生物反应器内构建组织工程心脏瓣膜  

Fabrication of tissue engineering heart valves in a bioreactor system

作　　者：史峰[1] 邓诚[1] 胡行健[1] 史嘉玮[1] 董念国[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院心血管外科,武汉430022

出　　处：《中华实验外科杂志》2013年第2期315-316,共2页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30872540）;国家863计划资助项目（2009AA032420）

摘　　要：目的去细胞瓣膜上种植人主动脉瓣膜间质细胞，生物反应器内构建组织工程心脏瓣膜。方法PEG交联去细胞瓣，GRGDSPC多肽共价修饰。RGD—PEG-去细胞瓣上种植人主动脉瓣膜问质细胞，缝制于生物反应器内。接种2、4、8h后，细胞计数观察细胞黏附情况。体外培养10d后行形态学观察和DNA含量测定。结果免疫荧光检测显示，GRGDSPC多肽与PEG一去细胞瓣有效共价接枝。接种2、4、8h后，RGD—PEG一去细胞瓣组和单纯去细胞瓣组细胞计数分别为：（4．98±0．46）X107／L比（3．35±0．15）×107／L，（5．71±0．29）×104个／ml比（3．554-0．18）×10’／L，（5．97±0．45）×10’／L比（3．62±0．18）×107／L，P〈0．05。RGD—PEG．去细胞瓣组较单纯去细胞瓣组DNA含量明显增高，（25．98±2．45）¨∥瓣叶比（19．61±1．94）ug/瓣叶，差异有统计学意义（P〈0．05）。结论生物反应器内，GRGDSPC接枝的PEG-去细胞瓣复合支架，可促进种子细胞的黏附，改善瓣膜支架生物学性能。Objective Human aortic valve interstitial cells （HAVICs） were seeded onto decellu- larized valves in order to fabricate tissue engineering heart valves in a bioreactor system. Methods Decellu- larized valves were cross-linked by polyethylene glycol. PEGylated deeellularized valves were covalently mod- ified by GRGDSPC peptides. The modified effect of GRGDSPC peptides was determined by using immunofiu- orescence. HAVICs were seeded onto PEGylated decellularized valves modified by GRGDSPC peptides. Then the valves were sutured into a bioreactor system. Cell adhesion was tested respectively after 2, 4 and 8 h. Af- ter 10 days of culture, hematoxylin-eosin staining, scanning electron microscopy and DNA content test were performed. Results Immunofluorescence showed that PEGylated decellularized vlves could be effectively modified by GRGDSPC peptides through covalent bonding. As compared with decellularized valves, PEGylat- ed decellularized valves modified by GRGDSPC peptides significantly promoted the adhesion of HAVICs. Number of adherent cells after 2, 4 and 8 h was respectively （4. 98 ±0. 46） 107/L vs. （3.35 _＋0. 15） x 10T/L, （5.71 ±0.29） xl07/L vs. （3.55 ＋0.18） x107/L, （5,97±0.45） ~ 107/L vs. （3.62 ＋ 0. 18） x 107/L,P 〈0. 05. The DNA content of PEGylated decellularized valves modified by GRGDSPC pep- tides was significantly higher than that of decellularized valves [ （25.98 ± 2.45 ）ug/valve vs. （ 19. 61 ± 1.94） ug/valve,P 〈 0. 05 ]. Conclusion Modification of PEGylated decellularized valves by GRGDSPC peptides can promote the adhesion of cells seeded on valves in a bioreactor system.

关 键 词：去细胞瓣 聚乙二醇 生物反应器 黏附 

分 类 号：R318.08[医药卫生—生物医学工程]