Promotion of osteogenesis through β-catenin signaling by desferrioxamine

doi:10.1016/j.bbrc.2008.03.092

Biochemical and Biophysical Research Communications

Volume 370, Issue 2, 30 May 2008, Pages 332-337

https://doi.org/10.1016/j.bbrc.2008.03.092 Get rights and content

Abstract

Desferrioxamine, an iron chelator with “hypoxia-mimetic” activity, promotes bone mineralization when used in aluminum-overloaded dialysis patients. However, the effect of desferrioxamine on osteoblastic differentiation from pluripotent mesenchymal stem cells (MSCs) has not been reported. In this study, pluripotent human MSCs and murine mesenchymal C3H10T1/2 cells were simultaneously treated with desferrioxamine and bone morphogenetic protein-2 (BMP2). In BMP2-treated MSCs, desferrioxamine levels of 15 μΜ were found to increase alkaline phosphatase (ALP) activity and calcium deposition, which were the markers of osteoblastic differentiation. These effects of desferrioxamine were accompanied by promoted phosphorylation of glycogen synthase kinase 3β (GSK-3β) and increased β-catenin protein content, a direct GSK-3β substrate. Knockdown of β-catenin by RNA interference eliminates this positive effect of desferrioxamine on ALP activity. Taken together, these data demonstrate that desferrioxamine plays a direct role in the differentiation of mesenchymal stem cells by activating β-catenin signaling cascades.

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Materials and methods

Cell culture and treatment. Human bone mesenchymal stem cells (hBMSCs) were isolated and expanded using the previously modified methods reported [10]. Bone marrow aspirates were obtained during routine orthopedic surgical procedures. Aspirates each amounting to 20 ml of bone marrow were harvested from three individual healthy male donors (A, 40 years old; B, 35 years old; and C, 28 years old) using a bone marrow biopsy needle inserted through the cortical bone. The aspirates were immediately

Nontoxic DFO increases BMP2-induced osteoblastic differentiation

We initially tried to determine the nontoxic concentrations of DFO on the adv-BMP2 transfected hBMSCs cells. The cells were transduced with adv-BMP2 or adv-β gal, and then were treated with various concentrations of DFO ranging from 5 μM to 100 μM for 2 days. Cell proliferation was determined by MTT assay. DFO, at a concentration ranging from 0 μM to 100 μM, inhibited cell growth in a dose-dependent manner (Fig. 1A). Cell proliferation rate was significantly lower when treated with DFO than when

Acknowledgments

This work was supported by the National Basic Research Program (973 Program) (Grant No. 2005CB522700), National Natural Science Foundation of China (Grant No. 30700402), and the Science and Technology Commission of Shanghai Municipality (Grant Nos. 05JC14034 and 07QA14062).

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Promotion of osteogenesis through β-catenin signaling by desferrioxamine

Abstract

Section snippets

Materials and methods

Nontoxic DFO increases BMP2-induced osteoblastic differentiation

Acknowledgments

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Bone

Biomaterials

Biochem. Biophys. Res. Commun.

Bone

Biochem. Biophys. Res. Commun.

Exp. Cell Res.

Biochem. Biophys. Res. Commun.

Effect of desferrioxamine and metals on the hydroxylases in the oxygen sensing pathway

FASEB J.