Effect of fluoride was assessed on molars during and after mineralization. Two groups of 7 sheep each were dosed orally with 3.5 mg of fluoride/kg of body weight daily for 4 months (from 5 to 9 months after birth). Sheep of the first group were slaughtered immediately after fluoride administration; those of the second group were slaughtered 4 months later at the age of 13 months. Three control groups of 7 sheep each were slaughtered at 5 months (to determine the state of the teeth at the beginning of fluoride administration), and at 9 and 13 months. During fluoride administration, plasma fluoride concentration rapidly increased to about 0.50 micrograms/ml; after fluoride administration, it stabilized at 0.20 micrograms/ml in treated sheep, whereas controls had concentration of 0.10 micrograms/ml (P < 0.01). Parts of the molars that were in the process of mineralization during fluoride administration (mainly second molars) had thinning enamel, with pits, mainly close to the apex, marked decrease in hardness throughout the layer (< 100 Vickers U, compared with 240 Vickers U), and fluoride accumulation twice as high as that in controls (1,000 to 2,500 mg(kg [dry weight]). Fluoride accumulation was higher in dentine (2,700 to 4,200 mg/kg), but hardness was less affected. On parts of the molars that were already mineralized mostly, the first molar), changes in the appearance of enamel and cementum, decreased hardness (less important than in teeth during mineralization) affecting outer enamel more than inner enamel, high fluoride concentration (4,000 to 5,500 mg(kg [dry weight]) in outer enamel extending over 200 Km were observed. Thus, in sheep, fluoride has a substantial postsecretory effect that may be explained by a slower maturation phase of enamel in this species. Because molar wear is correlated to enamel hardness (dentine at the occlusal surface has low resistance--30 Vickers U), abnormal abrasion of molar teeth that have mineralized before and during fluoride intakes can be observed.

Effect of fluoride was assessed on molars during and after mineralization. Two groups of 7 sheep each were dosed orally with 3.5 mg of fluoride/kg of body weight daily for 4 months (from 5 to 9 months after birth). Sheep of the first group were slaughtered immediately after fluoride administration; those of the second group were slaughtered 4 months later at the age of 13 months. Three control groups of 7 sheep each were slaughtered at 5 months (to determine the state of the teeth at the beginning of fluoride administration), and at 9 and 13 months. During fluoride administration, plasma fluoride concentration rapidly increased to about 0.50 micrograms/ml; after fluoride administration, it stabilized at 0.20 micrograms/ml in treated sheep, whereas controls had concentration of 0.10 micrograms/ml (P < 0.01). Parts of the molars that were in the process of mineralization during fluoride administration (mainly second molars) had thinning enamel, with pits, mainly close to the apex, marked decrease in hardness throughout the layer (< 100 Vickers U, compared with 240 Vickers U), and fluoride accumulation twice as high as that in controls (1,000 to 2,500 mg(kg [dry weight]). Fluoride accumulation was higher in dentine (2,700 to 4,200 mg/kg), but hardness was less affected. On parts of the molars that were already mineralized mostly, the first molar), changes in the appearance of enamel and cementum, decreased hardness (less important than in teeth during mineralization) affecting outer enamel more than inner enamel, high fluoride concentration (4,000 to 5,500 mg(kg [dry weight]) in outer enamel extending over 200 Km were observed. Thus, in sheep, fluoride has a substantial postsecretory effect that may be explained by a slower maturation phase of enamel in this species. Because molar wear is correlated to enamel hardness (dentine at the occlusal surface has low resistance--30 Vickers U), abnormal abrasion of molar teeth that have mineralized before and during fluoride intakes can be observed.

Studies were undertaken to assess the chicken embryo and newly hatched chicken as models for studying the effects of bone-active agents. Initially, 1,25-dihydroxycholecaliferol (1,25[OH]2D3), sodium fluoride (NaF), parathyroid extract, epidermal growth factor, and prostaglandin E2, were tested for lethality over a broad dose range. One or 3 injections of 1,25(OH)2D3 into the yolk sac of chicken embryos resulted in death of embryos given greater than 0.1 ng/injection, whereas 0.01 ng was tolerated by the embryos. Administering 1,25(OH)2D3 intraperitoneally to newly hatched chickens as a single injection or weekly for 3 weeks resulted in no deaths at doses up to 50 ng. One or 3 IV injections of less than 400 micrograms were tolerated by the embryo. Giving chickens feed and water containing 2.4 g of NaF/kg was lethal but no deaths occurred when chickens were given feed containing less than 1.2 g of NaF/kg. Mortality associated with the administration of epidermal growth factor to embryos was inconsistent, in that death occurred in embryos given a single injection of greater than 250 ng, but no deaths occurred in embryos given 3 injections at similar doses. Parathyroid extract and prostaglandin F2 were not lethal when administered to embryos and chickens in a single-injection or multiple-injection regimen. Overall, lethality in chicken embryos given a particular agent reflected the dose of bone-active agent injected, rather than the number of injections. Three of the bone-active agents were selected to characterize their microscopic bone effects in chicken embryos and chickens. Administration of 1,25(OH)2D3 to embryos on day 14 at doses of 100, 10, 1, and 0.1 ng led to subperiosteal hyperosteoidosis in all 5 of the tibiotarsi examined from the high-dose (100 ng) group necropsied on day 18 of incubation. Three of 5 of the tibiotarsi from the 10-ng treatment group were similarly affected. Bone effects were noticed in chickens hatched from the aforementioned treatment groups or in chickens given 1,25(OH)2D3 intraperitoneally and examined at 3 and 6 weeks of age. Administration of NaF to chicken embryos on the 10, 12th, and 14th days of incubation via the IV route at doses of 160, 80, 40 and 20 micrograms/embryo led to subperiosteal hyperosteoidosis in tibiotarsi from 3 of 10 embryos (examined at 18 days of incubation) from the 2 high-dose groups. Tibiotarsi of chickens from this treatment group were microscopically normal at 3 weeks after hatching. When newly hatched chickens were given a diet containing NaF at dosages of 1.2 g/kg, 0.6 g/kg, and 0.3 g/kg, a dose-dependent increase in osteoid was seen at 3 and 6 weeks. In addition, cortical thinning and expansion of the medullary canal were observed only at 3 weeks. In contrast to the effects observed with 1,25(OH)2D3 and NaF, parathyroid extract caused no microscopic bone alterations when given to embryos or chickens. Overall, the bone alterations in the embryo were attributed to increased subperiosteal osteoid formation and defective mineralization. These findings were consistent with known effects of NaF and 1,25(OH)2D3 on bone, and they establish the chicken embryo as a sensitive model for studying bone-active agents.