Total fluoride intake and implications for dietary fluoride supplementation.

This paper reviews the history and validity of recommended "optimal" levels of systemic fluoride intake and the available information on levels of fluoride intake in young children from foods and beverages (including water), dentifrices, dietary fluoride supplements, mouthrinses, and gels. Most of the studies emphasize the substantial variation in ingestion among individuals. Often, a substantial proportion of individuals received fluoride well beyond the mean exposure reported in the study. Limitations in the existing data make it difficult to determine the total distribution of fluoride intake from all sources. Therefore, hypothetical combinations of possible daily fluoride intake from the three main sources (diet, dentifrices, and supplements) are presented for those aged 6, 12, 24, and 36 months, with associated mean intake per kg body weight. Findings suggest that some children exceed the "optimal" level of fluoride intake from single sources alone, while others can from a combination of sources. Moreover, if current recommended "optimal" levels, which have been derived on an empirical basis, are actually lower than what has been quoted in the literature, then more children could be ingesting excessive amounts of fluoride, which could increase their risk of developing objectionable dental fluorosis. The variation and complexity of fluoride ingestion from all sources should be considered in the evaluation of recommendations for use of dietary fluoride supplements.

Mechanism of inhibition of acid production in Streptococcus mutans by sodium ions under strictly anaerobic conditions.

Acids excreted and intracellular levels of glycolytic intermediates during glucose metabolism in streptococcus mutans NCTC 10449 under strictly anaerobic conditions were quantified in an attempt to understand the effect of sodium ions on bacterial acid production. In the presence of NaCl (0.15-0.30 M), the total amount of individual carboxylic acids excreted was inhibited by up to 31%. The intracellular level of fructose 1,6-bisphosphate increased by 58% and levels of 3-phosphoglycerate and pyruvate decreased by 46% and 12%, respectively. Sodium ions directly inhibited the activities of fructose 1,6-phosphate aldolase and triose phosphate isomerase. This indicated that the glycolytic enzymes responsible for the catalysis of fructose 1,6-bisphosphate to 3-phosphoglycerate were inhibited. However, in spite of the expected reduction in acid production intracellularly, the intracellular pH actually decreased in the presence of sodium ions. It is possible that the low intracellular pH inhibits the activity of the glycolytic enzymes involved in the breakdown of fructose 1,6-bisphosphate to 3-phosphoglycerate.

Effect of sodium and potassium ions on intracellular pH and proton excretion in glycolyzing cells of Streptococcus mutans NCTC 10449 under strictly anaerobic conditions.

The effect of sodium and potassium ions on intracellular acid production and acid excretion by glycolyzing cells of Streptococcus mutans was examined. S. mutans NCTC 10449 grown under glucose-limited and strictly anaerobic conditions in a continuous culture system was loaded with bis(carboxyethyl)-carboxyfluorescein, a pH-sensitive fluorescent dye, washed and suspended in 0.00-0.30 M NaCl/KCl solution. The dye allowed for the continuous monitoring of intracellular pH while proton excretion was measured simultaneously with a pH-stat. Sodium ions inhibited and potassium ions, at low pH, accelerated the amount of measurable acid excreted extracellularly. In the presence of both NaCl and KCl, proton excretion following the addition of glucose was slightly higher or similar to that observed in the presence of 0.15 M KCl alone. Sodium and potassium ions did not affect the proton-ATPase enzyme or the intracellular level of ATP, suggesting that these ions did not directly effect proton pumping activity itself. The inhibition of proton excretion by sodium ions was considered to have probably occurred as a result of an indirect inhibition of proton-ATPase activity by the low intracellular pH induced by sodium ions.

Inhibition of purified enolases from oral bacteria by fluoride.

