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The Dual-Effects of LaCl3 on the Proliferation, Osteogenic Differentiation, and Mineralization of MC3T3-E1 Cells

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Abstract

A series of experimental methods including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, alkaline phosphatase (ALP) activity measurement, alizarin red S stain and measurement, quantitative real-time reverse transcriptase polymerase chain reaction, and Western blot analysis were employed to assess the effects of LaCl3 on the proliferation, osteogenic differentiation, and mineralization of a murine preosteoblast cell line MC3T3-E1 at cell and molecular levels. The results indicated that LaCl3 had dual effects on the proliferation, osteogenic differentiation, and mineralization of MC3T3-E1 cells. First, LaCl3 promoted the proliferation, osteogenic differentiation, and mineralization of MC3T3-E1 cells at lower concentrations, then had no effects and further turned to inhibit the proliferation, osteogenic differentiation, and mineralization of MC3T3-E1 cells with increasing concentrations. The expression of runt-related transcription factor 2 (Runx2), bone morphogenetic protein 2 (BMP2), ALP, bone sialoprotein (BSP), collagen I (Col I), and osteocalcin (OCN) genes was upregulated in the presence of 0.0001 and 0.1 μM LaCl3, but these genes were downregulated in the MC3T3-E1 cells treated with 1,000 μM LaCl3. In addition, the expression of BMP2, Runx2, and OCN proteins was promoted by LaCl3 at the concentration of 0.0001 μM, but these proteins were downregulated after 1,000 μM LaCl3 treatment. The results suggest that LaCl3 likely up- or downregulates the expression of Runx2, which subsequently up- or downregulates osteoblasts marker genes Col I and BMP2 at early stages and ALP and OCN at later stages of differentiation, thus causes to promote or inhibit the proliferation, osteogenic differentiation and mineralization of MC3T3-E1 cells. The results will be helpful for understanding the mechanisms of bone metabolism and application of lanthanum-based compounds in the future.

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Abbreviations

ARS:

Alizarin red S

ALP:

Alkaline phosphatase

α-MEM:

Alpha minimum essential medium

BMP2:

Bone morphogenetic protein 2

BSP:

Bone sialoprotein

Col I:

Collagen I

cDNA:

Complementary DNA

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

ECL:

Enhanced chemiluminescene

EDTA:

Ethylenediaminetetraacetic acid tetrasodium salt

FBS:

Fetal bovine serum

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

La:

Lanthanum

OD:

Optical density

OBs:

Osteoblasts

OCN:

Osteocalcin

OS:

Osteogenetic induction supplement

Q-PCR:

Quantitative real-time reverse transcriptase polymerase chain reaction

Runx2:

Runt-related transcription factor 2

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (no. 20971034), Natural Science Key Foundation of Hebei Province (no. B2009000161), Research Fund for the Doctoral Program of Higher Education of China (no.20111301110004).

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Authors and Affiliations

  1. College of Chemistry & Environmental Science, Chemical Biology Key Laboratory of Hebei Province, Hebei University, Baoding, People’s Republic of China

    Dandan Liu, Jinchao Zhang, Xiaolong Liu & Shuxiang Wang

  2. School of Medicine, Hebei University, Baoding, People’s Republic of China

    Guifang Wang

  3. Department of Biology and Chemistry, City University of Hong Kong, Hong Kong, China

    MengSu Yang

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  1. Dandan Liu
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  2. Jinchao Zhang
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  3. Guifang Wang
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  4. Xiaolong Liu
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  5. Shuxiang Wang
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  6. MengSu Yang
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Correspondence to Jinchao Zhang.

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Liu, D., Zhang, J., Wang, G. et al. The Dual-Effects of LaCl3 on the Proliferation, Osteogenic Differentiation, and Mineralization of MC3T3-E1 Cells. Biol Trace Elem Res 150, 433–440 (2012). https://doi.org/10.1007/s12011-012-9486-6

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  • DOI: https://doi.org/10.1007/s12011-012-9486-6

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