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Polyethylene glycol crosslinked decellularized single liver lobe scaffolds with vascular endothelial growth factor promotes angiogenesis in vivo

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Hepatobiliary & Pancreatic Diseases International 2023年第6期22卷 622-631页

作者： Jian-Se Zhang Zhi-Bin Wang Zhi-Zhen Lai Jing-Wen Yang Wen-Jing Song Yu-Bing Wei Jin Mei Jian-Guang Wang Anatomy Department School of Basic Medical SciencesWenzhou Medical UniversityWenzhou 325000China Institute of Bioscaffold Transplantation and Immunology Wenzhou Medical UniversityWenzhou 325000China Institute of Hypoxic Medicine School of Basic Medical SciencesWenzhou Medical UniversityWenzhou 325000China Intensive Care Unit Tongde Hospital of Zhejiang ProvinceHangzhou 310012China Department of Geriatric Medicine The First Affiliated HospitalWenzhou Medical UniversityWenzhou 325000China Department of Microbiology and Immunology Wenzhou Medical UniversityWenzhou 325000China Medical Research Center Ningbo City First HospitalNingbo 315000China Department of Biochemistry School of Basic Medical SciencesWenzhou Medical UniversityWenzhou 325000China

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 vascular 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 subcutaneous 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 translucent 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 i

关键词： Decellularization single liver lobe Polyethylene glycol Angiogenesis liver tissue engineering

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