在磁场作用下载硫酸软骨素酶ABC纳米微粒可影响许旺细胞的迁移  

Effect of nanoparticles carrying chondroitin sulfate ABC on the migration of Schwann cells in a magnetic field

作　　者：高楗勃 夏冰 李胜友 杨雨洁 马腾 于芃 罗卓荆 黄景辉 Gao Jianbo;Xia Bing;Li Shengyou;Yang Yujie;Ma Teng;Yu Peng;Luo Zhuojing;Huang Jinghui(Department of Orthopedics,Xijing Hospital of the Fourth Military Medical University,Xi'an 710032,Shaanxi Province,China;Department of Cardiology,Xijing Hospital of the Fourth Military Medical University,Xi'an 710032,Shaanxi Province,China)

机构地区：[1]空军军医大学西京医院骨科,陕西省西安市710032 [2]空军军医大学西京医院心内科,陕西省西安市710032

出　　处：《中国组织工程研究》2020年第28期4526-4532,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81672148),项目负责人:黄景辉.

摘　　要：背景:由于缺乏轴突再生微环境以及神经胶质瘢痕的形成,使脊髓损伤修复的临床效果通常不尽如人意。许旺细胞作为神经再生的支持细胞,因其在星形胶质细胞丰富的中枢神经系统的迁移能力差而限制了其对轴突再生的作用。目的:将载有硫酸软骨素酶ABC(chondroitinase ABC,ChABC)的超顺磁性纳米微粒导入许旺细胞,探讨在外部磁场的作用下对许旺细胞在星形胶质细胞区域迁移的影响。方法:从出生1-3d的SD大鼠坐骨、臂丛神经和大脑皮质分别提取许旺细胞和星形胶质细胞并纯化,通过免疫荧光染色鉴定其纯度。以超顺磁性纳米微粒(PEI-SPIONs)最佳转染质量浓度2 mg/L且与ChABC最佳质量比为1∶4对许旺细胞进行转染,同时给予外部磁场强度1.4 T作用2 d,用细胞迁移实验评估单纯许旺细胞、PEI-SPIONs/ChABC转染的许旺细胞以及外部磁场作用下PEI-SPIONs/ChABC转染的许旺细胞在星形胶质细胞区域的迁移能力。结果与结论:①许旺细胞和星形胶质细胞的纯度分别达到(91.7±1.2)%和(93.3±2.2)%;②与许旺细胞组相比,PEI-SPIONs/ChABC转染的许旺细胞进入星形胶质细胞区域的数量明显增加(P<0.05);③与许旺细胞组相比,在外部磁场的作用下PEI-SPIONs/ChABC转染的许旺细胞进入星形胶质细胞区域的数量和迁移距离显著增加(P<0.005);④结果表明,PEI-SPIONs/ChABC转染可以增强许旺细胞突破胶质瘢痕的能力,且增加了与星形胶质细胞的融合;同时在外部磁场的引导下显著增强了许旺细胞的迁移能力,该方法可能成为治疗脊髓损伤后轴突再生的一种新策略。BACKGROUND: The clinical effect of spinal cord injury is usually unfavorable due to the lack of axon regeneration and the formation of glial scar. Schwann cells, as the support cells for nerve regeneration, have poor migration ability in the central nervous system with abundant astrocytes, which limit its effect on axon regeneration. OBJECTIVE: To explore the effect on the migration of Schwann cells containing superparamagnetic nanoparticles loaded with chondroitinase ABC(ChABC) in the region of astrocytes in the external magnetic field. METHODS: Schwann cells and astrocytes were extracted from sciatic nerves and brachial plexus and cerebral cortex of Sprague-Dawley rats of postnatal day 1 to 3. Cell purity was identified by immunofluorescence staining. The toxicity of superparamagnetic nanoparticles(PEI-SPIONs) to Schwann cells was analyzed by live/dead cell staining. Schwann cells were transfected with PEI-SPIONS in an external magnetic field of 1.4 Td for 2 days. The optimal transfection concentration of PEI-SPIONS used was 2 mg/L and the optimal mass ratio of PEI-SPIONS to ChABC was 1:4. Cell migration test was used to evaluate the migration ability of not-treated Schwann cells, PEI-SPIONs/ChABC transfected Schwann cells, and PEI-SPIONs/ChABC transfected Schwann cells in an external magnetic field. RESULTS AND CONCLUSION: The purity of Schwann cells and astrocytes reached to(91.7±1.2)% and(93.3±2.2)%, respectively. Compared with the Schwann cells group, the number of PEI-SPIONs/ChABC-transfected Schwann cells that entered the region of astrocytes was significantly increased(P < 0.05). Under the external magnetic field, the number of PEI-SPIONs/ChABC-transfected Schwann cells that entered the region of astrocytes and the cell migration distance were significantly increased as compared with the Schwann cells group(P < 0.005). In summary, PEI-SPIONs/ChABC transfection can enhance the ability of Schwann cells to break the glial scar, and increase the fusion of astrocytes. Under the guidance of external magnetic

关 键 词：许旺细胞 星形胶质细胞 磁场 超顺磁性纳米微粒 迁移 硫酸软骨素酶ABC 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]