晚期脊髓损伤患者嗅鞘细胞移植术后交感神经皮肤反应变化的临床研究
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摘要
目的:
观察晚期脊髓损伤患者嗅鞘细胞移植术后交感神经皮肤反应的波幅、潜伏期的变化及感觉、运动ASIA评分变化,分析其术后疗效及价值。
方法:
①对象:自2010年1月至2010年12月收治的43例晚期脊髓损伤患者(病史>6个月),男39例,女4例,平均年龄(30.7±10.3)岁,平均病程(4.7±3.5)年;完全性17例,不完全性26例,受伤节段:C1-813例,T1-10 14例,T11及以下16例;受伤原因:车祸、摔伤、砸伤。双下肢均有不同程度的出汗减少,皮肤颜色灰暗,脱屑症状。②嗅鞘细胞的培养及鉴定:选择4～6个月中期引产胚胎(水囊引产),对其嗅球剥膜后机械性剪碎,然后培养消化细胞并制成单细胞悬液,浓度为1×1010 L-1左右,其纯度在96%左右。嗅鞘细胞免疫组织化学染色,表现为p75抗体阳性。③嗅鞘细胞移植手术:术前脊髓病变部位MRI检查,根据具体的脊髓损伤情况,准确进行定位。手术采用后正中入路,要把准确定位的损伤节段的棘突和椎板咬除,打开硬脊膜,暴露损伤部位的脊髓,在显微镜下,把嗅鞘细胞悬液多点注入脊髓病变部位上下端,和正常脊髓交界的部位,数量约约100μL,细胞数约1×106个嗅鞘细胞,关闭切口。未用免疫抑制剂。④功能评定:在相同的条件下术前1天和术后2周行SSR检测及ASIA运动、感觉进行评分,比较前后的变化。
结果:
所有患者术后2周其下肢运动、感觉、植物神经功能均有不同程度的改善,植物神经功能改善表现为皮肤均有不同程度出汗增加,颜色好转,脱屑减少。SSR测定术后其潜伏期及波幅优于手术前,其术后运动感觉ASIA评分都有明显提高.
结论:
晚期脊髓损伤患者通过嗅鞘细胞移植治疗,其神经功能都有不同程度改善,包括运动、感觉及植物神经功能,运动感觉可以通过ASIA功能表评价,SSR检测能较为客观的反映脊髓术后交感神经功能恢复情况,尤其是出汗的变化,可以作为ASIA功能评价的补充,对脊髓植物神经功能进行评价,能更准确全面地反映细胞移植后脊髓神经功能的变化。
Objective:
To Observe in patients with advanced spinal cord injury after transplantation of olfactory ensheathing cells of sympathetic skin response latency changes and amplitude changes in ASIA scores, analysis and value of its after effects.
Method:
①: PARTICIPANTS: 43 patients with advanced spinal cord injury patients, 36 males and 4 females, mean age (30.7±10.3) years, mean disease duration (4.7±3.5) years; complete in 17 cases, 26 incomplete cases, injury segments: cervical 13 cases, 14 cases of thoracic and lumbar 16; cause of injury: car accidents, falls, injured. preoperative can lead to SSR. of both lower extremities to reduce sweating in varying degrees, dark skin color, scaling symptoms.②Cell culture and identification: Take 4 to 6 months embryo induction medium (through the Hospital Ethics Committee approval) of the olfactory bulb, after stripping membrane mechanical manipulation, cultured in the presence of 10% -15% fetal calf serum D/F-12 Training liquid (GIBICO products) in, 3d after the purification of serum-free DMEM culture medium was changed every 3d semi-liquid 1, continue to develop 7-10 d. Digestive cells and made into single cell suspension, the cell concentration of 1×1010L-1 or so. The purified p75 antibodies in cultured cells selected for immunohistochemical staining, positive for olfactory ensheathing cells, the purity of 95% to 98%.③Surgery and cell transplantation: preoperative routine MRI, according to MRI to determine the site of spinal cord lesions. Segmental lesions anesthesia after midline approach to remove the spinous process and lamina of the lesion, dura cut to expose the spinal cord. With the operating microscope in the spinal cord and spinal cord lesions at the junction of the normal part of multi-point injection disposal of approximately 100μL suspension of olfactory ensheathing cells, about 1×106个olfactory ensheathing cells, tightly close the dura mater and suture layers. Given after surgery to stop bleeding, anti-infection treatment④Functional evaluation: the same conditions in the downstream SSR detection, using ASIA motor and sensory table rate before and after surgery were compared before and after the change.
