3D打印支架载重编程iPSCs定向分化NSCs移植在修复急性脊髓损伤中的应用研究  

3D printing stent load programming iPSCs directed differentiation NSCs transplantation applied research in the repair of acute spinal cord injury

作　　者：李长明[1] 邓小梅 方燕芬[1] 邵荣学[3] LI Changming;DENG Xiaomei;FANG Yanfen;SHAO Rongxue(Department of Spinal Surgery,Hangzhou Xiaoshan District Hospital of Traditional Chinese Medicine,Hangzhou 311201,China;Department of Endocrinology,Hangzhou Xiaoshan District Hospital of Traditional Chinese Medicine,Hangzhou 311201,China;Department of Orthopedics,Hangzhou Traditional Chinese Medicine Hospital,Hangzhou 311201,China)

机构地区：[1]杭州市萧山区中医院脊柱外科,浙江杭州311201 [2]杭州市萧山区中医院内分泌科,浙江杭州311201 [3]杭州市中医院骨科,浙江杭州311201

出　　处：《中国现代医生》2021年第26期45-51,I0001,共8页China Modern Doctor

基　　金：浙江省医药卫生科技计划项目(2018KY654);浙江省杭州市萧山区科技计划项目(2017209)。

摘　　要：目的 探讨3D打印支架载重编程iPSCs定向分化神经干细胞(NSCs)移植在修复急性脊髓损伤中的应用。方法 将40只健康成年雌性大鼠随机分为A组(对照)、B组(支架)、C组(支架+NSCs)、D组(支架+iPSCs-NSCs),每组各10只。原代NSCs分离、培养、传代及鉴定;构建并鉴定重编程iPSCs;携带i PSCs基因的逆转录病毒感染NSCs;培养支架+NSCs、支架+iPSCs-NSCs。结果 支架与NSCs和i PSCs-NSCs相容性较好;D组2、4、6、8周后BBB评分显著高于另外三组,4、6、8周后B、C组BBB评分显著高于A组,C组BBB评分高于B组(P<0.05);8周后A组Tarlov和Rivlin评分显著低于另外三组,B组低于C组,C组低于D组(P<0.05);8周后A组运动和感觉潜伏期显著常于另外三组,B组长于C组,C组长于D组(P<0.05);A组运动和感觉振幅显著低于另外三组,B组低于C组,C组低于D组(P<0.05);A组纤维断裂,另外三组病变周围脊髓纤维连续中断,C、D组损伤部位纤维数量显著多于B组,D组多于C组(P<0.05);8周后D组存活细胞数量、分化率显著高于C组(P<0.05)。结论 3D打印支架载重编程iPSCs定向分化NSCs可有效修复ASCI。Objective To explore the application of 3D printed scaffold loaded with induced pluripotent stem cells(iPSCs)and directedly differentiated neural stem cells(NSCs)transplantation in the repair of acute spinal cord injury(SSCI).Methods Forty healthy adult female rats were randomly divided into group A(control),group B(stent),group C(stent+NSCs)and group D(stent+iPCS-NSCs),with 10 rats in each group.Primary NSCs were isolated,cultured,passed on and identified.The reprogrammed iPSCs were constructed and identified.NSCs were infected with retrovirus carrying iPSCs gene.The stent+NSCs and stent+PSCs-NSCs were cultured.Results The stent was compatible with NSCs and iPSCs-NSCs.The BBB scores of group D after 2,4,6 and 8 weeks were significantly higher than those of the other three groups.The BBB scores of group B and C were significantly higher than those of group A after 4,6 and 8 weeks,and BBB score of group C was higher than that of group B(P<0.05).Eight weeks later,Tarlov and Rivlin scores in group A were significantly lower than those in the other three groups,while those in group B were lower than group C,and those in group C were lower than those in group D(P<0.05).Eight weeks later,the latencies of movement and sensation in group A were significantly higher than those in the other three groups,while those in group B were longer than those in group C.The latencies of movement and sensation in group C were longer than those in group D(P<0.05).The amplitudes of motion and sensation in group A were significantly lower than those in the other three groups.The indexes in group B were lower than those in group C,which were lower than those in group D(P<0.05).In group A,the fibers were broken,and in the other three groups,the spinal cord fibers around the lesions were continuously interrupted.The number of fibers at the injured sites in groups C and D was significantly higher than that in group B,and that in group D was more than that in group C(P<0.05).After 8 weeks,the number and differentiation rate of viable cells in

关 键 词：3D打印 诱导性多能干细胞 神经干细胞 急性脊髓损伤 

分 类 号：R651.2[医药卫生—外科学]