3D打印胶原/壳聚糖支架改善大鼠脊髓损伤后神经功能恢复  

Three-dimensional printed collagen/chitosan scaffold improves neurological recovery after spinal cord injury in rats

作　　者：史新宇 李晓红 叶益超 王景景 孙小喆 张艳龙 段敬豪 魏孟广 张赛 Shi Xinyu;Li Xiaohong;Ye Yichao;Wang Jingjing;Sun Xiaozhe;Zhang Yanlong;Duan Jinghao;Wei Mengguang;Zhang Sai(Graduate School of Tianjin Medical University,Tianjin 300070,China;Academy of Medical Engineering and Translational Medicine,Tianjin University,Tianjin 300072,China;lnstitute of Traumatic Brain Injury and Neuroscience,Characteristic Medical Center of Chinese People's Armed Police Force,Tianjin 300162,China)

机构地区：[1]天津医科大学研究生院,天津市300070 [2]天津大学医学工程与转化医学研究院,天津市300072 [3]武警特色医学中心脑创伤与神经疾病研究所,天津市300162

出　　处：《中国组织工程研究》2020年第22期3474-3479,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81771352),项目负责人:李晓红。

摘　　要：背景:3D打印技术可以根据需求制备出满足脊髓植入形状、大小和表面形态要求的生物支架。目的:观察3D打印胶原/壳聚糖支架对脊髓损伤大鼠神经功能恢复的影响。方法:将胶原和壳聚糖按2∶1的质量比混合,采用冷冻干燥法制备普通胶原/壳聚糖支架,采用3D打印机制备3D打印胶原/壳聚糖支架,分别测量两种支架的孔隙率和弹性模量,电镜观察支架形态。将神经干细胞分别与3D打印胶原/壳聚糖支架、普通胶原/壳聚糖支架共培养,进行扫描电镜观察与CCK-8检测。将40只雌性SD大鼠(由中国人民解放军医学科学院军事科学院提供)随机分成4组:假手术组、脊髓损伤组、普通胶原/壳聚糖支架组和3D打印胶原/壳聚糖支架组,后3组制作脊髓全横断损伤模型,普通胶原/壳聚糖支架组和3D打印胶原/壳聚糖支架组损伤处填充对应的支架材料,术后相应时间点进行后肢功能BBB评分、斜坡实验、神经电生理检测与磁共振平扫。实验方案经天津市神经创伤重点实验室伦理委员会批准。结果与结论:①扫描电镜显示,3D打印胶原/壳聚糖支架具有互连的多孔结构,普通胶原/壳聚糖支架内部结构紊乱;②神经干细胞在3D打印胶原/壳聚糖支架表面生长良好,完全伸展,且3D打印胶原/壳聚糖支架表面神经干细胞的活性显著高于普通胶原/壳聚糖支架组(P<0.05);③3D打印胶原/壳聚糖支架的孔隙率与弹性模量均高于普通胶原/壳聚糖支架组(P<0.05);④3D打印胶原/壳聚糖支架组术后3-8周的BBB评分高于脊髓损伤组、普通胶原/壳聚糖支架组(P<0.05),术后4,6,8周的斜坡实验角度大于脊髓损伤组、普通胶原/壳聚糖支架组(P<0.05);⑤3D打印胶原/壳聚糖支架组术后8周的运动诱发电位振幅、体感诱发电位振幅大于脊髓损伤组与普通胶原/壳聚糖支架组(P<0.05),运动诱发电位潜伏期、体感诱发电位潜伏期短于脊髓损伤组与普BACKGROUND:Three-dimensional(3D)printing technology can be used to prepare bioscaffolds to meet the requirements of shape,size and surface morphology of spinal cord implantation.OBJECTIVE:To investigate the effect of 3D printed collagen/chitosan scaffolds on the recovery of neurological function in rats with spinal cord injury.METHODS:The collagen/chitosan scaffolds were prepared by freeze-drying method with a mass ratio of 2∶1.The 3D printed collagen/chitosan scaffolds were prepared by 3D printer.The porosity and elastic modulus of the scaffolds were measured,and the morphology of the scaffolds was observed by electron microscope.Neural stem cells were co-cultured with 3D printed collagen/chitosan scaffold,and common collagen/chitosan scaffold,respectively,for scanning electron microscope and cell counting kit-8 assay.Forty female Sprague-Dawley rats(provided by Academy of Military Sciences of PLA)were randomly divided into four groups:sham-operated,spinal cord injury,common collagen/chitosan scaffold,and 3D printed collagen/chitosan scaffold groups.The rats in the latter three groups were used to prepare complete spinal cord transected injury model,followed by filled with the corresponding scaffold materials.Basso,Beattie,Bresnahan scores of the posterior limb,slope test,neuroelectrophysiological detection and MRI were conducted at each time point after surgery.The study was approved by the Animal Ethics Committee of Tianjin Key Laboratory of Traumatic Brain Injury.RESULTS AND CONCLUSION:(1)Scanning electron microscope showed that 3D printed collagen/chitosan scaffolds had interconnected porous structure,and the internal structure of common collagen/chitosan scaffolds was disordered.(2)Neural stem cells grew well on the surface of scaffolds and fully extended.The activity of neural stem cells on the surface of 3D printed collagen/chitosan scaffolds was significantly higher than that of the common collagen/chitosan scaffold group(P<0.05).(3)The porosity and modulus of elasticity in the 3D printed collagen/chit

关 键 词：3D打印 脊髓损伤 胶原 壳聚糖 支架 修复 神经电生理 磁共振 

分 类 号：R459.9[医药卫生—治疗学] R649.6[医药卫生—临床医学]