Inhibition of Streptococcus mutans Adsorption to Hydroxyapatite by Low-molecular-weight Chitosans

Abstract

The role of Streptococcus mutans in the initiation of dental caries has been recognized and attributed, at least in part, to its ability to colonize the tooth surface. Therefore, factors which prevent S. mutans attachment to hydroxyapatite (HA) are of considerable interest for the prophylaxis of this infectious disease. Chitosan, a chitin derivative by N-deacetylation, is an interesting candidate in this respect, since it stimulates the ordered regeneration of oral soft tissues, prevents the deleterious action of organic acid, and exhibits bactericidal action against several pathogens. In the present work, the efficacy of a low-molecular-weight chitosan (LMWC) and its derivatives N-carboxymethyl chitosan (NCMC) and imidazolyl chitosan (IMIC) in preventing S. mutans attachment to HA beads was assessed. The effects of chitosan on both sucrose-dependent and -independent adherence were evaluated. In both cases, when saliva-coated or uncoated HA beads were treated with any of the chitosans, a reduction in S. mutans adsorption ranging from 47 to 66% was observed. When HA beads were coated with saliva after the treatment with chitosan, neither carbohydrate caused a statistically significant reduction in S. mutans adsorption, suggesting that saliva deposition restores HA binding properties. Bacteria grown in the presence of chitosan sub-minimal inhibitory concentrations (sub-MICs) ranging from 12 to 500 ug mL^-1 adsorbed poorly to HA and exhibited a lower affinity toward xylene than untreated controls. In the presence of chitosan sub-MICs up to 60 ug mL^-1, an increase in the percentage of detached bacteria from two- to nine-fold was observed. The desorptive effect of chitosan was weaker when S. mutans had adhered to saliva-coated HA in the presence of sucrose. These results demonstrate that the presence of minor amounts of modified chitosans prevents S. mutans adsorption to HA and suggest that colonization of the tooth surface might be impaired by the use of toothpastes, mouthrinses, or chewing gums containing any of the tested polysaccharides.

Keywords

Streptococcus mutans adsorption chitosan hydroxyapatite.

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References

Bleiweis AS , Oyston PC , Brady IJ (1992). Molecular, immunological and functional characterization of the major surface adhesin of Streptococcus mutans. In: Genetically engineered vaccines. Ciardi JE , et al., editors. New York: Plenum Press, pp. 229-241.

Bowen WH , Schilling K. , Giertsen E. , Pearson S. , Lee SF , Bleiweis A. , et al. (1991). Role of a cell surface-associated protein in adherence and dental caries. Infect Immun 59:4606-4609.

De Leo C. , Coppola RC , Blasi G. , Eftimiadi C. , Salvarani M. , Molina AM (1990). Prevalence of Streptococcus mutans and dental decay in schoolchildren living in Genoa (Italy). Eur J Epidemiol 6:166-174.

Demuth DR , Lammey MS , Huck M. , Lally ET , Malamud D. (1990). Comparison of Streptococcus mutans and Streptococcus sanguis receptors for human salivary agglutinins. Microb Pathog 9:199-211.

Gibbons RJ (1984). Adherent interactions which may affect microbial ecology in the mouth. J Dent Res 63:378-385.

Gibbons RJ , Etherden I. (1985). Albumin as a blocking agent in studies of streptococcal adsorption to experimental salivary pellicles. Infect Immun 50:592-594.

Gibbons RJ , van Houte J. (1980). Bacterial adherence and the formation of dental plaque. In: Bacterial adherence. Beachey EH , editor. London: Chapman and Hall, pp. 61-104.

Gibbons RJ , Cohen I. , Hay DI (1986). Strains of Streptococcus mutans and Streptococcus sobrinus attach to different pellicle receptors. Infect Immun 52:555-561.

Hamada S. , Slade HD (1980). Biology, immunology and cariogenicity of Streptococcus mutans. Microbiol Rev 44:331-384.

10.

Hasty DL , Ofek I. , Courtney HS , Doyle RJ (1992). Multiple adhesins of streptococci. Infect Immun 60:2147-2152.

11.

Hoppenbrouwers Pmm , Borggreven JM , Van der Hoeven JS (1984). Adherence of oral bacteria to chemically modified hydroxyapatite. Caries Res 18:1-6.

12.

Jenkinson HF (1994). Adherence and accumulation of oral streptococci . Trends Microbiol 2:209-212.

13.

Koga T. , Asakawa H. , Okahashi N. , Haada S. (1986). Sucrose-dependent cell adherence and cariogenicity of serotype c Streptococcus mutans. J Gen Microbiol 132:2873-2883.

14.

Kolenbrander PE (1988). Intergeneric coaggregation among human oral bacteria and ecology of dental plaque. Ann Rev Microbiol 42:627-656.

15.

Kuramitsu HK (1993). Virulence factors of mutans streptococci: role of molecular genetics. Crit Rev Oral Biol Med 4:159-176.

16.

Lamberts BL , Pederson ED , Simonson LG (1985). The effects of basic and acidic synthetic polypeptides on the adherence of the oral bacteria Streptococcus mutans and S. sanguis to hydroxyapatite. Arch Oral Biol 30:295-298.

17.

Loesche WJ (1986). Role of Streptococcus mutans in human dental decay . Microbiol Rev 50:353-380.

