Under normal physiological conditions, mature human coronary artery smooth muscle cells (hCASMCs) exhibit a “contractile” phenotype marked by low rates of proliferation and protein synthesis, but these cells possess the remarkable ability to dedifferentiate into a “synthetic” phenotype when stimulated by conditions of pathologic stress. A variety of polyelectrolyte multilayer (PEMU) films are shown here to exhibit bioactive properties that induce distinct responses from cultured hCASMCs. Surfaces terminated with Nafion or poly(styrenesulfonic acid) (PSS) induce changes in the expression and organization of intracellular proteins, while a hydrophilic, zwitterionic copolymer of acrylic acid and 3-[2-(acrylamido)-ethyl dimethylammonio] propane sulfonate (PAA-co-PAEDAPS) is resistant to cell attachment and suppresses the formation of key cytoskeletal components. Differential expression of heat shock protein 90 and actin is observed, in terms of both their magnitude and cellular localization, and distinct cytoplasmic patterns of vimentin are seen. The ionophore A23187 induces contraction in confluent hCASMC cultures on Nafion-terminated surfaces. These results demonstrate that PEMU coatings exert direct effects on the cytoskeletal organization of attaching hCASMCs, impeding growth in some cases, inducing changes consistent with phenotypic modulation in others, and suggesting potential utility for PEMU surfaces as a coating for coronary artery stents and other implantable medical devices. 1. Introduction Vascular smooth muscle cells (VSMCs) are implicated as key contributors to numerous vascular pathologies, including atherosclerosis and the restenosis of angioplasty-treated blood vessels in the presence or absence of coronary stents, given their remarkable capacity for phenotypic modulation in response to pathological stressors [1–7]. Rather than achieving a state of terminal differentiation upon maturity, VSMCs are capable of dedifferentiation through apparently reversible pathways, as they transition between a “contractile” state marked by low rates of proliferation and protein synthesis, and a “synthetic” state marked by an increase in these parameters. An unknown number of transitional states likely reside between these two extremes, and the possibility that the transitory pathways between these two phenotypic states may not be identical lends additional complexity to this scenario [7]. There is also evidence that phenotypically heterogeneous subpopulations of VSMCs may exist within the arterial media itself [2, 6], but when placed into in vitro culture
References
[1]
S. M. Schwartz, G. R. Campbell, and J. H. Campbell, “Replication of smooth muscle cells in vascular disease,” Circulation Research, vol. 58, no. 4, pp. 427–444, 1986.
[2]
M. G. Frid, E. P. Moiseeva, and K. R. Stenmark, “Multiple phenotypically distinct smooth muscle cell populations exist in the adult and developing bovine pulmonary arterial media in vivo,” Circulation Research, vol. 75, no. 4, pp. 669–681, 1994.
[3]
N. F. Worth, B. E. Rolfe, J. Song, and G. R. Campbell, “Vascular smooth muscle cell phenotypic modulation in culture is associated with reorganisation of contractile and cytoskeletal proteins,” Cell Motility and the Cytoskeleton, vol. 49, no. 3, pp. 130–145, 2001.
[4]
M. Sata, A. Saiura, A. Kunisato et al., “Hematopoietic stem cells differentiate into vascular cells that participate in the pathogenesis of atherosclerosis,” Nature Medicine, vol. 8, no. 4, pp. 403–409, 2002.
[5]
D. Simper, P. G. Stalboerger, C. J. Panetta, S. Wang, and N. M. Caplice, “Smooth muscle progenitor cells in human blood,” Circulation, vol. 106, no. 10, pp. 1199–1204, 2002.
[6]
H. Hao, G. Gabbiani, and M.-L. Bochaton-Piallat, “Arterial smooth muscle cell heterogeneity: implications for atherosclerosis and restenosis development,” Arteriosclerosis, Thrombosis, and Vascular Biology, vol. 23, no. 9, pp. 1510–1520, 2003.
[7]
G. K. Owens, M. S. Kumar, and B. R. Wamhoff, “Molecular regulation of vascular smooth muscle cell differentiation in development and disease,” Physiological Reviews, vol. 84, no. 3, pp. 767–801, 2004.
