0
Research Papers

A Shearing-Stretching Device That Can Apply Physiological Fluid Shear Stress and Cyclic Stretch Concurrently to Endothelial Cells

[+] Author and Article Information
Daphne Meza, Louie Abejar, David A. Rubenstein

Department of Biomedical Engineering,
Stony Brook University,
Stony Brook, NY 11794

Wei Yin

Department of Biomedical Engineering,
Stony Brook University,
Stony Brook, NY 11794
e-mail: wei.yin@stonybrook.edu

1Corresponding author.

Manuscript received June 12, 2015; final manuscript received January 10, 2016; published online February 5, 2016. Assoc. Editor: Kristen Billiar.

J Biomech Eng 138(3), 031007 (Feb 05, 2016) (8 pages) Paper No: BIO-15-1297; doi: 10.1115/1.4032550 History: Received June 12, 2015; Revised January 10, 2016

Endothelial cell (EC) morphology and functions can be highly impacted by the mechanical stresses that the cells experience in vivo. In most areas in the vasculature, ECs are continuously exposed to unsteady blood flow-induced shear stress and vasodilation-contraction-induced tensile stress/strain simultaneously. Investigations on how ECs respond to combined shear stress and tensile strain will help us to better understand how an altered mechanical environment affects EC mechanotransduction, dysfunction, and associated cardiovascular disease development. In the present study, a programmable shearing and stretching device that can apply dynamic fluid shear stress and cyclic tensile strain simultaneously to cultured ECs was developed. Flow and stress/strain conditions in the device were simulated using a fluid structure interaction (FSI) model. To characterize the performance of this device and the effect of combined shear stress–tensile strain on EC morphology, human coronary artery ECs (HCAECs) were exposed to concurrent shear stress and cyclic tensile strain in the device. Changes in EC morphology were evaluated through cell elongation, cell alignment, and cell junctional actin accumulation. Results obtained from the numerical simulation indicated that in the “in-plane” area of the device, both fluid shear stress and biaxial tensile strain were uniform. Results obtained from the in vitro experiments demonstrated that shear stress, alone or combined with cyclic tensile strain, induced significant cell elongation. While biaxial tensile strain alone did not induce any appreciable change in EC elongation. Fluid shear stress and cyclic tensile strain had different effects on EC actin filament alignment and accumulation. By combining various fluid shear stress and cyclic tensile strain conditions, this device can provide a physiologically relevant mechanical environment to study EC responses to physiological and pathological mechanical stimulation.

FIGURES IN THIS ARTICLE
<>
Copyright © 2016 by ASME
Your Session has timed out. Please sign back in to continue.

