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Bone response to fluoride exposure is influenced by genetics.

Kobayashi CA, Leite AL, Peres-Buzalaf C, Carvalho JG, Whitford GM, Everett ET, Siqueira WL, Buzalaf MA - PLoS ONE (2014)

Bottom Line: Genetic factors influence the effects of fluoride (F) on amelogenesis and bone homeostasis but the underlying molecular mechanisms remain undefined.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil.

ABSTRACT
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.

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Related in: MedlinePlus

Western blot analysis of collagen type I.(A) Representative immunoblot showing collagen type I levels (upper) in A/J and 129P3/J mice after treatment with 0, 10 or 50 ppmF in drinking water for 8 weeks. The level of β-tubulin was used as control of sample loading (lower). (B) Values are mean ± SD of arbitrary values of four independent experiments quantified by densitometry analysis using the Image J Software (NIH Image). Arbitrary values from control A/J were standardized as 1. **Represents significant differences between strains for each treatment (p<0.01). No statistical differences were found for each strain after receiving F.
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pone-0114343-g005: Western blot analysis of collagen type I.(A) Representative immunoblot showing collagen type I levels (upper) in A/J and 129P3/J mice after treatment with 0, 10 or 50 ppmF in drinking water for 8 weeks. The level of β-tubulin was used as control of sample loading (lower). (B) Values are mean ± SD of arbitrary values of four independent experiments quantified by densitometry analysis using the Image J Software (NIH Image). Arbitrary values from control A/J were standardized as 1. **Represents significant differences between strains for each treatment (p<0.01). No statistical differences were found for each strain after receiving F.

Mentions: Analysis of collagen type I expression in mouse femur corroborated the quantitative proteomics data, confirming that there were no statistically significant differences among the F treatments for either strain (F = 1.45, p = 0.262) but there was a significant increase in the level of collagen type I in 129P3/J compared to A/J strain (F = 9.54, p = 0.006) for all F treatment groups (Fig. 5A,B). However, it is worth mentioning that quantitative proteomic analysis identified a significant difference between the strains only for control group.


Bone response to fluoride exposure is influenced by genetics.

Kobayashi CA, Leite AL, Peres-Buzalaf C, Carvalho JG, Whitford GM, Everett ET, Siqueira WL, Buzalaf MA - PLoS ONE (2014)

Western blot analysis of collagen type I.(A) Representative immunoblot showing collagen type I levels (upper) in A/J and 129P3/J mice after treatment with 0, 10 or 50 ppmF in drinking water for 8 weeks. The level of β-tubulin was used as control of sample loading (lower). (B) Values are mean ± SD of arbitrary values of four independent experiments quantified by densitometry analysis using the Image J Software (NIH Image). Arbitrary values from control A/J were standardized as 1. **Represents significant differences between strains for each treatment (p<0.01). No statistical differences were found for each strain after receiving F.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263599&req=5

pone-0114343-g005: Western blot analysis of collagen type I.(A) Representative immunoblot showing collagen type I levels (upper) in A/J and 129P3/J mice after treatment with 0, 10 or 50 ppmF in drinking water for 8 weeks. The level of β-tubulin was used as control of sample loading (lower). (B) Values are mean ± SD of arbitrary values of four independent experiments quantified by densitometry analysis using the Image J Software (NIH Image). Arbitrary values from control A/J were standardized as 1. **Represents significant differences between strains for each treatment (p<0.01). No statistical differences were found for each strain after receiving F.
Mentions: Analysis of collagen type I expression in mouse femur corroborated the quantitative proteomics data, confirming that there were no statistically significant differences among the F treatments for either strain (F = 1.45, p = 0.262) but there was a significant increase in the level of collagen type I in 129P3/J compared to A/J strain (F = 9.54, p = 0.006) for all F treatment groups (Fig. 5A,B). However, it is worth mentioning that quantitative proteomic analysis identified a significant difference between the strains only for control group.

Bottom Line: Genetic factors influence the effects of fluoride (F) on amelogenesis and bone homeostasis but the underlying molecular mechanisms remain undefined.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, SP, Brazil.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus