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
Koo, Hyun; Yamada, Kenneth M (2016) Dynamic cell-matrix interactions modulate microbial biofilm and tissue 3D microenvironments. Curr Opin Cell Biol 42:102-112
Wang, Yifei; Singh, Ajay P; Hurst, William J et al. (2016) Influence of Degree-of-Polymerization and Linkage on the Quantification of Proanthocyanidins using 4-Dimethylaminocinnamaldehyde (DMAC) Assay. J Agric Food Chem 64:2190-9
Wang, Yifei; Han, Alex; Chen, Eva et al. (2015) The cranberry flavonoids PAC DP-9 and quercetin aglycone induce cytotoxicity and cell cycle arrest and increase cisplatin sensitivity in ovarian cancer cells. Int J Oncol 46:1924-34
Klein, Marlise I; Hwang, Geelsu; Santos, Paulo H S et al. (2015) Streptococcus mutans-derived extracellular matrix in cariogenic oral biofilms. Front Cell Infect Microbiol 5:10
Fears, Kenan P; Gonzalez-Begne, Mireya; Love, Corey T et al. (2015) Surface-induced changes in the conformation and glucan production of glucosyltransferase adsorbed on saliva-coated hydroxyapatite. Langmuir 31:4654-62
Kim, Dongyeop; Hwang, Geelsu; Liu, Yuan et al. (2015) Cranberry Flavonoids Modulate Cariogenic Properties of Mixed-Species Biofilm through Exopolysaccharides-Matrix Disruption. PLoS One 10:e0145844
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, José A; Quivey Jr, Robert G; Koo, Hyun et al. (2013) Streptococcus mutans: a new Gram-positive paradigm? Microbiology 159:436-45
Koo, H; Falsetta, M L; Klein, M I (2013) The exopolysaccharide matrix: a virulence determinant of cariogenic biofilm. J Dent Res 92:1065-73
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

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