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The Effects of Mn2+ on the Proliferation, Osteogenic Differentiation and Adipogenic Differentiation of Primary Mouse Bone Marrow Stromal Cells

Abstract

The effects of Mn2+ on the proliferation, osteogenic and adipogenic differentiation of BMSCs were evaluated by employing MTT, ΔΨm, cell cycle, ALP activity, collagen production, ARS and oil red O stain assays. The results indicated that Mn2+ decreased the viability at most concentrations for 24 h, but the viability was increased with prolonging incubation time. Mn2+ at the concentrations of 1 × 10-7 and 1 × 10-6 mol/L decreased ΔΨm in the BMSCs for 48 h. Mn2+ induced G2/M phase cell cycle arrest at tested concentrations. On day 7 and 10, the effect of Mn2+ on the osteogenic differentiation depended on concentration, but it inhibited osteogenic differentiation at all tested concentrations for 14 d. The effect of Mn2+ on the synthesis of collagen of BMSCs depended on concentration for 7 d, but Mn2+ inhibited the synthesis of collagen at all tested concentrations for 10 d. On day 14, Mn2+ inhibited the formation of mineralized matrix nodules of BMSCs at all tested concentrations, the inhibitory effect turned to be weaker with prolonging incubation time. Mn2+ promoted the adipogenic differentiation of BMSCs at all tested concentrations for 10 d, but had no effect with prolonging incubation time. These findings suggested the effects of Mn2+ on the proliferation, osteogenic differentiation and adipogenic differentiation of BMSCs are very complicated, concentration and incubation time are key factors for switching the biological effects of Mn2+ from damage to protection.

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Abbreviations

ALP:

Alkaline phosphatase

ARS:

Alizarin red S

BMSCs:

Bone marrow stromal cells

BSP:

Bone sialo protein

DMSO:

Dimethylsulfoxide

MTT:

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

DMEM:

Dulbecco’s modified eagle’s medium

DLS:

Dynamic light scattering

KM:

Kun ming

ΔΨm:

Mitochondrial membrane potential

NBS:

Neonatal bovine serum

OD:

Optical density

OBs:

Osteoblasts

OS:

Osteogenic induction supplement

PTH:

Parathyroid hormone

PBS:

Phosphate buffer solution

PI:

Propidium iodide

Rh123:

Rhodamine 123

SD:

Standard deviation

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 20971034) and the Doctoral Program Foundation of Institutions of Higher Education of China (No. 20111301110004).

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Correspondence to Jinchao Zhang.

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Zhang, J., Zhang, Q., Li, S. et al. The Effects of Mn2+ on the Proliferation, Osteogenic Differentiation and Adipogenic Differentiation of Primary Mouse Bone Marrow Stromal Cells. Biol Trace Elem Res 151, 415–423 (2013). https://doi.org/10.1007/s12011-012-9581-8

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  • DOI: https://doi.org/10.1007/s12011-012-9581-8

Keywords

  • Osteoporosis
  • Mn2+
  • Proliferation
  • Differentiation
  • Bone marrow stromal cells