract disorder (Le gastritis, duodenal and gastric ulcer) use of narcotics, analgesics, anti - hypertensive, anti-depressant, and antibiotics and also due to stress. (Burket W Lister, 1961, Rosenberg M, Me Cullogh CAG, 1992, Rosenberg et ai, 1991, Mc Namara Thomas RF, PhD, et ai, 1992, Neiders Mirdza DDS et ai, 1999 in Darwita et al 2001). 5

Halitosis can be measured with Chromatographic measurement such as Halimeter Model RH -17K with Chart Recorder type 101, and with analytic unit measurement: concentration of sulphide in Volatile Sulphur Compounds (VSC). RR Darwita, Anton H and Alexander Agung, 2001) stated that the result of VSC concentration measurement on 48 subjects of research was follows: @ Prior to tooth brushing with dentifrice containing enzyme: Average of VSC was 112.7446 ppb ( Sd = 159. 4321). after tooth brushing with dentifrice containing enzyme: Average of VSC 65.2176 ppb( d = 69.9934).; So there is a decrease of 42.15 % ( P = 0.030) in VSC concentration. @. While after tooth brushing with non-enzyme dentifrice the decrease of VSC concentration is only 40.61 % ( P = 0.063) DISCUSSION From various researchers, it can be concluded that dentifrice containing enzyme has significant on population of microorganism in oral cavity, Recurrent Aphthous Stomatitis and Halittosis. Saliva as host in oral cavity, has various 'organic and antibacterial components, so saliva and oral microorganism can be classified as main factor in maintaining oral health.. Microorganism which develop and grow in oral cavity can become the cause of diseases if body's defense mechanism declines, it is call opportunistic infection. Dental caries and periodontal disease are disease, due to activity of oral cavity bacteria, which can not be overcome by body's defense mechanism Bacteria which have main role in dental caries are Streptococcus mutans. Dental caries due to activity of those bacteria is found in Kelapa and Panggang Islands population ( Soeherwin Mangundjaja, 1993, Soeherwin Mangundjaja, 1995, Soeherwin Mangundjaja et ai, 1995), while (Maya AY, 1998) reported the activity of mutans streptococci in periodontal disease. The most important aspect of oral and dental health is maintaining equilibrium of eco system in oral cavity, and it will involve antibacterial system in saliva as well from outside the body. Besides antibacterial factor such as Lysozime, saliva also contains antibacterial system, including Lactoperoxidase, Thiocyanate and Hydrogen peroxide system which influences the growth of bacteria in oral cavity. Host and oral microorganism are inter related in dynamic inter action, so to maintain oral cavity's eco system equilibrium, supporting factors ate needed. One of those factors is enzyme for maintaining oral cavity's health. From reference study I it is found out that dentifrice containing Amiloglucosidase and Glucoseoxidase enzymes can protect enamel by preventing dental plaque formation, inhibiting enzymes produced by Streptococcus mutans in dental plaque. So the bacteria can not make colony on the surface of teeth. In general, tooth brushing with dentifrice containing enzyme can influence population of microorganism in oral cavity, so it can reduce the probability of various oral diseases. 6

CONCLUSION Use of dentifrice containing enzyme can maintain equilibrium of oral bacteria's eco system, so it can reduce the probability of various oral diseases. REFERENCES Burket W Lester. Oral Medicine and Treatment 4th edtion. JB Lippincott Company Philadelphia, 1961: 182:.185 (In RR. Darwita et al 2001) Donatsky 0, Bendixen G (1972). In vitro demonstration of cellular hypersensitivity to Strep A2 in recurrent aphthous stomatitis by means of the leukocyte Migration Test.Acta Allergol1972; 27: 137 ( In Burket's Oral Medicine Diagnosis and Treatment. Lynch et ai, 1994) Effendi.l, (1986). Year Report of Government of Health. Ministry of Health Republic of Indonesia. 1986 Hendrik Doorn (1974). The effect of Lactoperoxidase - Thiocyanate - hydrogen peroxide on the metabolism of cariogenic microorganism in vitro and in the oral caviy, Doctor Dissertation Tenchnishe ogeschool Delft. 1974 Lynch, M. A (1994). Burket's oral medicine diagnosis and treatment. 9* ed. JB. Lippinctt Company, Philadelphia. Lehninger AL,(1975) Biochemistry, 2nd ed, Worth Pubt, New York, 1975 Mayes PA(1983) Metabolism of carbohydrate, Harfer review of Biochemistry 19* ed. Singapore. Lange Manizen Asian. 1983; 162-7. Maya AY Lewerissa (1998). Comparison salivary mutans streptococci population between gingivitis and periodontal disease's patients. Specialist Education Program. Faculty of Dentistry University of Indonesia 1998. Miller. T. E et al. (1991).' Effect of an anti microbial mouth rinse on recurrent Aphthous ulcerations. Oral Surgery Oral Mad. Oral Pathol. 72:559-561 Mc Namara ThomasF,PhD et al. The Role of Microorganism in the production of Oral Malador. Oral surgery, Oral Medicine Oral Pathology 34; 4148, 1972. ( In RR. Darwita et al., 2001) Newburn E.(1989) CariologYJ 3rd ed.quintessence Publ, Chicago,1989:20-21 76-84 Neides Mirdza DDS, MS, Ramos Brigette, DDS (1999) Operation of Bad breath Clinics. Quistessence International 5; 295-301, 1999. ( In RR. Darwita et ai, 2001 ) Rosenber M, McCullouch CAG(1992). Measurement of Oral malador current methods of future prospect. J Periodonal63, 776-782, 1992 (In RR. Darwita et ai, 2001) Roesenberg M et al.(1991) Halitosis measurement by an industrial sulphidemonitor. J. Periodontial 62;487-489,1991 (In RR. Darwita et ai, 2001 ) RR. Darwita Anton Rahardjo, L.A Agung (2001). Use of dentifrice containing enzyme in people of Tebet Jakarta Indonesia. Dental Public Health of the Faculty Dentistry University of Indonesia and Research & Development Department of PT Indonesia dental Industries. Jakarta Indonesia. Suwelo, 1.S.(1988). Karies gigi sulung dan urutan besar peranan faktor risiko terjadinya karies. Kajian pad a anak pra sekolah di Jakarta dan.sekitarnya. Dissertation Thesis. University of Gajah Mada. 1988 Indonesia 7

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