References

Cunningham, K. S. , and Gotlieb, A. I. , 2004, “ The Role of Shear Stress in the Pathogenesis of Atherosclerosis,” Lab. Invest., 85(1), pp. 9–23. [CrossRef]
Chiu, J.-J. , Usami, S. , and Chien, S. , 2009, “ Vascular Endothelial Responses to Altered Shear Stress: Pathologic Implications for Atherosclerosis,” Ann. Med., 41(1), pp. 19–28. [CrossRef] [PubMed]
Alexander, R. W. , 1995, “ Hypertension and the Pathogenesis of Atherosclerosis Oxidative Stress and the Mediation of Arterial Inflammatory Response: A New Perspective,” Hypertension, 25(2), pp. 155–161. [CrossRef] [PubMed]
Frangos, S. G. , Gahtan, V. , and Sumpio, B. , 1999, “ Localization of Atherosclerosis: Role of Hemodynamics,” Arch. Surg., 134(10), pp. 1142–1149. [CrossRef] [PubMed]
Dewey, C. F., Jr. , Bussolari, S. R. , Gimbrone, M. A., Jr. , and Davies, P. F. , 1981, “ The Dynamic Response of Vascular Endothelial Cells to Fluid Shear Stress,” ASME J. Biomech. Eng., 103(3), pp. 177–185. [CrossRef]
Hunt, B. J. , and Jurd, K. M. , 1998, “ Endothelial Cell Activation. A Central Pathophysiological Process,” BMJ, 316(7141), pp. 1328–1329. [CrossRef] [PubMed]
Sipkema, P. , van der Linden, P. J. , Westerhof, N. , and Yin, F. C. , 2003, “ Effect of Cyclic Axial Stretch of Rat Arteries on Endothelial Cytoskeletal Morphology and Vascular Reactivity,” J. Biomech., 36(5), pp. 653–659. [CrossRef] [PubMed]
Barron, V. , Brougham, C. , Coghlan, K. , McLucas, E. , O'Mahoney, D. , Stenson-Cox, C. , and McHugh, P. E. , 2007, “ The Effect of Physiological Cyclic Stretch on the Cell Morphology, Cell Orientation and Protein Expression of Endothelial Cells,” J. Mater. Sci. Mater. Med., 18(10), pp. 1973–1981. [CrossRef] [PubMed]
Michel, J. B. , 2003, “ Anoikis in the Cardiovascular System: Known and Unknown Extracellular Mediators,” Arterioscler., Thromb., Vasc. Biol., 23(12), pp. 2146–2154. [CrossRef]
Lacolley, P. , Challande, P. , Boumaza, S. , Cohuet, G. , Laurent, S. , Boutouyrie, P. , Grimaud, J. A. , Paulin, D. , Lamaziere, J. M. , and Li, Z. , 2001, “ Mechanical Properties and Structure of Carotid Arteries in Mice Lacking Desmin,” Cardiovas. Res., 51(1), pp. 178–187. [CrossRef]
Korff, T. , Aufgebauer, K. , and Hecker, M. , 2007, “ Cyclic Stretch Controls the Expression of CD40 in Endothelial Cells by Changing Their Transforming Growth Factor-Beta1 Response,” Circulation, 116(20), pp. 2288–2297. [CrossRef] [PubMed]
Benbrahim, A. , L'Italien, G. J. , Kwolek, C. J. , Petersen, M. J. , Milinazzo, B. , Gertler, J. P. , Abbott, W. M. , and Orkin, R. W. , 1996, “ Characteristics of Vascular Wall Cells Subjected to Dynamic Cyclic Strain and Fluid Shear Conditions In Vitro,” J. Surg. Res., 65(2), pp. 119–127. [CrossRef] [PubMed]
Peng, X. , Recchia, F. A. , Byrne, B. J. , Wittstein, I. S. , Ziegelstein, R. C. , and Kass, D. A. , 2000, “ In Vitro System to Study Realistic Pulsatile Flow and Stretch Signaling in Cultured Vascular Cells,” Am. J. Physiol. Cell Physiol., 279(3), pp. C797–C805. [PubMed]
Zhao, S. , Suciu, A. , Ziegler, T. , Moore, J. E. , Bürki, E. , Meister, J.-J. , and Brunner, H. R. , 1995, “ Synergistic Effects of Fluid Shear Stress and Cyclic Circumferential Stretch on Vascular Endothelial Cell Morphology and Cytoskeleton,” Arterioscler., Thromb., Vasc. Biol., 15(10), pp. 1781–1786. [CrossRef]
Benbrahim, A. , L'Italien, G. J. , Milinazzo, B. B. , Warnock, D. F. , Dhara, S. , Gertler, J. P. , Orkin, R. W. , and Abbott, W. M. , 1994, “ A Compliant Tubular Device to Study the Influences of Wall Strain and Fluid Shear Stress on Cells of the Vascular Wall,” J. Vasc. Surg., 20(2), pp. 184–194. [CrossRef] [PubMed]
Azuma, N. , Duzgun, S. A. , Ikeda, M. , Kito, H. , Akasaka, N. , Sasajima, T. , and Sumpio, B. E. , 2000, “ Endothelial Cell Response to Different Mechanical Forces,” J. Vasc. Surg., 32(4), pp. 789–794. [CrossRef] [PubMed]
Moore, J. E., Jr. , Bürki, E. , Suciu, A. , Zhao, S. , Burnier, M. , Brunner, H. R. , and Meister, J.-J. , 1994, “ A Device for Subjecting Vascular Endothelial Cells to Both Fluid Shear Stress and Circumferential Cyclic Stretch,” Ann. Biomed. Eng., 22(4), pp. 416–422. [CrossRef] [PubMed]
