辣根过氧化物酶逆行示踪评价组织工程人工神经修复大鼠坐骨神经缺损.docx

辣根过氧化物酶逆行示踪评价组织工程人工神经修复大鼠坐骨神经缺损张彩顺1，吕刚2辽宁医学院附属第一医院，1手足外科，2骨科，辽宁省锦州市121001Tissue engineered artificial nerves repair sciatic nerve defect in rats: an evaluation using horseradish peroxidase retrograde tracer techniqueZhang Cai-shun1, Lv Gang21Department of Hand and Foot Surgery, 2Department of Orthopedics, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning Province, China摘要 背景：作者前期制备了无细胞神经移植物，并将骨髓间充质干细胞种植其内，成功构建了组织工程人工神经。目的：应用辣根过氧化物酶神经逆行示踪技术，评价无细胞神经移植物复合骨髓间充质干细胞构建的组织工程人工神经桥接大鼠坐骨神经缺损后对感觉神经元的保护作用。方法：成年清洁级健康雄性SD大鼠，随机分成3组：实验组采用复合骨髓间充质干细胞的无细胞神经移植物桥接大鼠坐骨神经缺损；空白对照组采用无细胞神经移植物桥接大鼠坐骨神经缺损；自体神经对照组采用自体神经移植桥接大鼠坐骨神经缺损。移植后12周应用辣根过氧化物酶神经逆行示踪技术，对脊髓后根神经节感觉神经元的再生进行评价。结果与结论：移植12周后根神经节感觉神经元再生评价结果提示，实验组与自体神经对照组相比差异无显著性意义，优于空白对照组；足底皮肤S-100免疫组织化学染色结果显示，各组皮下均见棕色阳性反应。提示无细胞神经移植物复合骨髓间充质干细胞构建组织工程人工神经修复大鼠坐骨神经缺损，对大鼠脊髓后根神经节感觉神经元具有保护作用，可以促进感觉功能的恢复。中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：窗体顶端关键词 ： 组织构建； 组织工程； 辣根过氧化物酶； 骨髓间充质干细胞； 无细胞神经移植物； 神经缺损； 后根神经节； Abstract： BACKGROUND: We have previously prepared acellular nerve graft and implanted bone marrow mesenchymal stem cells into the graft, to successfully construct tissue engineered artificial nerves.OBJECTIVE: Horseradish peroxidase nerve retrograde tracer technique was used to evaluate protective effects on sensory neurons following sciatic nerve defect bridging with tissue engineered artificial nerves constructed by acellular nerve graft and bone marrow mesenchymal stem cells in rats.METHODS: Adult, clean, healthy, male Sprague-Dawley rats were randomly assigned to three group: (1) Experimental group: Rat sciatic nerve detect was bridged by acellular nerve graft combined with bone marrow mesenchymal stem cells; (2) Blank control group: Rat sciatic nerve defect was bridged by acellular nerve graft; (3) Autologous nerve control group: Rat sciatic nerve defect was bridged by autologous nerve transplantation. Regeneration of sensory neurons in the spinal dorsal root ganglia was assessed using horseradish peroxidase nerve retrograde tracer technique at 12 weeks following surgery.RESULTS AND CONCLUSION: Sensory neuron regeneration in the spinal dorsal root ganglia at 12 weeks following surgery was better in the experimental group compared with blank control group. No significant difference was detected between experimental group and autologous nerve control group. S-100 immunohistochemical staining in plantar skin showed brown positive reaction in each group. These findings indicate that tissue engineered artificial nerves constructed by acellular nerve graft and bone marrow mesenchymal stem cells have protective effects on sensory neurons in the spinal dorsal root ganglia, and can promote the recovery of sensory function and repair sciatic nerve defect in rats.