Results:
All patients after 2 weeks of their lower limb motor, sensory, autonomic function were improved to some extent, improve the performance of autonomic function in varying degrees of skin increased sweating, color improvement, scaling reduced. SSR latency and amplitude measured after its better than before surgery, the incubation period amplitude .The spinal cord motor and sensory function after surgery have significantly improved scores .
Conclusion:
Olfactory ensheathing cell transplantation for the treatment of spinal cord injury patients, the patients had improved motor and sensory autonomic functions, SSR detection can be more objectively reflect autonomic functional recovery after spinal cord can be used as supplementary ASIA functional assessment of the spinal cord Evaluation of autonomic function. More accurate and comprehensive reflection of cell transplantation for spinal cord nerve function changes.
观察晚期脊髓损伤患者嗅鞘细胞移植术后交感神经皮肤反应的波幅、潜伏期的变化及感觉、运动ASIA评分变化,分析其术后疗效及价值。
方法:
①对象:自2010年1月至2010年12月收治的43例晚期脊髓损伤患者(病史>6个月),男39例,女4例,平均年龄(30.7±10.3)岁,平均病程(4.7±3.5)年;完全性17例,不完全性26例,受伤节段:C1-813例,T1-10 14例,T11及以下16例;受伤原因:车祸、摔伤、砸伤。双下肢均有不同程度的出汗减少,皮肤颜色灰暗,脱屑症状。②嗅鞘细胞的培养及鉴定:选择4～6个月中期引产胚胎(水囊引产),对其嗅球剥膜后机械性剪碎,然后培养消化细胞并制成单细胞悬液,浓度为1×1010 L-1左右,其纯度在96%左右。嗅鞘细胞免疫组织化学染色,表现为p75抗体阳性。③嗅鞘细胞移植手术:术前脊髓病变部位MRI检查,根据具体的脊髓损伤情况,准确进行定位。手术采用后正中入路,要把准确定位的损伤节段的棘突和椎板咬除,打开硬脊膜,暴露损伤部位的脊髓,在显微镜下,把嗅鞘细胞悬液多点注入脊髓病变部位上下端,和正常脊髓交界的部位,数量约约100μL,细胞数约1×106个嗅鞘细胞,关闭切口。未用免疫抑制剂。④功能评定:在相同的条件下术前1天和术后2周行SSR检测及ASIA运动、感觉进行评分,比较前后的变化。
结果:
所有患者术后2周其下肢运动、感觉、植物神经功能均有不同程度的改善,植物神经功能改善表现为皮肤均有不同程度出汗增加,颜色好转,脱屑减少。SSR测定术后其潜伏期及波幅优于手术前,其术后运动感觉ASIA评分都有明显提高.
结论:
晚期脊髓损伤患者通过嗅鞘细胞移植治疗,其神经功能都有不同程度改善,包括运动、感觉及植物神经功能,运动感觉可以通过ASIA功能表评价,SSR检测能较为客观的反映脊髓术后交感神经功能恢复情况,尤其是出汗的变化,可以作为ASIA功能评价的补充,对脊髓植物神经功能进行评价,能更准确全面地反映细胞移植后脊髓神经功能的变化。
Objective:
To Observe in patients with advanced spinal cord injury after transplantation of olfactory ensheathing cells of sympathetic skin response latency changes and amplitude changes in ASIA scores, analysis and value of its after effects.
