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TECHNICAL PAPERS

Fluid Shear Stress-Induced Alignment of Cultured Vascular Smooth Muscle Cells

[+] Author and Article Information
Ann A. Lee, Dionne A. Graham, Sheila Dela Cruz, Anthony Ratcliffe

Advanced Tissue Sciences, Inc., La Jolla, CA 92037

William J. Karlon

Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093

J Biomech Eng 124(1), 37-43 (Oct 02, 2001) (7 pages) doi:10.1115/1.1427697 History: Received April 09, 2000; Revised October 02, 2001
Copyright © 2002 by ASME
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References

Liu,  S. Q., 1998, “Influence of Tensile Strain on Smooth Muscle Cell Orientation in Rat Blood Vessels,” ASME J. Biomech. Eng., 120, No. 3 pp. 313–320.
Clark,  J. M., and Glagov,  S., 1985, “Transmural Organization of the Arterial Media,” Arteriosclerosis (Dallas), 5, No. 1, pp. 19–34.
Canham,  P. B., Finlay,  H. M., and Boughner,  D. R., 1997, “Contrasting Structure of the Saphenous Vein and Internal Mammary Artery Used as Coronary Bypass Vessels,” Cardiovasc. Res., 34, No. 3, pp. 557–567.
Peters,  M. W., Canham,  P. B., and Finlay,  H. M., 1983, “Circumferential Alignment of Muscle Cells in the Tunica Media of the Human Brain Artery,” Blood Vessels, 20, No. 5, pp. 221–233.
Wolinsky,  H., and Glagov,  S., 1964, “Structural Basis for the Static Mechanical Properties of the Aortic Media,” Circ. Res., 14, pp. 400–413.
Tranquillo,  R. T., Girton,  T. S., Bromberek,  B. A., Triebes,  T. G., and Mooradian,  D. L., 1996, “Magnetically Orientated Tissue-Equivalent Tubes: Application to a Circumferentially Orientated Media-Equivalent,” Biomaterials, 17, pp. 349–357.
L’Heureux,  N., Germain,  L., Labbe,  R., and Auger,  F. A., 1993, “In Vitro Construction of a Human Blood Vessel From Cultured Vascular Cells: A Morphologic Study,'In Vitro Construction of a Human Blood Vessel From Cultured Vascular Cells: A Morphologic Study,'’ J. Vasc. Surg., 17, pp. 499–509.
Taber,  L. A., 1998, “A Model for Aortic Growth Based on Fluid Shear and Fiber Stresses,” ASME J. Biomech. Eng., 120, No. 3, pp. 348–354.
Lee,  A. A., Delhaas,  T. D., Waldman,  L. K., MacKenna,  D. A., Villarreal,  F. J., and McCulloch,  A. D., 1996, “An Equibiaxial Strain System for Cultured Cells,” American Journal of Physiology (Cell Physiology), 271, pp. C1400–C1408.
Chiu,  J. J., Wang,  D. L., Chien,  S., Skalak,  R., and Usami,  S., 1998, “Effects of Disturbed Flow on Endothelial Cells,” ASME J. Biomech. Eng., 120, No. 1, pp. 2–8.
Papadaki,  M., Tilton,  R. G., Eskin,  S. G., and McIntire,  L. V., 1998, “Nitric Oxide Production by Cultured Human Aortic Smooth Muscle Cells: Stimulation by fluid Flow,” American Journal of Physiology (Heart Physiology), 274, pp. H616–H626.
Kanda,  K., Matsuda,  T., and Oda,  T., 1992, “Two-Dimensional Orientational Response of Smooth Muscle Cells to cyclic Stretching,” ASAIO J., 38, pp. M382–M385.
