Curcumin suppresses cell growth and attenuates fluoride-mediated Caspase-3 activation in ameloblast-like LS8 cells.

The trace element fluoride can be beneficial for oral health by preventing dental caries. However, fluoride is also known as an environmental pollutant. Fluoride pollution can lead to fluoride over-ingestion and can cause health issues, including dental fluorosis. Curcumin attenuated fluoride-induced toxicity in animal models, however the molecular mechanisms of how curcumin affects fluoride toxicity remain to be elucidated. We hypothesized that curcumin attenuates fluoride toxicity through modulation of Ac-p53. Here we investigated how curcumin affects the p53-p21 pathway in fluoride toxicity. LS8 cells were treated with NaF with/without curcumin. Curcumin significantly increased phosphorylation of Akt [Thr308] and attenuated fluoride-mediated caspase-3 cleavage and DNA damage marker γH2AX expression. Curcumin-mediated attenuation of caspase-3 activation was reversed by Akt inhibitor LY294002 (LY). However, LY did not alter curcumin-mediated γH2AX suppression. These results suggest that curcumin inhibited fluoride-mediated apoptosis via Akt activation, but DNA damage was suppressed by other pathways. Curcumin did not suppress/alter fluoride-mediated Ac-p53. However, curcumin itself significantly increased Ac-p53 and upregulated p21 protein levels to suppress cell proliferation in a dose-dependent manner. Curcumin suppressed fluoride-induced phosphorylation of p21 and increased p21 levels within the nuclear fraction. However, curcumin did not reverse fluoride-mediated cell growth inhibition. These results suggest that curcumin-induced Ac-p53 and p21 led to cell cycle arrest, while curcumin attenuated fluoride-mediated apoptosis via activation of Akt and suppressed fluoride-mediated DNA damage. By inhibiting DNA damage and apoptosis, curcumin may potentially alleviate health issues caused by fluoride pollution. Further studies are required to better understand the mechanism of curcumin-induced biological effects on fluoride toxicity.

Histone acetyltransferase promotes fluoride toxicity in LS8 cells.

Previously we demonstrated that fluoride increased acetylated-p53 (Ac-p53) in LS8 cells that are derived from mouse enamel organ epithelia and in rodent ameloblasts. However, how p53 is acetylated by fluoride and how the p53 upstream molecular pathway responds to fluoride is not well characterized. Here we demonstrate that fluoride activates histone acetyltransferases (HATs) including CBP, p300, PCAF and Tip60 to acetylate p53. HAT activity is regulated by post-translational modifications such as acetylation and phosphorylation. HAT proteins and their post-translational modifications (p300, Acetyl-p300, CBP, Acetyl-CBP, Tip60 and phospho-Tip60) were analyzed by Western blots. p53-HAT binding was detected by co-immunoprecipitation (co-IP). Cell growth inhibition was analyzed by MTT assays. LS8 cells were treated with NaF with/without HAT inhibitors MG149 (Tip60 inhibitor) and Anacardic Acid (AA; inhibits p300/CBP and PCAF). MG149 or AA was added 1 h prior to NaF treatment. Co-IP results showed that NaF increased p53-CBP binding and p53-PCAF binding. NaF increased active Acetyl-p300, Acetyl-CBP and phospho-Tip60 levels, suggesting that fluoride activates these HATs. Fluoride-induced phospho-Tip60 was decreased by MG149. MG149 or AA treatment reversed fluoride-induced cell growth inhibition at 24 h. MG149 or AA treatment decreased fluoride-induced p53 acetylation to inhibit caspase-3 cleavage, DNA damage marker γH2AX expression and cytochrome-c release into the cytosol. These results suggest that acetylation of p53 by HATs contributes, at least in part, to fluoride-induced toxicity in LS8 cells via cell growth inhibition, apoptosis, DNA damage and mitochondrial damage. Modulation of HAT activity may, therefore, be a potential therapeutic target to mitigate fluoride toxicity in ameloblasts.

Removal of matrix-bound zoledronate prevents post-extraction osteonecrosis of the jaw by rescuing osteoclast function.

