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DENT 5302 TOPICS IN DENTAL BIOCHEMISTRY 4 April 2007 Objectives: • Role of fluoride on dental caries • Role of fluoride on the prevalence of dental fluorosis • The effect of fluoride on plaque bacteria Outline Overview of fluoride Fluoride, dental caries, and fluorosis: Historical perspective How does fluoride work? (anticaries mechanisms) Effect of fluoride on plaque bacteria Structural-bound fluoride Calcium fluoride-like material Fluoride in saliva and dental plaque Fluoride Fluoride: ionic form of fluorine Fluorine: Halogen group The most electronegative element 13th most abundant in the crust of the earth Found in virtually all inanimate and living things F- + H+ HF Hydrofluoric acid (pKa ~ 3.4) Fluoride: Potent inhibitor of many enzymes Elimination by kidneys Avid calcified tissue seeker Inhibit and even reverse the formation of dental caries F: The cornerstone of modern preventive dentistry Remarkable decline in dental caries Systemic and topical fluoride Water fluoridation Top 10 great public health achievements of the 20th century (CDC) Fluoride is a hazardous substance when large doses are taken acutely when lower doses are taken chronically Dental fluorosis Reversible gastric disturbances Skeletal fluorosis Death Fluoride, Dental Caries, and Fluorosis Mottled enamel Endemic in several regions of the southwestern USA Colorado Springs McKay FS. The relation of mottled enamel to caries. J Am Dent Assoc 15:1429-1437, 1928. “…these mottled enamel cases…are singularly free from caries.” Mottled enamel = Dental fluorosis F ? substance in drinking water ? Regular consumption of drinking water with fluoride Reduction of dental caries Dean et al: various levels of fluoride in most water supplies Water fluoride level ~ 1 ppm Caries prevention Low prevalence of dental fluorosis Dean HT et al., Domestic water and dental caries: V. Additional studies of the relation of fluoride domestic waters to dental caries experience in 4425 white children, aged 12-14 years, of 13 cities in 4 states. Public Health Rep 57:1155-1179, 1942. Dental Fluorosis Hypomineralization of enamel from excessive fluoride ingestion during tooth development Mild fluorosis (common) Minor cosmetic defect Severe fluorosis (rare) Increased caries risk: Pitting & Loss outer enamel Overintake of fluoride from 0-6 year old Generally less in primary teeth (develop prenatally) Most critical between 15-30 months of age: maxillary central incisors Why are we concerned about dental fluorosis? The prevalence of fluorosis has increased Optimal water F level (~ 1 ppm) 20 % prevalence of very mild or mild fluorosis Dentistry: Mild fluorosis is an acceptable tradeoff for caries prevention Esthetic: ‘mild’ cosmetic defect ? No fluorosis: 27 % dissatisfied with their tooth color Mild fluorosis: 50% dissatisfied Water Fluoridation 1945 Grand Rapids Michigan 1948: Grand Rapids had 60% less DMFT than Muskegon ‘control’ city Optimal level: 0.7 - 1.2 ppm F Colder climates drink less water need higher fluoride level EPA (Environmental Protection Agency) Max. Contaminant Level (Primary Drinking Water Standards) = 4 ppm F Naturally existed F in some municipal water & wells : 4-8 ppm F or higher USA: 67% (170 millions) on public water system receive fluoridated water Centers for Disease Control and Prevention, 2002 The most cost-effective community-based approach for caries prevention Direct annual cost of fluoridation: $ 0.68-3.0 per person per year $1 invested in water fluoridation saves > $38 in treatment costs Halo effect Persons not residing in fluoridated communities Foods and beverages produced with fluoridated water Discussion: (group of 6-8) 1. The optimal level of fluoride in water as determined by this graph is 1 ppm. What are the rationales? 2. Why in Minnesota the level of added fluoride in water varies from 0.7 to 1.2 ppm? How does fluoride work? Historical perspective Cariostatic effect of fluoride Systemic incorporation into enamel during development ‘More perfect’ enamel crystals Less acid soluble The more fluoride incorporated, the better cariostatic effect Shark enamel (100% fluoroapatite) can develop caries lesion! Øgaard B et al. Scand J Dent Res 96:209, 1988. Microradiographic study of demineralization of shark enamel in a human caries model. Current philosophy The caries-reducing effect of fluoride is primarily achieved by its presence during active caries development at the plaque/enamel interface where it directly alters the dynamics of mineral dissolution and reprecipitation, and to some extent, affect plaque bacteria. Cariostatic Mechanism of Fluoride Change tooth morphology Controversial and not universally accepted Effect on plaque bacteria Debatable: need much higher concentration of fluoride to be effective Inhibit demineralization and enhance remineralization process Goal: Try to maximize benefit with minimal adverse effects MORE IS NOT NECESSARILY BETTER! Effect of fluoride on plaque bacteria 1940: Fluoride