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Effects of Fluoride on the Interactions between Amelogenin and Apatite Crystals

¹ Department of Orofacial Sciences, University of California, San Francisco, 513 Parnasuss Avenue, San Francisco, CA 94143, USA;
² Department of Preventive and Restorative Dental Sciences, University of California, San Francisco, CA, USA

View larger version (23K): [in this window] [in a new window]   Figure 1. Affinity of rh174 to apatites. (A) Dissolution of mineral and measurement of the bound protein by Bradford assay showed a similar pattern of rh174 binding to carbonated hydroxyapatite (CHAP) as compared with that to fluoride-containing carbonated hydroxyapatite (FCHAP). (B) Within the initial 3 min, rh174 bound more rapidly to carbonated hydroxyapatite as compared with fluoride-containing hydroxyapatite. (C) Total rh174 bound to fluoride-containing hydroxyapatite (98 ppm fluoride, 1101.6 ± 89.2 µg/m2; 2021 ppm fluoride, 1253.6 ± 101.5 µg/m2) was significantly greater than rh174 bound to hydroxyapatite (843.3 ± 68.3 µg/m2) or carbonated hydroxyapatite (858.5 ± 69.5 µg/m2), as shown by the Bradford analysis. N (for each sample) = 3. Results represent the means ± SD. *Significance, P < 0.01.

View larger version (13K): [in this window] [in a new window]   Figure 2. Effect of NaF on the affinity of rh174 to apatites. (A) Bradford assay showed that there was no significant difference in binding of rh174 to carbonated hydroxyapatite and fluoride-containing carbonated hydroxyapatite (98 ppm F–) in the presence of a physiological (0.1 ppm) concentration of fluoride. (B) Significantly less rh174 bound to carbonated hydroxyapatite in the presence of NaF (329.9 ± 32.2 µg/m2 and 274.2 ± 35.3 µg/m2 at 1000 ppm F and 2000 ppm F, respectively) as compared with the NaCl control (397.9 ± 12.1 µg/m2 and 378.9 ± 12.8 µg/m2 at 1000 ppm Cl and 2000 ppm Cl, respectively). (C) There was a similar reduction in the binding of rh174 to fluoride-containing carbonated hydroxyapatite (98 ppm F–) (380.7 ± 41.5 µg/m2 and 247.9 ± 24.4 µg/m2 at 1000 ppm F and 2000 ppm F, respectively) in the presence of NaF in solution (380.7 ± 41.5 µg/m2 and 247.9 ± 24.4 µg/m2 at 1000 ppm F and 2000 ppm F, respectively) as compared with the NaCl (control) at > 1000 ppm F in solution (601.8 ± 68.3 µg/m2 and 598.9 ± 32.2 µg/m2 at 1000 ppm Cl and 2000 ppm Cl, respectively). N (for each sample) = 3. Results represent the means ± SD. *Significance, P < 0.01. NS, no significance.

View larger version (14K): [in this window] [in a new window]   Figure 3. Effect of NaF pre-treatment on the affinity of rh174 to apatites. (A) Pre-incubation of fluoride-containing carbonated hydroxyapatite (98 ppm F–) with NaF (500–5000 ppm) showed that NaF pre-treatment inhibited rh174 binding in dose-dependent patterns (369.7 ± 23.6 µg/m2, 247.6 ± 5.3 µg/m2, and 243.3 ± 38.2 µg/m2 at 1000 ppm F–, 2000 ppm F–, and 5000 ppm F–, respectively) as compared with the NaCl-pre-treated controls (569.3 ± 45.2 µg/m2, 538.4 ± 23.6 µg/m2, and 564.7 ± 26.9 µg/m2 at 1000 ppm Cl–, 2000 ppm Cl–, and 5000 ppm Cl–, respectively). (B) Binding of rh174 on carbonated hydroxyapatite or fluoride-containing carbonated hydroxyapatite (98 ppm F–) was inhibited by NaF (2000 ppm) pre-treatment (270.8 ± 27.4 µg/m2 and 309.3 ± 31.9 µg/m2, respectively) as compared with the NaCl-pre-treated controls (388.2 ± 30.2 µg/m2 and 540.4 ± 33.2 µg/m2, respectively). N (for each sample) = 3. Results represent the means ± SD. *Significance, P < 0.01.

View larger version (67K): [in this window] [in a new window]   Figure 4. AFM images of rh174 on fluorapatite glass-ceramic substrates. (A) AFM imaging showed both ceramic glass and typical hexagonal (001) faces of fluorapatite crystals, which appear as a dark, hexagonal structure. (B) When the fluorapatite crystal embedded in glass ceramic was pre-treated with NaCl, rh174 bound to the glass surface as well as to the apatite. (C) When the disk was pre-treated with fluoride, less rh174 bound to the apatite surface (note appearance of the dark apatite shape, C), as compared with the NaCl-pre-treated disk.

Journal of Dental Research, Vol. 87, No. 1, 39-44 (2008)
DOI: 10.1177/154405910808700106


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