In vivo liver decellularization has become a promising strategy to study in vivo liver engineering. However, long-term survival after in vivo liver decellularization has not yet been achieved due to anatomical and technical challenges. This study aimed at establishing a survival model of in vivo partial liver lobe perfusion-decellularization in rats. We compared three decellularization protocols (1% Triton X100 followed by 1% sodium dodecyl sulfate [SDS], 1% SDS vs. 1% Triton X100, n = 6/group). Using the optimal one as judged by macroscopy, histology and DNA content, we characterized the structural integrity and matrix proteins by using histology, scanning electron microscopy, computed tomography scanning, and immunohistochemistry (IHC). We prevented contamination of the abdominal cavity with the corrosive detergents by using polyvinylidene chloride (PVDC) film + dry gauze in comparison to PVDC film + dry gauze + aspiration tube (n = 6/group). Physiological reperfusion was assessed by histology. Survival rate was determined after a 7-day observation period. Only perfusion with 1% SDS resulted in an acellular scaffold (fully translucent without histologically detectable tissue remnants, DNA concentration is <2% of that in native lobe) with remarkable structural and ultrastructural integrity as well as preservation of main matrix proteins (IHC positive for collagen IV, laminin, and elastin). Contamination of abdominal organs with the potentially toxic SDS solution was achieved by placing a suction tube in addition to the PVDC film + dry gauze and allowed a 7-day survival of all animals without severe postoperative complications. On reperfusion, the liver turned red within seconds without any leakage from the surface of the liver. About 12 h after reperfusion, not only blood cells but also some clots were visible in the portal vein, sinusoidal matrix network, and central vein, suggesting physiological perfusion. In conclusion, our results of this study show the first available data on generation of a survival model of in vivo parenchymal organ decellularization, creating a critical step toward in vivo organ engineering.

中文翻译：

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体内部分肝叶脱细胞的存活模型，以实现体内肝脏工程。

体内肝脏去细胞化已成为研究体内肝脏工程的有前途的策略。然而，由于解剖和技术上的挑战，体内肝脏脱细胞后的长期存活尚未实现。本研究旨在建立大鼠体内部分肝叶灌注-脱细胞存活模型。我们比较了三种脱细胞方案（1% Triton X100，然后是 1% 十二烷基硫酸钠 [SDS]、1% SDS 与 1% Triton X100，n = 6/组）。使用通过肉眼观察、组织学和 DNA 含量判断的最佳蛋白，我们通过组织学、扫描电子显微镜、计算机断层扫描和免疫组织化学 (IHC) 来表征结构完整性和基质蛋白。我们通过使用聚偏二氯乙烯 (PVDC) 薄膜 + 干纱布与 PVDC 薄膜 + 干纱布 + 抽吸管 ( n = 6/组）。通过组织学评估生理再灌注。在 7 天的观察期后确定存活率。仅用 1% SDS 灌注产生无细胞支架（完全半透明，没有组织学可检测的组织残余，DNA 浓度小于天然叶中的 2%）具有显着的结构和超微结构完整性以及主要基质蛋白的保存（IHC 阳性IV 型胶原蛋白、层粘连蛋白和弹性蛋白）。除了 PVDC 薄膜 + 干纱布外，还通过放置吸管实现了潜在有毒 SDS 溶液对腹部器官的污染，并允许所有动物存活 7 天，而没有严重的术后并发症。再灌注时，肝脏在几秒钟内变红，肝脏表面没有任何渗漏。再灌注后约 12 h，在门静脉、正弦基质网络和中央静脉中不仅可以看到血细胞，还可以看到一些凝块，表明生理灌注。总之，我们的这项研究结果显示了关于生成生存模型的第一个可用数据体内实质器官去细胞化，为体内器官工程迈出了关键一步。