Zhao,  S., Suciu,  A., Ziegler,  T., Moore,  J. E., Burki,  E., Meister,  J. J., and Brunner,  H. R., 1995, “Synergistic Effects of Fluid Shear Stress and Cyclical Circumferential Stretch on Vascular Endothelial Cell Morphology and Cytoskeleton,” Arterioscler., Thromb., Vasc. Biol., 15, No. 10, pp. 1781–1786.
Dewey,  C. F. J., Bussolari,  S. R., Gimbrone,  M. A. J., and Davies,  P. F., 1981, “The Dynamic Response of Vascular Endothelial Cells to Fluid Shear Stress,” ASME J. Biomech. Eng., 103, pp. 177–185.
Levesque,  M. J., and Nerem,  R. M., 1985, “The Elongation and Orientation of Cultured Endothelial Cells in Response to Shear Stress,” ASME J. Biomech. Eng., 107, No. 4, pp. 341–7.
Malek,  A. M., and Izumo,  S., 1996, “Mechanism of Endothelial Cell Shape Change and Cytoskeletal Remodeling in Response to Fluid Shear Stress,” J. Cell. Sci., 109, pp. 713–726.
Galbraith,  C. G., Skalak,  R., and Chien,  S., 1998, “Shear Stress Induces Spatial Reorganization of the Endothelial Cell Cytoskeleton,” Cell Motil. Cytoskeleton, 40, No. 4, pp. 317–330.
Sterpetti,  A. V., Cucina,  A., Napoli,  F., Shafer,  H., Cavallaro,  A., and D’Angelo,  L. S., 1992, “Growth Factor Release by Smooth Muscle Cells is Dependent on Hemodynamic Factors,” Eur. J. Vasc. Surg., 6, No. 6, pp. 636–638.
Papadaki,  M., McIntire,  L. V., and Eskin,  S. G., 1996, “Effects of Shear Stress on the Growth Kinetics of Human Aortic Smooth Muscle Cells in Vitro,” Biotechnol. Bioeng., 50, pp. 555–561.
Rhoads,  D. N., Eskin,  S. G., and McIntire,  L. V., 2000, “Fluid Flow Releases Fibroblast Growth Factor-2 From Human Aortic Smooth Muscle Cells,” Arterioscler., Thromb., Vasc. Biol., 20, pp. 416–421.
Landeen, L. K., Zeltinger, J., Lee, A. A., Alexander, H. G., Graham, D. A., Ratcliffe, A., and Naughton, G. K. (2000) Stent Graft Update, Vossoughi, J., ed, pp. in press.
Frangos,  J. A., McIntire,  L. V., and Eskin,  S. G., 1988, “Shear Stress Induced Stimulation of Mammalian Cell Metabolism,” Biotechnol. Bioeng., 323, pp. 1053–1060.
Karlon,  W. J., Covell,  J. W., McCulloch,  A. D., Hunter,  J. J., and Omens,  J. H., 1998, “Automated Measurement of Myofiber Disarray in Transgenic Mice with Ventricular Expression of Rras,” Anat. Rec., 252, pp 612–625.
Karlon,  W. J., Hsu,  P. P., Li,  S., Chien,  S., McCulloch,  A. D., and Omens,  J. H., 1999, “Measurement of Orientation and Distribution of Cellular Alignment and Cytoskeletal Organization,” Ann. Biomed. Eng., 27, No. 6, pp. 712–729.
Chaudhuri,  B. B., Kundu,  P., and Sarkar,  N., 1993, “Detection and Gradation of Oriented Texture,” Pattern Recogn. Lett., 14, No. 2, pp. 147–153.
Palmer,  B. M., and Bizios,  R., 1997, “Quantitative Characterization of Vascular Endothelial Cell Morphology and Orientation Using Fourier Transform Analysis,” ASME J. Biomech. Eng., 119, pp. 159–165.
Buck,  R. C., 1983, “Behavior of Vascular Smooth Muscle Cells during repeated Stretching of the Substratum in Vitro,” Artherosclerosis 46, pp. 217–223.
Mills,  I., Cohen,  C. R., Kamal,  K., Li,  G., Shin,  T., Du,  W., and Sumpio,  B. E., 1997, “Strain Activation of Bovine Aortic Smooth Muscle Cell Proliferation and Alignment: Study of Strain Dependency and the Role of Protein Kinase A and C Signaling Pathways,” J. Cell Physiol., 170, No. 3, pp. 228–234.