[8]
C. Boccardi, A. Cecchettini, A. Caselli et al., “A proteomic approach to the investigation of early events involved in vascular smooth muscle cell activation,” Cell and Tissue Research, vol. 328, no. 1, pp. 185–195, 2007.
[9]
G. Decher, “Fuzzy nanoassemblies: toward layered polymeric multicomposites,” Science, vol. 277, no. 5330, pp. 1232–1237, 1997.
[10]
G. Decher and J. B. Schlenoff, Eds., Multilayer Thin Films—Sequential Assembly of Nanocomposite Materials, Wiley-VCH, Weinheim, Germany, 2003.
[11]
S. Y. Yang, J. D. Mendelsohn, and M. F. Rubner, “New class of ultrathin, highly cell-adhesion-resistant polyelectrolyte multilayers with micropatterning capabilities,” Biomacromolecules, vol. 4, no. 4, pp. 987–994, 2003.
[12]
S. Kidambi, I. Lee, and C. Chan, “Controlling primary hepatocyte adhesion and spreading on protein-free polyelectrolyte multilayer films,” Journal of the American Chemical Society, vol. 126, no. 50, pp. 16286–16287, 2004.
[13]
S. P. Forry, D. R. Reyes, M. Gaitan, and L. E. Locascio, “Facilitating the culture of mammalian nerve cells with polyelectrolyte multilayers,” Langmuir, vol. 22, no. 13, pp. 5770–5775, 2006.
[14]
C. Boura, P. Menu, E. Payan et al., “Endothelial cells grown on thin polyelectrolyte mutlilayered films: an evaluation of a new versatile surface modification,” Biomaterials, vol. 24, no. 20, pp. 3521–3530, 2003.
[15]
C. Boura, S. Muller, D. Vautier et al., “Endothelial cell—interactions with polyelectrolyte multilayer films,” Biomaterials, vol. 26, no. 22, pp. 4568–4575, 2005.
[16]
N. Berthelemy, H. Kerdjoudj, C. Gaucher et al., “Polyelectrolyte films boost progenitor cell differentiation into endothelium-like monolayers,” Advanced Materials, vol. 20, no. 14, pp. 2674–2678, 2008.
[17]
H. Kerdjoudj, N. Berthelemy, S. Rinckenbach et al., “Small vessel replacement by human umbilical arteries with polyelectrolyte film-treated arteries. In vivo behavior,” Journal of the American College of Cardiology, vol. 52, no. 19, pp. 1589–1597, 2008.
[18]
N. Berthelemy, H. Kerdjoudj, P. Schaaf et al., “O2 level controls hematopoietic circulating progenitor cells differentiation into endothelial or smooth muscle cells,” PloS one, vol. 4, no. 5, article e5514, 2009.
[19]
D. S. Salloum, S. G. Olenych, T. C. S. Keller, and J. B. Schlenoff, “Vascular smooth muscle cells on polyelectrolyte multilayers: hydrophobicity-directed adhesion and growth,” Biomacromolecules, vol. 6, no. 1, pp. 161–167, 2005.
[20]
S. G. Olenych, M. D. Moussallem, D. S. Salloum, J. B. Schlenoff, and T. C. S. Keller, “Fibronectin and cell attachment to cell and protein resistant polyelectrolyte surfaces,” Biomacromolecules, vol. 6, no. 6, pp. 3252–3258, 2005.
[21]
M. D. Moussallem, S. G. Olenych, S. L. Scott, T. C. S. Keller III, and J. B. Schlenoff, “Smooth muscle cell phenotype modulation and contraction on native and cross-linked polyelectrolyte multilayers,” Biomacromolecules, vol. 10, no. 11, pp. 3062–3068, 2009.
[22]
S. J. Assinder, J.-A. L. Stanton, and P. D. Prasad, “Transgelin: an actin-binding protein and tumour suppressor,” International Journal of Biochemistry and Cell Biology, vol. 41, no. 3, pp. 482–486, 2009.