Unlike other antiresorptive medications, bisphosphonate molecules accumulate in the bone matrix. Previous studies of side-effects of anti-resorptive treatment focused mainly on systemic effects. We hypothesize that matrix-bound bisphosphonate molecules contribute to the pathogenesis of bisphosphonate-related osteonecrosis of the jaw (BRONJ). In this study, we examined the effect of matrix-bound bisphosphonates on osteoclast differentiation in vitro using TRAP staining and resorption assay, with and without pretreatment with EDTA. We also tested the effect of zoledronate chelation on the healing of post-extraction defect in rats. Our results confirmed that bisphosphonates bind to, and can be chelated from, mineralized matrix in vitro in a dose-dependent manner. Matrix-bound bisphosphonates impaired the differentiation of osteoclasts, evidenced by TRAP activity and resorption assay. Zoledronate-treated rats that underwent bilateral dental extraction with unilateral EDTA treatment showed significant improvement in mucosal healing and micro-CT analysis on the chelated sides. The results suggest that matrix-bound bisphosphonates are accessible to osteoclasts and chelating agents and contribute to the pathogenesis of BRONJ. The use of topical chelating agents is a promising strategy for the prevention of BRONJ following dental procedures in bisphosphonate-treated patients.

4-phenylbutyrate Mitigates Fluoride-Induced Cytotoxicity in ALC Cells.

Chronic fluoride over-exposure during pre-eruptive enamel development can cause dental fluorosis. Severe dental fluorosis is characterized by porous, soft enamel that is vulnerable to erosion and decay. The prevalence of dental fluorosis among the population in the USA, India and China is increasing. Other than avoiding excessive intake, treatments to prevent dental fluorosis remain unknown. We previously reported that high-dose fluoride induces endoplasmic reticulum (ER) stress and oxidative stress in ameloblasts. Cell stress induces gene repression, mitochondrial damage and apoptosis. An aromatic fatty acid, 4-phenylbutyrate (4PBA) is a chemical chaperone that interacts with misfolded proteins to prevent ER stress. We hypothesized that 4PBA ameliorates fluoride-induced ER stress in ameloblasts. To determine whether 4PBA protects ameloblasts from fluoride toxicity, we analyzed gene expression of Tgf-ß1, Bcl2/Bax ratio and cytochrome-c release in vitro. In vivo, we measured fluorosis levels, enamel hardness and fluoride concentration. Fluoride treated Ameloblast-lineage cells (ALC) had decreased Tgf-ß1 expression and this was reversed by 4PBA treatment. The anti-apoptotic Blc2/Bax ratio was significantly increased in ALC cells treated with fluoride/4PBA compared to fluoride treatment alone. Fluoride treatment induced cytochrome-c release from mitochondria into the cytosol and this was inhibited by 4PBA treatment. These results suggest that 4PBA mitigates fluoride-induced gene suppression, apoptosis and mitochondrial damage in vitro. In vivo, C57BL/6J mice were provided fluoridated water for six weeks with either fluoride free control-chow or 4PBA-containing chow (7 g/kg 4PBA). With few exceptions, enamel microhardness, fluorosis levels, and fluoride concentrations of bone and urine did not differ significantly between fluoride treated animals fed with control-chow or 4PBA-chow. Although 4PBA mitigated high-dose fluoride toxicity in vitro, a diet rich in 4PBA did not attenuate dental fluorosis in rodents. Perhaps, not enough intact 4PBA reaches the rodent ameloblasts necessary to reverse the effects of fluoride toxicity. Further studies will be required to optimize protocols for 4PBA administration in vivo in order to evaluate the effect of 4PBA on dental fluorosis.

Biomarkers of chronic fluoride exposure in groundwater in a highly exposed population.

This study examined the relation between fluoride (F-) concentrations in fingernail clippings and urine and the prevalence and severity of enamel fluorosis (EF) among Ethiopian Rift Valley populations exposed to high levels of F- in drinking water. The utility of fingernail clippings as a biomarker for F- exposure and EF was also assessed for the first time in a high-F- region. The study recorded the EF status of 386 individuals (10 to 50years old), who consume naturally contaminated groundwater with widely varying F- concentration (0.6-15mg/L). The mean F- concentrations among residents of communities with primary reliance on groundwater were 5.1mg/kg (range: 0.5-34mg/kg) in fingernails and 8.9mg/L (range: 0.44-34mg/L) in urine. We show strong positive correlations between F- in drinking water and 12-hour urinary excretion (r=0.74, p<0.001, n=287), fingernail F- content (r=0.6, p<0.001, n=258), and mean individual measures of EF severity as measured using the Thylstrup and Fejerskov (TF) Index (r=0.42, p<0.001, n=316). The data indicate that both fingernail and urine measures are good biomarkers for F- exposure and EF outcomes, the latter being slightly more sensitive. Cases of moderate/severe EF were significantly more common among younger subjects (10 to 15years old) than older subjects (mostly >25years old) (p<0.001), consistent with their greater exposure to F- during early childhood, which is the only period of life the enamel is at risk of fluorosis. In this younger population, EF may be useful as a biomarker for identifying individuals with other potential health effects that depend on a specific age window of susceptibility. The finding of exceptionally high F- concentrations in water, fingernail clippings and urine in this region should motivate further investigations of other potential health consequences such as bone disease and abnormalities in the function of the neurological and endocrine systems.