中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：Key words： stem cells; mesenchymal stem cells; sciatic nerve; ganglia; neuronal tract-trancer; 窗体底端 中图分类号:R318 基金资助: 辽宁省高等学校优秀人才支持计划资助项目(LJQ2013090)；辽宁医学院校长基金资助项目(XZJJ20130245)，课题名称：组织工程人工神经修复周围神经缺损的研究通讯作者: 吕刚，教授，博士生导师，辽宁医学院附属第一医院骨科，辽宁省锦州市 121001 作者简介: 张彩顺，男，1973年生，辽宁省丹东市人，汉族，2007年中国医科大学毕业，博士，副教授，主要从事周围神经组织工程方面的研究。引用本文: 张彩顺,吕 刚. 辣根过氧化物酶逆行示踪评价组织工程人工神经修复大鼠坐骨神经缺损J. 中国组织工程研究, 2014, 18(29): 4658-4662. Zhang Cai-shun,Lv Gang. Tissue engineered artificial nerves repair sciatic nerve defect in rats: an evaluation using horseradish peroxidase retrograde tracer techniqueJ. Chinese Journal of Tissue Engineering Research, 2014, 18(29): 4658-4662. 摘要 背景：作者前期制备了无细胞神经移植物，并将骨髓间充质干细胞种植其内，成功构建了组织工程人工神经。目的：应用辣根过氧化物酶神经逆行示踪技术，评价无细胞神经移植物复合骨髓间充质干细胞构建的组织工程人工神经桥接大鼠坐骨神经缺损后对感觉神经元的保护作用。方法：成年清洁级健康雄性SD大鼠，随机分成3组：实验组采用复合骨髓间充质干细胞的无细胞神经移植物桥接大鼠坐骨神经缺损；空白对照组采用无细胞神经移植物桥接大鼠坐骨神经缺损；自体神经对照组采用自体神经移植桥接大鼠坐骨神经缺损。移植后12周应用辣根过氧化物酶神经逆行示踪技术，对脊髓后根神经节感觉神经元的再生进行评价。结果与结论：移植12周后根神经节感觉神经元再生评价结果提示，实验组与自体神经对照组相比差异无显著性意义，优于空白对照组；足底皮肤S-100免疫组织化学染色结果显示，各组皮下均见棕色阳性反应。提示无细胞神经移植物复合骨髓间充质干细胞构建组织工程人工神经修复大鼠坐骨神经缺损，对大鼠脊髓后根神经节感觉神经元具有保护作用，可以促进感觉功能的恢复。中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：窗体顶端关键词 ： 组织构建； 组织工程； 辣根过氧化物酶； 骨髓间充质干细胞； 无细胞神经移植物； 神经缺损； 后根神经节； Abstract： BACKGROUND: We have previously prepared acellular nerve graft and implanted bone marrow mesenchymal stem cells into the graft, to successfully construct tissue engineered artificial nerves.OBJECTIVE: Horseradish peroxidase nerve retrograde tracer technique was used to evaluate protective effects on sensory neurons following sciatic nerve defect bridging with tissue engineered artificial nerves constructed by acellular nerve graft and bone marrow mesenchymal stem cells in rats.METHODS: Adult, clean, healthy, male Sprague-Dawley rats were randomly assigned to three group: (1) Experimental group: Rat sciatic nerve detect was bridged by acellular nerve graft combined with bone marrow mesenchymal stem cells; (2) Blank control group: Rat sciatic nerve defect was bridged by acellular nerve graft; (3) Autologous nerve control group: Rat sciatic nerve defect was bridged by autologous nerve transplantation. Regeneration of sensory neurons in the spinal dorsal root ganglia was assessed using horseradish peroxidase nerve retrograde tracer technique at 12 weeks following surgery.RESULTS AND CONCLUSION: Sensory neuron regeneration in the spinal dorsal root ganglia at 12 weeks following surgery was better in the experimental group compared with blank control group. No significant difference was detected between experimental group and autologous nerve control group. S-100 immunohistochemical staining in plantar skin showed brown positive reaction in each group. These findings indicate that tissue engineered artificial nerves constructed by acellular nerve graft and bone marrow mesenchymal stem cells have protective effects on sensory neurons in the spinal dorsal root ganglia, and can promote the recovery of sensory function and repair sciatic nerve defect in rats.中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质