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3 134 Group No. 1 (Control) Group No. 2 (M: 1.0unit) Group No. 3 (M: 5.0units) Group (D: No unit) Group No. 5 (D: 5.0units) (M+D: Group No unit+1.0unit) (M+D: Group No unit +0.2unit) (M+D: Group No unit+1.0unit) Fig. 2Photographs experiment of 1. the lower right molars ( ~8.4) 26 of representative rats from each group in
4 135 Table 1Effect of mutanase, dextranase, and their combination on the inhibition of dental caries in rats.
5 136 b a C d Fig. 28 3
6 137 b a d c Fig. 29 4
7 138 b a c Fig d
8 Fig. 3Scanning electron micrographs of the lower left molars of representative rats from each group. E: Enamel, D: Dentin a: group 1 (control group), lower left third molar. b: group 2 (M: 1.0unit), lower left second molar. C: group 4 (D: 1. Ounit), lower left second molar. d: group 6 (M+D: 1.0unit+1.0unit), lower left second molar. a-d: ~30 Fig. 4 Scanning electron micrographs of the occlusal surfaces of enamel (portion of E in Fig. 3). a: group 1 (control group) b: group 2 (M: 1.0unit) C: group 4 (D: 1.0unit) d: group 6 (M+D: 1. Ounit +1. Ounit) a-d: ~3000 Fig. 5 Scanning electron micrographs of the occlusalsurfaces of dentin (portion of D in Fig. 3). a: group 1 (control group) b: group 2 (M: 1.0unit) C: group 4 (D: 1.0unit) d: group 6 (M+D: 1. Ounit+1.Ounit) a-d: ~3000
9 140 Group No. 1 (37-day-old control) Group No. 2 (62-day-old control) Group No. 3 (M: 1.0unit) Group (D: No.4 1.0unit) (M+D: Group No unit+1.0unit) (M+D: Group No unit+0.2unit) Group (M+D: Fig. 7 No unit+1.0unit) Photographs experiment of 2. the lower right molars ( ~8.4) 32 of representative rats from each group in
10 Table 2 Effect of mutanase, dextranse, and their combination on the inhibition of progress of dental caries in rats.
11 2) Konig, K. G. and Guggenheim, B.: In vivo effect of dextranase on plaque and caries, HeIv. Odont. Acta, 12: 48, ) Guggenheim, B., Konig, K. G., Miihleman, H. R. and Regolati, B.: Effect of dextranase on caries in rats harbouring an indigenous cariogenic bacterial flora, Archs. Oral. Biol., 14: 555, ) Guggenheim, B., Regolati, B. and Muhleman, H. R.: Caries and plaque inhibition by mutanase in rats, Caries Res., 6: 289, ) Guggenheim, B., Regolati, B., Schmid, R. and Miihleman, H. R.: Effect of the 'topical application of mutanase on rat caries, Caries
12 17) Schachtele, C. F., Statt, R. H. and Harlander, S. K.: Dextranase from oral barteria: Inhibition of water-insoluble glucan production and adherence to smooth surfaces by streptococcus mutans, Infect. Immun., 12: 309, ) Hamada, S., Mizuno, J., Murayama, Y., 0oshima, T., Masuda, N. and Sobue, S.: Effect of dextranase on the extracellular polysaccha.- ride synthesis of streptococcus mutans: che- and scanning electron microscopy studi- mical es. Infect. Immun.. 12: ) Keyes, P. H.: A method of recording and scoring gross carious lesions in the molar teeth of syrian hamster, J. Dent. Res., 23: 439, ) Fitzgerald, R. J., Keyes, P. H., Staudt, T. H. and Spinell, D. M.: The effects of a dextranase preparation on plaque and caries in hamster, a preliminary report, J. Amer. Dent. Assoc., 76: 301, ) Bloch, P. I., Pooley, C. L. and Howe, E. E.: The retardation of spontaneous periodental disease and the prevention of caries in hamsters with dextranase, J. Periodont., 41: 105, ) Hamada, S., Ooshima, T., Masuda, N. and Sobue, S.: Effect of dextranase prepared from spicaria violacea on dental caries in the hamster, J. Dent. Res., 55: 552, ) Guggenheim, B.: Enzymatic hydrolysis and structure of water-insoluble glucan produced by glucosyltransferases from a strain of Streptococcus mutans, Hely. Odont. Acta, 14: 89, ) Waker, G. J.: Some properties of a dextran glucosidase isolated from oral streptococci and its use in studies on dextran synthesis, J. Dent. Res., 51: 909, ) Futakami, K., shibuya, K., Yamazaki, Y., Igarashi, M. and Niwa,M.: The effect of dextranase on the release of reducing sugar from dental plaque, J. Dent. Res., 26: 193, 1976.
13 Abstract: Inhibitory effects of mutanase and dextranase on caries formation. II. Effect on experimental dental caries in rats. Masami SUDO*, Kazuo SHIMADA*, Masataka AKIYAMA*, Takashi KUBO*, Tsunehisa AMANO** and Kozo INOUE**. The preventive effect of mutanase (from Pseudomonas sp.) and dextranase (from Corynebacterium pseudodiphtheriticum) on the formation and progress of experimental dental caries in rats was examined. When the enzymes were given immediately after the infection with Streptococcus mutans OMZ 176, the statistical caries score indicated that both enzymes had a significant preventive effect on the formation of dental caries in rats. The effect was especially pronounced in a combination treatment (1 unit each enzyme/g diet or ml drinking water). In observations of the tooth surfaces under a scanning electron microscope, thick plaque adhered to the enamel and the dentin of the control group, but the plaque could not be observed and the fine structure of the enamel and the dentin of the treated group (with mutanase 1 unit+dextranase 1 unit) were normal. After experimental dental caries caused by infection with S. mutans OMZ 176 and then by Diet 2000, the rats were treated with a single or a combination dose of the two enzymes. The combination of the two enzymes (1 unit each enzymeig diet or ml drinking water) was the most effective for the inhibition of caries progression. From these results, the two enzymes, which have different substrate specificity, have a synergistically preventive effect on the formation and progress of dental caries in rats, suggesting the possibility of using dextranase in combination with mutanase for the prevention of human dental caries. Key words: Mutanase, Dextranase, Inhibition of caries formation, Streptococcus mutans
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