We propose a novel chemotherapeutic approach to reduce the incidence of dental caries, the most prevalent and costly oral infectious disease. Fluoride is the mainstay for the prevention of dental caries;however, as currently used, (1) fluoride offers incomplete protection against caries, and (2) does not address effectively the infectious aspect of the disease. Elevated amounts of bacterial-derived extracellular polysaccharides-EPS (Streptococcus mutans, a key contributor) and persistent low pH environment in the matrix of the biofilms are prime factors associated with the pathogenesis of dental caries and offer major targets for novel therapeutic agents. We propose a new concept of anti-caries therapy using specific combinations of cranberry flavonoids (1) to disrupt the formation of EPS and acidification of the biofilms matrix, (2) which will enhance the caries preventive effect of fluoride. Data from our USPHS/NIH supported studies have shown that aqueous extract of cranberry effectively reduced the formation of EPS and acidogenicity of S. mutans within biofilms in vitro, and the development of dental caries in vivo without displaying biocidal activity. Cranberries harbor a complex mixture of active, non-active and potentially detrimental compounds, such as organic acids. Further bioassay- guided purification studies have identified five major bioactive and structurally distinctive flavonoids in cranberries: a flavonol (quercetin-3-arabinofuranoside) and four A-type proanthocyanidins (procyanidin A2, tetramer, hexamer and nonamer). Elucidation of the particular mechanisms of action by which these compounds disrupt the ability of S. mutans to utilize sucrose to form EPS and acids is critical to further enhance their therapeutic effects;such information will guide us in selecting precisely the agents to be used in combination with fluoride for improved cariostatic activity. Thus, we propose a step-wise research plan as follows: (1) characterization of the biological actions of individual compounds and fluoride by studies on: (i) expression of S. mutans genes required for the EPS-matrix synthesis, (ii) catabolic pathway of S. mutans by specific assays on sugar uptake system and glycolytic enzymes, and (iii) retention time within biofilm;(2) identification of specific combinations of cranberry flavonoids based on their mode of action and pharmacological targets, potency and retention within biofilms;and whether the addition of fluoride can enhance the overall effects on EPS and acidogenicity, (3) testing the effects of combinations of agents on formation, biochemical, inorganic and microbial composition of multi-species biofilms on saliva-coated hydroxyapatite surface, and further selection of most effective therapies, and (4) evaluation of the effectiveness of selected combination therapies in disrupting establishment of S. mutans infection, and caries development using our rodent model. A comprehensive program ranging from basic laboratory studies to in vivo investigations in animals is offered that could result in providing new therapeutic agents working in concert with fluoride while simultaneously enhancing our understanding of the pathogenesis of dental caries.

Public Health Relevance

This proposal will elucidate the particular mechanisms by which cranberry flavonoids disrupt the pathogenesis of dental caries, and use the information to develop a novel chemotherapeutic approach to reduce the incidence of caries, the most prevalent and costly oral infectious diseases in the United States and worldwide.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE016139-06
Application #
8282940
Study Section
Oral, Dental and Craniofacial Sciences Study Section (ODCS)
Program Officer
Lunsford, Dwayne
Project Start
2004-01-01
Project End
2015-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
6
Fiscal Year
2012
Total Cost
$334,843
Indirect Cost
$68,273
Name
University of Rochester
Department
Dentistry
Type
Schools of Dentistry
DUNS #
041294109
City
Rochester
State
NY
Country
United States
Zip Code
14627
Hwang, Geelsu; Klein, Marlise I; Koo, Hyun (2014) Analysis of the mechanical stability and surface detachment of mature Streptococcus mutans biofilms by applying a range of external shear forces. Biofouling 30:1079-91
Lemos, Jose A; Quivey Jr, Robert G; Koo, Hyun et al. (2013) Streptococcus mutans: a new Gram-positive paradigm? Microbiology 159:436-45
Bueno-Silva, B; Koo, H; Falsetta, M L et al. (2013) Effect of neovestitol-vestitol containing Brazilian red propolis on accumulation of biofilm in vitro and development of dental caries in vivo. Biofouling 29:1233-42
Feng, Guoping; Klein, Marlise I; Gregoire, Stacy et al. (2013) The specific degree-of-polymerization of A-type proanthocyanidin oligomers impacts Streptococcus mutans glucan-mediated adhesion and transcriptome responses within biofilms. Biofouling 29:629-40
Koo, H; Falsetta, M L; Klein, M I (2013) The exopolysaccharide matrix: a virulence determinant of cariogenic biofilm. J Dent Res 92:1065-73
Xiao, Jin; Klein, Marlise I; Falsetta, Megan L et al. (2012) The exopolysaccharide matrix modulates the interaction between 3D architecture and virulence of a mixed-species oral biofilm. PLoS Pathog 8:e1002623
Bowen, W H; Koo, H (2011) Biology of Streptococcus mutans-derived glucosyltransferases: role in extracellular matrix formation of cariogenic biofilms. Caries Res 45:69-86
Jeon, J-G; Rosalen, P L; Falsetta, M L et al. (2011) Natural products in caries research: current (limited) knowledge, challenges and future perspective. Caries Res 45:243-63
Koo, H; Duarte, S; Murata, R M et al. (2010) Influence of cranberry proanthocyanidins on formation of biofilms by Streptococcus mutans on saliva-coated apatitic surface and on dental caries development in vivo. Caries Res 44:116-26
Koo, H; Jeon, J G (2009) Naturally occurring molecules as alternative therapeutic agents against cariogenic biofilms. Adv Dent Res 21:63-8

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