Abstract
MSCs (mesenchymal stem cells) may be promising seed cells for tissue regeneration because of their self-renewal and multi-differentiation potential. Shh (sonic hedgehog) is involved in the skeletal formation during embryo development and skeletal regeneration. However, how Shh regulates the biological characteristics of BM-MSCs (bone marrow-derived MSCs) is poorly understood. We have investigated the effect of rShh-N (recombinant N-terminal Shh) on the proliferation and osteogenic differentiation of rBM-MSCs (rat BM-MSCs) in vitro. rBM-MSCs were treated with rShh-N at concentrations up to 200 ng/ml. Proliferation and colony-forming ability of rBM-MSCs were increased in a dose-dependent manner. rShh-N increased the ratio of cells in S and G2/M phase, as well as the number of Ki-67+ cells. In addition, ALP (alkaline phosphatase) activity and matrix mineralization were enhanced by 200 ng/ml rShh-N. Real-time PCR showed that rShh-N (200 ng/ml) up-regulated the expression of genes encoding Cbfa-1 (core-binding factor α1), osteocalcin, ALP and collagen type I in rBM-MSCs. This information reveals some potential of rShh-N in the therapeutics of bone-related diseases.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alkaline Phosphatase / genetics
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Alkaline Phosphatase / metabolism
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Animals
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Bone Marrow Cells / cytology
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Bone Marrow Cells / drug effects*
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Bone Regeneration / drug effects
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Bone Regeneration / physiology
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Cell Cycle / drug effects
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Cell Differentiation / drug effects
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Cell Proliferation / drug effects
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Cells, Cultured
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Collagen Type I / genetics
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Collagen Type I / metabolism
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Core Binding Factor Alpha 1 Subunit / genetics
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Core Binding Factor Alpha 1 Subunit / metabolism
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Core Binding Factors / genetics
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Core Binding Factors / metabolism
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Dose-Response Relationship, Drug
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Gene Expression Regulation, Developmental / drug effects
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Hedgehog Proteins / pharmacology
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Hedgehog Proteins / physiology*
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Ki-67 Antigen / genetics
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Ki-67 Antigen / metabolism
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Mesenchymal Stem Cells / cytology
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Mesenchymal Stem Cells / drug effects*
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Osteocalcin / genetics
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Osteocalcin / metabolism
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Osteogenesis / drug effects*
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Osteogenesis / physiology
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Rats
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Recombinant Proteins / pharmacology
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Signal Transduction / drug effects
Substances
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Collagen Type I
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Core Binding Factor Alpha 1 Subunit
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Core Binding Factors
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Hedgehog Proteins
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Ki-67 Antigen
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Recombinant Proteins
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Runx2 protein, rat
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Shh protein, rat
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Osteocalcin
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Alkaline Phosphatase