Abstract
Human bone marrow-derived mesenchymal stem cells (MSCs) have potential applications for tissue engineering because they can differentiate into numerous cell lineages. Mechanical forces and mechanotransduction are important factors influencing cell responses, although such data are limited for MSCs. We investigated the gene expression response of integrin and extracellular matrix (ECM) genes in MSCs exposed to different magnitudes (1, 5, and 10 dyn/cm2) and durations (10 min, 1 h, and 24 h) of fluid flow-induced shear stress. Gene expression was examined using microarray and quantitative real-time RT-PCR analysis. In response to different magnitudes and durations of shear stress, we observed significant differential gene expression for two integrin genes: consistent up-regulation of integrin α2 subunit (ITGA2) [2- to 18-fold] and consistent down-regulation of integrin β8 subunit (ITGB8) [2- to 9-fold]. There was also evidence of ECM gene down-regulation, namely collagen type XI α1 (COL11A1), COL14A1, and COL21A1 (2- to 4-fold for each), and laminin α4 (LAMA4) [2- to 5-fold]. This indicates specific modulation of integrin gene expression in response to shear stress, supporting altered cell–ECM interactions and integrin-mediated mechanotransduction. These findings further our understanding of how mechanical stimuli regulate MSC behavior, which will be important in the development of mechanical conditioning strategies for tissue engineering.
Similar content being viewed by others
References
Bab, I., C. R. Howlett, B. A. Ashton, and M. E. Owen. Ultrastructure of bone and cartilage formed in vivo in diffusion chambers. Clin. Orthop. Relat. Res. Jul–Aug:243–254, 1984.
Bruder, S. P., D. J. Fink, and A. I. Caplan. Mesenchymal stem cells in bone development, bone repair, and skeletal regeneration therapy. J. Cell. Biochem. 56:283–294, 1994.
Burridge, K., and M. Chrzanowska-Wodnicka. Focal adhesions, contractility and signaling. Annu. Rev. Cell Dev. Biol. 12:463–518, 1996.
Bustin, S. A., V. Benes, T. Nolan, T. Nolan, and M. W. Pfaffl. Quantitative real-time RT-PCR—a perspective. J. Mol. Endocrinol. 34:597–601, 2005.
Calderwood, D. A. Integrin activation. J. Cell Sci. 117:657–666, 2004.
Cambier, S., D. Z. Mu, D. O’Connell, K. Boylen, W. Travis, W. H. Liu, V. C. Broaddus, and S. L. Nishimura. A role for the integrin alphavbeta8 in the negative regulation of epithelial cell growth. Cancer Res. 60:7084–7093, 2000.
Caplan, A. I. Mesenchymal stem cells. J. Orthop. Res. 9:641–650, 1991.
Chastain, S. R., A. K. Kundu, S. Dhar, J. W. Calvert, and A. J. Putnam. Adhesion of mesenchymal stem cells to polymer scaffolds occurs via distinct ECM ligands and controls their osteogenic differentiation. J. Biomed. Mater. Res. A 78:73–85, 2006.
Chen, K.-D., Y.-S. Li, M. Kim, S. Li, S. Yuan, S. Chien, and J. Y. Shyy. Mechanotransduction in response to shear stress. Roles of receptor tyrosine kinases, integrins and Shc. J. Biol. Chem. 274:18393–18400, 1999.
Chiquet, M. Regulation of extracellular matrix gene expression by mechanical stress. Matrix Biol. 18:417–426, 1999.
Davies, P. F., A. Robotewskyj, and M. L. Griem. Endothelial cell adhesion in real time. Measurements in vitro by tandem scanning confocal image analysis. J. Clin. Invest. 91:2640–2652, 1993.
Davies, P. F., A. Robotewskyj, and M. L. Griem. Quantitative studies of endothelial cell adhesion. Directional remodeling of focal adhesion sites in response to flow forces. J. Clin. Invest. 93:2031–2038, 1994.
Docheva, D., C. Popov, W. Mutschler, and M. Schieker. Human mesenchymal stem cells in contact with their environment: surface characteristics and the integrin system. J. Cell. Mol. Med. 11:21–38, 2007.