Ives, C. , Eskin, S. , and McIntire, L. , 1986, “ Mechanical Effects on Endothelial Cell Morphology: In Vitro Assessment,” In Vitro Cell. Dev. Biol., 22(9), pp. 500–507. [CrossRef] [PubMed]
Tarbell, Y. Q. J. M. , 2000, “ Interaction Between Wall Shear Stress and Circumferential Strain Affects Endothelial Cell Biochemical Production,” J. Vasc. Res., 37(3), pp. 147–157. [CrossRef] [PubMed]
Berardi, D. E. , and Tarbell, J. M. , 2009, “ Stretch and Shear Interactions Affect Intercellular Junction Protein Expression and Turnover in Endothelial Cells,” Cell. Mol. Bioeng., 2(3), pp. 320–331. [CrossRef] [PubMed]
Toda, M. , Yamamoto, K. , Shimizu, N. , Obi, S. , Kumagaya, S. , Igarashi, T. , Kamiya, A. , and Ando, J. , 2008, “ Differential Gene Responses in Endothelial Cells Exposed to a Combination of Shear Stress and Cyclic Stretch,” J. Biotechnol., 133(2), pp. 239–244. [CrossRef] [PubMed]
Estrada, R. , Giridharan, G. A. , Nguyen, M. D. , Roussel, T. J. , Shakeri, M. , Parichehreh, V. , Prabhu, S. D. , and Sethu, P. , 2011, “ Endothelial Cell Culture Model for Replication of Physiological Profiles of Pressure, Flow, Stretch, and Shear Stress In Vitro,” Anal. Chem., 83(8), pp. 3170–3177. [CrossRef] [PubMed]
Van Dyke, W. S. , Sun, X. , Richard, A. B. , Nauman, E. A. , and Akkus, O. , 2012, “ Novel Mechanical Bioreactor for Concomitant Fluid Shear Stress and Substrate Strain,” J. Biomech., 45(7), pp. 1323–1327. [CrossRef] [PubMed]
Maeda, E. , Hagiwara, Y. , Wang, J. H. , and Ohashi, T. , 2013, “ A New Experimental System for Simultaneous Application of Cyclic Tensile Strain and Fluid Shear Stress to Tenocytes In Vitro,” Biomed. Microdevices, 15(6), pp. 1067–1075. [CrossRef] [PubMed]
Breen, L. T. , McHugh, P. E. , McCormack, B. A. , Muir, G. , Quinlan, N. J. , Heraty, K. B. , and Murphy, B. P. , 2006, “ Development of a Novel Bioreactor to Apply Shear Stress and Tensile Strain Simultaneously to Cell Monolayers,” Rev. Sci. Instrum., 77(10), p. 104301. [CrossRef]
Owatverot, T. B. , Oswald, S. J. , Chen, Y. , Wille, J. J. , and Yin, F. C. , 2005, “ Effect of Combined Cyclic Stretch and Fluid Shear Stress on Endothelial Cell Morphological Responses,” ASME J. Biomech. Eng., 127(3), pp. 374–382. [CrossRef]
Thacher, T. N. , Silacci, P. , Stergiopulos, N. , and da Silva, R. F. , 2010, “ Autonomous Effects of Shear Stress and Cyclic Circumferential Stretch Regarding Endothelial Dysfunction and Oxidative Stress: An Ex Vivo Arterial Model,” J. Vasc. Res., 47(4), pp. 336–345. [CrossRef] [PubMed]
Yin, W. , and Rubenstein, D. , 2009, “ Dose Effect of Shear Stress on Platelet Complement Activation in a Cone and Plate Shearing Device,” Cell. Mol. Bioeng., 2(2), pp. 274–280. [CrossRef]
Banes, A. J. , Gilbert, J. , Taylor, D. , and Monbureau, O. , 1985, “ A New Vacuum-Operated Stress-Providing Instrument That Applies Static or Variable Duration Cyclic Tension or Compression to Cells In Vitro,” J. Cell Sci., 75, pp. 35–42. [PubMed]
Vande Geest, J. P. , Di Martino, E. S. , and Vorp, D. A. , 2004, “ An Analysis of the Complete Strain Field Within Flexercell Membranes,” J. Biomech., 37(12), pp. 1923–1928. [CrossRef] [PubMed]
Ethier, C. R. , and Simmons, C. A. , 2007, “ Cellular Biomechanics,” Introductory Biomechanics: From Cells to Organisms, Cambridge University Press, New York, pp. 82–86.
Blackman, B. R. , Barbee, K. A. , and Thibault, L. E. , 2000, “ In Vitro Cell Shearing Device to Investigate the Dynamic Response of Cells in a Controlled Hydrodynamic Environment,” Ann. Biomed. Eng., 28(4), pp. 363–372. [CrossRef] [PubMed]
Yin, W. , Shanmugavelayudam, S. K. , and Rubenstein, D. A. , 2011, “ The Effect of Physiologically Relevant Dynamic Shear Stress on Platelet and Endothelial Cell Activation,” Thromb. Res., 127(3), pp. 235–241. [CrossRef] [PubMed]
Fung, Y. C. , 1977, A First Course in Continuum Mechanics, Prentice-Hall, Englewood Cliffs, NJ.
Maria, Z. , Yin, W. , and Rubenstein, D. A. , 2014, “ Combined Effects of Physiologically Relevant Disturbed Wall Shear Stress and Glycated Albumin on Endothelial Cell Functions Associated With Inflammation, Thrombosis and Cytoskeletal Dynamics,” J. Diabetes Invest., 5(4), pp. 372–381. [CrossRef]
