Journal
JOURNAL OF ORTHOPAEDIC RESEARCH
Volume 31, Issue 9, Pages 1360-1365Publisher
WILEY
DOI: 10.1002/jor.22374
Keywords
cell density; chondrogenesis; matrix elasticity; mesenchymal stem cells; osteogenesis
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Funding
- National Institutes of Health [HL104402, HL106579, HL108735]
- Innovation and Attracting Talents Program for College and University of China (111 Project) [B06023]
- Directorate For Engineering
- Div Of Civil, Mechanical, & Manufact Inn [1120795] Funding Source: National Science Foundation
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Human mesenchymal stem cells (hMSCs) can differentiate into various cell types, including osteogenic and chondrogenic cells. The matrix elasticity and cell seeding density are important factors in hMSCs differentiation. We cultured hMSCs at different seeding densities on polyacrylamide hydrogels with different stiffness corresponding to Young's moduli of 1.6 +/- 0.3 and 40 +/- 3.6kPa. The promotion of osteogenic marker expression by hard gel is overridden by a high seeding density. Cell seeding density, however, did not influence the chondrogenic marker expressions induced by soft gel. These findings suggest that interplays between cell-matrix and cell-cell interactions contribute to hMSCs differentiation. The promotion of osteogenic differentiation on hard matrix was shown to be mediated through the Ras pathway. Inhibition of Ras (RasN17) significantly decreased ERK, Smad1/5/8 and AKT activation, and osteogenic markers expression. However, constitutively active Ras (RasV12) had little effect on osteogenic marker expression, suggesting that the Ras pathways are necessary but not sufficient for osteogenesis. Taken together, our results indicate that matrix elasticity and cell density are important microenvironmental cues driving hMSCs proliferation and differentiation. (c) 2013 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 31:1360-1365, 2013
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