Simvastatin induces osteoblastic differentiation and inhibits adipocytic differentiation in mouse bone marrow stromal cells

doi:10.1016/S0006-291X(03)01408-6

Biochemical and Biophysical Research Communications

Volume 308, Issue 3, 29 August 2003, Pages 458-462

https://doi.org/10.1016/S0006-291X(03)01408-6 Get rights and content

Abstract

To clarify the mechanism of the stimulatory effect of statins on bone formation, we investigated the effect of simvastatin, a widely used statin, on osteoblastic and adipocytic differentiation in primary cultured mouse bone marrow stromal cells (BMSCs). Simvastatin treatment enhanced the expression level of mRNA for osteocalcin and protein for osteocalcin and osteopontin, and increased alkaline phosphatase activity significantly (p<0.05). After BMSCs were exposed to an adipocyte differentiation agonist, Oil Red O staining, fluorescence activated cell sorting, and decreased expression level of lipoprotein lipase mRNA showed that treatment with simvastatin significantly inhibits adipocytic differentiation compared to controls that did not receive simvastatin (p<0.05). Lastly, we found that simvastatin induces high expression of BMP₂ in BMSCs. These observations suggested that simvastatin acts on BMSCs to enhance osteoblastic differentiation and inhibits adipocytic differentiation; this effect is at least partially mediated by inducing BMP₂ expression in BMSCs.

Section snippets

Materials and methods

Cell cultures. Culture medium was DMEM, supplemented with 15% fetal bovine serum (FBS), 100 U/mL penicillin, 100 μg/mL streptomycin, 10 mM β-glycerophosphate sodium, and 50 μg/mL ascorbate acid sodium (Sigma). Female BALB-C mice (8 weeks) were killed by cervical dislocation, metaphyses of femurs and tibias were cut aseptically, diaphysis cavities were repeatedly flushed with culture medium, and bone marrow cells were collected and plated. Culture medium was replaced every 2–3 days, red blood cells

Effect of simvastatin on osteoblastogenesis of BMSCs

After BMSCs were treated with simvastatin for 72 h, expression level of mRNA for OCN (Fig. 1A) and protein for OCN and OPN increased in a concentration-dependent manner (Fig. 2). BMSCs exhibited increased ALP activity in response to simvastatin; ALP activity increased 2–3-fold above basal levels (Fig. 3). Taken together, results showed that treatment of BMSCs with simvastatin induces osteoblastogenesis of BMSCs.

Effect of simvastatin on adipogenesis of BMSCs

Results showed that treatment of simvastatin inhibits adipocytic differentiation of

Discussion

Studies have demonstrated that BMSCs can differentiate into multiple cell types, including osteoblasts, myoblasts, chondrocytes, and adipocytes [19], [20]. Adipocytic and osteogenic cells are reciprocal cell types that are dominant in marrow. It is thought that changes in the ratios of these cells are involved in bone volume decreases associated with osteoporosis, such as that seen in cases of ovariectomy [1], immobilization [2], treatment with glucocorticoids [3], and age-related osteopenia [4]

Acknowledgements

The authors thank Professor L.W. Fisher for generous gift of primary antibody. We also thank Dr. Ming Gong and Dr. Shuyan Li for their technical assistance on molecular biology.

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Citation Excerpt :
It suppressed the TNF-α-to-Ras/Rho/MAPK pathway while enhancing BMP-2, alkaline phosphatase (ALP) activity, osteopontin (OPN) and osteocalcin (OCN) expression level during osteogenesis [20,24,25]. Although many reports have demonstrated the important role of SIM in degenerative bone disease treatment [26–28], a significant challenge remains in selecting a suitable carrier which can not only ensure long-term controllable drug release of SIM but also provide sufficient mechanical support. Although MBGs have been used effectively in delivering drugs, such as dexamethasone (DEX) [14] and dimethyloxallyl glycine (DMOG) [29], to enhance osteogenesis in bone regeneration [19], up to now, very little attention has been paid to MBGs as a carrier of SIM, let alone nanotubular MBG scaffolds.
Nanofibrous scaffolds with drug delivery functions are highly demanded in promoting tissue regeneration. Herein, we report a novel type of three-dimensional (3D) mesoporous bioactive glass (MBG) scaffolds. Such MBG scaffolds are consisted of 3D MBG nanotubes with highly ordered mesopores. The MBG nanotubes were prepared by template-assisted sol-gel procedure using bacterial cellulose (BC) as template and the mesopores were obtained using nonionic block copolymer (P123) as structure-directing agent. To improve bone regeneration capability, we loaded the nanotubular MBG scaffolds with simvastatin (SIM), as a representative small molecule, rather than commonly used growth factors which are large-sized, expensive, and fragile and may lead to serious side effects. By seeding human bone marrow stromal cells (hBMSCs) on the SIM-loaded nanotubular MBG scaffolds, the biocompatibility and bone regeneration were assessed by observing cell morphology, measuring cell viability and osteogenic-related gene expression. We found that SIM was successfully loaded into the nanotubular MBG scaffolds with a high loading efficiency of approximately 50%, and the nanotubular MBG scaffolds exhibited a sustained release of SIM. More importantly, the SIM-loaded nanotubular MBG scaffolds exhibited improved proliferation, ALP activity and osteogenic-related gene expression of hBMSCs over the bare MBG scaffold. We believe that the SIM-loaded nanotubular MBG scaffolds have great potential application in bone tissue regeneration.

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View full text

Simvastatin induces osteoblastic differentiation and inhibits adipocytic differentiation in mouse bone marrow stromal cells

Abstract

Section snippets

Materials and methods

Effect of simvastatin on osteoblastogenesis of BMSCs

Effect of simvastatin on adipogenesis of BMSCs

Discussion

Acknowledgements

Bone

Eur. J. Pharmacol.

FEBS Lett.

Exp. Cell Res.

Bone

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

Bone marrow fat content in relation to bone remodeling and serum chemistry in intact and ovariectomized dogs

Calcif. Tissue Int.

Marrow changes in paraplegic patients

Calcif. Tissue Int.

Fat cell changes as a mechanism of avascular necrosis in the femoral head in cortisone-treated rabbits

J. Bone Jt. Surg. A

Age related variations of fat tissue fraction in normal bone marrow depend both on size and number of adipocytes: a stereological study

Exp. Hematol.

Evidence for an inverse relationship between the differentiation of adipocytic and osteogenic cells in rat marrow stromal cell cultures

J. Cell Sci.