Functional modification of fibrous PCL scaffolds with fusion protein HGFI-VEGF enhanced cellularization and vascularization
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1
Nankai University, State Key Laboratory of Medicinal Chemical Biology, College of Life Science, China
The major challenge of tissue engineering is the establishment of efficient vascularization. Vascular endothelial growth factor (VEGF) is known to activate the proliferation, migration, and survival pathways of endothelial cells through phosphorylation of VEGF receptor-2 (VEGFR-2). VEGF has been widely utilized to induce vascularization. Hydrophobin HGFI, which is a member of amphiphilic proteins, can form a self-assembly layer on the surface of polymer scaffolds and convert their wettability. In this study, a fusion protein consisting of hydrophobin HGFI and VEGF was prepared by Pichia pastoris (P. pastoris) expression system. SDS-PAGE and Western blotting confirmed that the fusion protein VEGF-HGFI was successfully isolated and purified. Transmission electron microscope (TEM) and water contact angle (WCA) measurement demonstrated that VEGF-HGFI could form a coating layer on electrospun PCL fibers and increased their hydrophilicity. The VEGF-HGFI layer which was of about 25 nm in thickness was approved stable in PBS under shaking for at least one week. MTT assay showed that human umbilical vein endothelial cells (HUVECs) grew faster on the VEGF-HGFI modified PCL scaffolds than the HGFI modified or bare PCL scaffolds. Subcutaneous implantation in rat revealed that cellularization and vascularization were significantly improved in VEGF-HGFI modified PCL scaffolds. Bioluminescence imaging (BLI) showed that VEGF-HGFI could activate VEGFR2 receptors, which is a predominant VEGF receptor in angiogenic signaling. These results suggest that this fusion protein represents a new, simple and effective technique for functional modification of tissue engineering scaffolds to improve cellularization and vascularization, and may have a broad application in regenerative medicine.
Keywords:
Scaffold,
Surface modification,
Tissue Regeneration,
biofunctionalization
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Regenerative medicine: biomaterials for control of tissue induction
Citation:
Kong
D,
Wang
K,
Zhao
L,
Ma
S,
Pan
Y and
Qiao
M
(2016). Functional modification of fibrous PCL scaffolds with fusion protein HGFI-VEGF enhanced cellularization and vascularization.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.01842
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.