Method:
①: PARTICIPANTS: 43 patients with advanced spinal cord injury patients, 36 males and 4 females, mean age (30.7±10.3) years, mean disease duration (4.7±3.5) years; complete in 17 cases, 26 incomplete cases, injury segments: cervical 13 cases, 14 cases of thoracic and lumbar 16; cause of injury: car accidents, falls, injured. preoperative can lead to SSR. of both lower extremities to reduce sweating in varying degrees, dark skin color, scaling symptoms.②Cell culture and identification: Take 4 to 6 months embryo induction medium (through the Hospital Ethics Committee approval) of the olfactory bulb, after stripping membrane mechanical manipulation, cultured in the presence of 10% -15% fetal calf serum D/F-12 Training liquid (GIBICO products) in, 3d after the purification of serum-free DMEM culture medium was changed every 3d semi-liquid 1, continue to develop 7-10 d. Digestive cells and made into single cell suspension, the cell concentration of 1×1010L-1 or so. The purified p75 antibodies in cultured cells selected for immunohistochemical staining, positive for olfactory ensheathing cells, the purity of 95% to 98%.③Surgery and cell transplantation: preoperative routine MRI, according to MRI to determine the site of spinal cord lesions. Segmental lesions anesthesia after midline approach to remove the spinous process and lamina of the lesion, dura cut to expose the spinal cord. With the operating microscope in the spinal cord and spinal cord lesions at the junction of the normal part of multi-point injection disposal of approximately 100μL suspension of olfactory ensheathing cells, about 1×106个olfactory ensheathing cells, tightly close the dura mater and suture layers. Given after surgery to stop bleeding, anti-infection treatment④Functional evaluation: the same conditions in the downstream SSR detection, using ASIA motor and sensory table rate before and after surgery were compared before and after the change.
Results:
All patients after 2 weeks of their lower limb motor, sensory, autonomic function were improved to some extent, improve the performance of autonomic function in varying degrees of skin increased sweating, color improvement, scaling reduced. SSR latency and amplitude measured after its better than before surgery, the incubation period amplitude .The spinal cord motor and sensory function after surgery have significantly improved scores .
Conclusion:
Olfactory ensheathing cell transplantation for the treatment of spinal cord injury patients, the patients had improved motor and sensory autonomic functions, SSR detection can be more objectively reflect autonomic functional recovery after spinal cord can be used as supplementary ASIA functional assessment of the spinal cord Evaluation of autonomic function. More accurate and comprehensive reflection of cell transplantation for spinal cord nerve function changes.
引文
[1]郑望苟,潘卫红,郭卫春.脊髓损伤后脊髓自由基和超氧化物岐化酶的动态变化.中国骨伤杂志[J],2004,17(7):391-392
[2]李盛华,郭平德,王文晶.脊髓损伤的治疗现状与进展中国骨伤[J].2010,23(1):70-72
[3] Ramon-Cueo A, Avila. Olfactory ensheathing glia properties and function[ J] .Brain ResBull , 1998, 46( 3) : 175-187
