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人脐带静脉内皮细胞与骨髓间充质干细胞共培养于肝素化聚己内酯/明胶共纺纳米纤维上以促进内皮细胞重构。

Co-culture of mesenchymal stem cells and human umbilical vein endothelial cells on heparinized polycaprolactone/gelatin co-spun nanofibers for improved endothelium remodeling.

机构信息

Department of Chemical Engineering, Faculty of Engineering, Graduate School, Kyushu University, 744 Motooka, Nishi-Ku, Fukuoka 819-0395, Japan.

Department of Chemical Engineering, Faculty of Engineering, Graduate School, Kyushu University, 744 Motooka, Nishi-Ku, Fukuoka 819-0395, Japan.

出版信息

Int J Biol Macromol. 2020 May 15;151:186-192. doi: 10.1016/j.ijbiomac.2020.02.163. Epub 2020 Feb 16.

Abstract

Endothelization of a tissue-engineered substrate is important for its application as an artificial vascular graft. Despite recent advancements in artificial graft fabrication, a graft of <5 mm is difficult to fabricate owing to insufficient endothelization that results in thrombosis after transplantation. We aimed to perform a co-culture of adipose-derived mesenchymal stem cells (MSCs) with human umbilical vein endothelial cells (HUVECs) on antithrombogenic polycaprolactone (PCL)/heparin-gelatin co-spun nanofibers to evaluate the role of co-culturing in promoting quick endothelization of vascular substrates without surface modification by growth factors or other ECM proteins that trigger the endothelization process. Using a co-axial electrospinning technique, we attempted to fabricate our scaffold balancing between mechanical properties and biocompatibility. Antithrombogenic characteristics were then imparted to the fabricated nanofiber substrate by grafting of heparin. Finally, we performed a co-culture of MSCs and HUVECs on the fabricated co-spun nanofiber substrate to obtain proper endothelization of our material under the in-vitro culture. Staining for CD-31 at seven days of culture revealed enhanced CD-31 expression under the co-culture condition; actin staining revealed healthy cobblestone HUVEC morphology, suggesting that MSCs can aid in proper endothelization. Hence, we conclude that co-culture is effective for quick endothelization of vascular substrates.

摘要

组织工程化基底的内皮化对于其作为人工血管移植物的应用非常重要。尽管人工移植物制造技术最近取得了进展,但由于内皮化不足导致移植后血栓形成,因此难以制造 <5mm 的移植物。我们旨在在抗血栓聚己内酯(PCL)/肝素-明胶共纺纳米纤维上进行脂肪间充质干细胞(MSCs)与人脐静脉内皮细胞(HUVECs)的共培养,以评估共培养在促进血管基底的快速内皮化而无需表面改性生长因子或其他 ECM 蛋白以触发内皮化过程中的作用。我们使用同轴电纺技术试图在机械性能和生物相容性之间平衡制造我们的支架。通过肝素接枝赋予了制造的纳米纤维基底抗血栓形成特性。最后,我们在制造的共纺纳米纤维基底上进行 MSCs 和 HUVEC 的共培养,以在体外培养下获得我们材料的适当内皮化。培养 7 天后 CD-31 的染色显示共培养条件下 CD-31 表达增强;肌动蛋白染色显示健康的鹅卵石状 HUVEC 形态,表明 MSCs 可以帮助实现适当的内皮化。因此,我们得出结论,共培养对于快速内皮化血管基底是有效的。

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