Wang,  H., Ip,  W., Boissy,  R., and Grood,  E. S., 1995, “Cell Orientation Response to Cyclically Deformed Substrates: Experimental Validation of a Cell Model,” J. Biomech., 28, No. 12, pp. 1543–1552.
Feng,  Y., Yang,  J. H., Huang,  H., Kennedy,  S. P., Turi,  T. G., Thompson,  J. F., Libby,  P., and Lee,  R. T., 1999, “Transcriptional Profile of Mechanically Induced Genes in Human Vascular Smooth Muscle Cells,” Circ. Res., 85, No. 12, pp. 1118–1123.
Redmond,  E. M., Cahill,  P. A., and Sitzmann,  J. V., 1998, “Flow-Mediated Regulation of G-Protein Expression in Cocultured Vascular Smooth Muscle and Endothelial Cells,” Arterioscler., Thromb., Vasc. Biol., 18, No. 1, pp. 75–83.
Sterpetti,  A. V., Cucina,  A., Santoro D’Angelo,  L., Cardillo,  B., and Cavallaro,  A., 1992, “Response of arterial Smooth Muscle Cells to Laminar Flow,” J. Cardiovasc. Surg., 33, pp. 619–624.
Ueba,  J., Kawakami,  M., and Yaginuma,  T., 1997, “Shear Stress as an Inhibitor of Vascular Smooth Muscle Cell Proliferation. Role of Transforming Growth Factor-β1 and Tissue-Type Plasminogen Activator,” Arterioscler., Thromb., Vasc. Biol., 17, pp. 1512–1516.
Chapman,  G. B., Durante,  W., Hellums,  J. D., and Schafer,  A. I., 2000, “Physiological Cyclic Stretch Causes Cell Cycle Arrest in Cultured Vascular Smooth Muscle Cells,” American Journal of Physiology (Heart and Circulation Physiology), 278, No. 3, pp. H748–H754.
Helmlinger,  G., Geiger,  R. V., Schreck,  S., and Nerem,  R. M., 1991, “Effects of Pulsatile Flow on Cultured Vascular Endothelial Cell Morphology,” ASME J. Biomech. Eng., 113, No. 2, pp. 123–31.
Wang,  D. M., and Tarbell,  J. M., 1995, “Modeling Interstitial Flow in an Artery Wall Allows Estimation of Wall Shear Stress on Smooth Muscle Cells,” ASME J. Biomech. Eng., 117, No. 3, pp. 358–363.
Wagner,  C. T., Durante,  W., Christodoulides,  N., Hellums,  J. D., and Schafer,  A. I., 1997, “Hemodynamic Forces Induce the Expression of Heme Oxygenase in Cultured Vascular Smooth Muscle Cells,” J. Clin. Invest., 100, No. 3, pp. 589–596.
Barocas,  V. H., Girton,  T. S., and Tranquillo,  R. T., 1998, “Engineered Alignment in Media Equivalents: Magnetic Prealignment and Mandrel Compaction,” ASME J. Biomech. Eng., 120, No. 5, pp. 660–666.
L’Heureux,  N., Paquet,  S., Labbe,  R., Germain,  L., and Auger,  F. A., 1998, “A Completely Biological Tissue-Engineered Human Blood Vessel,” FASEB J., 12, No. 1, pp. 47–56.
Shinoka,  T., Shum-Tim,  D., Ma,  P. X., Tanel,  R. E., Isogai,  N., Langer,  R., Vacanti,  J. P., and Mayer,  J. E., 1998, “Creation of Viable Pulmonary Artery Autografts Through Tissue Engineering,” J. Thorac. Cardiovasc. Surg., 115, No. 3, pp. 536–45; discussions pp. 55–546.
Niklason,  L. E., Gao,  J., Abbott,  W. M., Hirschi,  K. K., Houser,  S., Marini,  R., and Langer,  R., 1999, “Functional Arteries Grown in Vitro,” Science, 284, pp. 489–493.