[23]
A. J. Engler, C. Carag-Krieger, C. P. Johnson et al., “Embryonic cardiomyocytes beat best on a matrix with heart-like elasticity: scar-like rigidity inhibits beating,” Journal of Cell Science, vol. 121, no. 22, pp. 3794–3802, 2008.
[24]
C. L. McCormick and L. C. Salazar, “Water soluble copolymers: 46. Hydrophilic sulphobetaine copolymers of acrylamide and 3-(2-acrylamido-2-methylpropanedimethylammonio)-1-propanesulphonate,” Polymer, vol. 33, no. 21, pp. 4617–4624, 1992.
[25]
R. M. Ji Sr., H. H. Rmaile, and J. B. Schlenoff, “Hydrophobic and ultrahydrophobic multilayer thin films from perfluorinated polyelectrolytes,” Angewandte Chemie—International Edition, vol. 44, no. 5, pp. 782–785, 2005.
[26]
S. Diziain, J. Dejeu, L. Buisson, D. Charraut, F. Membrey, and A. Foissy, “Investigations in the initial build-up stages of polyelectrolyte multilayers by laser reflectometry and atomic force microscopy,” Thin Solid Films, vol. 516, no. 1, pp. 1–7, 2007.
[27]
S. Mehrotra, S. C. Hunley, K. M. Pawelec et al., “Cell adhesive behavior on thin polyelectrolyte multilayers: cells attempt to achieve homeostasis of its adhesion energy,” Langmuir, vol. 26, no. 15, pp. 12794–12802, 2010.
[28]
P. L. Faries, D. I. Rohan, H. Takahara et al., “Human vascular smooth muscle cells of diabetic origin exhibit increased proliferation, adhesion, and migration,” Journal of Vascular Surgery, vol. 33, no. 3, pp. 601–607, 2001.
[29]
D. Picard, “Hsp90 invades the outside,” Nature Cell Biology, vol. 6, no. 6, pp. 479–480, 2004.
[30]
M. E. Fultz, C. Li, W. Geng, and G. L. Wright, “Remodeling of the actin cytoskeleton in the contracting A7r5 smooth muscle cell,” Journal of Muscle Research and Cell Motility, vol. 21, no. 8, pp. 775–787, 2000.
[31]
S. Koyasu, E. Nishida, T. Kadowaki, et al., “Two mammalian heat shock proteins, HSP90 and HSP100, are actin-binding proteins,” Proceedings of the National Academy of Sciences of the United States of America, vol. 83, no. 21, pp. 8054–8058, 1986.
[32]
F. K. Gyoeva and V. I. Gelfand, “Coalignment of vimentin intermediate filaments with microtubules depends on kinesin,” Nature, vol. 353, no. 6343, pp. 445–448, 1991.
[33]
C. Picart, J. Mutterer, L. Richert et al., “Molecular basis for the explanation of the exponential growth of polyelectrolyte multilayers,” Proceedings of the National Academy of Sciences of the United States of America, vol. 99, no. 20, pp. 12531–12535, 2002.
[34]
L. Richert, P. Lavalle, E. Payan et al., “Layer by layer buildup of polysaccharide films: physical chemistry and cellular adhesion aspects,” Langmuir, vol. 20, no. 2, pp. 448–458, 2004.
[35]
C. Heitner-Wirguin, “Recent advances in perfluorinated ionomer membranes: structure, properties and applications,” Journal of Membrane Science, vol. 120, no. 1, pp. 1–33, 1996.
[36]
W. K. Ward, L. B. Jansen, E. Anderson, G. Reach, J.-C. Klein, and G. S. Wilson, “A new amperometric glucose microsensor: in vitro and short-term in vivo evaluation,” Biosensors and Bioelectronics, vol. 17, no. 3, pp. 181–189, 2002.
[37]
X. Kang, Z. Mai, X. Zou, P. Cai, and J. Mo, “A sensitive nonenzymatic glucose sensor in alkaline media with a copper nanocluster/multiwall carbon nanotube-modified glassy carbon electrode,” Analytical Biochemistry, vol. 363, no. 1, pp. 143–150, 2007.
