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Fluorosis: A New Model and New Insights
J.D. Bartlett,
S.E. Dwyer,
E. Beniash,
Z. Skobe and
T.L. Payne-Ferreira*
Department of Biomineralization and Department of Cytokine Biology, The Forsyth Institute, and Department of Oral and Developmental Biology, Harvard School of Dental Medicine, 140 The Fenway, Boston, MA 02115, USA;
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Figure 1. Phenotypic effect of fluoride treatment. (A) Ventral view of Alcian-blue-stained cartilages of a 7-dpf zebrafish larva. The first few teeth are located on cb5. (B-D) Adult alizarin-red-stained cb5 arches and higher-magnification images of ankylosed teeth. (E) SEM tooth images from 2-month-old fish that were exposed to the indicated fluoride concentrations for 2 mos prior to tooth removal. Characteristic enameloid pitting is seen at concentrations of 38 ppm F and higher. Top, 20,000X; bottom, 2000X magnification. Abbreviations: cb, ceratobranchial; d, dentin; D, dorsal tooth; dpf, days post-fertilization; e, enameloid; M, Meckel’s cartilage; MD, mediodorsal tooth; p, pulp; V, ventral tooth.
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Figure 2. FTIR spectra of the control (solid line) and fluoride-treated (dashed line) zebrafish teeth. As a measure of mineral content, the ratios between the maximum height of an amide I absorption band (a major protein absorption) at ~ 1645 cm–1 and a phosphate absorption band at ~ 1020 cm–1 were calculated. The amide I/ 3 PO4 ratio in the control sample was 0.33, whereas this ratio in fluoride-treated teeth was 0.6. These results suggest that the fluoride-treated teeth have a much higher organic content than do normal teeth. No significant differences in the mineral phase of the control and experimental teeth were observed.
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Figure 3. Apoptosis assay. Two-month-old fish were placed in either 0-ppm-F water (A,B,C) or 38-ppm-F water (D,E,F) for 4 wks. After 4 wks, ssDNA MAB was used to detect apoptotic nuclei (red asterisks) in the paraffin sections. (Panels A, D) Low magnification of ssDNA-stained paraffin sections. Red box indicates cb5 and its associated pharyngeal teeth. Dark brown nuclear staining indicates apoptosis. (Panels B-C, E-F) 20x magnification of control and NaF-treated fish, respectively. (Panels B'-C') 100x magnification of control (0 ppm) tooth germs. (Panels E'-F') 100x magnification of 38-ppm-F-treated tooth germs. Abbreviations: AM, ameloblast-like IEE cell; d, dentin; G, gut; GR, gill rays; PT, pharyngeal teeth.
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Figure 4. Paraffin sections of 4-month-old zebrafish pharyngeal teeth stained with anti-Alk8 polyclonal antibody, and E-cadherin monoclonal antibody. (A,C) Untreated. (B,D) 38-ppm-fluoride-treated. Alk8 is present on the surfaces of the IEE cells, as is E-cadherin. The arrowhead in panels A and B indicates later-stage IEE cells, and the asterisks in the E-cadherin sections (C and D) indicate early-stage tooth germs and more uniform staining of the less mature IEE cells. Abbreviations: am, ameloblast-like IEE cell; ce, crypt epithelium; D, dentin; T, bell-stage tooth.
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Journal of Dental Research, Vol. 84, No. 9,
832-836 (2005)
DOI: 10.1177/154405910508400910
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