Friedenstein, A. J., R. K. Chailakhyan, and U. V. Gerasimov. Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers. Cell Tissue Kinet. 20:263–272, 1987.
Goessler, U. R., K. Biebeck, P. Bugert, T. Heller, H. Sadick, K. Hormann, and F. Riedel. In vitro analysis of integrin expression during chondrogenic differentiation of mesenchymal stem cells and chondrocytes upon dedifferentiation in cell culture. Int. J. Mol. Med. 17:301–307, 2006.
Goessler, U. R., P. Bugert, K. Bieback, J. Stern-Straeter, G. Bran, K. Hormann, and F. Riedel. Integrin expression in stem cells from bone marrow and adipose tissue during chondrogenic differentiation. Int. J. Mol. Med. 21:271–279, 2008.
Huang, H., R. D. Kamm, and R. T. Lee. Cell mechanics and mechanotransduction: pathways, probes, and physiology. Am. J. Physiol. Cell Physiol. 287:C1–C11, 2004.
Hynes, R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69:11–25, 1992.
Ishida, T., T. E. Peterson, N. L. Kovach, and B. C. Berk. MAP kinase activation by flow in endothelial cells. Circ. Res. 79:310–316, 1996.
Jalali, S., M. A. del Pozo, K.-D. Chen, H. Miao, Y.-S. Li, M. A. Schwartz, J. Y. Shyy, and S. Chien. Integrin-mediated mechanotransduction requires its dynamic interaction with specific extracellular matrix (ECM) ligands. Proc. Natl. Acad. Sci. USA 98:1042–1046, 2001.
Kanehisa, M., M. Araki, S. Goto, M. Hattori, M. Hirakawa, M. Itoh, T. Katayama, S. Kawashima, S. Okuda, T. Tokimatsu, and Y. Yamanishi. KEGG for linking genomes to life and the environment. Nucleic Acids Res. 36(Database issue):D480–D484, 2008.
Kanehisa, M., and S. Goto. KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res. 28:27–30, 2000.
Kanehisa, M., S. Goto, M. Hattori, K. F. Aoki-Kinoshita, M. Itoh, S. Kawashima, T. Katayama, M. Araki, and M. Hirakawa. From genomics to chemical genomics: new developments in KEGG. Nucleic Acids Res 34(Database issue):D354–D357, 2006.
Katsumi, A., W. Orr, E. Tzima, and M. A. Schwartz. Integrins in mechanotransduction. J. Biol. Chem. 279:12001–12004, 2004.
Kundu, A. K., and A. J. Putnam. Vitronectin and collagen I differentially regulate osteogenesis in mesenchymal stem cells. Biochem. Biophys. Res. Commun. 347:347–357, 2006.
Langholz, O., D. Rockel, C. Mauch, E. Kozlowska, I. Bank, T. Krieg, and B. Eckes. Collagen and collagenase gene expression in three-dimensional collagen lattices are differentially regulated by alpha 1 beta 1 and alpha 2 beta 1 integrins. J. Cell Biol. 131:1903–1915, 1995.
Majumdar, M. K., M. Keane-Moore, D. Buyaner, W. B. Hardy, M. A. Moorman, K. R. McIntosh, and J. D. Mosca. Characterization and functionality of cell surface molecules on human mesenchymal stem cells. J. Biomed. Sci. 10:228–241, 2003.
Mardon, H. J., J. Bee, K. von der Mark, and M. E. Owen. Development of osteogenic tissue in diffusion chambers from early precursor cells in bone marrow of adult rats. Cell Tissue Res. 250:157–165, 1987.
Mauch, C., C. B. Adelmann-Grill, A. Hatamochi, and T. Krieg. Collagenase gene expression in fibroblasts is regulated by a three-dimensional contact with collagen. FEBS Lett. 250:301–305, 1989.
Milner, R., X. Huang, J. Wu, S. Nishimura, R. Pytela, D. Sheppard, and C. Ffrench-Constant. Distinct roles for astrocyte alphavbeta5 and alphavbeta8 integrins in adhesion and migration. J. Cell Sci. 112:4271–4279, 1999.
