Abstract
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.
Highlights
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 [1,2,3,4,5,6,7]
Very few cells attached to the (PAH/poly(acrylic acid) (PAA)-co-PAEDAPS)2 surface, and those that did were almost exclusively rounded as opposed to flat, with numerous filopodia-like extensions of the cell membrane extending in all directions (Figure 1(d))
This study demonstrates that polyelectrolyte multilayer (PEMU) surfaces can be used to alter many characteristics of human coronary artery smooth muscle cells (hCASMCs) in culture, and that some of these alterations are consistent with changes linked to phenotypic modulation
Summary
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 [1,2,3,4,5,6,7]. 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]. Phenotypic modulation of VSMCs has been reported in vitro utilizing serum deprivation and readministration [8], or growth surfaces coated with type IV collagen at specific passages and time points of primary VSMC cultures [3]. The potential utility of thin polyelectrolyte multilayer (PEMU) films in the induction of phenotypic modulation in such cells is explored. The PEMUs incorporated into this study were generated utilizing layer-bylayer deposition [9, 10], in which a substrate is moved back and forth between dilute baths of positively and negatively
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