Abstract
Arterial restenosis associated with intimal hyperplasia is the major cause of long-term failure of vascular interventions. Endothelium injury and the proliferation and migration of smooth muscle cells (SMC) are key events in the development of intimal hyperplasia. The objectives of this study were to develop an ex vivo artery injury model for studying endothelial cell (EC) migration and to compare it with an in vitro co-culture arterial wall injury model in terms of the effect of flow on EC migration and its effect on SMC migration and proliferation. Our results demonstrated that shear flow improves reendothelialization in the injured area by promoting EC migration. The migration distance of ECs is much smaller in the arteries than in an in vitro cell culture model (3.57 ± 1.29 mm vs. 5.2 ± 1.4 cm, p < 0.001). SMC proliferation was significantly less in the EC intact and reendothelialization areas than in the EC denuded areas indicating that reendothelialization suppresses SMC proliferation. Our models provide a new approach to study techniques to enhance endothelium healing.
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Albuquerque, M. L., C. M. Waters, U. Savla, H. W. Schnaper, and A. S. Flozak. Shear stress enhances human endothelial cell wound closure in vitro. Am. J. Physiol. 279(1):H293–H302, 2000.
Bailey, S. R. Endovascular stents: update on stents in practice. J. Long Term Eff. Med. Implants 10(1–2):7–18, 2000.
Bardy, N., G. J. Karillon, R. Merval, J. L. Samuel, and A. Tedgui. Differential effects of pressure and flow on DNA and protein synthesis and on fibronectin expression by arteries in a novel organ culture system. Circ. Res. 77(4):684–694, 1995.
Chesler, N. C., D. N. Ku, and Z. S. Galis. Transmural pressure induces matrix-degrading activity in porcine arteries ex vivo. Am. J. Physiol. 277(5 Pt 2):H2002–H2009, 1999.
Chien, S. Mechanotransduction and endothelial cell homeostasis: the wisdom of the cell. Am. J. Physiol. Heart Circ. Physiol. 292(3):H1209–H1224, 2007.
Chien, S., S. Li, Y. T. Shiu, and Y. S. Li. Molecular basis of mechanical modulation of endothelial cell migration. Front. Biosci. 10:1985–2000, 2005.
Christen, T., M.-L. Bochaton-Piallat, P. Neuville, S. Rensen, M. Redard, G. van Eys, and G. Gabbiani. Cultured porcine coronary artery smooth muscle cells: a mew model with advanced differentiation. Circ. Res. 85(1):99–107, 1999.
Clerin, V., R. J. Gusic, J. O’Brien, P. M. Kirshbom, R. J. Myung, J. W. Gaynor, and K. J. Gooch. Mechanical environment, donor age, and presence of endothelium interact to modulate porcine artery viability ex vivo. Ann. Biomed. Eng. 30(9):1117–1127, 2002.
Conklin, B. S., D. S. Zhong, W. Zhao, P. H. Lin, and C. Chen. Shear stress regulates occludin and VEGF expression in porcine arterial endothelial cells. J. Surg. Res. 102(1):13–21, 2002.
Davis, N. P., H. C. Han, B. Wayman, and R. Vito. Sustained axial loading lengthens arteries in organ culture. Ann. Biomed. Eng. 33(7):867–877, 2005.
Edelman, E. R., and C. Rogers. Pathobiologic responses to stenting. Am. J. Cardiol. 81(7A):4E–6E, 1998.
El Hamamsy, I., L. M. Stevens, P. M. Vanhoutte, and L. P. Perrault. Injury of the coronary endothelium at implantation increases endothelial dysfunction and intimal hyperplasia after heart transplantation. J. Heart Lung Transplant. 24(3):251–258, 2005.
Gleason, R. L., E. Wilson, and J. D. Humphrey. Biaxial biomechanical adaptations of mouse carotid arteries cultured at altered axial extension. J. Biomech. 2006.
Guerin, P., F. Rondeau, G. Grimandi, M. F. Heymann, D. Heymann, P. Pillet, O. Al Habash, G. Loirand, P. Pacaud, and D. Crochet. Neointimal hyperplasia after stenting in a human mammary artery organ culture. J. Vasc. Res. 41(1):46–53, 2004.
