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1 Division of Cardiology, Department of Experimental and Clinical Medicine, Magna Graecia University, Catanzaro, Italy
2 Division of Cardiology, "Federico II" University, Naples, Italy; Genecor Foundation, Naples, Italy
3 Department of Experimental and Clinical Medicine, "Magna Graecia" University, Catanzaro, Italy
4 Division of Cardiology, "Federico II" University, Naples, Italy
5 Division of Cardiology, Department of Experimental and Clinical Medicine, Magna Graecia University, Catanzaro, Italy; Genecor Foundation, Naples, Italy
* To whom correspondence should be addressed. E-mail: indolfi{at}unicz.it.
Synthetic polymers, like methacrylate (MA) compounds, have been clinically introduced as inert coatings to locally deliver drugs inhibiting restenosis after stenting. The aim of this study was to evaluate the effects of MA-coating alone on VSMC growth in vitro. Stainless steel stents were coated with MA at the following doses: 0.3 ml, 1.5 ml, and 3 ml. Uncoated/bare metal stents were used as controls. VSMCs were cultured in dishes and a MA-coated stent or an uncoated bare metal stent was gently added in each well. VSMC proliferation was assessed by bromodeoxyuridine incorporation. Apoptosis was analyzed by three distinct approaches: a) Annexin-V/propidium iodide fluorescence detection; b) DNA laddering; c) caspase-3 activation and PARP cleavage. MA-coated stents induced a significant decrease of BrdU incorporation compared to uncoated stents both at the low and high concentrations. In VSMCs incubated with MA-coated stents, Annexin-V/propidium iodide fluorescence detection show a significant increase in apoptotic cells which was corroborated by the typical DNA laddering. Apoptosis of VSMCs after incubation with MA-coated stents was characterized by caspase-3 activation and PARP cleavage. MA-coated stent induced VSMC growth arrest by inducing apoptosis in a dosedependent manner. Thus, methacrylate is not an inert platform for eluting drugs because is biologically active per se. This effect should be taken in account evaluating an association of this coating with anti-proliferative agents for in-stent restenosis prevention.
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