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Mechanistic analysis of in vivo rapid endothelialization of biofunctionalized small-diameter acellular graft

机译：生物功能化小直径脱细胞移植物体内快速内皮化的机理分析

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Introduction: The clinical efficiency of small-diameter synthetic vascular grafts has been limited by rapid thrombosis and poor patency. We recently achieved excellent patency of the small-diameter decellularized vascular grafts with the inner diameter of 2 mm and the length of 20-30 cm in porcine femoral-femoral crossover bypass model (FF bypass) by modifying the luminal surface with the neointima-inducing peptide modifier. This modification was found to reduce the early stage thrombus formation, which is one of the most important mechanisms. In this study, the neointima formation process in the minipig model was monitored from 1 hr to 1 week, and the cells participated in the luminal endothelialization was identified by scanning electron microscopic (SEM) analysis and histological staining. Material and Method: Ostrich carotid artery was decellularized by ultra-high hydrostatic pressure treatment. The graft was immersed with 10 mM peptide solution for peptide modification. The peptide-modified and unmodified grafts were transplanted into pig femoral artery as FF bypass and orthotopic transplantation. After transplantation, the luminal surface was observed by SEM. Endothelialization was evaluated by histological staining using HE, vWF, as well as primary antibody against vimentin, CD31, CD105, CD34, and Flk-1. Results: SEM and HE staining revealed that the luminal surface of the grafts in FF bypass at the center part was completely covered with cells in three days. These cells expressed CD34 and Flk-1, which were progenitor makers. Cellular uptake of acetylated-low density lipoprotein was also confirmed. After orthotopic transplantation for three month, the cell layer expressed vWF, CD31, and CD105 without CD34 and Flk-1 expression. Discussion: Some researchers have been proposing that the progenitor cells for endothelial cells exist in the peripheral blood. In our experimental results clarified that the endothelial-like progenitor cells were captured onto the luminal surface of peptide-modified grafts in 3 days. These cells formed the matured endothelial cell layer with the histological feature similar to the native blood vessels after three months. Conclusion: The progenitor cells, which exist in the peripheral blood flow, were captured rapidly by the peptide-modified surface, and it was found that the good patency and suppressed thrombogenicity were achieved due to the rapid endothelialization mechanism.
机译：简介：小直径合成血管移植物的临床疗效受到快速血栓形成和通畅性的限制。我们最近通过用新内膜诱导修饰管腔表面，在猪股-股交叉旁路模型（FF旁路）中获得了内径为2 mm，长度为20-30 cm的小直径脱细胞血管移植物的出色通畅性。肽修饰剂。发现这种修饰减少了早期血栓形成，这是最重要的机制之一。在这项研究中，从1小时到1周监测了小型猪模型中的新内膜形成过程，并通过扫描电子显微镜（SEM）和组织学染色鉴定了参与腔内内皮化的细胞。材料与方法：鸵鸟颈动脉经超高压静水处理脱细胞。将移植物浸入10 mM肽溶液中以进行肽修饰。经FF旁路和原位移植将肽修饰的和未修饰的移植物移植到猪股动脉中。移植后，通过SEM观察腔表面。使用HE，vWF以及针对波形蛋白，CD31，CD105，CD34和Flk-1的一抗通过组织学染色评估内皮化。结果：SEM和HE染色显示，在3天的FF旁路中，移植物的腔表面在中心部分被细胞完全覆盖。这些细胞表达了CD34和Flk-1，它们是祖细胞的制造者。还证实了乙酰化低密度脂蛋白的细胞摄取。原位移植三个月后，细胞层表达vWF，CD31和CD105，而没有CD34和Flk-1表达。讨论：一些研究人员已经提出，内皮细胞的祖细胞存在于外周血中。在我们的实验结果中阐明，内皮样祖细胞在3天内被捕获到了肽修饰的移植物的腔表面上。这些细胞在三个月后形成了成熟的内皮细胞层，其组织学特征类似于天然血管。结论：外周血中存在的祖细胞可通过肽修饰的表面快速捕获，并发现通过快速内皮化机制可获得良好的通畅性和抑制的血栓形成性。

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