Han, H. C., and D. N. Ku. Contractile responses in arteries subjected to hypertensive pressure in seven-day organ culture. Ann. Biomed. Eng. 29(6):467–475, 2001.
Han, H. C., D. N. Ku, and R. P. Vito. Arterial wall adaptation under elevated longitudinal stretch in organ culture. Ann. Biomed. Eng. 31(4):403–411, 2003.
Han, H. C., S. Marita, and D. N. Ku. Changes of opening angle in hypertensive and hypotensive arteries in 3-day organ culture. J. Biomech. 39(13):2410–2418, 2006.
Haudenschild, C. C., and S. M. Schwartz. Endothelial regeneration. II. Restitution of endothelial continuity. Lab. Invest. 41(5):407–418, 1979.
Hsu, P. P., S. Li, Y. S. Li, S. Usami, A. Ratcliffe, X. Wang, and S. Chien. Effects of flow patterns on endothelial cell migration into a zone of mechanical denudation. Biochem. Biophys. Res. Commun. 285(3):751–759, 2001.
Ku, D. N. Blood flow in arteries. Annu. Rev. Fluid Mech. 29(1):399–434, 1997.
Labadie, R. F., J. F. Antaki, J. L. Williams, S. Katyal, J. Ligush, S. C. Watkins, S. M. Pham, and H. S. Borovetz. Pulsatile perfusion system for ex vivo investigation of biochemical pathways in intact vascular tissue. Am. J. Physiol. 270(2 Pt 2):H760–H768, 1996.
Lee, Y. U., D. Drury-Stewart, R. P. Vito, and H. C. Han. Morphologic adaptation of arterial endothelial cells to longitudinal stretch in organ culture. J. Biomech. 41(15):3274–3277, 2008.
Lehoux, S., and A. Tedgui. Cellular mechanics and gene expression in blood vessels. J. Biomech. 36(5):631–643, 2003.
Lloyd-Jones, D., R. Adams, M. Carnethon, G. De Simone, T. B. Ferguson, K. Flegal, E. Ford, K. Furie, A. Go, K. Greenlund, N. Haase, S. Hailpern, M. Ho, V. Howard, B. Kissela, S. Kittner, D. Lackland, L. Lisabeth, A. Marelli, M. McDermott, J. Meigs, D. Mozaffarian, G. Nichol, C. O’Donnell, V. Roger, W. Rosamond, R. Sacco, P. Sorlie, R. Stafford, J. Steinberger, T. Thom, S. Wasserthiel-Smoller, N. Wong, J. Wylie-Rosett, and Y. Hong. Heart disease and stroke statistics—2009 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee. Circulation 119(3):e21–e181, 2009.
Losordo, D. W., J. M. Isner, and L. J. Diaz-Sandoval. Endothelial recovery: the next target in restenosis prevention. Circulation 107(21):2635–2637, 2003.
Mitra, A. K., and D. K. Agrawal. In stent restenosis: bane of the stent era. J. Clin. Pathol. 59(3):232–239, 2006.
Mohan, S., N. Mohan, A. J. Valente, and E. A. Sprague. Regulation of low shear flow-induced HAEC VCAM-1 expression and monocyte adhesion. Am. J. Physiol. Cell Physiol. 276(5):C1100–C1107, 1999.
Ong, A. T., J. Aoki, M. J. Kutryk, and P. W. Serruys. How to accelerate the endothelialization of stents. Arch. Mal. Coeur Vaiss. 98(2):123–126, 2005.
Palmaz, J. C. Intravascular stents in the last and the next 10 years. J. Endovasc. Ther. 11(Suppl 2):II200–II206, 2004.
Redmond, E. M., J. P. Cullen, P. A. Cahill, J. V. Sitzmann, S. Stefansson, D. A. Lawrence, and S. S. Okada. Endothelial cells inhibit flow-induced smooth muscle cell migration: role of plasminogen activator inhibitor-1. Circulation 103(4):597–603, 2001.
Rhee, K., and J. M. Tarbell. A study of the wall shear rate distribution near the end-to-end anastomosis of a rigid graft and a compliant artery. J. Biomech. 27(3):329–338, 1994.
Schwartz, R. S., and T. D. Henry. Pathophysiology of coronary artery restenosis. Rev. Cardiovasc. Med. 3(Suppl 5):S4–S9, 2002.