Skeletal Fluorosis Due To Inhalation Abuse of a Difluoroethane-Containing Computer Cleaner.

Skeletal fluorosis (SF) is endemic in many countries and millions of people are affected worldwide, whereas in the United States SF is rare with occasional descriptions of unique cases. We report a 28-year-old American man who was healthy until 2 years earlier when he gradually experienced difficulty walking and an abnormal gait, left hip pain, loss of mobility in his right wrist and forearm, and progressive deformities including enlargement of the digits of both hands. Dual-energy X-ray absorptiometry (DXA) of his lumbar spine, femoral neck, total hip, and the one-third forearm revealed bone mineral density (BMD) Z-scores of +6.2, +4.8, +3.0, and -0.2, respectively. Serum, urine, and bone fluoride levels were all elevated and ultimately explained by chronic sniffing abuse of a computer cleaner containing 1,1-difluoroethane. Our findings reflect SF due to the unusual cause of inhalation abuse of difluoroethane. Because this practice seems widespread, particularly in the young, there may be many more such cases. © 2016 American Society for Bone and Mineral Research.

Mode of drinking fluoridated milk: effect on intraoral fluoride concentrations.

OBJECTIVE: To determine the effect of the mode of drinking fluoridated milk on salivary and plaque fluoride concentrations. METHODS: Fluoridated milk was ingested by 32 children in three ways: (a) directly from the container (1.0 and 5.0 mg F/litre), (b) through a straw with the tip between the lips (5.0 mg F/litre), and (c) with the tip deep in the oral cavity (5.0 mg F/litre). Saliva was collected at baseline and 2, 15, and 40 min and plaque at baseline and 20 min after drinking. Fluoride concentrations were determined using the electrode after HMDS-facilitated diffusion. RESULTS: The mode of drinking did not affect fluoride concentrations in saliva or plaque. The average 2-min salivary concentrations were 65 ng F/mL for the 1.0 mg F/litre group and 276 ng F/mL for the three 5.0 mg F/litre groups (P < 0.01). The average of the 15- and 40-min salivary concentrations was 22 ng F/mL for the 1.0 mg F/litre group and 41 ng F/mL for the 5.0 mg F/litre groups (P < 0.01). Plaque concentrations showed the same patterns as in saliva, that is, they were higher in the three 5.0 mg F/litre groups than in the 1.0 mg F/litre group and the differences among the 5.0 mg F/litre groups were not statistically significant. CONCLUSION: Salivary and plaque fluoride concentrations were independent of the mode of drinking but directly related to milk fluoride concentrations.

Effect of exercise on fluoride metabolism in adult humans: a pilot study.

An understanding of all aspects of fluoride metabolism is critical to identify its biological effects and avoid fluoride toxicity in humans. Fluoride metabolism and subsequently its body retention may be affected by physiological responses to acute exercise. This pilot study investigated the effect of exercise on plasma fluoride concentration, urinary fluoride excretion and fluoride renal clearance following no exercise and three exercise intensity conditions in nine healthy adults after taking a 1-mg Fluoride tablet. After no, light, moderate and vigorous exercise, respectively, the mean (SD) baseline-adjusted i) plasma fluoride concentration was 9.6(6.3), 11.4(6.3), 15.6(7.7) and 14.9(10.0) ng/ml; ii) rate of urinary fluoride excretion over 0-8 h was 46(15), 44(22), 34(17) and 36(17) µg/h; and iii) rate of fluoride renal clearance was 26.5(9.0), 27.2(30.4), 13.1(20.4) and 18.3(34.9) ml/min. The observed trend of a rise in plasma fluoride concentration and decline in rate of fluoride renal clearance with increasing exercise intensity needs to be investigated in a larger trial. This study, which provides the first data on the effect of exercise with different intensities on fluoride metabolism in humans, informs sample size planning for any subsequent definitive trial, by providing a robust estimate of the variability of the effect.

