京尼平交联音猬因子复合纤维蛋白支架修复大鼠脊髓损伤  

Genipin crosslinked sonic hedgehog composite fibrin scaffolds for the repair of spinal cord injury in rats

作　　者：陈平波 王晶 孙勇 徐晓峰 陈谦[3] 张志坚[3] Chen Pingbo;Wang Jing;Sun Yong;Xu Xiaofeng;Chen Qian;Zhang Zhijian(Department of Orthopedics,Yijishan Hospital of Wannan Medical College,Wuhu 241000,Anhui Province,China;Department of Pain,Zhenjiang First People’s Hospital,Zhenjiang 212001,Jiangsu Province,China;Medical College of Jiangsu University,Zhenjiang 212001,Jiangsu Province,China)

机构地区：[1]皖南医学院弋矶山医院骨科,安徽省芜湖市241000 [2]镇江市第一人民医院疼痛科,江苏省镇江市212001 [3]江苏大学医学院,江苏省镇江市212001

出　　处：《中国组织工程研究》2022年第21期3345-3350,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金地区科学基金项目(81571830),项目负责人:张志坚,国家自然科学基金地区科学基金项目(81501077),项目负责人:陈谦;皖南医学院中青年自然基金(WK2019F16),项目负责人:陈平波。

摘　　要：背景:组织工程支架移植是修复脊髓损伤最有应用前景的治疗手段之一,通过组织工程支架募集内源性神经干细胞并促进其分化为神经细胞,进而修复脊髓损伤。目的:观察京尼平交联的音猬因子复合纤维蛋白支架移植到大鼠脊髓损伤处的修复作用,探讨其作用机制。方法:分别制备纤维蛋白胶、音猬因子复合纤维蛋白支架、京尼平交联音猬因子复合纤维蛋白支架,检测京尼平交联音猬因子复合纤维蛋白支架的体外降解率与缓释性能。将120只雌性SD大鼠随机分为4组,每组30只,A组T10-T11脊髓损伤部位不植入任何材料,B组T10-T11脊髓损伤部位植入纤维蛋白胶,C组T10-T11脊髓损伤部位植入音猬因子复合纤维蛋白支架,D组T10-T11脊髓损伤部位植入京尼平交联音猬因子复合纤维蛋白支架,术后每周对大鼠进行运动功能BBB评分,术后12周取脊髓损伤处及周围脊髓组织,进行组织形态与WesternBlot检测。结果与结论:(1)音猬因子复合纤维蛋白支架14 d基本完全降解,京尼平交联的纤维蛋白支架14 d的降解率不超过20%;音猬因子复合纤维蛋白支架7 d基本完全释放了音猬因子;京尼平交联音猬因子复合纤维蛋白支架在1-3 d爆发释放音猬因子,4-14 d内缓慢释放音猬因子,具有良好的缓释性能;(2)随着时间的延长,各组大鼠的运动功能BBB评分升高,B、C组术后12周的BBB评分高于A组(P<0.05),D组术后12周的BBB评分高于B、C组(P<0.05);(3)苏木精-伊红染色显示,A组脊髓损伤处恢复最差,D组恢复最好;(4)免疫组化染色与WesternBlot检测显示,A组巢蛋白、神经生长相关蛋白43、神经丝蛋白200、髓鞘碱性蛋白表达量最低,胶质纤维酸性蛋白表达最高;D组巢蛋白、神经生长相关蛋白43、神经丝蛋白200、髓鞘碱性蛋白表达量最高,胶质纤维酸性蛋白表达最低;(5)结果表明,京尼平交联音猬因子复合纤维蛋白支架具有良好的缓释性能,�BACKGROUND:Tissue engineering scaffold transplantation is one of the most promising therapeutic methods for repairing spinal cord injury.Endogenous neural stem cells are recruited through tissue engineering scaffolds and promoted to differentiate into nerve cells to repair spinal cord injury.OBJECTIVE:To observe the effect of transplantation of genipin crosslinked sonic hedgehog(Shh)composite fibrin scaffolds into the injured spinal cord of rats on the repair of spinal cord injury and to explore the underlying mechanism.METHODS:Fibrin glue,Shh composite fibrin scaffold,and genipin crosslinked Shh composite fibrin scaffold were prepared separately.In vitro degradation rate and sustained release performance of genipin crosslinked Shh composite fibrin scaffold were detected.Totally 120 female Sprague-Dawley rats were randomly divided into four groups(n=30).No material was implanted at the T_(10)-T_(11)spinal cord injury site in group A.Fibrin glue was implanted in the T_(10)-T_(11)spinal cord injury site of group B.T_(10)-T_(11)spinal cord injury site was implanted with Shh composite fibrin scaffold in group C.Genipin crosslinked Shh composite fibrin scaffold was implanted at the T_(10)-T_(11)spinal cord injury site in group D.Atier surgery,the rats were scored with BBB motor function every week.At 12 weeks atier surgery,spinal cord tissue at and around the injury site was harvested and used for morphological observation and western blot assay.RESULTS AND CONCLUSION:(1)Shh composite fibrin scaffold was basically completely degraded at 14 days.The degradation rate of the genipin crosslinked fibrin scaffold at 14 days did not exceed 20%.The Shh composite fibrin scaffold basically released the Shh at 7 days.Genipin crosslinked Shh composite fibrin scaffold released Shh explosively in 1-3 days,and slowly released Shh within 4-14 days,which had a good slow-release property.(2)As time went by,the BBB scores of motor function of rats increased in each group.The BBB scores of groups B and C were higher than those of group A

关 键 词：京尼平 音猬因子 纤维蛋白 缓释脊髓损伤 交联 支架 移植 修复 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]