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Research Papers

Mechanical Stimulation Mediates Gene Expression in MC3T3 Osteoblastic Cells Differently in 2D and 3D Environments

[+] Author and Article Information
Matthew J. Barron

Department of Biomedical Engineering, Michigan Technological University, 1400 Townsend Drive, Houghton, MI 49849mjbarron@mtu.edu

Chung-Jui Tsai

Department of Genetics and School of Forestry and Natural Resources, University of Georgia, 111 Riverbend Road, Athens, GA 30602cjtsai@warnell.uga.edu

Seth W. Donahue1

Department of Biomedical Engineering, Michigan Technological University, 1400 Townsend Drive, Houghton, MI 49849swdonahu@mtu.edu

1

Corresponding author.

J Biomech Eng 132(4), 041005 (Mar 17, 2010) (6 pages) doi:10.1115/1.4001162 History: Received October 13, 2009; Revised January 20, 2010; Posted February 02, 2010; Published March 17, 2010; Online March 17, 2010

Successful bone tissue engineering requires the understanding of cellular activity in three-dimensional (3D) architectures and how it compares to two-dimensional (2D) architecture. We developed a perfusion culture system that utilizes fluid flow to mechanically load a cell-seeded 3D scaffold. This study compared the gene expression of osteoblastic cells in 2D and 3D cultures, and the effects of mechanical loading on gene expression in 2D and 3D cultures. MC3T3-E1 osteoblastlike cells were seeded onto 2D glass slides and 3D calcium phosphate scaffolds and cultured statically or mechanically loaded with fluid flow. Gene expression of OPN and FGF-2 was upregulated at 24 h and 48 h in 3D compared with 2D static cultures, while collagen 1 gene expression was downregulated. In addition, while flow increased OPN in 2D culture at 48 h, it decreased both OPN and FGF-2 in 3D culture. In conclusion, gene expression is different between 2D and 3D osteoblast cultures under static conditions. Additionally, osteoblasts respond to shear stress differently in 2D and 3D cultures. Our results highlight the importance of 3D mechanotransduction studies for bone tissue engineering applications.

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Copyright © 2010 by American Society of Mechanical Engineers
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Figures

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Figure 1

Bioreactor setup—see text for details

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Figure 2

OPN gene expression at 48 h in MC3T3 pre-osteoblasts after 1 h of oscillatory fluid flow (per day) followed by 23 h of static culture. Samples were loaded two times—once at the beginning of each 24 h period. Represented as means (n=5) with standard error bars.

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Figure 3

FGF-2 is downregulated at 48 h in 3D, but not 2D when osteoblasts are exposed to 1 h of oscillatory fluid flow (per day) followed by 23 h of static culture. Represented as means (n=5) with standard error bars.

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Figure 4

COX-2 is upregulated significantly at 48 h in 3D, but not 2D when osteoblasts are exposed to 1 h of oscillatory fluid flow each day. Represented as means (n=5) with standard error bars.

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