State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China.
Center for General Practice Medicine, Department of Gastroenterology, Zhejiang Provincial People's Hospital, Affiliated People's Hospital, Hangzhou Medical College, Hangzhou 310059, Zhejiang, China.
ACS Biomater Sci Eng. 2023 Apr 10;9(4):1940-1951. doi: 10.1021/acsbiomaterials.2c01213. Epub 2023 Mar 13.
Functional bioengineered livers (FBLs) are promising alternatives to orthotopic liver transplantation. However, orthotopic transplantation of FBLs has not yet been reported. This study aimed to perform the orthotopic transplantation of FBLs in rats subjected to complete hepatectomy. FBLs were developed using rat whole decellularized liver scaffolds (DLSs) with human umbilical vein endothelial cells implanted via the portal vein, and human bone marrow mesenchymal stem cells (hBMSCs) and mouse hepatocyte cell line implanted via the bile duct. FBLs were evaluated in terms of endothelial barrier function, biosynthesis, and metabolism and orthotopically transplanted into rats to determine the survival benefit. The FBLs with well-organized vascular structures exhibited endothelial barrier function, with reduced blood cell leakage. The implanted hBMSCs and hepatocyte cell line were well aligned in the parenchyma of the FBLs. The high levels of urea, albumin, and glycogen in the FBLs indicated biosynthesis and metabolism. Orthotopic transplantation of FBLs achieved a survival time of 81.38 ± 4.263 min in rats ( = 8) subjected to complete hepatectomy, whereas control animals ( = 4) died within 30 min ( < 0.001). After transplantation, CD90-positive hBMSCs and the albumin-positive hepatocyte cell line were scattered throughout the parenchyma, and blood cells were limited within the vascular lumen of the FBLs. In contrast, the parenchyma and vessels were filled with blood cells in the control grafts. Thus, orthotopic transplantation of whole DLS-based FBLs can effectively prolong the survival of rats subjected to complete hepatectomy. In summary, this work was the first to perform the orthotopic transplantation of FBLs, with limited survival benefits, which still has important value for the advancement of bioengineered livers.
功能化生物工程肝脏（FBL）是肝移植的一种很有前途的替代方法。然而，FBL 的原位移植尚未见报道。本研究旨在对接受完全肝切除术的大鼠进行 FBL 的原位移植。通过门静脉植入人脐静脉内皮细胞，胆管植入人骨髓间充质干细胞（hBMSC）和鼠肝细胞系，在大鼠全去细胞化肝脏支架（DLS）上构建 FBL。从内皮屏障功能、生物合成和代谢等方面评估 FBL，并将其原位移植到大鼠体内，以确定其生存获益。具有组织良好血管结构的 FBL 表现出内皮屏障功能，减少了血细胞渗漏。植入的 hBMSC 和肝细胞系在 FBL 的实质中排列整齐。FBL 中高浓度的尿素、白蛋白和糖原表明其具有生物合成和代谢功能。FBL 原位移植可使接受完全肝切除术的大鼠的生存时间延长至 81.38±4.263 分钟（n=8），而对照组大鼠（n=4）在 30 分钟内死亡（<0.001）。移植后，CD90 阳性 hBMSC 和白蛋白阳性肝细胞系散在分布于实质中，血细胞局限于 FBL 的血管腔中。相比之下，对照组移植物的实质和血管中充满了血细胞。因此，基于全 DLS 的 FBL 的原位移植可以有效延长接受完全肝切除术的大鼠的生存时间。总之，这是首次进行 FBL 的原位移植，其生存获益有限，但对生物工程肝脏的发展仍具有重要价值。