Ng, C. P. , Hinz, B. , and Swartz, M. A. , 2005, “ Interstitial Fluid Flow Induces Myofibroblast Differentiation and Collagen Alignment In Vitro,” J. Cell Sci., 118(20), pp. 4731–4739. [CrossRef] [PubMed]
Sternberg, S. R. , 1983, “ Biomedical Image Processing,” Computer, 16(1), pp. 22–34. [CrossRef]
Zhang, J. , Betson, M. , Erasmus, J. , Zeikos, K. , Bailly, M. , Cramer, L. P. , and Braga, V. M. , 2005, “ Actin at Cell-Cell Junctions Is Composed of Two Dynamic and Functional Populations,” J. Cell Sci., 118(Pt 23), pp. 5549–5562. [CrossRef] [PubMed]
Caille, N. , Thoumine, O. , Tardy, Y. , and Meister, J. J. , 2002, “ Contribution of the Nucleus to the Mechanical Properties of Endothelial Cells,” J. Biomech., 35(2), pp. 177–187. [CrossRef] [PubMed]
Martinelli, R. , Zeiger, A. S. , Whitfield, M. , Sciuto, T. E. , Dvorak, A. , Van Vliet, K. J. , Greenwood, J. , and Carman, C. V. , 2014, “ Probing the Biomechanical Contribution of the Endothelium to Lymphocyte Migration: Diapedesis by the Path of Least Resistance,” J. Cell Sci., 127(Pt 17), pp. 3720–3734. [CrossRef] [PubMed]
Tojkander, S. , Gateva, G. , and Lappalainen, P. , 2012, “ Actin Stress Fibers—Assembly, Dynamics and Biological Roles,” J. Cell Sci., 125(Pt 8), pp. 1855–1864. [CrossRef] [PubMed]
Liu, Z. , Tan, J. L. , Cohen, D. M. , Yang, M. T. , Sniadecki, N. J. , Ruiz, S. A. , Nelson, C. M. , and Chen, C. S. , 2010, “ Mechanical Tugging Force Regulates the Size of Cell-Cell Junctions,” Proc. Natl. Acad. Sci. U.S.A., 107(22), pp. 9944–9949. [CrossRef] [PubMed]
Chen, C. S. , Tan, J. , and Tien, J. , 2004, “ Mechanotransduction at Cell-Matrix and Cell-Cell Contacts,” Annu. Rev. Biomed. Eng., 6(1), pp. 275–302. [CrossRef] [PubMed]
Chatzizisis, Y. S. , Coskun, A. U. , Jonas, M. , Edelman, E. R. , Feldman, C. L. , and Stone, P. H. , 2007, “ Role of Endothelial Shear Stress in the Natural History of Coronary Atherosclerosis and Vascular Remodeling: Molecular, Cellular, and Vascular Behavior,” J. Am. Coll. Cardiol., 49(25), pp. 2379–2393. [CrossRef] [PubMed]
Maalej, N. , and Folts, J. D. , 1996, “ Increased Shear Stress Overcomes the Antithrombotic Platelet Inhibitory Effect of Aspirin in Stenosed Dog Coronary Arteries,” Circulation, 93(6), pp. 1201–1205. [CrossRef] [PubMed]
Hasan, M. , Rubenstein, D. A. , and Yin, W. , 2013, “ Effects of Cyclic Motion on Coronary Blood Flow,” ASME J. Biomech. Eng., 135(12), p. 121002. [CrossRef]
Zhao, S. , Suciu, A. , Ziegler, T. , Moore, J. E., Jr. , Burki, E. , Meister, J. J. , and Brunner, H. R. , 1995, “ Synergistic Effects of Fluid Shear Stress and Cyclic Circumferential Stretch on Vascular Endothelial Cell Morphology and Cytoskeleton,” Arterioscler., Thromb., Vasc. Biol., 15(10), pp. 1781–1786. [CrossRef]
Chien, S. , 2007, “ Mechanotransduction and Endothelial Cell Homeostasis: The Wisdom of the Cell,” Am. J. Physiol. Heart Circ. Physiol., 292(3), pp. H1209–1224. [CrossRef] [PubMed]
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(4), pp. 341–347. [CrossRef]
Potter, C. M. , Schobesberger, S. , Lundberg, M. H. , Weinberg, P. D. , Mitchell, J. A. , and Gorelik, J. , 2012, “ Shape and Compliance of Endothelial Cells After Shear Stress In Vitro or From Different Aortic Regions: Scanning Ion Conductance Microscopy Study,” PloS One, 7(2), p. e31228. [CrossRef] [PubMed]
Levesque, M. J. , Liepsch, D. , Moravec, S. , and Nerem, R. M. , 1986, “ Correlation of Endothelial Cell Shape and Wall Shear Stress in a Stenosed Dog Aorta,” Arteriosclerosis, 6(2), pp. 220–229. [CrossRef] [PubMed]
Ohashi, T. , and Sato, M. , 2005, “ Remodeling of Vascular Endothelial Cells Exposed to Fluid Shear Stress: Experimental and Numerical Approach,” Fluid Dyn. Res., 37(1), pp. 40–59. [CrossRef]
Wang, J. H.-C. , Goldschmidt-Clermont, P. , Wille, J. , and Yin, F. C.-P. , 2001, “ Specificity of Endothelial Cell Reorientation in Response to Cyclic Mechanical Stretching,” J. Biomech., 34(12), pp. 1563–1572. [CrossRef] [PubMed]

