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Fluoride Affects Calcium Homeostasis and Osteogenic Transcription Factor Expressions Through L-type Calcium Channels in Osteoblast Cell Line

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Abstract

Osteoblast L-type voltage-dependent calcium channels (VDCC) play important roles in maintaining intracellular homeostasis and influencing multiple cellular processes. In particular, they contribute to the activities and functions of osteoblasts (OBs). In order to study how L-type VDCC modulate calcium ion (Ca2+) homeostasis and the expression of osteogenic transcription factors in OBs exposed to fluoride, MC3T3-E1 cells were exposed to a gradient of concentrations of fluoride (0, 2.0, 5.0, 10.0 mg/L) in combination with 10 μM nifedipine, a specific inhibitor of VDCC, for 48 h. We examined messenger RNA (mRNA) and protein levels of Cav1.2, the main subunit of VDCC, and c-fos, c-jun, runt-related transcription factor 2 (Runx2), osterix (OSX), and intracellular free Ca2+ ([Ca2+]i) concentrations in MC3T3-E1 cells. Our results showed that [Ca2+]i levels increased in a dose-dependent manner with increase in concentration of fluoride. Meantime, results indicated that lower concentrations of fluoride (less than 5 mg/L, especially 2 mg/L) can lead to high expression of Cav1.2 and enhance osteogenic function, while high concentration of fluoride (10 mg/L) can induce decreased Cav1.2 and osteogenic transcriptional factors in MC3T3E1 cells exposed to fluoride. However, the levels of [Ca2+]i, Cav1.2, c-fos, c-jun, Runx2, and OSX induced by fluoride were significantly altered and even reversed in the presence of nifedipine. These results demonstrate that L-type calcium channels play a crucial role in Ca2+ homeostasis and they affect the expression of osteogenic transcription factors in fluoride-treated osteoblasts.

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Acknowledgments

This work was supported by a grant for skeletal fluorosis research from the National Natural Science Foundation of China (81172608).

Conflict of Interest

All authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Rehabilitation Medicine of the Second Hospital Norman Bethune, JiLin University, Changchun, 130041, People’s Republic of China

    Xiao-qin Duan & Huan Wang

  2. Institute of Endemic Disease, Jilin University, 1163 Xinmin Street, Changchun, 130021, People’s Republic of China

    Zhi-tao Zhao, Xiu-yun Zhang, Da-wei Liu, Guang-sheng Li & Ling Jing

  3. The First Hospital Norman Bethune, JiLin University, Changchun, 130021, People’s Republic of China

    Ying Wang

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  1. Xiao-qin Duan
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  2. Zhi-tao Zhao
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  4. Ying Wang
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  5. Huan Wang
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Correspondence to Ling Jing.

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Duan, Xq., Zhao, Zt., Zhang, Xy. et al. Fluoride Affects Calcium Homeostasis and Osteogenic Transcription Factor Expressions Through L-type Calcium Channels in Osteoblast Cell Line. Biol Trace Elem Res 162, 219–226 (2014). https://doi.org/10.1007/s12011-014-0118-1

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  • DOI: https://doi.org/10.1007/s12011-014-0118-1

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