[38]
S. Inaba, K. Nibu, H. Takano et al., “Potassium-adsorption filter for RBC transfusion: a phase III clinical trial,” Transfusion, vol. 40, no. 12, pp. 1469–1474, 2000.
[39]
B. K. Eustace, T. Sakurai, J. K. Stewart et al., “Functional proteomic screens reveal an essential extracellular role for hsp90α in cancer cell invasiveness,” Nature Cell Biology, vol. 6, no. 6, pp. 507–514, 2004.
[40]
M. J. Sierevogel, G. Pasterkamp, D. P. V. de Kleijn, and B. H. Strauss, “Matrix metalloproteinases: a therapeutic target in cardiovascular disease,” Current Pharmaceutical Design, vol. 9, no. 13, pp. 1033–1040, 2003.
[41]
L. J. Feldman, M. Mazighi, A. Scheuble et al., “Differential expression of matrix metalloproteinases after stent implantation and balloon angioplasty in the hypercholesterolemic rabbit,” Circulation, vol. 103, no. 25, pp. 3117–3122, 2001.
[42]
G. Burgstaller and M. Gimona, “Podosome-mediated matrix resorption and cell motility in vascular smooth muscle cells,” American Journal of Physiology—Heart and Circulatory Physiology, vol. 288, no. 6, pp. H3001–H3005, 2005.
[43]
D. Brown, A. Dykes, J. Black, S. Thatcher, M. E. Fultz, and G. L. Wright, “Differential actin isoform reorganization in the contracting A7r5 cell,” Canadian Journal of Physiology and Pharmacology, vol. 84, no. 8-9, pp. 867–875, 2006.
[44]
B. M. Jockusch, C.-A. Schoenenberger, J. Stetefeld, and U. Aebi, “Tracking down the different forms of nuclear actin,” Trends in Cell Biology, vol. 16, no. 8, pp. 391–396, 2006.
[45]
E. Fuchs and K. Weber, “Intermediate filaments: structure, dynamics, function, and disease,” Annual Review of Biochemistry, vol. 63, pp. 345–382, 1994.
[46]
F. Terzi, D. Henrion, E. Colucci-Guyon et al., “Reduction of renal mass is lethal in mice lacking vimentin: role of endothelin-nitric oxide imbalance,” Journal of Clinical Investigation, vol. 100, no. 6, pp. 1520–1528, 1997.
[47]
N. Wang and D. Stamenovic, “Mechanics of vimentin intermediate filaments,” Journal of Muscle Research and Cell Motility, vol. 23, no. 5-6, pp. 535–540, 2002.
[48]
B. Eckes, D. Dogic, E. Colucci-Guyon et al., “Impaired mechanical stability, migration and contractile capacity in vimentin deficient fibroblasts,” Journal of Cell Science, vol. 111, no. 13, pp. 1897–1907, 1998.
[49]
J. J. Murray, P. W. Reed, and F. S. Fay, “Contraction of isolated smooth muscle cells by ionophore A23187,” Proceedings of the National Academy of Sciences of the United States of America, vol. 72, no. 11, pp. 4459–4463, 1975.
[50]
M. A. Kolber and D. H. Haynes, “Fluorescence study of the divalent cation-transport mechanism of ionophore A23187 in phospholipid membranes,” Biophysical Journal, vol. 36, no. 2, pp. 369–391, 1981.
[51]
C. J. Chapman, A. K. Puri, R. W. Taylor, and D. R. Pfeiffer, “General features in the stoichiometry and stability of ionophore A23187-cation complexes in homogeneous solution,” Archives of Biochemistry and Biophysics, vol. 281, no. 1, pp. 44–57, 1990.
[52]
C. M. Deber and D. R. Pfeiffer, “Ionophore A23187. Solution conformations of the calcium complex and free acid deduced from proton and carbon-13 nuclear magnetic resonance studies,” Biochemistry, vol. 15, no. 1, pp. 132–141, 1976.
