组织诱导性神经导管的制备及性能评价  

The Preparation and Evaluation of Tissue Inducible Nerve Guide Conduit

作　　者：赵红斌[1] 刘兴炎[1] 葛宝丰[1] 郭超[1] 甄平[1] 

机构地区：[1]兰州军区兰州总医院骨科研究所,兰州730050

出　　处：《生物医学工程学杂志》2012年第2期315-322,共8页Journal of Biomedical Engineering

基　　金：兰州军区医药卫生科研项目资助(LXH-2007007)

摘　　要：本研究的目的是构建一种新型的组织诱导性神经导管并评价其生物学性能。壳聚糖包被中药川芎嗪制备微球,利用体外缓释方法检测壳聚糖/川芎嗪微球的缓释效果;壳聚糖/川芎嗪微球与胶原蛋白复合构建组织诱导性神经导管,2%京尼平交联导管;万能材料试验机评价交联前后神经导管的力学特征;体外降解试验分析交联前后神经导管的降解性能;应用织物手感评价仪检测神经导管的柔韧性;扫描电镜(SEM)观察神经导管交联前后空间结构及微球的分布;光学显微镜(LMS)和免疫荧光方法观察并评价壳聚糖微球/胶原蛋白神经导管与大鼠间充质干细胞(MSCs)共培养对MSCs向神经细胞定向分化的影响;SEM和细胞免疫荧光方法分别评价神经导管与MSCs的复合情况及对细胞定向分化的影响。结果显示:壳聚糖微球具有良好的缓释效果;交联前后神经导管的最大载荷和断裂载荷分别为(0.23±0.09)N、(0.76±0.15)N和(0.20±0.12)N、(0.69±0.17)N,两者相比具有统计学意义(P<0.05);体外降解实验表明,交联前后导管的平均失重率分别为(58.62±7.59)mg和(9.23±2.47)mg,两者相比具有统计学意义(P<0.01);干湿态神经导管的平均线性度分别为(0.597±0.012)LC和(0.333±0.015)LC,两者相比差异具有统计学意义(P<0.01),湿态神经导管的柔韧性较干态好;交联后神经导管中胶原蛋白排列紧密,微球均匀分布于胶原支架材料;神经导管与MSCs共培养后,微球缓释的川芎嗪能促进MSCs表达神经细胞相关标志分子NSE和MAP2;神经导管与MSCs复合培养,通过缓释的川芎嗪促进MSCs增殖和NSE的表达。构建的组织工程化神经导管具有良好的组织相容性和组织诱导性功能。The objective of this research was to fabricate a novel tissue inducible nerve guide conduit,and to evaluate its biologic property.The microspheres were prepared with chitosan that encapsulated ligustrazine.The drug release of the chitosan microspheres was detected with application of the controlled release method in vitro.Chitosan microsphereswere mixed with collagen to fabricate the tissue inducible nerve conduit,which were crosslinked with 2% genipin for 24h.Mechanical properties of the nerve guide conduit samples,including maximum load and breaking load,were measured using an Instron Series IX Automated Materials Testing System.The flexibility of the nerve guide conduit was determined with the texture evaluation instrument.Different methods,such as scanning electron microscope（SEM）,light microscope（LMS） and immunofluorescence were used to analyze the spatial structure of the nerve guide conduit,the distribution of the microspheres,the state of the nerve duct combined with mesenchymal stem cells（MSCs）,and the effect of the ligustrazine that released from chitosan microsphere on MSCs differentiation into nerve cells,respectively.The results showed that the chitosan microspheres had better releasing effect.The mechanical properties resultant nerve guide conduit were determined.The maximum load and breaking load of the genipin crosslinked samples were significantly higher than that observed with the non-crosslinkers,increasing to（0.76±0.15）N and（0.69±0.17）N from（0.23±0.09）N and（0.20±0.12）N for the non-crosslinkers（P0.01）.The degradation rates of non-crosslinked and crosslinked by genipin were（58.62±7.59）mg and（9.23±2.47）mg,respectively.This had a statistical significance（P0.01）.The average linearities in dry and hygrometric state of the nerve guide conduit were（0.597±0.012）LC and（0.333±0.015）LC,respectively,which also had statistical significance（P0.01）.The flexibility in the hygrometric state of the nerve guide conduit was better than that of the dr

关 键 词：胶原蛋白 壳聚糖微球 川芎嗪 组织诱导性神经导管 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]