Histomorphometric profile of bone fluorosis induced by prolonged ingestion of Vichy Saint-Yorre water. Comparison with bone fluorine levels

Nine transiliac bone biopsies from 7 patients with skeletal fluorosis due to prolonged ingestion of often high quantities of Vichy Saint-Yorre water were analyzed. Four of these patients also suffered from a chronic renal failure. A histomorphometric study was possible in 8 out of the 9 biopsies. The measurement of bone fluoride content, and a microradiographic examination, were performed on all bone samples. The radiologically evident osteosclerosis observed in each patient was confirmed by the significant increase of trabecular bone volume. Furthermore, the osteoid surfaces were very extended but the thickness of osteoid seams was normal in 6 out of 8 cases. Two biopsies demonstrated a morphological evidence of osteomalacia with abnormally thick osteoid seams. Calcification rate, measured in one of these 2 cases after tetracycline double labeling, was extremely low (less than 0.20 micron/d). The bone fluoride content was significantly high in each specimen (greater than 0.40 bone ash%) and correlated with relative osteoid volume (r' = 0.91) and thickness index of osteoid seams (r' = 0.83). Histologically, bone tissue showed modifications classically reported in the various types of skeletal fluorosis (formation defects, mottled bone with mottled periosteocytic lacunae). In conclusion, the prolonged administration of Vichy Saint-Yorre water containing 8.5 mg of fluoride ion per liter, provokes a skeletal fluorosis. This intoxication appeared very quickly if the patient suffered from an even mild renal failure. Once again, it is shown that a disturbed renal function predisposes to an excessive retention of fluoride.     

Uniform partial dissolution of bone mineral by using fluoride and phosphate ions combination

Mineral content is one of the main predictors of the mechanical properties of bone tissue. The contribution of the bone mineral phase to the mechanical properties of bone has been investigated by reducing the mineral content of bone with different in vitro treatment techniques such as hydrochloric acid (HCl), ethylenedinitrilo tetraacetic acid (EDTA), and fluoride ion treatment. In this study, we propose a new treatment technique which combines fluoride and phosphate ions. Bovine femur specimens were used to determine the mechanical properties of cortical bone after different fluoride phosphate ion combination treatments. The treatment solutions, which contain different fluoride and phosphate ion concentrations, dissolved part of the bone mineral in a uniform manner throughout the bone samples. Dissolution by products, which precipitated in the bone tissue, contained calcium fluoride with phosphate ions (CaF(2)/P) and fluorapatite/fluorhydroxyapatite-type material (FAp/FHAp) and acted as filler. Depending on the fluoride and phosphate concentration in a treatment solution, the precipitated material's ratio of FAp/FHAp to total fluoride containing phase (FAp/FHAp + CaF(2)/P) in bone tissue also changed. High fluoride ion content in treatment solutions generated more CaF(2)/P type of precipitate, and low fluoride ion concentration generated more FAp/FHAp type precipitates as compared to high fluoride concentration treatments. These experiments show that phosphate ions are another important parameter of a treatment solution, in addition to ionic strength, pH, and the duration of treatment. In vitro, phosphate fluoride combinations partially dissolve bone mineral content in a wider range than fluoride treatment alone in a uniform manner. With this new technique one can control more precisely the partial dissolution of the bone mineral and mineral phase's contribution to mechanical properties of bone tissue.     

Varying the mechanical properties of bone tissue by changing the amount of its structurally effective bone mineral content

The effect of fluoride ions on the mechanical properties of bone tissue in tension was investigated with an in vitro model. Structurally effective Bone Mineral Content (BMC) of bovine bone tissue was changed by fluoride ion treatment. First, bovine cortical bone specimens were treated with a detergent solution in order to increase the diffusion rates of the treatment ions across the samples. After the initial treatment, different ion solutions were used to treat the tension samples (fluoride, sodium and chloride). Ionic strength and pH were varied. Experimental results showed that the sodium chloride solutions of different ionic strengths, at physiological and high pH, do not affect the mechanical properties of bone tissue in tension. However, uniform fluoride treatment across the samples reduced the mechanical strength of bone tissue by converting small amounts of bone mineral to mostly calcium fluoride. This action reduces the structurally effective BMC and also possibly effects the interface bonding between the bone mineral and the organic matrix of the bone tissue.     