Enolase activity in strains of oral streptococci previously has been found to be inhibited by 50% (Ki) by fluoride concentrations ranging from 50 to 300 microM or more in the presence of 0.5 to 1.0 mM inorganic phosphate ions. In this study, enolase was extracted and partly purified by a two-step process from five oral bacterial species and the effect of fluoride on the kinetics of enolase examined. The molecular weight of the putative enolase proteins was 46-48 kDa. The Vmax values ranged from 20 to 323 IU/mg and K(m) for glycerate-2-phosphate from 0.22 to 0.74 mM. Enolase activity was inhibited competitively by fluoride, with Ki values ranging from 16 to 54 microM in the presence of 5 mM inorganic phosphate ions. Ki values for phosphate ranged from 2 to 8 mM. The enolase from Streptococcus sanguis ATCC 10556 was more sensitive to fluoride (Ki = 16 +/- 2) than was enolase from Streptococcus salivarius ATCC 10575 (Ki = 19 +/- 2) or Streptococcus mutans NCTC 10449 (Ki = 40 +/- 4) and all three streptococcal strains were more sensitive to fluoride than either Actinomyces naeslundii WVU 627 (Ki = 46 +/- 6) or Lactobacillus rhamnosus ATCC 7469 (Ki = 54 +/- 6) enolases. The levels of fluoride found to inhibit the streptococcal enolases in this study are much lower than previously reported and are likely to be present in plaque, especially during acidogenesis, and could exert an anti-glycolytic effect.

Effect of low levels of fluoride on proton excretion and intracellular pH in glycolysing streptococcal cells under strictly anaerobic conditions.

The effect of low levels of fluoride on intracellular acid production and proton excretion in Streptococcus mutans NCTC 10449 at different growth and extracellular pH (pHo) levels was monitored under strictly anaerobic conditions. The sensitivity of S. mutans to fluoride increased as pHo decreased. Cells grown under acidic (pH 6.0 and 5.5) conditions were more resistant to fluoride than cells grown at a neutral pH. Under acidic extracellular conditions (pHo = 5.0), 0.025 mM fluoride inhibited proton excretion by approximately 50% in cells grown at pH 7.0. Slightly higher levels of fluoride (0.05-0.07 mM) were required for 50% or more inhibition in cells exposed to alkaline extracellular conditions or cells grown at acidic pH. Such levels of fluoride are about 10-20 times lower than that reported previously. Therefore, it is possible that as pH falls during initial bacterial glycolysis, sufficient amounts of anionic fluoride may be released, from its bound form in plaque, to cause significant inhibition of net proton movement out of the bacterial cell during further glycolysis, especially under the anaerobic environment of the deep layers of plaque. In addition to proton excretion, fluoride was also found to inhibit intracellular acid production.

Total fluoride intake in children aged 3 to 4 years--a longitudinal study.

Several previous studies using food consumption tables or diet records have estimated that children aged 1 to 12 years resident in fluoridated (1 ppm) areas receive, on average, between 0.05 and 0.07 mg fluoride/kg body weight from foods and drinks alone. In this study, the duplicate-diet approach, which is a more accurate method of determining nutrient intake, was used to determine if levels of fluoride intake from foods and drinks are similar to those estimated from food consumption tables or diet records. Duplicate portions of all foods and drinks consumed over 24 hours by 66 children aged 3 to 4 years resident in fluoridated and low-fluoride areas of New Zealand were collected on three separate days over a period of 12 months and analyzed for fluoride. Fluoride intake from the use and ingestion of toothpastes and fluoride supplements was also determined for each child. It was hypothesized that the total amount of fluoride received by children in low-fluoride areas from diet, toothpastes, and fluoride supplements was similar to that received by children in fluoridated areas from diet and toothpastes. The mean fluoride intake from foods and drinks alone in the low-fluoride areas was 0.008 +/- 0.003 mg/kg body weight (0.15 +/- 0.06 mg/day; n = 34) and in the fluoridated areas was 0.019 +/- 0.009 mg/kg body weight (0.36 +/- 0.17 mg/day; n = 32). The mean fluoride intake from foods and drinks and toothpastes in the low-fluoride areas was 0.027 +/- 0.012 mg/kg body weight (0.49 +/- 0.25 mg/day) and in the fluoridated areas was 0.036 +/- 0.015 mg/kg body weight (0.68 +/- 0.27 mg/day). Fluoride intake from diet alone did not exceed 0.04 mg/kg body weight (0.74 mg/day), and fluoride intake from diet and toothpaste did not exceed 0.07 mg/kg body weight (1.31 mg/day). The results suggest that levels of fluoride intake from foods and drinks alone as estimated by the duplicate-diet approach are much lower than previously estimated from food consumption tables or diet records. It was calculated that if all children in the low-fluoride areas were to take currently recommended dosages of fluoride tablets, which have been based on dietary surveys and diet records, then the total fluoride intake of some children in the low-fluoride areas would exceed that of their counterparts in the fluoridated areas. The results suggest that currently recommended dosages of fluoride tablets need to be further reduced if dental fluorosis in children is to be avoided.

