Heterotopic vascularization and functionalization of implantable bio engineered hepatic tissue alleviates liver injury in rats
Heterotopic vascularization and functionalization of implantable bio engineered hepatic tissue alleviates liver injury in rats
复制标题
可植入生物工程肝组织的异位血管化和功能化减轻大鼠肝损伤
DOI：
10.1111/liv.14267
复制
发表时间：
2019-10-13
影响因子：
6.7
通讯作者：
Bao, Ji
中科院分区：
文献类型：
--
作者：
Li, Yi;Wu, Qiong;Bao, Ji
背景生物工程肝的挑战在于维持高质量肝细胞的数量和血液灌注的血管系统。我们的特点是肝素化的猪脱细胞肝支架（DLS）作为载体，以支持肝细胞血管生成，从而开发功能和血管化的肝组织，可用于治疗肝损伤。方法采用去除细胞成分的方法获得猪DLS，并采用末端附着法进行肝素化处理。用大鼠肝细胞球再细胞化肝素化的脱细胞层构建工程化肝组织。将肝组织异位植入四氯化碳（CCl4）诱导的大鼠肝损伤模型的大网膜内。结果肝素化DLS中的肝细胞球体在体内至少可存活10周。整个支架充满肝细胞，排列良好。肝素化DLS组的体积扩大超过400倍。肝脏特异性功能如白蛋白合成、糖原储存和细胞色素P3A4活性在肝组织中高度表达。此外，内皮细胞被招募，如CD31染色所示，新血管形成，如异硫氰酸荧光素标记的葡聚糖活体共聚焦显微镜观察。肝素化生物工程肝组织通过调节细胞外基质的沉积和降解，减轻CCl4诱导的肝损伤。结论原代肝细胞球体在肝素化DLS上可长期存活并扩增形成血管化、功能化的生物工程肝组织，可减轻大鼠肝损伤。
Background The challenge of using bioengineered liver lies in sustaining the quantity of high-quality hepatocytes and the vasculature for blood perfusion. We characterized the heparinization of a porcine decellularized liver scaffold (DLS) as a carrier to support hepatocyte angiogenesis, thereby developing functional and vascularized hepatic tissue useful to treat liver injury. Method The porcine DLS was obtained by the removal of cellular components and then subjected to heparinization by the end-point attachment technique. The heparinized DLSs were recellularized with rat hepatocyte spheroids to construct engineered hepatic tissue. The hepatic tissue was heterotopically implanted in the omentum majus of a rat model with liver injury induced by carbon tetrachloride (CCl4). Results Hepatocyte spheroids in the heparinized DLS remained viable for at least 10 weeks in vivo. The entire scaffold was populated with hepatocytes and arranged well. The volume of the heparinized DLS group was expanded over 400-fold. Liver-specific functions such as albumin synthesis, glycogen storage and cytochrome P 3A4 activity were highly expressed in the hepatic tissue. In addition, endothelial cells were recruited, as shown by CD31 staining, and new blood vessels formed, as visualized by fluorescein isothiocyanate-labelled dextran intravital confocal microscopy. The heparinized bioengineered hepatic tissue alleviated CCl4-induced liver injury by regulating the deposition and degradation of the extracellular matrix. Conclusion Primary hepatocyte spheroids survived for an extended time on the heparinized DLS and expanded to generate vascularized and functional bioengineered hepatic tissue that can alleviate liver injury in rats.