[4] Ramon-Cueto A, Valverde F. Olfactory bulb ensheathing glia:a unique cell type with axonal growt hpromoting properties[J] .Gli a, 1995, 14( 3) : 163-173
[5] Bartolomei JC, Greer CA. Olfactory ensheathing cells: bridging the gap in spinal cord injury[ J ]. Neurosurgery, 2000, 47(5): 1057 -1069
[6]陈莉发,余华荣,李兵仓.嗅鞘细胞在神经系统损伤修复中的作用[J].创伤外科杂志2010,12(1):91-93
[7] Masanori S, Joel AB, Karen LL, etal. Molecular reconstruction of nodes of ranvier after remyelination by transplanted olfactory ensheathing cells in the demyelinated spinal cord. J Neurosci, 2006, 26(6): 1803-1812
[8]王广志,刘明娜.接受嗅鞘细胞移植脊髓损伤大鼠电生理及后肢功能变化[ J ].中国组织工程研究与临床康复,2010,14(1):112-115
[9]黄红云,陈琳等,嗅鞘细胞移植改善晚期脊髓损伤患者神经功能的近期效果[ J ].中国临床康复2006,10(13):25
[10]黄红云,陈琳等.嗅鞘细胞移植治疗晚期脊髓损伤安全性评价38个月磁共振随访结果[ J ].中国修复重建外科杂志,2006,4:153-157
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[13] Levy DM, Reid G, Rowley DA. et al. Quantitative measures of sympathetic skin response in diabetes: relation to sudomotor and neurological function. J Neurol Neurosurg Psychiatry[J], 1992, 55: 902
[14]王建军,张贞浏.交感神经皮肤反应.临床脑电学杂志[J].1999,2:55-57
[15]高瑞,刘超,郑遵成.人胚胎嗅鞘细胞的分离培养与纯化.中国组织工程研究与临床康复[J].2007,11(46),9285-9288
[16]黄红云.嗅鞘细胞移植[M].北京:科学出版社,2007:74
[17]任玉水,闫军浩,王洪美.人胚嗅球嗅鞘细胞原代培养的实验研究,解剖科学进展[J].2010,16(2):97~100
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[20]孙天胜,任继鑫,史建国.嗅鞘细胞移植促进脊髓损伤的修复作用.中国医学科学院学报[J] .2005,27(2):143-147
[21]魏开斌,郑遵成.嗅鞘细胞移植治疗脊髓损伤的临床思考[J].中国矫形外科杂志,2010,18(13) 1137-1140
[22]张磊,郑遵成,刘超等.嗅鞘细胞移植部位与剂量的选择对陈旧性脊损伤患者60例神经恢复的影响[J].中国民康医学2008,20(9)870-871
[23]郑遵成,刘超等.移植时间对嗅鞘细胞移植治疗脊髓损伤效果的影响中国组织工程研究与临床康复2008 ,12(3):583-585
[24]刘超,郑遵成等.嗅鞘细胞移植术后脊髓损伤患者功能的评价[ J ].中国组织工程研究与临床康复,2008;12(16):3037-3039
[25]焦玲.交感神经皮肤反应研究进展.临床神经电生理学杂志[J].2002,11(2),126-128
[26] Hoeldtke RD, Davis KM, Hshieh PB, et al. Autonomic surface potential analysis: assessmeent of reproducibility and sensitivity.Muscle Nerve[J], 1992, 15: 926
[27] Levy DM, Reid G, Rowley DA. et al. Quantitative measures of sympathetic skin response in diabetes: relation to sudomotor and neurological funct ion. J Neurol Neurosurg Psychiatry[J], 1992, 55: 902
[28]焦玲,吕如锋.交感神经皮肤反应的正常值和影响因素.贵阳医学院学报[J].2002,27(1):16-19
[29]焦玲,吕如锋,张文渊.脑梗死患者交感神经皮肤反应的临床研究[J].临床神经电生理学杂志, 2001, 10( 3) : 139~ 143
[30] Korpelainen JT, Tolonen U, Sotaniemi KA, et al. Supperssed sympathetic skin response in brain infarction . St roke , 1993, 24( 9) : 1389~ 1392.
[31]刘萍.体感及运动诱发电位检测对脑梗死的临床价值[J].医学综述.2008,22:100-103.
[32]谢莉红.交感神经皮肤反应检测在糖尿病性周围神经病诊断中的价值[J].宁夏医科大学学报.2009,1:38-40.
[33]彭超,许光旭,王红星等.陈文红脊髓损伤患者交感神经皮肤反应的变化[J] .中国脊柱脊髓杂志.2010,20(9):716-720.
[1]郑望苟,潘卫红,郭卫春.脊髓损伤后脊髓自由基和超氧化物岐化酶的动态变化[J].中国骨伤杂志,2004,17(7):391-392
[2]李盛华,郭平德,王文晶.脊髓损伤的治疗现状与进展[J].中国骨伤杂志,2010,23(1):70-72
[3]黎文,林志雄,陈艺等.无骨折脱位型颈髓损伤的手术治疗[J].中华创伤杂志,2005,21(7):499-501
[4]黄红云.嗅鞘细胞移植[M].北京:科学出版社,2007:74
[5] Bartolomei JC, Greer CA. Olfactory ensheathing cells: bridging the gap in spinal cordinjury[ J ]. Neurosurgery, 2000, 47(5): 1057 -1069.
[6]陈莉发,余华荣,李兵仓.嗅鞘细胞在神经系统损伤修复中的作用[J].创伤外科杂志,2010,12 (1):91-93.
[7]孙天胜,任继鑫,史建国.嗅鞘细胞移植促进脊髓损伤的修复作用[J].中国医学科学院学报,2005,27(2):143-147
[8]魏开斌,郑遵成.嗅鞘细胞移植治疗脊髓损伤的临床思考[J].中国矫形外科杂志,2010,18(13) 1137-1140.
