Scaffold-free tissue-engineered arterial grafts derived from human skeletal myoblasts.
Scaffold-free tissue-engineered arterial grafts derived from human skeletal myoblasts.
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来自人类骨骼成肌细胞的无支架组织工程动脉移植物。
DOI：
10.1111/aor.13930
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发表时间：
2021
期刊：
影响因子：
2.4
通讯作者：
and Ishikawa Y
中科院分区：
文献类型：
--
作者：
Saito J;Yokoyama U;Nakamura T;Kanaya T;Ueno T;Naito Y;Takayama T;Kaneko M;Miyagawa S;Sawa Y;and Ishikawa Y
组织工程血管移植（TEVGs）在成人和儿童患者中都有迫切的需求。虽然有几种方法利用血管平滑肌细胞（SMCs）和内皮细胞作为TEVGs的细胞来源，但这些细胞来源的增殖能力有限，导致无法重建新组织。骨骼肌母细胞因其具有高增殖能力而成为极具吸引力的细胞来源，并且已经在缺血性心肌病患者的临床试验中进行了测试。我们之前的研究表明，周期性静水加压（PHP）促进了血管SMCs中纤维连接蛋白的纤维形成，并且PHP诱导的细胞外基质（ECM）排列能够在不使用支架材料的情况下制造源自SMCs的植入式动脉移植物。我们评估了人类骨骼肌成细胞对PHP暴露的分子反应，并旨在通过暴露于PHP来制造成肌细胞的动脉移植物。为了检测PHP反应基因，我们对暴露于PHP后的人类骨骼肌母细胞进行了大量RNA测序。基因集富集分析显示，PHP暴露与血管发育相关基因之间存在显著正相关。实时聚合酶链反应（RT - PCR）显示，PHP显著上调胶原蛋白和弹性纤维形成相关基因的表达，如纤维连接蛋白、赖氨酸氧化酶、I型胶原α1、IV型胶原α1和tropoelastin。基于这些发现显示了PHP在血管形成中的潜在作用，我们通过重复细胞播种和每24小时暴露于PHP来制造动脉移植物。最终获得的15层成肌细胞移植物胶原含量高，抗拉断裂强度为899±104 mm Hg。将人骨成肌细胞移植物作为膜片植入免疫抑制大鼠的主动脉，在术后60天内发现其内皮化并完全通畅。植入的人成肌细胞逐渐被宿主来源的细胞取代，成功形成具有层状弹性纤维的血管新生组织。这些发现表明，在PHP培养条件下，人骨骼肌母细胞有可能成为无支架植入式动脉移植物的可行细胞来源。
Tissue‐engineered vascular grafts (TEVGs) are in urgent demand for both adult and pediatric patients. Although several approaches have utilized vascular smooth muscle cells (SMCs) and endothelial cells as cell sources for TEVGs, these cell sources have a limited proliferative capacity that results in an inability to reconstitute neotissues. Skeletal myoblasts are attractive cell sources as they possess high proliferative capacity, and they are already being tested in clinical trials for patients with ischemic cardiomyopathy. Our previous study demonstrated that periodic hydrostatic pressurization (PHP) promoted fibronectin fibrillogenesis in vascular SMCs, and that PHP‐induced extracellular matrix (ECM) arrangements enabled the fabrication of implantable arterial grafts derived from SMCs without using a scaffold material. We assessed the molecular response of human skeletal myoblasts to PHP exposure, and aimed to fabricate arterial grafts from the myoblasts by exposure to PHP. To examine the PHP‐response genes, human skeletal myoblasts were subjected to bulk RNA‐sequencing after PHP exposure. Gene‐set enrichment analysis revealed significant positive correlations between PHP exposure and vascular development‐related genes. Real‐time polymerase chain reaction (RT‐PCR) demonstrated that PHP significantly upregulated collagen and elastic fiber formation‐related gene expression, such as fibronectin, lysyl oxidase, collagen type I α1, collagen type IV α1, and tropoelastin. Based on these findings showing the potential role of PHP in vessel formation, we fabricated arterial grafts by repeated cell seeding and exposure to PHP every 24 hours. The resultant 15‐layered myoblast grafts had high collagen content, which provided a tensile rupture strength of 899 ± 104 mm Hg. Human skeletal myoblast grafts were implanted as patch grafts in the aorta of immunosuppressed rats and found to be endothelialized and completely patent until the endpoint of 60 postoperative days. Implanted human myoblasts were gradually replaced by host‐derived cells, which successfully formed vascular neotissues with layered elastic fibers. These findings suggest that human skeletal myoblasts have the potential to be a feasible cell source for scaffold‐free implantable arterial grafts under PHP culture conditions.