Simvastatin induces osteogenic differentiation of murine embryonic stem cells

Abstract

Statins are potent inhibitors of cholesterol synthesis. Several statins are available with different molecular and pharmacokinetic properties. Simvastatin is more lipophilic than pravastatin and has a higher affinity to phospholipid membranes than atorvastatin, allowing its passive diffusion through the cell membrane. In vitro studies on bone marrow stromal cells, osteoblast-like cells, and embryonic stem cells have shown statins to have cholesterol-independent anabolic effects on bone metabolism; alas, statins were supplemented in osteogenic medium, which does not facilitate elucidation of their potential osteoinductive properties. Embryonic stem cells (ESCs), derived from the inner cell mass of the blastocyst, are unique in that they enjoy perpetual self-proliferation, are pluripotent, and are able to differentiate toward all the cellular lineages composing the body, including the osteogenic lineage. Consequently, ESCs represent a potentially potent cell source for future clinical cellular therapies of various bone diseases, even though there are several hurdles that still need to be overcome. Herein we demonstrate, for the first time to our knowledge, that simvastatin induces murine ESC (mESC) differentiation toward the osteogenic lineage in the absence of osteoinductive supplements. Specifically, we found that a simvastatin concentration in the micromolar range and higher was toxic to the cells and that an effective concentration for osteoinduction is 0.1 nM, as shown by increased alizarin red staining as well as increased osteocalcin and osetrix gene expression. These results suggest that in the future, lipophilic simvastatin may provide a novel pharmacologic agent for bone tissue engineering applications. ß

Simvastatin Maintains Osteoblastic Viability While Promoting Differentiation by Partially Regulating the Expressions of Estrogen Receptors α

“…Doses employed in this study were similar to those reported previously, since simvastatin dosed at 10 À6 to 10 À9 M had already been tested on MC3T3-E1 cells by other investigators [4,11]. It should be noted that the use of higher doses of statins was previously shown to yield a negative effect on cell viability [12].…”

“…However, there could be variations in doses to achieve the maximal effect, depending on the system model, the stage of differentiation of osteoblasts, and the culturing time period [2,14]. Moreover, simvastatin can induce murine embryonic stem cell (mESC) differentiation toward the osteogenic lineage in the absence of osteoinductive supplements, according a recently published report [12].…”

Controlled release of simvastatin from biodegradable hydrogels promotes odontoblastic differentiation

“…In this context, the activation and differentiation of DPSC may function positively for dentin regeneration. Recently, it has been reported that simvastatin (ST) has the potential to induce osteoblastic bone formation or enhance the expression of bone morphogenetic protein (BMP)-2 for several osteogenic cells [6][7][8] . Some investigators have demonstrated that DPSC can be differentiated into odontoblasts by BMP-2, resulting in the promotion of dentin formation [9][10][11] .…”

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

“…The majority of osteogenic protocols for ESCs direct cell differentiation by including factors in the culture medium, such as b-glycerophosphate (BGP), ascorbate, dexamethasone, simvastatin, retinoic acid, vitamin D 3 , and bone morphogenic proteins [3,[22][23][24][25][26][27][28][29][30]. Although traditional osteogenic differentiation strategies for ESCs leads to the formation of bone nodules and expression of osteogenic markers, little research has compared this to the in vitro differentiation of osteoblasts.…”