Abstract 373: Effects of Coronary Wall Mechanics on Smooth Muscle Cell Phenotypic Switch and CD44 + Mesenchymal Cell Repopulation in Saphenous Vein Grafts

Abstract

Background: Despite the preferred application of arterial conduits, the greater saphenous vein (SV) remains indispensable for bypass grafting, especially in multi-vessel coronary artery disease. Early remodeling induced by altered wall mechanics has been recognized to play a key role in SV graft disease. The mechanism remains, however, unknown. Aim: To investigate mechanical factors involved in early graft remodeling, we characterized SV-derived smooth muscle cells (SMCs) after both ex vivo coronary-like mechanical stimulation of SV segments and in vitro unidirectional strain. Methods: SV segments from patients receiving coronary artery bypass grafts were stimulated in a custom-made coronary pulse-duplicator bioreactor. After 7 (n=6) or 14 (n=5) days, stretched and control SVs were fixed and stained for immunofluorescence. Additionally, SMCs isolated from SVs of 7 patients undergoing saphenectomy were subjected to uniaxial cyclic strain (10% elongation, 1 Hz) for 24 or 72 hours using a Flexcell platform. SMCs analysis was performed by western blotting and mass spectrometry-based secretome analysis. Results: Coronary stimulation elevated apoptosis of SV medial cells after 7 days, and consistently reduced the percentage of cells positive for contractile markers α-SMA and calponin. Conversely, synthetic phenotype marker tropomyosin-4 (TM4) and early contractile marker SM22α were elevated at T14. Mesenchymal marker CD44 was markedly upregulated in cells populating the media after 14 days of stimulation. In accordance, strained SMCs displayed decreased α-SMA and SM22α, and increased TM4 protein expression after 72h. Analysis of the supernatant showed a significant increase of plasminogen activator inhibitor-1 and thrombospondin 1. Conclusions: Mechanical stimulation of SVs leads to apoptosis of medial cells and a decrease of contractile SMC markers, followed by repopulation with cells expressing the mesenchymal marker CD44. In addition, unidirectional strain induces a switch of SMC phenotype and secretion of proteins related to vascular remodeling. We are currently investigating whether CD44 + cells derive from SMCs undergoing phenotypic switch, or from progenitor cells localized in the adventitia.