硫酸肝素复合胶原蛋白支架材料桥接大鼠坐骨神经的形态学研究  

Morphology research of the rat sciatic nerve bridged by collage-heparin sulfate scaffold

作　　者：王树森[1] 胡蕴玉[1] 罗卓荆[1] 陈良为[2] 刘慧玲[2] 孟国林[1] 吕荣[1] 徐新 

机构地区：[1]第四军医大学西京医院全军骨科研究所,西安710032 [2]第四军医大学西京医院全军神经科学研究所,西安710032

出　　处：《中华外科杂志》2005年第8期531-534,共4页Chinese Journal of Surgery

基　　金：国家自然科学基金资助项目(30070761)

摘　　要：目的　以形态学方法观测硫酸肝素复合胶原蛋白支架材料桥接大鼠坐骨神经10mm缺损的疗效。方法　以硫酸肝素复合胶原蛋白经冷冻干燥技术制备新型神经组织支架材料,并用此材料桥接修复SD大鼠坐骨神经缺损10mm,术后36周分别以辣根过氧化物酶(HRP)逆行示踪,HE、甲苯氨蓝和镀银染色法,S100、生长相关蛋白43 (GAP 43)、神经丝蛋白(NF)免疫组化染色法、MBP免疫荧光染色法及透射电镜等形态学方法观测其引导神经再生的疗效。结果　硫酸肝素复合胶原蛋白支架材料在移植术后36周已神经化。除再生有髓神经纤维的密度低于自体移植,有髓神经纤维的面积和髓鞘厚度与自体移植组无明显差异。结论　以硫酸肝素复合胶原蛋白制备的神经组织工程支架材料,可以引导神经再生。Objective To observe the treating effect of collage-heparin sulfate after the 10 mm rat sciatic nerve defect was bridged by it. Methods A new kind of nervous tissue engineering scaffold was produced by freeze-drying technique from collagen-heparin sulfate. Thirt-two SD rats were randomly divided into A, B, C and D groups. Sciatic nerve defect in group A was bridged by collagen-heparin sulfate. In group B, sciatic nerve was bridged by auto-nerve transplantation. Group C was the blank control group. Animals in group D were normal. And 10 mm sciatic nerve defect was bridged in the experiment. Thirty-six weeks after the operation, the experimental animals were detected by HRP labeled retrograde trace, HE staining, toluidine staining,silvering staining, S100, GAP-43 and NF immunohistological staining, MBP immunofluorscence staining and transmission electron microscope to observe the nerve regeneration inducing effect of this new scaffold. Results Nine months after operation, the collage-heparin sulfate scaffold was replaced by newly regenerated nerve. The number of HRP labeled spinal cord anterior horn cells and the area of sensation nerve fiber at the posterior horn were similar with that was repaired by auto-nerve. GAP-43, NF and S100 labeled regenerated nerve fiber had passed the total scaffold and entered the distal terminal. The regenerated nerve fibers were paralleled, lineage arranged, coincide with the prearranged regenerating “channel” in the collagen-heparin sulfate scaffold. MBP immunofluorscence staining also proved that the newly regenerated nerve fiber could be ensheathed. In the experimental group, the area of myelinated nerve fiber and the thickness of the myelin sheath had no obvious difference with that of the group repaired by auto-nerve, except that the density of the regenerated myelinated sheath fiber was lower than that of the control group.Conclusion Nervous tissue engineering scaffold produced by collagen-heparin sulfate can guide the regeneration of nerve fibers. The nerve function

关 键 词：硫酸肝素 胶原蛋白 形态学研究 复合 桥接 组织工程支架材料 有髓神经纤维 辣根过氧化物酶 免疫组化染色法 免疫荧光染色法 形态学方法 冷冻干燥技术 坐骨神经缺损 生长相关蛋白 神经再生 自体移植 镀银染色法 神经丝蛋白 

分 类 号：R651.3[医药卫生—外科学]