Figures

Grahic Jump Location
Fig. 2

A constant angular velocity (18 rad/s) was used as the velocity input of the cone, and a cyclic vacuum pressure (magnitude at 30 kPa, frequency at 1.1 Hz) was used as the transient pressure input on the flexible membrane

Grahic Jump Location
Fig. 1

The schematic drawing of the shearing and stretching device. (a) The housing for a cell culture plate, (b) the six-well cell culture plate with flexible membranes, (c) shearing cones, (d) the piston pump, and (e) enlarged view of the flexible membrane and the supporting plate.

Grahic Jump Location
Fig. 3

(a) Velocity (m/s) vectors of the fluid phase as the vacuum pressure changed between 0 and 30 kPa in one stretching cycle. Simultaneously, the cone was rotating at a constant angular velocity of 18 rad/s. (b) Corresponding shear stress (Pa) distribution in the flow field when both the cone and the membrane were moving.

Grahic Jump Location
Fig. 4

(a) Nodal displacement along the radial direction in the in-plane area of the membrane at the maximum deformation (t = 0.6 s). (b) Circumferential and radial strain of a randomly chosen node (6 mm from the center of the membrane) change as a function of time during one stretching cycle.

Grahic Jump Location
Fig. 5

Measured circumferential and radial strain. (a) Radial strain did not vary significantly with angle (P > 0.3); (b) circumferential strain did not vary significantly with radial distance (P > 0.1). The circumferential and radial strains do not differ (P > 0.1), indicating strain field uniformity.

Grahic Jump Location
Fig. 6

Representative images of ECs following shear stress and/or tensile strain treatment. White arrows indicate flow direction and red arrows indicate the directions of tensile strain. Scale bars represent 25 μm.

Grahic Jump Location
Fig. 7

(a) Cell elongation and (b) nucleus elongation after HCAEC were treated with shear stress and/or cyclic tensile strain. Data are presented as mean + standard deviation (n = 24). *indicates significant difference (P < 0.05).

Grahic Jump Location
Fig. 8

Actin alignment was quantified using FFT. Significant difference was detected between ECs that were treated with shear stress alone, and those treated with tensile strain alone (P < 0.05, n = 12–19). Data are presented as mean + standard deviation.

Grahic Jump Location
Fig. 9

Actin accumulation along EC boundaries. Shear stress, alone or combined with tensile strain, induced significant increase in actin accumulation along cell boundaries (P < 0.05, n = 31–93). Biaxial tensile strain alone did not induce any changes in actin accumulation at cell junctions. Data are presented as mean + standard deviation.

Tables

Errata

Discussions

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging and repositioning the boxes below.

Related Journal Articles
Related eBook Content
Topic Collections

Sorry! You do not have access to this content. For assistance or to subscribe, please contact us:

  • TELEPHONE: 1-800-843-2763 (Toll-free in the USA)
  • EMAIL: asmedigitalcollection@asme.org
Sign In