摘要
目的探讨应用去细胞猪主动脉瓣支架(ace llu larized porc ine aortic va lve scaffo ld,APAV S)与兔骨髓干细胞(bone m arrow strom a l ce lls,BM SC s)体外构建组织工程瓣膜的可行性。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,并作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BM SC s,行形态学检查和免疫组织化学染色观察。结果光学显微镜、扫描及透射电子显微镜下可见猪主动脉瓣膜中的细胞成分可完全被去除,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(642±102g/mm2vs.636±127g/mm2)和断裂伸长率(62.2%±18.1%vs.54.4%±16.0%)差别无统计学意义(P>0.05)。种植的兔BM SC s在APAV S表面形成一层连续的细胞层。免疫组织化学检查α-平滑肌动蛋白抗体(+),CD 31(-)。结论种植兔BM SC s于APAV S上,可在体外构建组织工程心脏瓣膜。
Objective To explore the feasibility of tissue-engineered heart valve （TEHV） reconstructed on acellularized porcine aortic valve and rabbit bone marrow stromal cells （BMSCs） in vitro. Methods Acellularized was performed in porcine aortic valve by the detergent and enzymatic extraction process . Morphological and biomechanical properties were compared between the decellularized scaffolds and the fresh valves. Rabbit BMSCs were seeded on the scaffolds. The TEHV were analyzed by light microscopy, electron microscopy and immunohistochemistry. Results Almost complete removal of the cellular components and soluble protein of valves were observed , while the construction of matrix was properly maintained. Biomechanical tests demonstrated no statistically significant change in the breaking intensity （642 ± 102 g/mm^2 vs. 636 ± 127g/mm^2） and breaking extensibility （62. 2%± 18. 1% vs. 54. 4%±16. 0%） in the porcine values before and after decellularization. Subsequent seeding with rabbit BMSCs on the matrix was so successful that the surface of the scaffold had been covered with a continuous monolayer cells through light microscopy and electron microscopy. Positive of α-smooth muscle actin and negative of CD31 were observed after rabbit BMSCs seeded on the matrix through immunohistochemistry. Conclusion It is feasible to reconstruct TEHV in vitro on acellularized porcine aortic valve scaffold and rabbit BMSCs.
出处
《中国胸心血管外科临床杂志》
CAS
2005年第5期339-342,共4页
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery
基金
黑龙江省自然科学基金资助项目(D01-08)~~
关键词
去细胞支架
骨髓干细胞
心脏瓣膜
组织工程
Acellularized valve
Bone marrow stromal cell
Heart valve
Tissue engineering
