• 实验研究 • 上一篇    下一篇

组织工程心脏瓣膜体外构建的初步研究

阮昕华1,刘金鹏2,魏民新2,刘建实3,徐美林4   

  1. 1. 天津医科大学总医院心血管外科
    2. 天津医科大学总医院
    3. 天津市胸科医院心外科
    4. 天津市胸科医院病理科
  • 收稿日期:2010-01-27 修回日期:2010-11-16 出版日期:2011-03-15 发布日期:2011-03-15
  • 通讯作者: 阮昕华

Construction of Tissue engineering heart valves in vitro

  • Received:2010-01-27 Revised:2010-11-16 Published:2011-03-15 Online:2011-03-15

摘要:  目的:初步构建组织工程人工心脏瓣膜。方法:种子细胞选用人骨髓间充质干细胞。分别应用去垢剂、破膜剂、胰蛋白酶对新鲜猪心瓣膜进行去细胞,构建去细胞瓣膜支架。将人骨髓间充质干细胞接种于瓣膜支架上静态培养,观察生长状况。结果:曲拉通-SD法去细胞彻底,保持原有纤维结构。种植的人骨髓间充质干细胞生长于瓣膜支架的表面,电镜可见细胞生长于支架表面。结论 曲拉通法是一种较为理想的猪瓣膜脱细胞方法; 人骨髓间充质干细胞能在猪瓣膜支架上较好地黏附和生长。从而初步构建组织工程人工心脏瓣膜。

关键词:  组织工程心脏瓣膜, 骨髓间充质干细胞, 曲拉通, 去细胞猪瓣膜

Abstract: Objective : Construction of Tissue engineering heart valves in vitro. Methods: firstly, comparison of different methods of obtaining acellularized valve xenografts. Specimens of porcine aortic roots were treated with either Trypsin-sodium deoxycholate(SD)or sodium-dodecyl-sulfate (SDS) or Triton-X100-SD with specific concentrations. Tissue samples were then processed for scanning electron microscopy and Hematoxylin- Eosin and Masson’s trichrome. Observe cell-scaffold under light microscope and scanning electron microscopy. Then, bone marrow mesenchymal stem cells (MSCs) were harvested , cultured and expended and reseeded on the acellularized valve xenografts under static conditions for 7 days .Results: compared with the treatments of Trypsin -SD and SDS, Triton-SD achieved both complete decellularization and preservation of the matrix structure. Cell-scaffold after implantation formed three dimensional tissue, scanning electron microscopy show bone MSCs can grow on the surface of Scaffold. Conclusion: Techniques of decellularization are highly variable in efficiency and matrix preservation and was best achieved in our study with Triton-SD. Bone MSCs are good sources of seeding cells which can adhesion and grow well to decelluarized porcine heart valve in vitro environment.

Key words: Tissue engineering Heart valves, Bone marrow mesenchymal stem cells (MSCS), Triton, Decelluarized porcine heart valve