Outgrowth endothelial cells (OEC) isolated and expanded from human peripheral blood are an attractive cell source of autologous EC for applications in tissue engineering. On micrometric fibroin fibers OEC formed differentiated endothelial cell layers. After embedding in a wound healing matrix (fibrin hydrogel) OEC migrated from the fibroin scaffolds into the fibrin and formed a microvessel-like network. In this study we investigated the response of OEC to silk fibroin nets with fibers in the nanometric range, that is, the physical dimensions of fibers of the extracellular matrix. The viability of the cells, the formation of EC layers, focal contacts and cellular polarization on nanometric fibroin fibers were studied. The formation of cell contacts with the scaffolding material and cell migration on the fibers are highly important to understand and to be able to guide the process of endothelialization or vascularization of tissue engineered constructs based on newly designed biomaterials. On the nanometric nets OEC formed cell layers with focal adhesions on the single nanofibers (demonstrated by immunofluorescence for focal adhesion kinase). Other molecules relevant for endothelial cell polarization and response of endothelial cells, such as caveolin-1 and its phosphorylated form, were investigated in parallel and revealed polarized OEC, indicating that OEC can use the nanometric fibers as a matrix for cell polarization and migration. Further studies are currently underway to investigate the possible beneficial effects of nanometric fibroin nets on important endothelial cell functions. (This work was supported by grants HIPPOCRATES, EXPERTISSUES from the European Commission)

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