采用纳米支架材料的新型多层平板型生物人工肝的免疫安全性研究

目的：探讨新型生物人工肝（BAL）系统的免疫安全性。 方法：采用自制纳米支架材料的新型多层平板型生物反应器与猪肝细胞-骨髓间充质干细胞共培养体系构建新型的BAL系统。正常比格犬接受新型BAL体外循环灌注6 h，期间定时收集BAL系统中的血液及培养液。ELISA法检测培养液及术前、术中、术后血浆中IgG，IgM及CH50（C3）水平；检测外循环前后的血常规情况；免疫荧光法检测术后反应器内细胞材料表面以及犬心、肝、脾、肺、肾等器官免疫蛋白的沉积情况。 结果：经过6 h的体外循环灌注，犬体内IgG，IgM免疫抗体水平术中、术后均未出现明显的升高或下降，而术中补体CH50水平出现一过性下降，但很快恢复至正常水平，且并未发生严重的过敏反应及排斥反应，反应器内培养液中检出极少量的IgG和CH50；血常规检测显示，术后白细胞、血小板及淋巴细胞未出现明显的升高或降低；术后反应器内细胞材料表面仅有微量的IgG与C3沉积，各重要脏器均未检测到IgG，IgM及补体C3的沉积。 结论：新型BAL系统具有良好的免疫安全性。

Immunosafety assessment of a novel multilayer flat-plate bioartificial liver based on nanofiber scaffolds

Objective: To assess the immunosafety of a novel bioartificial liver (BAL) system. Methods: The novel BAL system was constructed with the multilayer flat-plate bioreactor based on nanofiber scaffolds and the coculture system consisted of porcine hepatocytes and bone marrow mesenchymal stem cells. Normal beagles underwent extracorporeal circulation perfusion with this novel BAL system for 6 hours, and the blood and medium samples were collected at regular intervals from the BAL system during operation. The levels of IgG, IgM and CH50 (C3) in the medium and the pre-, intra- and postoperative plasma were detected by ELISA assay. The routine blood tests of the beagles were performed before and after operation. The deposition of immune proteins in the cell material surface of the bioreactor and tissues from heart, liver, spleen, lung and kidney of the beagles were determined by immunofluorescence assay. Results: The levels of IgG and IgM of the beagles had no significant increase or decrease during and after the 6-hour extracorporeal circulation, while the level of CH50 had a transient decrease during operation, then quickly recovered to normal level, and no severe allergic or rejection reaction occurred. There was only a very small amount of IgG and CH50 detected in the medium of the bioreactor. No significant increase or decrease of white blood cells, platelets or lymphocytes after operation was found on routine blood tests. A very small deposition of IgG and C3 was detected in the cell material surface of the bioreactor but no deposition of IgG or IgM or C3 was detected in any of the important organs. Conclusion: The novel BAL system has a satisfactory immunosafety profile.