Fibroblast cell derived extracellular matrix containing electrospun scaffold as a hybrid biomaterial to promote in vitro endothelial cell expansion and functionalization

Biomaterials to be used for vascular tissue engineering must allow attachment, proliferation, and functionalization of vasoactive cells especially endothelial cells. In this study, decellularized L929 fibroblast cell-derived ECM containing electrospun scaffolds were fabricated and their biological response was investigated using rat glomerulus endothelial cells (rGECs). The L929 cells were grown for one week to get cell sheets on PCL membranes followed by decellularization of whole cell sheet-PCL membrane (PCL-ECM) using sodium dodecyl sulfate (SDS)/triton X-100 (TX) or freeze/thaw (F/T)/Deoxyribonuclease cycle to yield the corresponding mechanically stable scaffold. The nucleic acids and structural proteins quantification were performed on various membranes before and after decellularization process. Seeded rGECs on PCL, PCL-ECM (SDS/TX) and PCL-ECM (F/T) membranes were investigated through immunofluorescence and cell proliferation assay. The bio-macromolecules contents on decellularized scaffolds showed diverse outcome because of different decellularization methods used. The hydrophilic PCL-ECM (F/T) scaffold showed the best result by ensuring stability, good cytocompatibility, and interconnections among endothelial cells as was further confirmed by endothelial gene expression analysis. In short, the outcomes of this study may pave the way for the construction of new cell-derived ECM based vascular tissue engineering scaffolds as well as for the development of in vitro models to study endothelial cell function.