上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

聚电解质多层膜修饰脱细胞组织工程血管的实验研究

刘俊，张晓膺   

  1. 苏州大学附属第三医院心胸外科，常州 213003
  • 出版日期:2016-12-28 发布日期:2016-12-29
  • 通讯作者: 张晓膺，电子信箱：zhangxy6689996@163.com。
  • 作者简介:刘俊（1976—），男，副主任医师，博士生；电子信箱：liujun52@126.com。 [通信作者]

Experimental study on polyelectrolyte multilayer film-modified decellularized tissue engineered vascular grafts

LIU Jun, ZHANG Xiao-ying   

  1. Department of Cardiothoracic Surgery, the Third Affiliated Hospital, Soochow University, Changzhou 213003, China
  • Online:2016-12-28 Published:2016-12-29

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摘要：

目的 ·寻找制备组织工程血管的最佳脱细胞试剂，用肝素-SDF-1α聚电解质多层膜修饰脱细胞血管支架，评估其组织相容性。方法 ·分别用含有0.25%Triton-X100+0.25%SD（TS组）、1%SDS（SDS组）和0.5%Triton-X100 +0.5%SD+0.5%SDS（STS组）的PBS对大鼠主动脉进行脱细胞处理；以未脱细胞的大鼠主动脉作为对照组。通过苏木精-伊红染色、DNA定量和扫描电子显微镜验证最佳脱细胞试剂。用肝素-SDF-1α聚电解质多层膜修饰脱细胞血管表面，采用免疫荧光法明确聚电解质多层膜在血管表面的结合；同时，通过血小板激活测定及大鼠骨髓间充质干细胞黏附、迁移实验在体外验证其生物相容性。结果 · STS组大鼠主动脉脱细胞效果最完全，同时血管的三维结构得到了很好的保留。肝素-SDF1α聚电解质多层膜能牢固结合在脱细胞血管管壁上，肝素-SDF1α修饰后的脱细胞血管明显减少了血小板在血管表面的黏附激活，增强了大鼠骨髓间充质干细胞在血管表面的黏附增殖和迁移能力。结论 ·含有0.5%Triton-X100 +0.5%SD+0.5%SDS的PBS是最佳脱细胞试剂。肝素-SDF-1α聚电解质多层膜能很好地结合在脱细胞血管表面，其修饰的脱细胞血管具有较好的生物相容性。

关键词: 聚电解质多层膜, 组织工程血管, 脱细胞血管

Abstract:

Objective · To find the best acellular reagent for the preparation of tissue engineered vascular grafts (TEVGs) and to modify decellularized vascular grafts with heparin -SDF-1α polyelectrolyte multilayer film and evaluate the biocompatibility of TEVGs. Methods · We decellularized rat aortas with PBS solutions containing 0.25% Triton-X100 and 0.25% SD (the TS group), 1% SDS (the SDS group), and 0.5% Triton-X100, 0.5% SD, and 0.5% SDS (the STS group) with untreated rat aortas serving as controls. The optimal acellular reagent was identified with HE staining, DNA quantification, and scanning electron microscope. After modifying decellularized vascular grafts with heparin -SDF-1α polyelectrolyte multilayer film, we used immunofluorescence to verify the binding of TEVGs on vascular surface. The biocompatibility of modified TEVGs was evaluated with migration assay in vitro and the platelet activation assay was used to measure the adhesion of rat bone marrow mesenchymal stem cells. Results · The STS group had the best aorta decellularization with good preservation of the three-dimensional structure of blood vessels. Heparin -SDF1α polyelectrolyte multilayer film could bind firmly on the surface of TEVGs. Heparin -SDF-1α polyelectrolyte multilayer film-modified TEVGs significantly reduced the adhesion and activation of platelet on the surface of TEVGs and greatly enhanced proliferation and migration abilities of rat bone marrow mesenchymal stem cells. Conclusion · The PBS solution containing 0.5% SD, 0.5% SDS, and 0.5% Triton-X100 was the best acellular reagent. The heparin -SDF-1 polyelectrolyte multilayer film can favorably bind on the surface of acellular vessels, which have good biocompatibility.

Key words: polyelectrolyte multilayer film, tissue engineered vascular graft, decellularized vascular