Increased ash contents and estimation of dissolution from chemical changes due to in-vitro fluoride treatments

The in-vitro fluoride treatment technique has been introduced to investigate the composite behavior of bone tissue. Bone tissue with different mechanical properties can be obtained by varying the concentration, pH and immersion time in fluoride ion solutions. The chemical and physical changes in intact pieces of bone treated in-vitro with different concentrations of fluoride ions are studied. The amount of bone mineral that does not contribute to the mechanical behavior of bone tissue is estimated from the dissolution occurring in the fluoride treated bones. Cortical bones from 18-month-old steers were treated in-vitro with 0.145, 0.5 and 2.0 M sodium fluoride (NaF) solutions for three days. The dissolved bone mineral precipitates as calcium fluoride-like (CaF2/P with some phosphate [P] ions) and fluorapatite(FAp)/fluorhydroxyapatite(FHAp)-like materials within the bone tissue. The dissolution estimated from the presence of the precipitated fluoride phases is 5.6, 11.7, and 13.1% of the initial bone mineral content for the 0.145 M, 0.5 M, and 2.0 M NaF treatments respectively. Estimates of dissolution based on the measurements of phosphate and carbonate ions are lower and higher respectively when compared to the fluoride ion measurements. The wet and dry densities decreased slightly due to dissolution and re-precipitation while the ash content (ratio of the ash weight to dry weight) increased a small amount with increasing concentration of fluoride ion treatments. The increased ash content was due to the excess loss of water in the fluoride treated bones as compare to controls (untreated bone samples) during the drying process. The increased removal of water during the drying process may explain the increased ash contents in some in-vivo treatments.     

Uniform Partial Dissolution of Bone Mineral by Using Fluoride and Phosphate Ions Combination

Mineral content is one of the main predictors of the mechanical properties of bone tissue. The contribution of the bone mineral phase to the mechanical properties of bone has been investigated by reducing the mineral content of bone with different in vitro treatment techniques such as hydrochloric acid (HCl), ethylenedinitrilo tetraacetic acid (EDTA), and fluoride ion treatment. In this study, we propose a new treatment technique which combines fluoride and phosphate ions. Bovine femur specimens were used to determine the mechanical properties of cortical bone after different fluoride phosphate ion combination treatments. The treatment solutions, which contain different fluoride and phosphate ion concentrations, dissolved part of the bone mineral in a uniform manner throughout the bone samples. Dissolution by products, which precipitated in the bone tissue, contained calcium fluoride with phosphate ions (CaF₂/P) and fluorapatite/fluorhydroxyapatite-type material (FAp/FHAp) and acted as filler. Depending on the fluoride and phosphate concentration in a treatment solution, the precipitated material's ratio of FAp/FHAp to total fluoride containing phase (FAp/FHAp + CaF₂/P) in bone tissue also changed. High fluoride ion content in treatment solutions generated more CaF₂/P type of precipitate, and low fluoride ion concentration generated more FAp/FHAp type precipitates as compared to high fluoride concentration treatments. These experiments show that phosphate ions are another important parameter of a treatment solution, in addition to ionic strength, pH, and the duration of treatment. In vitro, phosphate fluoride combinations partially dissolve bone mineral content in a wider range than fluoride treatment alone in a uniform manner. With this new technique one can control more precisely the partial dissolution of the bone mineral and mineral phase's contribution to mechanical properties of bone tissue.     

Increased Ash Contents and Estimation of Dissolution from Chemical Changes due to In-Vitro Fluoride Treatments

