A novel bioscaffold with naturally-occurring extracellular matrix promotes hepatocyte survival and vessel patency in mouse models of heterologous transplantation
A novel bioscaffold with naturally-occurring extracellular matrix promotes hepatocyte survival and vessel patency in mouse models of heterologous transplantation
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一种具有天然细胞外基质的新型生物支架可促进异源移植小鼠模型中的肝细胞存活和血管通畅
DOI：
10.1016/j.biomaterials.2018.05.026
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发表时间：
2018
期刊：
影响因子：
14
通讯作者：
Lianhua Bai
中科院分区：
文献类型：
--
作者：
Wei Yang;Quanyu Chen;Renpei Xia;Yujun Zhang;Ling Shuai;Jiejuan Lai;Xiaolin You;Yan Jiang;Ping Bie;Leida Zhang;Hongyu Zhang;Lianhua Bai
背景：由于缺乏合适的细胞外基质（ECM）来提供“主动微环境”支持，基于天然脱细胞肝支架（nDLS）的组织工程受到了损害。目的：本研究旨在检测一种新的，具有活性ECM的再生DLS（rDLS）是否能提高原代肝细胞存活率并预防血栓形成。方法：rDLS取自30-55%肝部分切除术，在体内保存3-5天，然后在体外用洗涤剂灌注。与正常肝脏产生的nDLS相比，rDLS由于体内再生期而具有生物活性分子。通过Ki-67染色和台盼蓝排除法评价原代小鼠肝细胞存活率。通过免疫组织化学和离体稀释的全血灌注评估血栓形成。通过近红外激光多普勒血流仪和异位移植测定血液相容性。结果：再细胞化后，rDLS比nDLS含有更多的Ki-67阳性原代肝细胞。rDLS的血氧饱和度和血流速度高于nDLS，整合素αIIb和α4的表达低于nDLS。肿瘤坏死因子-α、肝细胞生长因子、白细胞介素-10、白细胞介素-6和白细胞介素-1 β在整个rDLS中高度表达，而I型胶原、IV型胶原和血小板生成素在rDLS中的表达低于nDLS。在体外和体内均观察到rDLS改善的血管通畅性。在小鼠中的结果在大型动物（猪）中得到证实。结论：rDLS是一种有效的DLS，具有支持原代肝细胞存活和促进血管通畅的“活性微环境”。这是第一项证明rDLS具有促进肝细胞存活和抵抗血栓形成的血管微血管网络的研究。
BACKGROUND:.Naïve decellularized liver scaffold (nDLS)-based tissue engineering has been impaired by the lack of a suitable extracellular matrix (ECM) to provide "active micro-environmental" support...AIM:.The present study aimed to examine whether a novel, regenerative DLS (rDLS) with an active ECM improves primary hepatocyte survival and prevents thrombosis...METHODS:.rDLS was obtained from a 30-55% partial hepatectomy that was maintained in vivo for 3-5 days and then perfused with detergent in vitro. Compared to nDLS generated from normal livers, rDLS possesses bioactive molecules due to the regenerative period in vivo. Primary mouse hepatocyte survival was evaluated by staining for Ki-67 and Trypan blue exclusion. Thrombosis was assessed by immunohistochemistry and ex vivo diluted whole-blood perfusion. Hemocompatibility was determined by near-infrared laser-Doppler flowmetry and heterotopic transplantation...RESULTS:.After recellularization, rDLS contained more Ki-67-positive primary hepatocytes than nDLS. rDLS had a higher oxygen saturation and blood flow velocity and a lower expression of integrin αIIb and α4 than nDLS. Tumor necrosis factor-α, hepatocyte growth factor, interleukin-10, interleukin-6 and interleukin-1β were highly expressed throughout the rDLS, whereas expression of collagen-I, collagen-IV and thrombopoietin were lower in rDLS than in nDLS. Improved blood vessel patency was observed in rDLS both in vitro and in vivo. The results in mice were confirmed in large animals (pigs)...CONCLUSION:.rDLS is an effective DLS with an "active microenvironment" that supports primary hepatocyte survival and promotes blood vessel patency. This is the first study to demonstrate a rDLS with a blood microvessel network that promotes hepatocyte survival and resists thrombosis.