Scott, N. A. Restenosis following implantation of bare metal coronary stents: pathophysiology and pathways involved in the vascular response to injury. Adv. Drug Deliv. Rev. 58(3):358–376, 2006.
Soyombo, A. A., G. D. Angelini, and A. C. Newby. Neointima formation is promoted by surgical preparation and inhibited by cyclic nucleotides in human saphenous vein organ cultures. J. Thorac. Cardiovasc. Surg. 109(1):2–12, 1995.
Sprague, E. A. Endothelial and smooth muscle cell injury in contrived models and natural disease. In: Comprehensive Toxicology, edited by S. P. Bishop and W. D. Kerns. Pergamon: Oxford, 1997, pp. 213–240.
Sprague, E. A., J. Luo, and J. C. Palmaz. Human aortic endothelial cell migration onto stent surfaces under static and flow conditions. J. Vasc. Interv. Radiol. 8(1 Pt 1):83–92, 1997.
Sprague, E. A., J. Luo, and J. C. Palmaz. Endothelial cell migration onto metal stent surfaces under static and flow conditions. J. Long Term Eff. Med. Implants 10(1–2):97–110, 2000.
Vorp, D. A., D. G. Peters, and M. W. Webster. Gene expression is altered in perfused arterial segments exposed to cyclic flexure ex vivo. Ann. Biomed. Eng. 27(3):366–371, 1999.
Walpola, P. L., A. I. Gotlieb, M. I. Cybulsky, and B. L. Langille. Expression of ICAM-1 and VCAM-1 and monocyte adherence in arteries exposed to altered shear stress. Arterioscler. Thromb. Vasc. Biol. 15(1):2–10, 1995.
Walter, D. H., M. Cejna, L. Diaz-Sandoval, S. Willis, L. Kirkwood, P. W. Stratford, A. B. Tietz, R. Kirchmair, M. Silver, C. Curry, A. Wecker, Y. S. Yoon, R. Heidenreich, A. Hanley, M. Kearney, F. O. Tio, P. Kuenzler, J. M. Isner, and D. W. Losordo. Local gene transfer of phVEGF-2 plasmid by gene-eluting stents: an alternative strategy for inhibition of restenosis. Circulation 110(1):36–45, 2004.
Weintraub, W. S. The pathophysiology and burden of restenosis. Am. J. Cardiol. 100(5, Suppl 1):S3–S9, 2007.
Werner, N., S. Junk, U. Laufs, A. Link, K. Walenta, M. Bohm, and G. Nickenig. Intravenous transfusion of endothelial progenitor cells reduces neointima formation after vascular injury. Circ. Res. 93(2):e17–e24, 2003.
Willis, A. I., D. Pierre-Paul, B. E. Sumpio, and V. Gahtan. Vascular smooth muscle cell migration: current research and clinical implications. Vasc. Endovasc. Surg. 38(1):11–23, 2004.
Womersley, J. R. Method for the calculation of velocity, rate of flow and viscous drag in arteries when the pressure gradient is known. J. Physiol. 127(3):553–563, 1955.
Zampetaki, A., J. P. Kirton, and Q. Xu. Vascular repair by endothelial progenitor cells. Cardiovasc. Res. 78(3):413–421, 2008.
Acknowledgments
This work was supported by the Advanced Research Program from the Texas Higher Educational Coordinating Board through grant # 003659-0014-2006. It was also partially supported by an MBRS-SCORE pilot grant from the National Institute of Health through grant # S06GM008194 and grant # 0602834 from the National Science Foundation. The authors thank the Granzins at New Braunfels and Wiatrek at Poth, TX for generously providing the arteries for this work. The authors also thank Mr. Kurtis Johnson for his help in this work and Ms. Patricia Navarro for proofreading the manuscript.
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Associate Editor Julia E. Babensee oversaw the review of this article.
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Lee, YU., Luo, J., Sprague, E. et al. Comparison of Artery Organ Culture and Co-culture Models for Studying Endothelial Cell Migration and Its Effect on Smooth Muscle Cell Proliferation and Migration. Ann Biomed Eng 38, 801–812 (2010). https://doi.org/10.1007/s10439-009-9877-9
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DOI: https://doi.org/10.1007/s10439-009-9877-9