Basic Fibroblast Growth Factor Coating and Endothelial Cell Seeding of a Decellularized Heparin‐coated Vascular Graft

Abstract

The objective of this study was to determine the effect of basic fibroblast growth factor (bFGF) coating on endothelial cell seeding and proliferation on a decellularized heparin coated vascular graft and to determine the retention of seeded cells on the graft under flow conditions. Disks of heparin coated decellularized grafts were incubated for 24 h as controls or with bFGF. Human microvascular endothelial cells (HMECs) or canine peripheral blood endothelial progenitor cells (CEPC) were seeded onto the disks and incubated for 96 h or 48 h, respectively. HMECs were also seeded onto the luminal surfaces of two heparin-coated decellularized grafts for 3 h. One graft was placed in a perfusion culture system and cultured for an additional 6 h with flow and pressure. After culturing, there were 4.7 +/- 1.4 cells/mm(2) HMECs on control grafts and 11.4 +/- 1.4 cells/mm(2) in bFGF treated grafts (P < 0.05). Likewise, with CEPCs, there were 14.8 +/- 4.8 cells/mm(2) in control grafts and 33.3 +/- 7.3 cells/mm(2) in bFGF treated grafts. After only 3 h of cell attachment, 60% of HMECs were retained in the intact graft exposed flow relative to the static control graft, which is an acceptable level. These data demonstrate that bFGF coating on the heparin bound decellularized grafts significantly increases both HMEC and dog EPC proliferation and that seeded cells are stable under perfusion conditions.

Endothelial progenitor cells: diagnostic and therapeutic considerations

“…Other VEGF, (92) GM-CSF, (82) TGF-beta1, (39) angiopoietin-1, (93) SDF-1, (94) constitutive hTERT, (34) HGF, (95) PDGF-B, (96) bFGF, (97) apoptotic bodies from EC, (98) laminar shear stress, (40) haemodialysis. (64) Smoking, (20,45,58) haemodialysis, (63) angiostatin.…”

Recent Advances in Decellularization and Recellularization for Tissue-Engineered Liver Grafts

“…The presence of these growth factors stems from either the maintenance of native growth factors during the decellularization process [Yagi et al, 2013], or from the additional immobilization of desired growth factors [Conklin et al, 2004;Zhou et al, 2009;Nakamura and Ijima, 2013]. A successful immobilization should yield a liver scaffold with sufficient concentrations and stable release of growth factors in order to enhance implanted cell survival and function.…”

Scaffolds and Cell-Based Tissue Engineering for Blood Vessel Therapy

“…Reseeding EPCs on a vein graft can prevent vein graft thrombosis [Brown et al, 2010]. Various conditions have been reported to accelerate the proliferation of EPCs, such as adding basic fibroblast growth factor [Conklin et al, 2004], exposing the cells to shear stress [Yamamoto et al, 2003] and seeding on fibronectin [Salmon et al, 2009]. …”