Figures

Grahic Jump Location
Fluid shear stress induces alignment in canine vascular SMC. Cells were cultured on fibronectin-coated glass slides under static conditions (A) or exposed to 20 dyn/cm2 shear stress for 48 hours (B). Fluorescent visualization of F-actin in cells cultured under static (C) or flow conditions (D). Arrows indicate the direction of fluid flow. An intensity gradient imaging technique was used to quantify the distribution and angular standard deviation of mean cell orientations for static (E) and flow (F) conditions.
Grahic Jump Location
Vascular SMC alignment is dependent on the magnitude of and exposure time to applied shear stress. Canine SMC were subjected to fluid shear stresses up to 20 dyn/cm2 for 0, 24, and 48 hours. Histograms of mean cell orientation angle are shown for 10, 15 and 20 dyn/cm2 . Mean cell angle measurements range from 0 to 180 deg, with 0 deg defined as the direction of fluid flow.
Grahic Jump Location
Summary of time- and magnitude-dependence of shear stress-induced SMC alignment. Cells subjected to 20 dyn/cm2 aligned more rapidly and to a higher degree than those cultured under lower flow conditions (*p=0.002 at 24 hours, compared with static control; at 48 hours, **p<0.0001, p=0.001, p=0.005, compared with 0, 10, and 15 dyn/cm2 , respectively). At 48 hours, alignment of SMC subjected to 10 and 15 dyn/cm2 were also significantly different compared with static control (p<0.05 for both conditions). Cells cultured in static and very low flow conditions (0 and 1 dyn/cm2 ) did not align in any preferred direction and remained randomly distributed throughout the time course.
Grahic Jump Location
Vascular SMC initiate the alignment process within hours of the onset of fluid flow. Images of cells subjected to fluid shear stress of 20 dyn/cm2 were acquired by time-lapse video over a 24-hour period. Images were digitized to quantify the increasing degree of cellular alignment, measured as the decreasing angular standard deviation of mean cell orientation.
Grahic Jump Location
Inhibition of intracellular calcium by quin-2 AM prevents the alignment of vascular SMC under fluid flow. After pretreatment with 10 μM quin-2 AM for 1 hour in static culture, cells were cultured under several conditions for 48 hours: “Static+Vehicle” (a), “Flow + Vehicle” (b), and “Flow+Quin-2 AM” (c). In the flow conditions, SMC were subjected to 20 dyn/cm2 shear stress with vehicle or 1 μM quin-2 AM in the circulating medium. Arrows indicate the direction of fluid flow.
Grahic Jump Location
Image analysis of the effects of quin-2 AM on flow-induced SMC alignment. Treatment with quin-2 AM was shown to significantly block flow-induced alignment, as measured by the angular standard deviation of mean cell angles (*p=0.007, compared with “Flow+Vehicle”; **p=0.001, compared with “Flow”).
Grahic Jump Location
Flow-induced SMC alignment was attenuated by disrupting actin filaments and microtubules. To examine the role of actin filaments in cell alignment, 40 nM cytochalasin D was used to pre-treat the cells for 1 hour (top row). Cells were subsequently cultured as static controls (a) or subjected to 20 dyn/cm2 for 48 hours with vehicle (b) or with 40 nM cytochalasin D in the circulating medium for 48 hours (c). To disrupt microtubules, cells were pretreated with 3 μM nocodazole for 1 hour (bottom row). Static controls (d) were compared with cells subjected to fluid flow for 48 hours under circulating DMSO vehicle (e) or nocodazole treatment (f).

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