去细胞肌肉组织工程支架与人羊膜上皮细胞的相容性研究_临床医学论文.docVIP

去细胞肌肉组织工程支架与人羊膜上皮细胞的相容性研究_临床医学论文 去细胞肌肉组织工程支架与人羊膜上皮细胞的相容性研究_临床医学论文 作者：薛辉 陈东 刘佳梅 刘颖 孟晓婷 殷迪 【摘要】 目的 制作去细胞肌肉组织工程支架，并检测其与人羊膜上皮细胞的生物相容性。方法 采用三硝基甲苯和十二烷基磺酸钠结合的化学萃取方法制作去细胞肌肉组织工程支架，冰冻切片观察其结构。将人羊膜上皮细胞种入支架培养7 d后，用免疫组化检测羊膜上皮细胞的增殖活性、NT及BDNF的表达,扫描电子显微镜观察其超微结构。结果 支架中细胞去除完全，其主要结构为平行排列的管状结构。细胞外基质的主要成分弹性纤维和胶原纤维保持完好。羊膜上皮细胞在支架里有增殖活性，并呈现NT、BDNF免疫反应阳性。扫描电镜显示，羊膜上皮细胞在支架中分布均匀，生长良好。结论 成功的制作了去细胞肌肉组织工程支架，其与人羊膜上皮细胞有良好的相容性。 【关键词】 去细胞肌肉;人羊膜上皮细胞;生物相容性 　　【Abstract】 Objective To produce the acellular muscle as biomaterial scaffold and to observe its biocompatibility with the human amniotic epithelial cell.　Methods The acellular muscle scaffolds were made by extraction with 3% Triton X amniotic cells were implanted into acellular muscle scaffolds. After cultured 1 w, the scaffolds with cells were sectioned, and BrdU，NT3 and BDNF of amniotic cells in scaffold. SEM was performed to examine the distribution of human amniotic epithelial cells growing in scaffold.　Results The cells in scaffold completely disappeared after the muscle had been extracted. The scaffold of acellular muscle was arranged in parallel tubules composed of collagen fibers and elastic fibers which were left intact. In scaffold the human amnion cells could proliferate and synthesize NT3 and BDNF. Scanning electron microscopy demonstrated a uniform distribution of cells in scaffold.　Conclusions The cells in rat skeletal muscle are successfully removed by chemical extraction. The acellular muscle scaffold made in this way possess good compatibility with human amniotic cells. 　　【Key words】 Acellular muscle; Human amniotic cell; Biocompatibility 　　 近年来组织工程研究的重要进展之一就是采用自体或异体移植物制作天然生物降解材料的组织工程支架。其中去细胞移植物与机体有良好的生物相容性。去细胞肌肉支架可作为生物工程支架支持神经细胞轴突再生。Mligiliche等〔1〕把去细胞肌肉移植入大鼠坐骨神经缺损处,4 w后发现有大量神经轴突长入去细胞肌肉支架中。由于单独应用去细胞肌肉支架治疗神经系统疾病的效果有限，去细胞肌肉支架要发挥更大的作用往往需要向支架中植入种子细胞〔2，3〕。研究表明羊膜上皮细胞可分泌多种神经因子〔4，5〕，促进神经元轴突的生长，是一种良好的治疗神经系统疾病的种子细胞。本研究利用化学去细胞的方法制成去细胞肌肉支架，并把羊膜上皮细胞种入去细胞肌肉支架内，探究两者的相容性，为开展组织工程治疗神经系统方面的疾病提供新的途径。 　　1 材料与方法 　　1.1 材料 　　1.1.1 实验动物 Wistar 大鼠由吉林大学白求恩医