Mizuno, M., R. Fujisawa, and Y. Kuboki. Type I collagen-induced osteoblastic differentiation of bone marrow cells mediated by collagen-alpha2beta1 integrin interaction. J. Cell. Physiol. 184:207–213, 2000.
Mizuno, M., and Y. Kuboki. Osteoblast-related gene expression of bone marrow cells during the osteoblastic differentiation induced by type I collagen. J. Biochem. 129:133–138, 2001.
Moyle, M., M. A. Napier, and J. W. McLean. Cloning and expression of a divergent integrin subunit beta 8. J. Biol. Chem. 266:19650–19658, 1991.
Mu, D., S. L. Cambier, L. Fjellbirkeland, J. L. Baron, J. S. Munger, H. Kawakatsu, D. Sheppard, V. C. Broaddus, and S. L. Nishimura. The integrin alpha v beta 8 mediates epithelial homeostasis through MT1-MMP-dependent activation of TGF-beta 1. J. Cell Biol. 157:493–507, 2002.
Nishimura, S. L., D. Sheppard, and R. Pytela. Integrin alpha v beta 8. Interaction with vitronectin and functional divergence of the beta 8 cytoplasmic domain. J. Biol. Chem. 269:28708–28715, 1994.
Pavalko, F. M., N. X. Chen, C. H. Turner, D. B. Burr, S. Atkinson, Y.-F. Hsieh, J. Qiu, and R. L. Duncan. Fluid shear-induced mechanical signaling in MC3T3-E1-E1 osteoblasts requires cytoskeleton-integrin interactions. Am. J. Physiol. 275:C1591–C1601, 1998.
Pittenger, M. F., A. M. Mackay, S. C. Beck, R. K. Jaiswal, R. Douglas, J. D. Mosca, M. A. Moorman, D. W. Simonetti, S. Craig, and D. R. Marshak. Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147, 1999.
Prockop, D. J. Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276:71–74, 1997.
Riddle, R. C., A. F. Taylor, D. C. Genetos, and H. J. Donahue. MAP kinase and calcium signaling mediate fluid flow-induced human mesenchymal stem cell proliferation. Am. J. Physiol. Cell Physiol. 290:C776–C784, 2006.
Schneider, G. B., R. Zaharias, and C. Stanford. Osteoblast integrin adhesion and signaling regulate mineralization. J. Dent. Res. 80:1540–1544, 2001.
Shyy, J. Y., and S. Chien. Role of integrins in endothelial mechanosensing of shear stress. Circ. Res. 91:769–775, 2002.
Staatz, W. D., K. F. Fok, M. M. Zutter, S. P. Adams, B. A. Rodriguez, and S. A. Santoro. Identification of a tetrapeptide recognition sequence for the alpha 2 beta 1 integrin in collagen. J. Biol. Chem. 266:7363–7367, 1991.
Takahashi, M., and B. C. Berk. Mitogen-activated protein kinase (ERK1/2) activation by shear stress and adhesion in endothelial cells. Essential role for a herbimycin-sensitive kinase. J. Clin. Invest. 98:2623–2631, 1996.
Tzima, E., M. A. del Pozo, S. J. Shattil, S. Chien, and M. A. Schwartz. Activation of integrins in endothelial cells by fluid shear stress mediates Rho-dependent cytoskeletal alignment. EMBO J. 20:4639–4647, 2001.
Wang, N., J. P. Butler, and D. E. Ingber. Mechanotransduction across the cell surface and through the cytoskeleton. Science 260:1124–1127, 1993.
Acknowledgments
We wish to thank Mr. Brian King for designing and manufacturing the slide chambers for cell lysis. This work was supported by a Research Grant (BB/D000548/1) from the Biotechnology and Biological Sciences Research Council (BBSRC), UK.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Glossop, J.R., Cartmell, S.H. Differential Gene Expression of Integrins Alpha 2 and Beta 8 in Human Mesenchymal Stem Cells Exposed to Fluid Flow. Cel. Mol. Bioeng. 2, 544–553 (2009). https://doi.org/10.1007/s12195-009-0083-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12195-009-0083-5