组织工程心脏瓣膜支架制备方法研究

Tissue-engineered heart valve leaflets: an effective method of obtaining acellularized pocine valve

目的 :探讨去垢剂、胰蛋白酶对于猪主动脉瓣膜去细胞过程的影响 ,研究心脏瓣膜组织工程生物支架的最佳制备方法。方法 :将新鲜猪主动脉瓣膜用氯化钠、十二烷基硫酸钠、曲拉通和胰蛋白酶等处理 ,去除内皮细胞和间质细胞 ,常规苏木精伊红和Masson染色 ,电子扫描显微镜检查 ,评价效果。结果 :NaCl-SDS对瓣膜超微结构损伤小 ,但去细胞不彻底 ,胰蛋白酶法去细胞彻底 ,对结构损害大 ,曲拉通法能有效去除细胞成分 ,对瓣膜结构损伤小。结论

Objective: Tissue engineering of heart valves should avoid the disadvantages of conventional prostheses. In this study we tested different decellularization procedures for their potential of cell removal and their ability to preserve the matrix. Methods:Specimens of porcine aortic roots were treated with either trypsin or NaCl and sodium-dodecyl-sulfate (SDS) or Triton-X 100 and RNase and DNase with specific concentrations. Tissue samples were then processed for scanning electron microscopy and Hematoxylin-Eosin and Masson's trichrome. Results:NaCl and SDS achieved only incomplete decellularization, In contrast trypsin removed cells completely but caused strong structural alterations. Treatment with Triton-X100 achieved both complete decellularization and preservation of the matrix structure. Conclusion:Techniques of decellularization are highly variable in efficiency and matrix preservation and was best achieved in our study with Triton-X100.