title: 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.
authors.affiliation: Biological Systems Engineering Laboratory, Department of Chemical Engineering, Imperial College London, London, United Kingdom.
authors.forename: Joseph
authors.initials: J
authors.lastname: Pagkalos
authors.forename: Jae Min
authors.initials: JM
authors.lastname: Cha
authors.forename: Yunyi
authors.initials: Y
authors.lastname: Kang
authors.forename: Manolis
authors.initials: M
authors.lastname: Heliotis
authors.forename: Eleftherios
authors.initials: E
authors.lastname: Tsiridis
authors.forename: Athanasios
authors.initials: A
authors.lastname: Mantalaris
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000880:Anthraquinones; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D056945:Hep G2 Cells; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D051379:Mice; D010012:Osteogenesis / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D019821:Simvastatin / Q000494:pharmacology*
chemical_list: D000880:Anthraquinones; D055396:Bone Morphogenetic Protein 2; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; C010078:alizarin; D019821:Simvastatin; D000469:Alkaline Phosphatase