采用多重标记物示踪化学去细胞异体神经修复大鼠面神经缺损  

Evaluation of the regeneration of chemically extracted acellular facial nerve allograft for the whole facial nerve defect in rat with multiple retrograde tracing

作　　者：杨昱[1,2] 彭江[1] 杨启友[2] 王玉[1] 赵斌[1] 眭翔[1] 张莉[1] 王鑫[1] 许文静[1] 卢世璧[1] 

机构地区：[1]解放军总医院骨科研究所,北京100853 [2]贵阳医学院附属医院骨科,贵阳550004

出　　处：《中国矫形外科杂志》2010年第8期659-662,共4页Orthopedic Journal of China

基　　金：国家863计划项目(编号:2009AA03Z312);军队"十一五"计划资助项目(编号:06Z057);国家支撑计划(编号:2009BAI78B02)

摘　　要：[目的]探索用荧光金、量子点、固蓝"三标"逆行示踪方法来评价化学去细胞异体神经移植修复大鼠面神经缺损后颊支、下颌缘支、颈支的再生及神经的物质运输功能。[方法]外科显微镜下解剖、分离出鼠的左侧颅外段面神经主干及各分支(颞支、颧支、颊支、下颌缘支、颈支),在出茎乳孔处离断面神经主干,分别在距该断点10 mm处离断5个分支,移植化学去细胞异体全面神经,术后2个月暴露面神经,在颊支、下颌缘支、颈支吻合口远端分别注射荧光金、量子点、固蓝,3 d后脑干取材,冰冻切片,并在荧光显微镜下观察脑干内3种示踪剂的分布情况。[结果]将发源于脑干的面神经核团进行冰冻切片,荧光显微镜下观察到被荧光金、量子点、固蓝标记的神经元分别显示出黄色、红色和蓝色。[结论]根据神经轴浆运输的原理,采用多种标记物示踪法评价异体神经移植修复面神经损伤后神经干及各分支连续性的恢复情况,操作简便,可靠易行,是一种理想的评价方法。[ Objective ] To evaluate the axonal transport function of the regenerate nerve of buceal branch, marginal mandibular branch and cervical branch ＇after chemically extracted aeellular facial nerve allograft to the whole facial nerve defect in rat using fluoro-gold（FG） ,quantum dot（QD） and fast blue （FB）. [ Method ] The extraeranial section of facial nerve and branches were dissected under light microscope, and 10-mm-long segments of facial nerve gap were made on the left side. The defect was repaired by acellular facial nerve allografls with epineural suture method. Two months later, fluoro-gold（FG）, quantum dot （QD） and fast blue （FB） were injected into distal bridged side of bueeal branch, marginal mandibular branch and cervical branch. After three days,the slices of brain stem were collected and the facial motoneuron was observed under fluorescences microscope. [ Result] There were three kinds of different fluorescences in the frozen sections of the facial nerve originated from the brainstem nuclei under the fluorescence microscope. Fluoro-gold （FG） , quantum dot （QD） and fast blue （FB） labeled neurons were shown yellow,red and blue colors,respectively. [ Conclusion] According to estimation of axonal branching by triple retrograde labeling, it was found that the regenerate facial nerve repaired the gap, and nerve continuity obtained restoration. The evaluation method of this study is simple, reliable and easy. It is an ideal evaluation method.

关 键 词：荧光金 量子点 快蓝 逆行示踪 面神经 神经移植 

分 类 号：R651.3[医药卫生—外科学]