基于迈克尔加成反应构建抗血小板黏附的去细胞猪主动脉瓣膜复合支架  

Constructing an anti-platelet adhesion composite scaffold for decellularized porcine aortic valve based on the Michael addition reaction

作　　者：周建良[1] 丁静丽[2] 聂彬恩[1] 胡时栋[1] 朱志刚[1] 徐建军[1] 

机构地区：[1]南昌大学第二附属医院心胸外科,330006 [2]南昌大学第二附属医院消化内科,330006

出　　处：《中华临床医师杂志（电子版）》2013年第24期197-200,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然科学基金(81260047);江西省青年科学基金(20122BAB215016);江西省卫生厅科技计划项目(20133064)

摘　　要：目的对去细胞猪主动脉瓣膜(DPAV)进行聚乙二醇(PEG)改性,构建血液相容性好的复合支架(PEG-DPAV)。方法基于固相有机合成理论对去细胞瓣进行巯基化修饰利用巯基-丙烯酸酯发生的迈克尔加成反应,使引入到DPAV中的巯基与4-arm-PEG-acrylate中的丙烯酸酯共价结合,全反射傅里叶变换红外光谱(ATR-FTIR)对PEG化的DPAV进行表征。复合支架血小板黏附实验评价其血液相容性,CCK-8法检测复合支架浸提液对大鼠内皮细胞增殖率的影响,评价其细胞毒性。结果 PEG-DPAV ATR-FTIR光谱:除出现DPAV特征峰外还出现PEG特征信号峰:1100,1342,C-O,-C-H2振动峰,证明DPAV成功接枝上PEG。复合支架和DPAV毒性评级均0级,和阴性对照组差异无统计学意义。大鼠内皮细胞经含浸提液的培养液培养后形态良好,增殖旺盛,复合支架表面血小板黏附数量较少,无聚集。结论利用迈克尔加成反应成功将PEG交联到DPAV上,反应条件温和,改性效果确切。复合支架表现出良好的血液相容性,无细胞毒性,适合组织工程心脏瓣膜支架的构建。Objective Decellularized porcine aortic valve （DPAV） was modified with PEG, constructing a good hemocompatibility acellular heart valve composite scaffold. Methods Thiol modification on decellularized porcine aortic valve was based on solid-phase organic synthesis theory. Using mercapto-acrylate produced by the Michael addition reaction, the thiolated DPAV was covalently bound to the PEG-acrylate ester of acrylic acid, and the DPAV was subsequently modified. Attenuated total reflectance-Fourier transform infrared （ATR-FTIR） was used to characterize the composite scaffold. Platelet adhesion assays, the preliminary evaluation of composite scaffold for blood compatibility, and a CCK-8 assay of composite scaffold leach liquor for mouse endothelial cell proliferation were used to determine cytotoxicity. Results ATR-FTIR showed that PEG peaks that were characteristic of signal peaks outside DPAV had also appeared, at the 1100, 1342, C-O, and -C-H2 vibration peaks, and these showed that DPAV successfully grafted with PEG. The toxicity of complex scaffold and acellular valve were 0 grade. No significant differences were observed as compared with a negative control group. Mouse endothelial cells that were treated with culture medium extracts showed physiological shapes and were highly proliferative, and the scaffold showed diminished platelet adhesion and no aggregation. Conclusions Michael addition reaction was successfully used to crosslink PEG to the DPAV, and the mild reaction conditions achieved an ideal environment for this modification. The composite scaffold showed good biocompatibility, it was non-cytotoxic, and it appears to be a suitable approach for heart valve scaffold tissue engineering.

关 键 词：聚乙二醇 支架改性 迈克尔加成反应 去细胞猪主动脉瓣 抗血小板黏附 

分 类 号：R318.08[医药卫生—生物医学工程]