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首页> 外文期刊>Annals of Plastic Surgery >Tissue-engineered blood vessel graft produced by self-derived cells and allogenic acellular matrix: a functional performance and histologic study.
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Tissue-engineered blood vessel graft produced by self-derived cells and allogenic acellular matrix: a functional performance and histologic study.

机译：自体细胞和同种异体脱细胞基质产生的组织工程血管移植物：功能性能和组织学研究。

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INTRODUCTION: Current prosthetic alternatives to autologous vascular grafts remain poor in terms of patency. Growing biologic blood vessels has been proposed as an alternative. In this work, the authors demonstrate a method for producing a tissue-engineered vascular graft (TEVG) with self-derived endothelial cells, smooth muscle cells, and allogenic acellular matrix in vitro. The aim of this study was to find out if the graft is suitable as the carotid artery substitute. METHODS: A canine model was developed for this study. Endothelial and smooth muscle cells were used as seeding cells, and allogenic acellular matrix was used as scaffold to produce the TEVG. Endothelial and smooth muscle cells from the saphenous vein were harvested by trypsin and collagenase digestion respectively. These isolated cells were cultured and expanded by routine cell culture technique. The common carotid artery was harvested from other fresh dog cadavers and processed by a multistep decellularizing technique to remove original cells and preserve elastic and collagen fibers. Then, the inner surface of the acellular matrix as a scaffold was sequentially seeded with cells. Smooth muscle cells were seeded onto the scaffold. It was placed in bioreactors filled Dulbecco modified Eagle medium supplemented with growth factors. After 4 weeks, the vessels were harvested from the bioreactors and seeded with endothelial cells at the lumen for 7 days. Finally, the cell-seeded graft was transplanted to the cell-donated dog to substitute part of the native common carotid artery (2 cm in length). All animals were followed up for 6 months. Twenty-four dogs were divided into 3 groups randomly: group A (native artery graft), group B (allogenic acellular matrix graft only), and group C (acellular allogenic matrix coated with endothelial and smooth muscle cells). These grafts were subjected to regular echocardiography at 1, 3, and 6 months postoperatively. Then, the TEVG were harvested for histologic evaluation at 6 months after transplantation. The vascular luminal surfaces were observed by electronic scanning microscopy. RESULTS: The TEVG showed good functional performance demonstrated by regular Doppler ultrasonography at 1, 3, and 6 months postoperatively, compared with that of native arteries. All vascular grafts in group A and C provided patent rates of 100%; however, the patency rate of group B was 50% at 3 and 6 months postoperatively. The TEVG had a similar macroscopic appearance to that of native vessels. Histologic and immunohistochemical analyses indicated the presence of high cell density and development of a highly organized structure of ECM. CONCLUSION: Cultured self-derived endothelial and smooth muscle cells could be used as seeding cells and allogenic acellularized matrix could be used as scaffold in producing the TEVG. The TEVG had histologic and functional properties consistent with native arteries.
机译：简介：自体血管移植物的当前修复方法在通畅性方面仍然较差。已经提出了生长生物血管作为替代方案。在这项工作中，作者演示了一种在体外生产具有自体内皮细胞，平滑肌细胞和同种异体脱细胞基质的组织工程血管移植物（TEVG）的方法。这项研究的目的是找出移植物是否适合作为颈动脉替代物。方法：为该研究开发了犬模型。内皮细胞和平滑肌细胞用作接种细胞，同种异体脱细胞基质用作支架生产TEVG。通过胰蛋白酶和胶原酶消化分别收集大隐静脉的内皮细胞和平滑肌细胞。这些分离的细胞通过常规细胞培养技术进行培养和扩增。颈总动脉取自其他新鲜的狗尸体，并通过多步脱细胞技术进行处理，以去除原始细胞并保留弹性和胶原纤维。然后，将无细胞基质的内表面作为支架顺序地接种细胞。将平滑肌细胞接种到支架上。将其置于填充有生长因子的Dulbecco改性Eagle培养基中的生物反应器中。 4周后，从生物反应器中收获血管，并在管腔内接种内皮细胞7天。最后，将播种细胞的移植物移植到捐赠细胞的狗中，以替代天然颈总动脉的一部分（长度为2 cm）。对所有动物进行随访6个月。 24只狗随机分为3组：A组（天然动脉移植物），B组（仅异基因无细胞基质移植物）和C组（涂有内皮和平滑肌细胞的无细胞异体基质）。在术后1、3和6个月对这些移植物进行常规超声心动图检查。然后，在移植后6个月收获TEVG用于组织学评估。通过电子扫描显微镜观察血管腔表面。结果：与常规动脉相比，TEVG在术后1、3和6个月通过常规多普勒超声检查显示出良好的功能表现。 A和C组的所有血管移植物的专利率均为100％;但是，B组术后3个月和6个月的通畅率为50％。 TEVG具有与天然血管相似的宏观外观。组织学和免疫组织化学分析表明高细胞密度的存在和高度组织化的ECM结构的发展。结论：培养的自身来源的内皮细胞和平滑肌细胞可作为种子细胞，同种异体脱细胞基质可作为制备TEVG的支架。 TEVG具有与天然动脉一致的组织学和功能特性。

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