The in-vitro fluoride treatment technique has been introduced to investigate the composite behavior of bone tissue. Bone tissue with different mechanical properties can be obtained by varying the concentration, pH and immersion time in fluoride ion solutions. The chemical and physical changes in intact pieces of bone treated in-vitro with different concentrations of fluoride ions are studied. The amount of bone mineral that does not contribute to the mechanical behavior of bone tissue is estimated from the dissolution occurring in the fluoride treated bones. Cortical bones from 18-month-old steers were treated in-vitro with 0.145, 0.5 and 2.0M sodium fluoride (NaF) solutions for three days. The dissolved bone mineral precipitates as calcium fluoride-like (CaF 2 /P with some phosphate [P] ions) and fluorapatite(FAp)/fluorhydroxyapatite-(FHAp)-like materials within the bone tissue. The dissolution estimated from the presence of the precipitated fluoride phases is 5.6, 11.7, and 13.1% of the initial bone mineral content for the 0.145M, 0.5M, and 2.0M NaF treatments respectively. Estimates of dissolution based on the measurements of phosphate and carbonate ions are lower and higher respectively when compared to the fluoride ion measurements. The wet and dry densities decreased slightly due to dissolution and re-precipitation while the ash content (ratio of the ash weight to dry weight) increased a small amount with increasing concentration of fluoride ion treatments. The increased ash content was due to the excess loss of water in the fluoride treated bones as compare to controls (untreated bone samples) during the drying process. The increased removal of water during the drying process may explain the increased ash contents in some in-vivo treatments.     

Surface coatings for improvement of bone cell materials and antimicrobial activities of Ti implants

Ti surface was modified to simultaneously improve bone cell materials and antimicrobial activities. Titanium surface was first anodized in sodium fluoride and sulfuric acid electrolytic solution to form titania nanotube on the surface to improve the biocompatibility of the surface. Silver was electrodeposited on the titania nanotube surface at 5 V. Silver added titania nanotube surface was tested for compatibility with bone-cell materials interactions using human osteoblast bone cells. The antibacterial effect was studied using Pseudomonas aeruginosa. Our results show that silver-treated titania nanotube surface may provide antibacterial properties to prevent implants against postoperative infections without interference to the attachment and proliferation of bone tissue on titanium, which is commonly used in dental and orthopedic surgical procedures.     

Surface properties and ion release from fluoride-containing bioactive glasses promote osteoblast differentiation and mineralization in vitro

Bioactive glasses (BG) are suitable for bone regeneration applications as they bond with bone and can be tailored to release therapeutic ions. Fluoride, which is widely recognized to prevent dental caries, is efficacious in promoting bone formation and preventing osteoporosis-related fractures when administered at appropriate doses. To take advantage of these properties, we created BG incorporating increasing levels of fluoride whilst holding their silicate structure constant, and tested their effects on human osteoblasts in vitro. Our results demonstrate that, whilst cell proliferation was highest on low-fluoride-containing BG, markers for differentiation and mineralization were highest on BG with the highest fluoride contents, a likely effect of a combination of surface effects and ion release. Furthermore, osteoblasts exposed to the dissolution products of fluoride-containing BG or early doses of sodium fluoride showed increased alkaline phosphatase activity, a marker for bone mineralization, suggesting that fluoride can direct osteoblast differentiation. Taken together, these results suggest that BG that can release therapeutic levels of fluoride may find use in a range of bone regeneration applications.     

The effect ofin vitro fluoride ion treatment on the ultrasonic properties of cortical bone

The mechanical properties of composites are influenced, in part, by the volume fraction, orientation, constituent mechanical properties, and interfacial bonding. Cortical bone tissue represents a short-fibered biological composite where the hydroxyapatite phase is embedded in an organic matrix composed of type I collagen and other noncollagenous proteins. Destructive mechanical testing has revealed that fluoride ion treatment significantly lowers theZ-axis tensile and compressive properties of cortical bone through a constituent interfacial debonding mechanism. The present ultrasonic data indicates that fluoride ion treatment significantly alters the longitudinal velocity in theZ-axis as well as the circumferential and radial axes of cortical bone. This suggests that the distribution of constituents and interfacial bonding amongst them may contribute to the anisotropic nature of bone tissue.     

In vivo expression of osteogenic markers and bone mineral density at the surface of fluoride-modified titanium implants

The aim of the present study was to investigate the biological mechanisms of the functional attachment of fluoride-modified titanium implants to cortical bone by studying the association of the pull-out test results with gene expression of osteoblast (runx2, osteocalcin, collagen-I and IGF-I), osteoclast (TRAP, H(+)-ATPase and calcitonin receptor) and inflammation (TNF-alpha, IL-6 and IL-10) markers from peri-implant bone tissue using real-time RT-PCR, following a 4- and 8-week healing period. After implant detachment, wound fluid from the implant site was collected for LDH and ALP activity analysis. A new method to study volumetric bone mineral density (vBMD) of sub-implant cortical bone was developed using micro-computed tomography. Our results show lower LDH activity and TRAP mRNA levels in fluoride implants after 4 weeks of healing, yet no differences were found either on the pull-out force or expression of bone formation marker genes. After 8 weeks of healing, both pull-out, vBMD and osteocalcin, runx2 and collage type I gene expression were higher in fluoride implants. In conclusion, fluoride-modified implants seem to modulate both inflammation and bone resorption/formation events at the bone-implant interface, suggesting that these biological effects are an intrinsic part of the clinical performance of this surface.     