Bone response to fluoride exposure is influenced by genetics.

Genetic factors influence the effects of fluoride (F) on amelogenesis and bone homeostasis but the underlying molecular mechanisms remain undefined. A label-free proteomics approach was employed to identify and evaluate changes in bone protein expression in two mouse strains having different susceptibilities to develop dental fluorosis and to alter bone quality. In vivo bone formation and histomorphometry after F intake were also evaluated and related to the proteome. Resistant 129P3/J and susceptible A/J mice were assigned to three groups given low-F food and water containing 0, 10 or 50 ppmF for 8 weeks. Plasma was evaluated for alkaline phosphatase activity. Femurs, tibiae and lumbar vertebrae were evaluated using micro-CT analysis and mineral apposition rate (MAR) was measured in cortical bone. For quantitative proteomic analysis, bone proteins were extracted and analyzed using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), followed by label-free semi-quantitative differential expression analysis. Alterations in several bone proteins were found among the F treatment groups within each mouse strain and between the strains for each F treatment group (ratio ≥1.5 or ≤0.5; p<0.05). Although F treatment had no significant effects on BMD or bone histomorphometry in either strain, MAR was higher in the 50 ppmF 129P3/J mice than in the 50 ppmF A/J mice treated with 50 ppmF showing that F increased bone formation in a strain-specific manner. Also, F exposure was associated with dose-specific and strain-specific alterations in expression of proteins involved in osteogenesis and osteoclastogenesis. In conclusion, our findings confirm a genetic influence in bone response to F exposure and point to several proteins that may act as targets for the differential F responses in this tissue.

Uncoupling protein-2 is an antioxidant that is up-regulated in the enamel organ of fluoride-treated rats.

Dental fluorosis is characterized by subsurface hypomineralization and retention of enamel matrix proteins. Fluoride (F(-)) exposure generates reactive oxygen species (ROS) that can cause endoplasmic reticulum (ER)-stress. We therefore screened oxidative stress arrays to identify genes regulated by F(-) exposure. Vitamin E is an antioxidant so we asked if a diet high in vitamin E would attenuate dental fluorosis. Maturation stage incisor enamel organs (EO) were harvested from F(-)-treated rats and mice were assessed to determine if vitamin E ameliorates dental fluorosis. Uncoupling protein-2 (Ucp2) was significantly up-regulated by F(-) (∼1.5 & 2.0 fold for the 50 or 100 ppm F(-) treatment groups, respectively). Immunohistochemical results on maturation stage rat incisors demonstrated that UCP2 protein levels increased with F(-) treatment. UCP2 down-regulates mitochondrial production of ROS, which decreases ATP production. Thus, in addition to reduced protein translation caused by ER-stress, a reduction in ATP production by UCP2 may contribute to the inability of ameloblasts to remove protein from the hardening enamel. Fluoride-treated mouse enamel had significantly higher quantitative fluorescence (QF) than the untreated controls. No significant QF difference was observed between control and vitamin E-enriched diets within a given F(-) treatment group. Therefore, a diet rich in vitamin E did not attenuate dental fluorosis. We have identified a novel oxidative stress response gene that is up-regulated in vivo by F(-) and activation of this gene may adversely affect ameloblast function.

Skeletal fluorosis from brewed tea.

