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| Polyethylene glycol crosslinked decellularized single liver lobe scaffolds with vascular endothelial growth factor promotes angiogenesis in vivo |
| Jian-Se Zhang a , b , c , # , Zhi-Bin Wang a , b , c , # , Zhi-Zhen Lai d , # , Jing-Wen Yang e , Wen-Jing Song f , Yu-Bing Wei a , Jin Mei b , g , Jian-Guang Wang h ,∗ |
a Anatomy Department, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325000, China
b Institute of Bioscaffold Transplantation and Immunology, Wenzhou Medical University, Wenzhou 325000, China
c Institute of Hypoxic Medicine, School of Basic Medical Science , Wenzhou Medical University, Wenzhou 325000, China
d Intensive Care Unit, Tongde Hospital of Zhejiang Province, Hangzhou 310012, China
e Department of Geriatric Medicine, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou 325000, China
f Department of Microbiology and Immunology, Wenzhou Medical University, Wenzhou 325000, China
g Medical Research Center, Ningbo City First Hospital, Ningbo 315000, China
h Department of Biochemistry, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325000, China
∗Corresponding author.
E-mail address: wz_wjg@163.com (J.-G. Wang).
# Contributed equally. |
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Abstract Background: Improving the mechanical properties and angiogenesis of acellular scaffolds before transplantation is an important challenge facing the development of acellular liver grafts. The present study aimed to evaluate the cytotoxicity and angiogenesis of polyethylene glycol (PEG) crosslinked decellularized single liver lobe scaffolds (DLSs), and establish its suitability as a graft for long-term liver tissue engineering.
Methods: Using mercaptoacrylate produced by the Michael addition reaction, DLSs were first modified using N-succinimidyl S-acetylthioacetate (SATA), followed by cross-linking with PEG as well as vascu- lar endothelial growth factor (VEGF). The optimal concentration of agents and time of the individual steps were identified in this procedure through biomechanical testing and morphological analysis. Subsequently, human umbilical vein endothelial cells (HUVECs) were seeded on the PEG crosslinked scaffolds to detect the proliferation and viability of cells. The scaffolds were then transplanted into the subcuta- neous tissue of Sprague-Dawley rats to evaluate angiogenesis. In addition, the average number of blood vessels was evaluated in the grafts with or without PEG at days 7, 14, and 21 after implantation.
Results: The PEG crosslinked DLS maintained their three-dimensional structure and were more translu- cent after decellularization than native DLS, which presented a denser and more porous network structure. The results for Young’s modulus proved that the mechanical properties of 0.5 PEG crosslinked DLS were the best and close to that of native livers. The PEG-VEGF-DLS could better promote cell proliferation and differentiation of HUVECs compared with the groups without PEG cross-linking. Importantly, the average density of blood vessels was higher in the PEG-VEGF-DLS than that in other groups at days 7, 14, and 21 after implantation in vivo.
Conclusions: The PEG crosslinked DLS with VEGF could improve the biomechanical properties of native DLS, and most importantly, their lack of cytotoxicity provides a new route to promote the proliferation of cells in vitro and angiogenesis in vivo in liver tissue engineering.
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2023, Vol. 22 
