自供氧光动力疗法联合Nafamostat修复脊髓损伤的研究

Spinal cord injury is a kind of highly disabling disease. Glial scar formed after injury acts as a barrier that hinders axonal regeneration and nerve repair. In previous work, the team tried to use photodynamic therapy to reduce the formation of glial scar and promote the repair of spinal cord injury. However, local oxygen supply deficiency limits the effect of photodynamic therapy, and inhibiting glial scar only is difficult to improve the micro-environment of the damaged area. Therefore, applicant is looking for a new method to overcome the limitations of oxygen deficiency and single repair effect in photodynamic therapy, optimize local micro-environment, promote axonal regeneration, and maximize repair capacity. On the basis of our previous research work, the applicant intends to prepare the multifunctional hollow gold nanosphere carrying Nafamostat, which has oxygen self-supported, photodynamic therapy and comprehensive improvement of micro-environment function, to apply to animal models of spinal cord injury. Morphological changes of spinal cord tissue are observed by light and electron microscope, axon regeneration is performed by antegrade tracer, Western blot is used to detect the expression of GFAP and inflammatory factors around the injured area, BBB score is used to evaluate the recovery of hind limb function, somatosensory and motor evoked potentials are used to evaluate spinal cord function, to explore the effectiveness and provide a new idea for clinical repair of spinal cord injury.

脊髓损伤是一种高致残性疾病，损伤后形成的胶质瘢痕作为一道屏障，阻碍了轴突再生与神经修复。本团队在前期工作中尝试应用光动力疗法减少胶质瘢痕生成，促进脊髓损伤修复。然而局部供氧不足限制了光动力疗法的效果，且单纯抑制胶质瘢痕难以起到改善损伤区微环境的综合修复作用。因此申请者拟寻求一种新的方法，克服光动力治疗过程中氧气缺乏与修复效果单一的局限，优化局部微环境，促进轴突再生，最大程度提高修复能力。结合前期工作基础，申请者拟制备携带Nafamostat，兼具自供氧、光动力治疗、综合改善微环境功能的多功能空心金纳米球，应用于脊髓损伤动物模型，通过光电镜观察脊髓组织形态学改变，顺行示踪标记轴突再生，Western Blot检测损伤区周围GFAP及炎症因子表达，BBB评分评价大鼠后肢功能恢复程度，体感、运动诱发电位综合评估脊髓功能恢复情况，探讨其修复脊髓损伤的有效性，为临床修复脊髓损伤提供新思路。

脊髓损伤是一种高致残性疾病，损伤后形成的胶质瘢痕作为一道屏障，阻碍了轴突再生与神经修复。本研究制备性质稳定、粒径分布均匀、氧富集特性良好，兼具自供氧、光动力治疗、综合修复作用的多功能空心金纳米球，克服光动力抑制瘢痕过程中氧气缺乏与修复效果单一的局限，优化局部微环境，促进轴突再生，最大程度提高修复能力。研究中将多功能空心金纳米球与大鼠星形胶质细胞共培养，经NIR激光照射后，应用CCK8法测定其细胞毒性，通过活-死细胞染色及ROS荧光检测，证实多功能空心金纳米球在660nm激光照射下对细胞的杀伤力更强。采用脊髓打击损伤的方法建立大鼠脊髓损伤模型，将制备的多功能空心金纳米球于动物模型制作1周后，局部注射于脊髓损伤处，经NIR激光照射，光电镜观察脊髓组织形态学改变，见其能够显著减少胶质瘢痕病变区域；免疫荧光及Western Blot检测损伤区周围GAP43、GFAP表达量及炎症因子表达量变化，表明多功能空心金纳米球能有效修复大鼠脊髓损伤；通过BBB评分与斜板实验验证其具有更优的功能恢复效果，证实其修复脊髓损伤的有效性，为临床修复脊髓损伤提供新思路。