BACKGROUND: High fluoride ion (F(-)) levels are found in many surface and well waters. Drinking F(-)-contaminated water typically explains endemic skeletal fluorosis (SF). In some regions of Asia, however, poor quality "brick tea" also causes this disorder. The plant source of brick, black, green, orange pekoe, and oolong tea, Camellia sinensis, can contain substantial amounts of F(-). Exposure to 20 mg F(-) per day for 20 yr of adult life is expected to cause symptomatic SF. High F(-) levels stimulate osteoblasts and enhance bone apposition but substitute for OH(-) groups in hydroxyapatite crystals and thereby result in skeletal fragility and perhaps lead to secondary hyperparathyroidism. Beginning in 2005, we showed that daily consumption of 1-2 gallons of instant tea made from this plant can lead to SF. AIM: We describe a 48-yr-old American woman who developed SF from brewed tea. PATIENT AND METHODS: Our patient had elevated bone mineral density revealed by dual-energy x-ray absorptiometry (spine Z-score, +9.9), severe chronic bone and joint pain, and kyphosis after consuming 1-2 gallons of brewed orange pekoe tea daily for more than three decades. F(-) levels were high in her serum, urine, and clippings of fingernails and toenails, as well as in our reproduction of her beverage. Renal function was normal. She had vitamin D deficiency. Elevated serum PTH levels were unresponsive to adequate vitamin D supplementation. Pain resolved over several months when she stopped drinking tea and continued ergocalciferol. CONCLUSION: Our patient shows that SF can result from chronic consumption of large volumes of brewed tea.

Evidence-based clinical recommendations regarding fluoride intake from reconstituted infant formula and enamel fluorosis: a report of the American Dental Association Council on Scientific Affairs.

BACKGROUND: This article presents evidence-based clinical recommendations regarding the intake of fluoride from reconstituted infant formula and its potential association with enamel fluorosis. The recommendations were developed by an expert panel convened by the American Dental Association (ADA) Council on Scientific Affairs (CSA). The panel addressed the following question: Is consumption of infant formula reconstituted with water that contains various concentrations of fluoride by infants from birth to age 12 months associated with an increased risk of developing enamel fluorosis in the permanent dentition? TYPES OF STUDIES REVIEWED: A panel of experts convened by the ADA CSA, in collaboration with staff of the ADA Center for Evidence-based Dentistry (CEBD), conducted a MEDLINE search to identify systematic reviews and clinical studies published since the systematic reviews were conducted that addressed the review question. RESULTS: CEBD staff identified one systematic review and two clinical studies. The panel reviewed this evidence to develop recommendations. CLINICAL IMPLICATIONS: The panel suggested that when dentists advise parents and caregivers of infants who consume powdered or liquid concentrate infant formula as the main source of nutrition, they can suggest the continued use of powdered or liquid concentrate infant formulas reconstituted with optimally fluoridated drinking water while being cognizant of the potential risks of enamel fluorosis development. These recommendations are presented as a resource to be considered in the clinical decision-making process. As part of the evidence-based approach to care, these clinical recommendations should be integrated with the practitioner's professional judgment and the patient's needs and preferences.

Fluoride excess and periostitis in transplant patients receiving long-term voriconazole therapy.

BACKGROUND: We describe a heart transplant patient with painful periostitis and exostoses who was receiving long-term therapy with voriconazole, which is a fluoride-containing medication. Elevated plasma and bone fluoride levels were identified. Discontinuation of voriconazole therapy led to improvement in pain and reduced fluoride and alkaline phosphatase levels. METHODS: To determine whether voriconazole is a cause of fluoride excess, we measured plasma fluoride levels in 10 adult post-transplant patients who had received voriconazole for at least 6 months and 10 post-transplant patients who did not receive voriconazole. To assess the effect of renal insufficiency on fluoride levels in subjects receiving voriconazole, half were recruited on the basis of a serum creatinine level of ≥1.4 mg/dL on their most recent measurement, whereas the other 5 subjects receiving voriconazole had serum creatinine levels <1.4 mg/dL. All control subjects had serum creatinine levels of ≥1.4 mg/dL. Patients were excluded from the study if they received a fluorinated pharmaceutical other than voriconazole. RESULTS: All subjects who received voriconazole had elevated plasma fluoride levels, and no subjects in the control group had elevated levels (14.32 µmol/L ± 6.41 vs 2.54 ± 0.67 µmol/L; P<.001). Renal function was not predictive of fluoride levels. Plasma fluoride levels remained significantly higher in the voriconazole group after adjusting for calcineurin inhibitor levels and doses. Half of the voriconazole group subjects had evidence of periostitis, including exostoses in 2 patients. Discontinuation of voriconazole therapy in patients with periostitis resulted in improvement of pain and a reduction in alkaline phosphatase and fluoride levels. CONCLUSIONS: Voriconazole is associated with painful periostitis, exostoses, and fluoride excess in post-transplant patients with long-term voriconazole use.

Dietary fluoride restriction does not alter femoral biomechanical strength in col1a2-deficient (oim) mice with type I collagen glomerulopathy.