Concentration dependence of fluorine impurity spin-lattice relaxation rate in bone mineral.

The concentration dependence of the fluoride ion spin-lattice relaxation rate has been observed by nuclear magnetic resonance experiments on samples of defatted and dried bone. The 19F spin-lattice relaxation rates increased linearly with the bone fluoride concentration. Different results were obtained from trabecular than from cortical bone. For the same macroscopic fluoride content per gram of bone calcium, the relaxation rate is significantly faster in cortical bone. Relaxation rates in cortical bone samples prepared from both rats and dogs were apparently controlled by the same species-independent processes. For samples from beagle dogs, the bulk fluoride concentrations measured by neutron activation analysis were 3.1 +/- 0.3 times greater in trabecular bone than in the corresponding cortical bone. The beagle spin-lattice relaxation data suggest that the microscopic fluoride concentrations in bone mineral were 1.8 +/- 0.4 times greater in trabecular bone than in cortical bone. It is concluded that the accumulation of fluoride impurities in bone mineral is non-uniform.     

Accumulation of Calcium,Phosphate, and Collagen in Bones and Accumulation of Creatine in Muscles of Mice with Acute Hepatic Intoxication during Shin Fracture Healing

The intensity of calcium and phosphate accumulation in regenerating bone tissue decreased significantly during the posttraumatic period in 54 male CBA mice with acute hepatic intoxication. The formation of organic components of the bone decreased to a lesser degree. Creatine metabolism in the muscles of damaged segment virtually did not change.     

燃煤污染型氟中毒家猪胸椎（T2）骨小梁结构的研究

长期摄入过量氟可导致慢性氟中毒。应用骨形态计量学方法对氟中毒家猪胸椎（Ｔ２）的结构进行了研究，测量了７种骨形态学参数，发现部分家猪椎体上部骨小梁体积增加，结节间小梁和结节闭环小梁的长度有增加的趋势，游离小粱和结节游离小粱减少，表现为小梁结构致密；部分家猪骨小梁体积减少，ＮＬ减少，ＦＦ和ＮＦ明显增加，结构破坏，这种骨病变的多样性与骨含氟量有关。     

The pathology of chronic bovine fluorosis: a review.

Clinical, pathologic, and analytical records from 200 cattle were reviewed to determine if long-term exposures to elevated fluorides resulted in previously unrecognized or unreported pathologic changes, especially skeletal neoplasia. Animals were part of comprehensive field and laboratory investigations of bovine fluorosis conducted by the Utah State University Agricultural Experiment Station over a 25-year period. Records indicated that over 170 cattle included in this review were exposed to dietary fluorides levels in excess of 25 ppm (dry wt), for most of their life span, and these animals exhibited bone fluoride concentrations ranging between 2,000 and 12,500 ppm (dry wt). Although dental and/or skeletal changes were present in most animals, significant soft tissue damage or neoplasia was not observed in any organ system. Renal degeneration and mineralization were slightly more prevalent in range cattle ingesting high fluoride levels, but these changes were not recognized in animals that received high experimental fluoride doses. The absence of significant soft tissue damage or neoplasia in these cattle combined with results of an extensive literature review suggests that environmental fluorides are not significant factors in mammalian carcinogenesis.     

Pathology of Clostridium welchii type D enterotoxaemia. 3. Basis of the hyperglycaemic response

It has been demonstrated that the hyperglycaemia, which is a prominent feature of Clostridium welchii type D epsilon toxin intoxication, results from rapid mobilization of hepatic glycogen.Liver glycogen stores are rapidly depleted in intoxicated animals and no elevation of blood glucose level occurs in animals in which hepatic glycogen has been depleted prior to intoxication.No detectable interference with tissue respiration could be demonstrated in tissues from intoxicated animals, suggesting that the hyperglycaemia is not the result of interference with glycolysis.     