inhibited carbohydrate metabolism in pure cultures of streptococci and lactobacilli. Bibby BG, van Kesteren M. The effect of fluoride on mouth bacterial. J Dent Res 1940:19;391-402. Fluoride affects oral bacteria and dental plaque ecology IMPLY a reduced risk of caries Effect on dental plaque bacteria F inhibits bacterial adsorption In vitro: 9500 ppm F in solution inhibit bacterial adsorption to hydroxyapatite Clinical: rinses & toothpaste with Sn or amine F reduce plaque deposit F reduces proportion of cariogenic bacteria in dental plaque Chemostat: 19 ppm F prevent MS from growing to a larger proportion Clinical: only high concentration of fluoride works! No difference in subjects from area upto 21 ppm F in water Reduced MS in plaque after daily use of APF gel (12,300 ppm F) F decreases acid production Fluoridated water or daily rinse with 0.2% (~900 ppm) NaF solution Reduce 0.1 - 0.2 unit in pH drop after a sucrose challenge No effect after 0.05 % (~200 ppm) rinse Antimicrobial effect of F HF forms when external pH is lower than pKa (3.4) H+ + F- HF Fluoride enters cell as HF (not F-) HF H+ + FAccumulation of fluoride Accumulation of H+ 1 2 Bound to enzymes Cytoplasmic acidification - Enolase - Proton-extruding ATPase ‘Fluoride has inhibitory effects on plaque metabolism’ To what extent do these effects contribute to caries prevention? How much fluoride is needed for antimicrobial effect? In vitro: 9500 ppm F in solution inhibit bacterial adsorption to hydroxyapatite Reduced MS in plaque after daily use of APF gel (12,300 ppm F) Fluoridated water or daily rinse with 0.2% (~900 ppm) NaF solution reduced 0.1 - 0.2 unit in pH drop after a sucrose challenge No effect in pH drop after 0.05 % (~200 ppm) NaF rinse No reduction of MS in plaque in subjects from area upto 21 ppm F in drinking water How much fluoride is needed to reduce enamel solubility? ppm F to reduce solubility << ppm F for antimicrobial effect At pH 4-5 Fluoride in solution reduces the amount of enamel dissolved Effective at a few ppm F The most important cariostatic mechanism of F: De- and Remineralization Fluoride in HAP crystal: Structurally-bound F F- substitute OHDecrease crystal dimension (F- is smaller) Strong attraction with calcium (F: the most electronegative) F OH Ca Hydroxyapatite lattice structure • F- fill vacancy • H-bond with O Stabilize the lattice structure Improve the crystallinity Lower dissolution rate ‘Fluorhydroxyapatite’ ‘Fluoridated hydroxyapatite’ ‘Fluoroapatite-like material’ H-bond F How much fluoride is in dental plaque? Range from 5-50 ppm wet weight 1% is available as fluoride ion 15-75% is ionizable Some firmly-bound fluoride (bacterial uptake?) Plaque matrix concentrate fluoride from saliva: +ve charges in matrix & on bacterial surface attract Ca2+ Ca2+ bind fluoride. Plaque fluid F and Saliva F after 1-min rinse with NaF ( 900 ppm) or MFP ( 1000 ppm) F decreases exponentially Elevated for ~ 3 hours Plaque fluid F Clinical study: F level in saliva and plaque remained for 18 h Saliva F F reservoir Calcium fluoride Oral mucosa? Ekstrand J. Enhancing effects of fluoride. Cariology for the nineties, 409-420. Discussion: (group of 6-8) What characters affect caries development? Low Caries-Active High Caries-Active 4.8 + 5 * 18.9 + 7.3 0* 3.8 + 3.1 Age Male / female Salivary flow (ml/min) 26 + 8 9 / 14 1.5 + 0.3 24 + 8 13 / 11 1.5 + 0.5 MS (log CFU/ml saliva) Lactobacilli (log CFU/ml saliva) Brushing time (min) Amount of toothpaste (g) 4.2 + 1 3.8 + 0.8 2.8 + 1.6 1.1 + 0.5 5.3 + 1 4.5 + 1.2 2.6 + 2 1.2 + 0.6 Rinse frequency 1.5 + 0.7 * 3.6 + 1.9 Amount of water to rinse (ml) 70 + 60 * 190 + 10 F in saliva (immediate) (mM) F in saliva (accumulate) (mM·min) 0.6 + 0.4 * 6.9 + 3.9 * 0.3 + 0.3 3.9 + 2.9 Characteristics DMFT Decay surface ten Cate JM, van Loveren C. Fluroide Mechanisms. Dent Clin N Am 1999;43:713-742. Adapted from Sjögren T, Birkhed D. Caries Res 1993;27:474. Discussion: (group of 6-8) I believe that the main anticaries effect of fluoride is by changing the equilibrium towards remineralization, not antimicrobial effect. Why? Recommended references 1. Ten Cate JM, van Loveren C. Fluoride Mechanisms. Dent Clin North Am 1999;43(4):713-742. 2. Featherstone JD. The science and practice of caries prevention. J Am Dent Assoc 2000;131:887-899. 3. Gordon Nikiforuk. Understanding Dental Caries 1. Etiology and Mechanisms, Basic and Clinical Aspects. Basel; New York: Karger 1985. Chapters 4. 4. Gordon Nikiforuk. Understanding Dental Caries 2. Prevention, Basic and Clinical Aspects. Basel; New York: Karger 1985. Chapters 3. 5. van Loveren C. Antimicrobial activity of fluoride and its in vivo importance: Identification of research questions. Caries Res 2001;35(suppl 1):65-70. 6. Fejerskov O. Changing paradigms in concepts on dental caries: Consequences for oral health care. Caries Res 2004;38:182-191. 7. Ten Cate JM. Review on fluoride, with special emphasis on calcium fluoride mechanisms in caries prevention. Eur J oral Sci 1997;105:461-465.