体外构建组织工程心脏瓣膜动物实验初步研究

目的探讨应用去细胞猪主动脉支架(decellularized porcine aortic valve scaffold，APAVS)与兔骨髓干细胞(rabbit bone marrow stromal cells，RBMSCs)体外构建组织工程心脏瓣膜的可行性。方法采用去垢剂-核酸酶消化法处理，去除猪主动脉瓣细胞成分，并做去细胞前后的形态学检查和生物力学测定；在去细胞支架上种植兔骨髓干细胞，行形态学检查和免疫组化测定。结果光镜及电镜证实，猪主动脉瓣膜中的细胞成分可完全去除，获得完整无细胞的纤维网状支架；瓣叶去细胞前后的断裂强度和断裂伸长率无明显变化；种植的RBMSCs可在ACPAV表面形成一层连续的细胞层。结论种植RBMSCs于ACPAV上，可体外构造组织工程人工心脏瓣膜。

Primary study on tissue-engineered heart valve in vitro

Objective To study the feasibility of tissue engineered heart valve reconstructed on acellularized porcine aortic valve and rabbit bone marrow stromal cells.Methods The porcine aortic valve was acellularized with the detergent and enzymatic extraction process.Then the decellularized scaffolds and the fresh valves were compared in the morphological and biomechanical properties.The scaffolds were seeded with RBMSCs for 3 times and cultured for another 5 days.The TEHVs were analyzed by light microscopy,electron microscopy and immunohistochemistry.Results The acellularization procedure resulted in an almost complete removal of the cellular components and soluble protein of valves while the construction of matrix was properly maintained;The subsequent seeding with RBMSCs on the matrix was so successful that the surface of the scaffold had been covered with a continuous monolayer of cells through LM and EM.Immunohistochemistry indicated,the RBMSCs seeded on the matrix were positive for ASMA,and negative for CD31.Conclusion It is feasible to reconstruct TEHVs in vitro on APAVS and RBMSCs.