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7d后,用免疫组化检测羊膜上皮细胞的增殖活性、NT-3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-3、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。
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-100 and 1% sodium dodecyl sulfate.The histological structure of acellular muscle was observed in frozen section.Then the cultured human amniotic cells were implanted into acellular muscle scaffolds.After cultured 1 w,the scaffolds with cells were sectioned,and BrdU,NT-3 and BDNF immunohistochemical staining were performed to observe the proliferating capacity and the expressions of NT-3 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.