Osteogenesis imperfecta (OI) is a clinically and genetically heterogeneous disease due primarily to mutations in the type I procollagen genes, COL1A1 and COL1A2, causing bone deformity and numerous lifetime fractures. OI murine (oim) model mice carry a mutation in the col1a2 gene causing aberrant production of homotrimeric type I collagen [α1(I)(3)], leading to bone fragility and glomerular accumulation of type I collagen. Previous studies demonstrated that heterozygous (+/oim) and homozygous (oim/oim) mice have elevated tibiae fluoride concentrations but reduced femoral biomechanics. However, it is unclear whether these 2 variables are causally related, because impaired renal function could reduce urinary fluoride excretion, thus elevating bone fluoride concentrations regardless of disease status. Our goal in this study was to determine whether dietary fluoride restriction would improve femoral biomechanics in oim mice. Wild-type, +/oim, and oim/oim mice were fed a control (5 mg/kg fluoride) or fluoride-restricted diet (0 mg/kg fluoride) for ∼13 wk, at which time plasma and femora were analyzed for fluoride concentrations and bone biomechanical properties. In wild-type, +/oim, and oim/oim mice, dietary fluoride restriction reduced femoral fluoride burden by 54-74%, respectively (P < 0.05), without affecting glomerular collagen deposition. Oim/oim mice fed the fluoride-restricted diet had reduced material tensile strength (P < 0.05) compared with oim/oim mice fed the control diet. However, dietary fluoride restriction did not affect stiffness or whole bone femoral breaking strength, regardless of genotype. These data suggest that oim mice have reduced bone strength due to homotrimeric type I collagen, independent of bone fluoride content.

Appetitive-based learning in rats: lack of effect of chronic exposure to fluoride.

BACKGROUND: Chronic ingestion of optimally fluoridated water (ca. 1.0 mg/L) has not been associated with any adverse health effects. Possible effects on the nervous system, however, have received little attention. One study with rats given high doses of fluoride reported subtle behavioral changes. The authors suggested that the ability of humans to learn might be reduced and recommended further study with humans and rats. The present study was done to provide data with which to assess this suggestion. METHODS: Weanling, female rats (n=32) were provided with water containing graded doses of fluoride (0, 2.9, 5.7, 11.5 mg/kg body weight/day) for eight months. While under restricted food access they were tested for their ability to learn an operant response for food and to adjust their responding under schedules of reinforcement requiring high rates of responding (5 days) and then low rates of responding (10 days). Bone, plasma and seven regions of brain were analyzed for fluoride. RESULTS: There were no significant differences among the groups in learning or performance of the operant tasks. Tissue fluoride concentrations were directly related to the levels of exposure. In the 11.5 mg/kg/day group the bone, plasma and brain concentrations were 99, 305 and 221 times higher, respectively, than those in the control group. The average brain-to-plasma fluoride concentration ratios in each of the seven brain sections fell within a narrow range and did not exceed 0.40. There was no consistent evidence for the preferential uptake of fluoride by any given brain section. CONCLUSION: Chronic ingestion of fluoride at levels up to 230 times more than that experienced by humans whose main source of fluoride is fluoridated water had no significant effect on appetitive-based learning.

Skeletal fluorosis from instant tea.

INTRODUCTION: Skeletal fluorosis (SF) can result from prolonged consumption of well water with >4 ppm fluoride ion (F(-); i.e., >4 mg/liter). Black and green teas can contain significant amounts of F(-). In 2005, SF caused by drinking 1-2 gallons of double-strength instant tea daily throughout adult life was reported in a 52-yr-old woman. MATERIALS AND METHODS: A 49-yr-old woman developed widespread musculoskeletal pains, considered fibromyalgia, in her mid-30s. Additionally, she had unexplained, increasing, axial osteosclerosis. She reported drinking 2 gallons of instant tea each day since 12 yr of age. Fluoxetine had been taken intermittently for 5 yr. Ion-selective electrode methodology quantitated F(-) in her blood, urine, fingernail and toenail clippings, tap water, and beverage. RESULTS: Radiographs showed marked uniform osteosclerosis involving the axial skeleton without calcification of the paraspinal, intraspinal, sacrotuberous, or iliolumbar ligaments. Minimal bone excrescences affected ligamentous attachments in her forearms and tibias. DXA Z-scores were +10.3 in the lumbar spine and +2.8 in the total hip. Her serum F(-) level was 120 microg/liter (reference range, 20-80 microg/liter), and a 24-h urine collection contained 18 mg F(-)/g creatinine (reference value, <3). Fingernail and toenail clippings showed 3.50 and 5.58 mg F(-)/kg (control means, 1.61 and 2.02, respectively; p(s) < 0.001). The instant tea beverage, prepared as usual extra strength using tap water with approximately 1.2 ppm F(-), contained 5.8 ppm F(-). Therefore, the tea powder contributed approximately 35 mg of the 44 mg daily F(-) exposure from her beverage. Fluoxetine provided at most 3.3 mg of F(-) daily. CONCLUSIONS: SF from habitual consumption of large volumes of extra strength instant tea calls for recognition and better understanding of a skeletal safety limit for this modern preparation of the world's most popular beverage.

