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Published in: Calcified Tissue International 4/2010

01-10-2010

Extracellular Matrix Produced by Osteoblasts Cultured Under Low-Magnitude, High-Frequency Stimulation is Favourable to Osteogenic Differentiation of Mesenchymal Stem Cells

Authors: Virginie Dumas, Benjamin Ducharne, Anthony Perrier, Carole Fournier, Alain Guignandon, Mireille Thomas, Sylvie Peyroche, Daniel Guyomar, Laurence Vico, Aline Rattner

Published in: Calcified Tissue International | Issue 4/2010

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Abstract

The effects of low-magnitude, high-frequency (LMHF) mechanical stimulation on osteoblastic cells are poorly understood. We have developed a system that generates very small (15–40 με), high-frequency (400 Hz, sine) deformations on osteoblast cultures (MC3T3-E1). We investigated the effects of these LMHF stimulations mainly on extracellular matrix (ECM) synthesis. The functional properties of this ECM after decellularization were evaluated on C3H10T1/2 mesenchymal stem cells (MSCs). LMHF stimulations were applied 20 min once daily for 1, 3, or 7 days in MC3T3-E1 culture (1, 3, or 7 dLMHF). Cell number and viability were not affected after 3 or 7 dLMHF. Osteoblast response to LMHF was assessed by an increase in nitric oxide secretion, alteration of the cytoskeleton, and focal contacts. mRNA expression for fibronectin, osteopontin, bone sialoprotein, and type I collagen in LMHF cultures were 1.8-, 1.6-, 1.5-, and 1.7-fold higher than controls, respectively (P < 0.05). In terms of protein, osteopontin levels were increased after 3 dLMHF and ECM organization was altered as shown by fibronectin topology after 7 dLMHF. After decellularization, 7 dLMHF-ECM or control ECM was reseeded with MSCs. Seven dLMHF-ECM improved early events such as cell attachment (2 h) and focal contact adhesion (6 h) and, later (16 h), modified MSC morphological parameters. After 5 days in multipotential medium, gene-expression changes indicated that 7 dLMHF-ECM promoted the expression of osteoblast markers at the expense of adipogenic marker. LMHF stimulations of osteoblasts are therefore efficient and sufficient to generate osteogenic matrix.
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Metadata
Title
Extracellular Matrix Produced by Osteoblasts Cultured Under Low-Magnitude, High-Frequency Stimulation is Favourable to Osteogenic Differentiation of Mesenchymal Stem Cells
Authors
Virginie Dumas
Benjamin Ducharne
Anthony Perrier
Carole Fournier
Alain Guignandon
Mireille Thomas
Sylvie Peyroche
Daniel Guyomar
Laurence Vico
Aline Rattner
Publication date
01-10-2010
Publisher
Springer-Verlag
Published in
Calcified Tissue International / Issue 4/2010
Print ISSN: 0171-967X
Electronic ISSN: 1432-0827
DOI
https://doi.org/10.1007/s00223-010-9394-8

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