18.

Murray PA , Prakobphol A. , Lee T. , Hoover CI , Fisher SJ (1992). Adherence of oral streptococci to salivary glycoproteins . Infect Immun 60:31-38.

19.

Muzzarelli Raa (1986). Dechelating ability of aminoacid glucans and sugar acid chitosans. In: Chitin in nature and technology. Muzzarelli RAA , Jeuniaux C , Gooday GW , editors. New York: Plenum Press, pp. 321-323.

20.

Muzzarelli Raa (1993). Biochemical significance of exogenous chitins and chitosans in animal and patients. Carbohydr Polym 20:7-16.

21.

Muzzarelli RA , Tanfani F. , Emanuelli M. , Mariotti S. (1982). N-carboxymethylidene chitosans and N-carboxymethyl chitosans. Carbohydr Res 107:199-219.

22.

Muzzarelli Raa , Biagini G. , Pugnaloni A. , Filippini 0 , Baldassare V. , Castaldini C. , et al. (1989). Reconstruction of periodontal tissue with chitosan Biomaterials 10:598-603.

23.

Muzzarelli R. , Tarsi R. , Filippini 0 , Giovanetti E. , Biagini G. , Varaldo PE (1990). Antimicrobial properties of N-carboxybutyl chitosan . Antimicrob Agents Chemother 34:2019-2023.

24.

Muzzarelli RA , Ilari P. , Petrarulo M. (1994a). Solubility and structure of N-carboxymethyl chitosan . Int J Biol Macromol 16:177-180.

25.

Muzzarelli RA , Mattioli-Belmonte M. , Tietz C. , Biagini R. , Ferioli G. , Brunelli MA , et al. (1994b). Stimulatory effect on bone formation exerted by a modified chitosan. Biomaterials 15:1075-1081.

26.

Muzzarelli Raa , Tomasetti M. , Ilari P. (1994c). Depolymerization of chitosans with the aid of papain. Enzyme Microbial Technol 16:110-114.

27.

Ohta H. , Kato H. , Okahashi N. , Takahashi I. , Hamada S. , Koga T. ( 1989). Characterization of a cell-surface protein antigen of hydrophilic Streptococcus mutans strain GS-5. J Gen Microbiol 135:981-988.

28.

Olsson J. , Emilson CG (1988). Implantation and cariogenicity in hamster of Streptococcus mutans with different hydrophobicity. Scand J Dent Res 96:85-90.

29.

Olsson J. , Carlen A. , Holmberg K. (1990). Modulation of bacterial binding to salivary pellicle by treatment with hydrophilizing compounds. Arch Oral Biol 35:137S-140S.

30.

Rosenberg M. , Gutnick D. , Rosenberg E. (1980). Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Letts 9:29-33.

31.

Rundegren J. , Olsson J. (1987). Interactions between salivary agglutinins and strains of Streptococcus mutans with varying degrees of surface hydrophobicity . FEMS Microbiol Letts 40:141-146.

32.

Russell MW , Mansson-Rahemtulla B. (1989). Interaction between surface protein antigens of Streptococcus mutans and human salivary components. Oral Microbiol Immunol 4:106-111.

33.

Sano H. , Matsukubo T. , Shibasaki K. , Itoi H. , Takaesu Y. (1991). Inhibition of adsorption of oral streptococci to saliva treated hydroxyapatite by chitin derivatives. Bull Tokyo Dent Coll 32:9-17.

34.

Shibasaki K. , Sano H. , Matsukubo T. , Takaesu Y. (1994). Effects of low molecular chitosan on pH changes in human dental plaque. Bull Tokyo Dent Coll 35:33-39.

35.

Shibl AM (1985). Effect of antibiotics on adherence of microorganisms to epithelial cell surfaces. Rev Infect Dis 7:51-65.

36.

Schilling KM , Bowen WH (1992). Glucan synthesized in situ in experimental salivary pellicle function as specific binding sites for Streptococcus mutans. Infect Immun 60:284-295.

37.

Staat RH , Langley SD , Doyle RJ (1980). Streptococcus mutans adherence: presumptive evidence for protein-mediated attachment followed by glucan-dependent cellular accumulation . Infect Immun 27:675-681.

38.

Svanberg M. , Westergren G. , Olsson J. (1984). Oral implantation in humans of Streptococcus mutans strains with different degrees of hydrophobicity. Infect Immun 43:817-821.

39.

Tellefson LM , Germaine GR (1986). Adherence of Streptococcus sanguis to hydroxyapatite coated with lysozyme and lysozyme supplemented saliva. Infect Immun 51:750-759.

40.

van Houte J. (1983). Bacterial adherence in the mouth. Rev Infect Dis 5 (Suppl 4):659-669.

41.

Visca P. , Berlutti F. , Vittorioso P. , Dalmastri C. , Thaller MC , Valenti P. (1989). Growth and adsorption of Streptococcus mutans 6715-13 to hydroxyapatite in the presence of lactoferrin. Med Microbiol Immunol 178:69-79.

42.

Zhanel GG , Nicolle LE (1992). Effect of subinhibitory antimicrobial concentrations (sub-MICs) on in vitro bacterial adherence to uroepithelial cells. J Antimicrob Agents Chemother 29:617-627.