摘要
目的 初步探讨组织工程心脏瓣叶体外生成的可行性。方法 杂种猪 6只 ,取主动脉。分离血管内皮细胞及成纤维、平滑肌细胞 ,予培养扩增。将成纤维、平滑肌细胞与内皮细胞按顺序种植在预湿处理的支架材料上 ,使用DMEM培养。以首次种植成纤维、平滑肌细胞为起点 ,2 8d送样本进行扫描电镜、组织学检查和羟脯氨酸测定。结果 扫描电镜检测显示 ,瓣叶支架上可见大量细胞粘附 ,支架表面细胞融合成片 ,表面尚可见细胞基质形成。HE染色显示材料上可见大量细胞粘附 ,已长入材料中部。弹性纤维染色显示有弹性纤维合成。羟脯氨酸测定显示瓣叶材料有羟脯氨酸合成。结论 使用体外分离、培养及传代的内皮细胞和成纤维、平滑肌细胞 ,经体外种植、培养 ,有可能生成具有活性的组织工程心脏瓣叶。
Objective: The experiment was designed to evaluate the in vitro creation of tissue engineered heart valve leaflets. Methods: Aortic artery was harvested from 6 hybrid young pigs. The endothelial and myofibroblast cells were isolated and cultured to get enough cells for seeding. The myofibroblast and endothelial cells were seeded on the scaffolds sequentially. The cellular scaffolds were cultured 28 days and sent for scanning electron microscopy (SEM), histological and hydroxyproline examination. Results: SEM showed that a large amount of cells attached on the scaffolds. The cell had covered a large area of scaffold with a lot of matrix synthesized. The HE stain showed large amount of cells growing on the scaffolds and some cells were found in the central part of scaffolds. Gomori stain showed that there was elastin synthesized on the scaffolds. Hydroxyproline content is (10.93±2.89) μg/g (scaffolds). Conclusion: With vascular cell, it is possible to seed them onto biodegradable scaffolds and create a viable tissue-engineered heart valve leaflets in vitro.
出处
《中华胸心血管外科杂志》
CSCD
北大核心
2004年第1期41-43,共3页
Chinese Journal of Thoracic and Cardiovascular Surgery
基金
广东省自然科学基金重点项目资助 (c 0 10 177)
关键词
组织工程
心脏瓣叶
体外生成
细胞培养
羟脯氨酸
人工心脏瓣膜
Heart valve prosthesis Cell culture Scanning electron microscopy Histology hydroxyproline Tissue engineered
