负载脑源性神经营养因子胶原/肝素硫酸支架促进颅脑创伤大鼠神经运动功能的恢复  

Collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor promote neurological and locomotor function recovery in rats after traumatic brain injury

作　　者：张健 陈淼 李伟鑫 叶益超 徐会友 马珂 陈旭义 孙洪涛 张赛 Zhang Jian;Chen Miao;Li Weixin;Ye Yichao;Xu Huiyou;Ma Ke;Chen Xuyi;Sun Hongtao;Zhang Sai(Logistics University of People's Armed Police Force,Tianjin 300300,China;lnstitute of Neurotrauma Repair,Characteristic Medical Center of Chinese People's Armed Police Force,Tianjin Key Laboratory of Neurotruama Repair,Tianjin 300171,China;The Fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830002,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]武警后勤学院,天津市300300 [2]武警特色医学中心神经创伤及修复研究所,天津市神经创伤修复重点实验室,天津市300171 [3]新疆医科大学第四临床医学院,新疆维吾尔自治区乌鲁木齐市830002

出　　处：《中国组织工程研究》2020年第34期5538-5544,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(11672332,11102235),项目负责人:陈旭义;国家自然科学基金(11932013),项目参与者:陈旭义;国家重点研发计划(2016YFC1101500),项目负责人:张赛;天津市自然科学基金项目(15JCYBJC28600),项目负责人:孙洪涛;天津市自然科学基金项目(17JCDJC5400),项目负责人:陈旭义;天津市科技支撑重点项目(17YFZCSY00620),项目负责人:孙洪涛;天津市科技军民融合重大项目(18ZXJMTG00260),项目参与者:陈旭义。

摘　　要：背景:研究表明胶原和硫酸肝素交联后可有效固定生长因子,显著改善脊髓损伤大鼠的神经运动功能恢复。目的:观察负载脑源性生长因子胶原/硫酸肝素支架移植修复颅脑创伤的效果。方法:制作胶原支架、胶原/硫酸肝素支架与负载脑源性神经营养因子的胶原/硫酸肝素支架,检测胶原支架、胶原/硫酸肝素支架的孔隙率、吸水率、压缩模量和压缩应力;检测载脑源性生长因子胶原/硫酸肝素支架的体外缓释性能;检测胶原/硫酸肝素支架与负载脑源性神经营养因子的胶原/硫酸肝素支架的细胞毒性与细胞相容性。将48只SD大鼠随机分4组干预:对照组开骨窗后缝合,空腔组仅制作颅脑创伤空腔模型,支架组、生长因子支架组颅脑创伤后空腔处分别植入胶原/硫酸肝素支架、负载脑源性神经营养因子的胶原/硫酸肝素支架,术后分别进行改良神经功能缺损评分、水迷宫测试与脑损伤形态学观察。动物实验获得武警特色医学中心伦理委员会批准。结果与结论:①胶原/硫酸肝素支架的孔隙率、压缩模量和压缩应力高于胶原支架(P<0.05),吸水率低于胶原支架(P<0.05);②胶原/硫酸肝素支架无细胞毒性,负载脑源性神经营养因子后更加有利于脑微血管内皮细胞的增殖;脑微血管内皮细胞在负载脑源性神经营养因子支架微孔内生长良好,分布均匀;③形态学观察显示,两支架组损伤灶处可见大量新生神经细胞及神经纤维,其中以生长因子支架组的新生神经细胞数量及神经纤维密度更高;④生长因子支架组大鼠的逃逸潜伏期短于空腔组、支架组(P<0.01,P<0.05),象限停留时间和平台穿越次数高于支架组、空腔组(P<0.01,P<0.05);⑤生长因子支架组术后3-7周的改良神经功能缺损评分低于支架组、空腔组(P<0.05,P<0.01);⑥结果表明,负载脑源性神经营养因子胶原/硫酸肝素支架移植可以促进大鼠颅脑创伤�BACKGROUND:The porous scaffold fabricated with collagen and heparin sulfate can effectively immobilize neurotrophic factor and greatly improve neurological and locomotor function recovery.OBJECTIVE:To investigate the effect of collagen/heparan sulfate scaffold loaded with brain-derived neurotrophic factor in the treatment of traumatic brain injury(TBI).METHODS:Collagen scaffolds,collagen/heparan sulfate scaffolds and collagen/heparan sulfate scaffolds loaded with brain-derived neurotrophic factor were prepared.The porosity,water absorption,compression modulus and compression stress of collagen scaffolds and collagen/heparan sulfate scaffolds were measured.The in vitro sustained-release performance of collagen/heparan sulfate scaffolds loaded with brain-derived neurotrophic factor was measured.The cytotoxicity and cytocompatibility of collagen/heparin sulfate scaffolds and collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor were detected.Forty-eight Sprague-Dawley rats were randomly assigned to undergo corresponding interventions:opening bone window followed by suture(control group),induction of craniocerebral trauma(model group),induction of craniocerebral trauma+implantation of collagen/heparin sulfate scaffolds(scaffold group),and induction of craniocerebral trauma+implantation of collagen/heparin sulfate scaffolds loaded with brain-derived neurotrophic factor(brain-derived neurotrophic factor group).After surgery,modified neurological severity scoring and Morris water maze test were performed to evaluate the recovery of neurological and locomotor function.Rat cerebral morphology was performed.This study was approved by the Animal Ethics Committee of Characteristic Medical Center of Chinese People’s Armed Police Force,China.RESULTS AND CONCLUSION:(1)The porosity,compression modulus and compression stress of the collagen/heparin sulfate scaffold were significantly higher than those of the collagen scaffold(P<0.05),and the water absorption rate of the collagen/heparin sulfate scaffo

关 键 词：颅脑创伤 胶原支架 胶原/硫酸肝素支架 脑源性神经营养因子 BDNF 水迷宫 脑微血管内皮细胞 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]