[9]刘超,郑遵成等.嗅鞘细胞移植术后脊髓损伤患者功能的评价[ J ].中国组织工程研究与临床康复,2008,12(16):3037-3039
[10]黄红云.嗅鞘细胞移植[M].北京:科学出版社,2007:74
[11] Stephen AR, Patricia EP. Mouse olfactory ensheathing glia enhanceaxon outgrow on a myelin substrate in vitro [ J ]. Exp Neurol, 2009,216(1): 95 - 104.
[12] Cosgaya JM, Chan JR, Wu YJ, et al. Neurotrophins are key media 2tors of the myelination program in the peripheral nervous system[ J ]. Proc Natl Acad Sci USA, 2001, 98(25): 14661 - 14668.
[13]谢莉红,刘南平.交感神经皮肤反应及其临床应用[J].宁夏医科大学学报, 2009,20(3) 135-138
[14]焦玲.交感神经皮肤反应研究进展.临床神经电生理学杂志[J].2002,11(2),126-128
[15] Hoeldtke RD, Davis KM, Hshieh PB, et al. Autonomic surface potential analysis: assessmeent of reproducibility and sensitivity.Muscle Nerve[J], 1992, 15: 926
[16] Levy DM, Reid G, Rowley DA. et al. Quantitative measures of sympathetic skin response in diabetes: relation to sudomotor and neurological function[J]. Neurol Neurosurg Psychiatry, 1992, 55: 902
[17]王建军,张贞浏.交感神经皮肤反应[J].临床脑电学杂志,1999,2,55-57
[18]焦玲,吕如锋.交感神经皮肤反应的正常值和影响因素[J].贵阳医学院学报,2002,27(1):16-19.
[19]焦玲,吕如锋,张文渊.脑梗死患者交感神经皮肤反应的临床研究[J].临床神经电生理学杂志, 2001, 10( 3) : 139~ 143
[20] Korpelainen JT, Tolonen U, Sotaniemi KA, et al. Supperssed sympathetic skin response in brain infarction . St roke , 1993, 24( 9) : 1389~ 1392.
[21]刘萍.体感及运动诱发电位检测对脑梗死的临床价值[J].医学综述,2008,22:100-103.
[22]谢莉红.交感神经皮肤反应检测在糖尿病性周围神经病诊断中的价值[J].宁夏医科大学学报,2009,1:38-40.
[23]彭超,许光旭,王红星等.脊髓损伤患者交感神经皮肤反应的变化[J] .中国脊柱脊髓杂志,2010,20(9):716-720.
[2]李盛华,郭平德,王文晶.脊髓损伤的治疗现状与进展中国骨伤[J].2010,23(1):70-72
[3] Ramon-Cueo A, Avila. Olfactory ensheathing glia properties and function[ J] .Brain ResBull , 1998, 46( 3) : 175-187
[4] Ramon-Cueto A, Valverde F. Olfactory bulb ensheathing glia:a unique cell type with axonal growt hpromoting properties[J] .Gli a, 1995, 14( 3) : 163-173
[5] Bartolomei JC, Greer CA. Olfactory ensheathing cells: bridging the gap in spinal cord injury[ J ]. Neurosurgery, 2000, 47(5): 1057 -1069
[6]陈莉发,余华荣,李兵仓.嗅鞘细胞在神经系统损伤修复中的作用[J].创伤外科杂志2010,12(1):91-93
[7] Masanori S, Joel AB, Karen LL, etal. Molecular reconstruction of nodes of ranvier after remyelination by transplanted olfactory ensheathing cells in the demyelinated spinal cord. J Neurosci, 2006, 26(6): 1803-1812
[8]王广志,刘明娜.接受嗅鞘细胞移植脊髓损伤大鼠电生理及后肢功能变化[ J ].中国组织工程研究与临床康复,2010,14(1):112-115
[9]黄红云,陈琳等,嗅鞘细胞移植改善晚期脊髓损伤患者神经功能的近期效果[ J ].中国临床康复2006,10(13):25
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