Vascular endothelial growth factor immobilized in collagen scaffold promotes penetration and proliferation of endothelial cells.
A key challenge in engineering functional tissues in vitro is the limited transport capacity of oxygen and nutrients into the tissue. Inducing vascularization within engineered tissues is a key strategy to improving their survival in vitro and in vivo. The presence of vascular endothelial growth factor (VEGF) in a three-dimensional porous collagen scaffold may provide a useful strategy to promote vascularization of the engineered tissue in a controlled manner. To this end, we investigated whether immobilized VEGF could promote the invasion and assembly of endothelial cells (ECs) into the collagen scaffolds. We conjugated VEGF onto collagen scaffolds using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride chemistry, and measured the concentrations of immobilized VEGF in collagen scaffolds by direct VEGF enzyme-linked immunosorbent assay. We demonstrated that immobilized VEGF (relative to soluble VEGF) promoted the penetration and proliferation of ECs in the collagen scaffold, based on results of cell density analysis in histological sections, immunohistochemistry, XTT proliferation assay, glucose consumption and lactate production. Furthermore, we observed increased viability of ECs cultured in scaffolds with immobilized VEGF relative to soluble VEGF. This research demonstrates that immobilization of VEGF is a useful strategy to promote the invasion and proliferation of ECs into a scaffold, which may in turn lead to a vascularized scaffold.
Shen
Yi Hao
YH
Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Rm. 407, Toronto, Ontario, Canada M5S 3G9.
Shoichet
Molly S
MS
Radisic
Milica
M
D000818:Animals; D002452:Cell Count; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D003094:Collagen / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology*; D005947:Glucose / Q000378:metabolism; D007150:Immunohistochemistry; D007211:Indoles; D019344:Lactic Acid / Q000378:metabolism; D051379:Mice; D008855:Microscopy, Electron, Scanning; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D016062:Porosity; D054457:Tissue Scaffolds; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D007211:Indoles; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D042461:Vascular Endothelial Growth Factor A; D019344:Lactic Acid; C007293:DAPI; D003094:Collagen; D005947:Glucose