摘要
目的用振荡法制备脊髓脱细胞支架修复同种异体大鼠脊髓缺损,观察术后大鼠行为学及组织再生情况。为脊髓缺损后修复提供新的研究思路。方法将30只SD大鼠脊髓分别用50 ml 3%聚乙二醇辛基苯基醚(Triton X-100)和2%脱氧胆酸钠溶液在摇床上做振荡处理,对比处理前后细胞残留情况及组织的空间结构,了解支架本身的组织构成。将90只SD大鼠随机分成空白对照组、单纯脊髓缺损组和支架移植组。切除单纯脊髓缺损组和支架移植组大鼠腰椎9~10节段,移植脱细胞支架至支架移植组大鼠。术后饲养12周,期间进行行为学评分观察,分别在4、8及12周时取大鼠损伤部位的脊髓进行HE染色及神经再生相关蛋白免疫荧光检测。结果通过HE、Masson、甲苯胺蓝染色显示,脱细胞处理后的脊髓脱细胞支架上神经细胞及轴突彻底清除,保留了脊髓细胞外基质。扫描电子显微镜观察发现,支架保留一定多孔网状支架结构。脱细胞支架在体实验中,Basso-Beattie-Bresnahan(BBB)评分显示,移植有脱细胞支架的大鼠后肢运动功能恢复优于单纯损伤组大鼠。组织学HE染色显示,脱细胞支架能够填补缺损的脊髓节段,加快损伤脊髓的修复过程。免疫荧光显示,支架移植组大鼠的损伤部位有一定轴突再生。结论脊髓脱细胞支架保留了细胞外基质并具有一定空间结构,能够一定程度加快脊髓缺损修复,对神经再生具有一定的促进作用。
Objective Exploring the effect of spinal cord decellularized scaffold on spinal cord defects and observing the behavior and regeneration of rats after operation.Methods The spinal cords of 30 SD rats were treated with3%Triton X-100 and 2%sodium deoxycholate on oscillator.The cell residue and the spatial structure of the tissue were compared before and after treatment,in order to understand the tissue structure of the stent itself.90 SD rats were randomly divided into control group,simple injury group and stent transplantation group.Excision of the spinal cord 9\|10 segments in the simple injury group and the stent graft group the acellular scaffold was transplanted to the stent graft group.Behavioral scores were observed postoperatively.At 4,8,and 12 weeks,the spinal cords of the injured part of the rats were taken for HE staining and immunofluorescence detection of nerve regeneration-related proteins.Results After decellularization of the spinal cord,the nerve cells and axons were completely removed,and the extracellular matrix of the spinal cord was preserved.Scanning electron microscopy revealed that the scaffold retained a certain porous network scaffold structure.In the experiment of decellularized scaffold in vivo,the Basso-Beattie-Bresnahan(BBB)score showed that the recovery of hindlimb motor function in rats with decellularized scaffolds was better than that in rats with simple injury.HE staining showed that the decellularized scaffold could fill the defect of the spinal cord segment and accelerate the repair process of the injured spinal cord.Immunofluorescence showed that there was a certain axonal regeneration in the injured part of the stent transplantation group.Conclusion The spinal cord decellularized scaffold retains the extracellular matrix and has a certain spatial structure,which can accelerate the process of spinal cord defect repair to a certain extent,and has a certain promoting effect on nerve regeneration.
作者
朱君毅
张宜家
杜胜虎
余方正
卢颖枫
李培丰
王志斌
梅劲
王健
ZHU Jun-yi;ZHANG Yi-jia;DU Sheng-hu;YU Fang-zheng;LU Ying-feng;LI Pei-feng;WANG Zhi-bin;MEI Jin;WANG Jian(Department of Hand Surgery and Peripheral Neurosurgery,the First Affiliated Hospital of Wenzhou Medical University,Zhejiang Wenzhou 325035,China;Department of Reproductive Center,the First Affiliated Hospital of Wenzhou Medical University,Zhejiang Wenzhou 325035,China;Department of Anatomy,Wenzhou Medical University,Zhejiang Wenzhou 325035,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2020年第4期502-506,共5页
Acta Anatomica Sinica
基金
温州市科技局基金(Y20180498)。
关键词
脊髓再生
脊髓损伤
脱细胞脊髓支架
轴突再生
苏木素-伊红染色
大鼠
Spinal cord regeneration
Spinal cord injury
Acellular spinal scaffold
Axonal regeneration
Hematoxylin\|eosin staining
Rat
