组织工程与重建外科杂志 ›› 2025, Vol. 21 ›› Issue (5): 458-.

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华通胶衍生静电纺丝制备人工血管支架在血管重建中的应用研究

  

  • 出版日期:2025-10-13 发布日期:2025-10-30

Application of Wharton's jelly-derived electrospun artificial vascular grafts in vascular reconstruction research

  • Online:2025-10-13 Published:2025-10-30

摘要:

目的 探讨以华通胶(Wharton's jelly,WJ)制备静电纺丝人工血管支架用于血管重建的可行性。方法 取自 愿捐赠新鲜脐带,分离WJ并行胰蛋白酶脱细胞处理,然后与聚己内酯(Polycaprolactone,PCL)混合,通过静电纺丝制备 WJ/PCL静电纺丝人工血管支架。同法制备PCL静电纺丝人工血管支架,作为对照。采用液体置换法测定孔隙率,静态 接触角测试法检测亲水性,BCA蛋白检测试剂盒评估蛋白吸附性,称重方法测定体外降解性能。与人脐静脉内皮细胞
(Human umbilical vein endothelial cells,hUVECs)共培养7 d,活/死细胞染色和CCK-8检测细胞相容性;同时行兔皮下免 疫反应评估(CD45和CD163免疫荧光染色)。取10只新西兰大白兔制备左侧颈动脉长1 cm缺损,分别使用PCL和WJ/PCL静电纺丝人工血管修复(n=5),术后 1个月多普勒超声和大体观察血管管径和通畅率、生物力学性能观测,以及组织学(HE 及 Masson 染色)、免疫组织化学染色[平滑肌细胞(Smooth muscle cell,SMC)、α 平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和血管内皮细胞(CD31)]评估血管结构。结果 实验成功制备管状PCL和WJ/PCL静电纺丝人工 血管支架,与PCL静电纺丝人工血管支架相比,WJ/PCL静电纺丝人工血管支架亲水性、蛋白吸附性、体外降解率、细胞相容性均更好(P<0.05),免疫反应更低(P<0.05)。动物体内实验显示,术后1个月,相较于PCL组,WJ/PCL组颈动脉通畅率更高、管径更大(P<0.05);大体观察显示较白的修复段血管组织;生物力学评估示更高的极限拉伸应力和断裂拉伸率;无严重肉芽增生,可见更加均质成熟的纤维结缔组织,SMC、α-SMA 和 CD31 表达均提升,差异有统计学意义(P<0.05)。结论 基于WJ制备的WJ/PCL静电纺丝人工血管能够有效促进兔颈动脉缺损的修复。

关键词: 华通胶, &emsp, 人工血管, &emsp, 静电纺丝, &emsp, 组织工程, &emsp, 血管重建

Abstract:

Objective To explore the feasibility of preparing electrospun artificial vascular grafts using Wharton's jelly
(WJ) for promoting vascular reconstruction. Methods Fresh umbilical cords were voluntarily donated, and WJ was isolated and decellularized with parallel trypsinization. The decellularized WJ was then mixed with polycaprolactone (PCL) and electrospun to prepare WJ/PCL electrospun artificial vascular scaffolds. PCL electrospun artificial vascular scaffolds were prepared using the same method as a control. Liquid displacement method was used to determine the porosity, static contact angle test was used to detect hydrophilicity, BCA protein assay kit was used to evaluate protein adsorption, and weighing method was used to determine in vitro degradation performance. Human umbilical vein endothelial cells (HUVECs) were cocultured for 7 days, and cell viability was assessed using live/dead cell staining and cell counting kit-8 (CCK-8). Subcutaneous immune response assessment was conducted in rabbits (immuno fluorescence staining for CD45 and CD163). Ten New Zealand white rabbits were used to prepare a 1 cm defect in the left carotid artery. PCL and WJ/PCL electrospun  artificial blood vessels were used for repair (n=5 each). Doppler ultrasound and gross observation were performed 1 month postoperatively to assess vessel lumen diameter and patency. Biomechanical properties, and histology (Hematoxylin and Eosin staining and Masson's trichrome staining), as well as immunohistochemistry staining [smooth muscle cells (SMCs), α-smooth muscle actin (α-SMA), and endothelial cells (CD31)] were used for evaluating vascular structure. Results The experiment successfully prepared tubular PCL and WJ/PCL electrospun artificial vascular scaffolds. Compared to PCL electrospun artificial vascular scaffolds, the WJ/PCL scaffolds exhibited better hydrophilicity, protein adsorption, in vitro degradation rate, and cell compatibility (P<0.05), with lower immune response (P<0.05). Animal experiments showed that 1 month postoperatively, compared to the PCL group, the WJ/PCL group had higher carotid artery patency rate and larger lumen diameter (P<0.05). Gross observation showed a whiter repaired segment of vascular vessel tissue. Biomechanical evaluation showed higher ultimate tensile stress and fracture elongation. There was no severe granulation tissue hyperplasia, and more homogeneous and mature fibrous connective tissue was observed. The expression of SMCs,α-SMA, and CD31 was increased, with statistically significant differences (P<0.05). Conclusion The WJ/PCL electrospun artificial vascular scaffolds prepared based on WJ effectively promotes the repair of rabbit carotid artery defects.

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