Value of the bone biopsy in the diagnosis of industrial fluorosis

Summary Iliac crest biopsies taken from 43 men with industrial fluorosis were compared with control bone samples. The bone fluoride content was determined, histological examinations were made on stained sections and microradiographs, and morphometric analysis performed on the microradiographs alone.In the subjects with fluorosis, the bone fluoride content (5617±2143 ppm) was found to be significantly higher (P<0.00005) than in control subjects (1036±627 ppm). It decreased slowly, however, after exposure had ceased (to about 50% in 20 years). The histological changes consisted of a nonspecific remodeling activity (resulting in increased trabecular bone volume and cortical porosity, as well as hypervascularization and linear formation defects) and modifications of the perilacunar walls (i.e., presence of mottled lacunae and enlarged lacunae). These histological changes were found more likely to occur when the bone fluoride content was high but no correlation between the two parameters was observed.Although certain clinical and radiological data associated with a high urine fluoride content can sometimes establish a diagnosis of skeletal fluorosis, many cases require the use of bone biopsy, which also provides a direct evaluation of the bone fluoride content and can establish the absence of any other bone disease.     

Estimation of bone turnover in climacteric women by the whole body retention of fluoride.

This paper reports the measurement of whole body retention using fluoride (WBRF) as an estimator of skeletal turnover in a group of climacteric women that received an oral dose of 700 mumol of sodium fluoride. WBRF is defined as 100(1-(urinary fluoride/fluoride load)). WBRF was significantly correlated with whole body retention of 99m-Tc-methylene-diphosphonate, the serum levels of the bone alkaline phosphatase and the urinary excretion of hydroxyproline. WBRF values ranged from 20% to 95% and were affected by calcium intake and the urinary calcium excretion. In normal subjects with high turnover, the measurement of serum alkaline phosphatase activity and/or urinary hydroxyproline excretion helps to distinguish these cases from patients with metabolic bone diseases due to metastases, Paget disease, etc. The fact that the fraction of fluoride not incorporated into bone is not further metabolized plus the accuracy, preciseness and rapidity of fluoride measurements in urine are the main advantages of this technique.     

Molecular basis of complement factor I (CFI) polymorphism: one of two polymorphic suballeles responsible for CFI A is Japanese-specific

Isoelectric focusing has revealed that human complement factor I (CFI) is controlled by two polymorphic alleles, CFI(*)A and CFI(*)B, and a few rare variant alleles. In this study the molecular basis of the CFI polymorphism was investigated in 174 Japanese. The CFI(*)A was divided into two suballeles, CFI(*)As (R201S) and CFI(*)Ah (R406H). CFI(*)Aj, a rare variant allele originating from CFI(*)Ah, had an additional mutation (R502L). The distribution of these three mutations and two registered SNPs was investigated in a total of 2,471 individuals in 20 populations from various areas, and six haplotypes were observed. Haplotype H3, which is characterized by CFI(*)As, was found only in Far East populations: the frequencies were about 0.03 in the main island of Japan and lower than 0.01 in Okinawa and Korea. Haplotype H5, characterized by CFI(*)Ah, prevailed almost exclusively in East Asians and was observed at the highest frequencies in southern Chinese Han and Thais. CFI(*)Ah must have arisen in a southeastern part of Asia and thereafter have spread to neighboring populations.     

Effect of fluoride ions on apatite crystal formation in rat hard tissues

Fluoride is widely believed to be a useful chemical substance for preventing dental caries. However, the mechanism underlying crystal perforation in the tooth enamel and the effect of fluoride on hard tissues are unclear. To clarify the mechanism of the biological action of fluoride in the mineralization process, we examined the hard tissues of rats having received water containing a relatively low fluoride level. Electron microscopy revealed that fluoride ions could interrupt the crystal nucleation process, resulting in crystal perforation in the developing tooth enamel and the presence of amorphous minerals in bone crystals. Furthermore, the results of enzymatic analyses indicated that fluoride directly interfered with the synthesis of carbonic anhydrase by the enamel-forming cells, rather than being directly involved in the crystal formation. From the results, we would like to provide a possible mechanism of crystal perforation in the enamel induced by fluoride intake. Also, we would like to suggest that regardless of its amount, fluoride intake has harmful effects on both tooth and bone formation.