Combination of olfactory ensheathing cell transplantation and repetitive transcranial magnetic stimulation for the treatment of spinal cord injury

doi:10.3969/j.issn.2095-4344.2017.01.018

Abstract

Abstract:

BACKGROUND: Conventional wisdom suggests that the mammalian brain and spinal cord cannot be regenerated after injury and the hope for functional recovery is slim after spinal cord injury.
OBJECTIVE: To test and analyze the therapeutic effect of olfactory ensheathing cell transplantation combined with repetitive transcranial magnetic stimulation (rTMS) in spinal cord injury rats.
METHODS: Eighty rats were used to make spinal cord injury models by T₁₀ laminectomy and then rat models were equivalently randomized into model, cell transplantation, rTMS, and combined treatment groups. Twenty-four hours after modeling, rats in the latter three groups were given olfactory ensheathing cell transplantation around the injured region, rTMS above the left anterior bregma, or their combination, respectively.
RESULTS AND CONCLUSION: At 4 weeks following transplantation, the motor function of rats in the combined treatment group was significantly improved, accompanied by increased Basso, Beattie and Bresnahan score and significantly improved sciatic nerve electrophysiological function. These findings indicate that the olfactory ensheathing cell transplantation combined with rTMS treatment achieves remarkable outcomes, and the combined treatment can significantly improve the axonal regeneration and functional recovery after spinal cord injury.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Spinal Cord Injuries, Transcranial Magnetic Stimulation, Tissue Engineering

CLC Number:

R394.2

Jiang Yong, Chi Xiao-fei, Zou Xi-jun, Ci Yuan, Yao Qi, Yang Mao-wei. Combination of olfactory ensheathing cell transplantation and repetitive transcranial magnetic stimulation for the treatment of spinal cord injury[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(1): 98-102.

Figures/Tables 1

2.1 嗅鞘细胞的形态培养7 d时双极细胞为主，细胞密度增加，胞体较亮，突起变长，细胞主要呈双极、三极，胞体立体感强，折光性好，背景清楚，细胞团聚状生长(图1A)，至14 d时，细胞更加密集，双极、三极细胞的胞体折光度稍有降低。胞体变得大小不一，细胞胞体突起可呈现平行排列(图1B)。 2.2 嗅鞘细胞的鉴定纯化培养10 d时，免疫荧光染色结果表明，培养板内双极和三极细胞呈p75阳性(图2)，证实为嗅鞘细胞，细胞纯度分别94.3%。 2.3 实验动物数量分析有3只大鼠术中死亡，其余大鼠在术后2 h完全清醒，2只前肢可爬行，均出现典型的截瘫综合征，术后1周内大鼠活动减少，饮食少、常常伴有腹部胀气、大便困难伴硬结、尿潴留、肌张力低、肌力0级、双后肢拖行，4周后逐渐开始恢复排便排尿。整个实验过程中各种原因(如麻醉意外、感染、衰竭等)死亡的大鼠共7只，死亡率19%，其中术中死亡3只(8%)，术后死亡4只(11%)；发生脑脊液漏3只(8%)；发生感染3只(8%)；发生血尿1只(3%)；发生尿潴留3只(8%)；发生下肢溃疡3只(8%)。补充大鼠至每组20只。 2.4 运动功能造模后2周，4组大鼠后肢均没有任何运动功能，BBB评分为0分；4周后4组大鼠BBB评分较2周时增高(P < 0.01)，且联合组大鼠BBB评分高于其余3组(P < 0.01)，而嗅鞘细胞移植组和重复经颅磁刺激组大鼠BBB评分接近(P > 0.05；表1)，提示重复经颅磁刺激联合嗅鞘细胞移植可明显促进脊髓损伤大鼠后肢运动功能恢复。 2.5 神经电生理学检查结果随着时间的延长，4组大鼠坐骨神经运动诱发电位的峰-峰值增加，且潜伏期缩短；且联合组大鼠运动诱发电位峰-峰值和潜伏期更为显著(P < 0.01；表2，3)。 2.6 脊髓病理形态 2.6.1 苏木精-伊红染色重复经颅磁刺激组和联合组大鼠脊髓损伤区域细胞走行方向较一致，且神经束明显增多，有神经束跨过损伤区域。胸髓横断损伤组和嗅鞘细胞移植组大鼠脊髓损伤区结构紊乱，纤维走行方向扭曲、不一致，且损伤区内细胞和纤维结构均较少，有许多大小不等的空洞。4组大鼠脊髓空洞和瘢痕面积差异有显著性意义(P < 0.01；表4)。 2.6.2 嗜银染色重复经颅磁刺激组和联合组大鼠脊髓损伤区内再生神经纤维较多，排列相对整齐；而胸髓横断损伤、嗅鞘细胞移植组的损伤区内神经纤维较少，排列紊乱、扭曲。4组大鼠脊髓损伤区神经纤维线密度差异有显著性(P < 0.01；表4)。 2.6.3 免疫组化染色重复经颅磁刺激组和联合组大鼠损伤脊髓组织中NF200平均灰度值高于胸髓横断损伤组和嗅鞘细胞移植组(P < 0.01)，重复经颅磁刺激组和联合组大鼠脊髓组织中神经纤维排列相对规律，而胸髓横断损伤组和嗅鞘细胞移植组神经纤维排列较乱。重复经颅磁刺激组和联合组大鼠脊髓损伤远端横切面上神经纤维数量明显多于胸髓横断损伤组和嗅鞘细胞移植组(P < 0.01；表4)。"

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