CAJ | 학술논문

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。
목적：대거세포저주동맥판막（DPAV）진행취을이순（PEG）개성，구건혈액상용성호적복합지가（PEG-DPAV）。방법기우고상유궤합성이론대거세포판진행구기화수식이용구기-병희산지발생적매극이가성반응，사인입도 DPAV 중적구기여4-arm-PEG-acrylate 중적병희산지공개결합，전반사부리협변환홍외광보（ATR-FTIR）대PEG화적DPAV진행표정。복합지가혈소판점부실험평개기혈액상용성, CCK-8법검측복합지가침제액대대서내피세포증식솔적영향，평개기세포독성。결과 PEG-DPAV ATR-FTIR광보：제출현DPAV특정봉외환출현PEG특정신호봉：1100，1342，C-O，-C-H2진동봉，증명DPAV성공접지상PEG。복합지가화DPAV독성평급균0급，화음성대조조차이무통계학의의。대서내피세포경함침제액적배양액배양후형태량호，증식왕성，복합지가표면혈소판점부수량교소，무취집。결론이용매극이가성반응성공장PEG교련도DPAV상，반응조건온화，개성효과학절。복합지가표현출량호적혈액상용성，무세포독성，괄합조직공정심장판막지가적구건。
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.