Metrologic evaluation of the surface of white MTA after the use of two endodontic irrigants.

This study examined the effects of calcium-depleting endodontic irrigants, ethylenediamine tetra-acetic acid (EDTA) and BioPure MTAD, on the dissolution, surface characteristics, and ultrastructural characteristics of white mineral trioxide aggregate (MTA). The latter was mixed in a 0.35 water-cement ratio, condensed into cylindrical wells created in Plexiglas platforms, and allowed to harden completely before initial treatment with 1.3% NaOCl and final treatments with either 17% EDTA for 5 minutes, or BioPure MTAD for 1, 3, or 5 minutes. Analysis of the mean depths of material removed using three-dimensional profilometry revealed significantly more material removed by BioPure MTAD. Although these minor depth changes (<10 microm) are unlikely to cause clinical concern, the BioPure MTAD-treated MTA surfaces exhibited higher surface roughness and with more calcium extracted when compared with EDTA treatment. Decomposition of particle-binding hydration phases by acid corrosion raises potential concern on the strength and sealing properties of MTA-repaired perforations following final irrigation by BioPure MTAD.

Fluoride concentrations in dental plaque and saliva after the use of a fluoride dentifrice preceded by a calcium lactate rinse.

Plaque fluoride concentrations ([F]) are directly related to plaque calcium concentrations [Ca]. Attempts to increase plaque F uptake from dentifrices or rinses have used methods designed to increase plaque [Ca] but with inconsistent results. This double-blind, double-crossover study tested the effect of a 150 mM calcium lactate rinse used prior to brushing with placebo or fluoridated dentifrices (1030 p.p.m. as NaF) on plaque and salivary [F] and [Ca]. Sixteen children (8-10 yr of age) were randomly assigned to four different groups according to the four treatments (placebo dentifrice or fluoridated dentifrice preceded by calcium lactate or deionized water prerinses). Plaque and saliva were collected 1 and 12 h after brushing on day 7 after starting to use the dentifrices. F was determined using the electrode and Ca was determined using atomic absorption spectrometry. Plaque and salivary [Ca] were not significantly increased after use of the calcium lactate prerinse, except for plaque [Ca] 1 h after the use of the placebo dentifrice. A significant increase in salivary [F] was associated with the calcium lactate prerinse only at 1 h after the use of the fluoridated dentifrice. The the calcium lactate prerinse did not significantly affect plaque [F] under any condition.

A reproducibility study for a fluoride assay in bone.

Osteosarcoma is a rare malignancy of largely unknown etiology. Although there is no consistent evidence for an association between fluoridation and cancer, some concerns remain about osteosarcoma. As part of the design of a collaborative study, bone samples were collected to allow for an evaluation of the association between osteosarcoma risk and individual fluoride exposure measured by levels of fluoride in bone. In this report, we provide the results of pilot experiments to consider issues that arose during the study design and to assess the reliability of the bone assays. Correlations of fluoride levels between normal bone near the affected area and iliac crest bone were strong and positive. The day-to-day laboratory analysis of fluoride in human and deer jaw bone yielded acceptable average coefficients of variation below 10% and an overall estimate of 5%. The intraclass correlation (ICC) is of particular importance to epidemiologists because it indicates the effect of measurement error on study results. Here, the estimated ICC is 0.86, and the estimated downward bias is only 14%. Hence, the ICC is strong enough so that the estimates of the relative risk will suffer little attenuation from lab measurements.