Assessing fluoride concentrations of juices and juice-flavored drinks.

Few studies have investigated fluoride exposures from juices and juice-flavored drinks manufactured with water. In this study, the authors analyzed 532 juices and juice drinks for fluoride. Fluoride ion concentrations ranged from 0.02 to 2.80 parts per million, in part because of variations in fluoride concentrations of water used in production. Children's ingestion of fluoride from juices and juice-flavored drinks can be substantial and a factor in the development of fluorosis.

The effect of fluorhydroxyapatite-derived fluoride on acid production by streptococci.

The effect of fluoride derived from fluorhydroxyapatite (FHAp) minerals on bacterial glycolysis under aerobic and strictly anaerobic conditions was studied to validate the claims that this mineral could be used as a reservoir of fluoride in plaque. To isolate the direct effect of fluoride on bacterial glycolysis from that of an indirect pH-buffering effect of hydroxyl or phosphate ions which are also dissolved from the mineral, we equalized the pH-fall time course of reactions by manually adding KOH or HCl. This ensured that pH effects on glycolysis were minimized. Under controlled pH-fall and strictly anaerobic conditions, fluoride derived from the dissolution of FHAp containing more than 30,100 ppm fluoride (i.e., when the substitution of OH by F in the mineral was greater than 80%) had a direct inhibitory effect on lactic acid production in Streptococcus mutans. Under free pH-fall and strictly anaerobic conditions, increasing amounts of fluoride in FHAp (starting as low as 2000 ppm fluoride), appeared to have a pronounced indirect inhibitory effect on lactic acid production. This was probably mediated through a reducing pH buffer effect of the mineral. Even in the presence of high-fluoride FHAp, only 0.01 to 0.025 mmol/L fluoride was found in the reaction mixtures, a probable result of non-stoichiometric dissolution of FHAp. In spite of such low levels of fluoride, marked inhibitory effects on bacterial glycolysis were demonstrated. The results of this study suggest that high-fluoride FHAp may serve as a reservoir of fluoride for the inhibition of anaerobic acid production by S. mutans.

Infants' fluoride intake from drinking water alone, and from water added to formula, beverages, and food.

In infants, the majority of total ingested fluoride is obtained from water, formula and beverages prepared with water, baby foods, and dietary fluoride supplements. Few studies have investigated the distribution of fluoride intake from these sources among young children at risk for dental fluorosis. The purpose of this study was to assess estimated water fluoride intake from different sources of water among a birth cohort studied longitudinally from birth until age 9 months. Parental reports were collected at 6 weeks, 3 months, 6 months, and 9 months of age for water, formula, beverage, and other dietary intake during the preceding week. Fluoride levels of home and child-care tap and bottled water sources were determined. This report estimates daily quantities of fluoride ingested only from water--both by itself and used to reconstitute formula, beverages, and food. Daily fluoride intake from water by itself ranged to 0.43 mg, with mean intakes < 0.05 mg. Water fluoride intake from reconstitution of concentrated infant formula ranged to 1.57 mg, with mean intakes by age from 0.18 to 0.31 mg. Fluoride intake from water added to juices and other beverages ranged to 0.67 mg, with means < 0.05 mg. Estimated total daily water fluoride intake ranged to 1.73 mg fluoride, with means from 0.29 to 0.38 mg.

Stoichiometry of fluoride release from fluorhydroxyapatite during acid dissolution.

Release of F from fluorhydroxyapatite (FHAp) during acid dissolution was studied to validate the use of this mineral as a plaque reservoir of F. FHAp minerals having a wide range of F concentrations were synthesised by aqueous precipitation, and samples repeatedly exposed to 50 mM lactic acid solution, pH 4.5, or similar lactic/acetic/formic acid mixtures, until dissolution was complete. While the Ca/P ratio in solution remained relatively constant and close to the ratio in the solid, the solution F/Ca ratio invariably changed during dissolution. During initial stages the F/Ca solution ratio was lower than in the solid but rose to reach a plateau higher than in the solid as dissolution progressed, an effect that was more pronounced with low-F FHAp. With these minerals the plateau F/Ca level never reached 0.2, suggesting that a F-enriched FHAp rather than pure fluorapatite precipitates during dissolution. It is concluded that a high-F FHAp mineral would best serve as an apatitic plaque reservoir of F.