不同来源嗅鞘细胞移植治疗脑出血的比较

Comparison of different genesis of olfactory ensheathing cell transplantation for intracerebral hemorrhage

背景:嗅鞘细胞是目前已知用于移植细胞中惟一可以跨越周围神经系统与中枢神经系统边界的细胞。在众多的国内外文献中,大多以嗅球来源嗅鞘细胞作为观察对象;但采用嗅黏膜嗅鞘细胞移植具有来源方便、损伤小、可自体取材等优势。目的:比较嗅球来源嗅鞘细胞和嗅黏膜嗅鞘细胞移植对脑出血后神经功能损伤恢复作用的差异。设计、时间及地点:免疫组织化学水平的随机对照动物实验,于2005-10/2007-10在无锡市第三人民医院细胞实验室完成。材料:选用健康雄性SD大鼠30只,按随机数字表法分为3组,对照组、嗅球来源嗅鞘细胞移植组及嗅黏膜嗅鞘细胞移植组各10只。方法:分别获取人鼻中隔黏膜、人胚嗅球,进行嗅鞘细胞的分离培养纯化,取培养10d的细胞用作细胞移植。取SD大鼠制备尾状核出血模型,嗅球来源嗅鞘细胞移植组及嗅黏膜嗅鞘细胞移植组各取10μL细胞悬液在立体定向下引导微量注射器向大鼠脑组织内匀速注射(1μL/min),对照组注入等量培养基,注射点为右侧尾状核。主要观察指标:观察两种来源嗅鞘细胞的体外培养情况。嗅鞘细胞移植后对动物行定期行为学评定,功能损伤越重,得分越高;并观察组织切片靶区域免疫细胞化学分析结果。结果:从体外培养结果看,无论在形态上还是表型上,两者无明显区别。嗅球来源嗅鞘细胞移植组及嗅黏膜嗅鞘细胞移植组大鼠的Longa5分制法及前肢放置试验评分在移植后第14,28,56天显著低于对照组(P<0.05),两移植组差异无显著性意义(P>0.05),移植组各时间段差异无显著性意义(P>0.05)。结论:用于移植修复神经功能的两种嗅鞘细胞在细胞特性、移植效果方面均无明显差异。

BACKGROUND： Olfactory ensheathing cells can cross the boundary of the peripheral nervous system and the central nervous system during cell transplantation. Previous studies observed the olfactory bulb-derived olfactory ensheathing cells （OB-OECs）. However, the transplantation of olfactory mucosa-derived OECs （OM-OECs） is convenient to obtain, with a few injuries. OBJECTIVE： To compare the differences of OB-OEC or OM-OEC transplantation on repairing neurological function after intracerebral hemorrhage. DESIGN, TIME AND SETTING： The immunohistochemistry, randomized, controlled animal study was performed at the Cell Laboratory of Wuxi Third People＇s Hospital from October 2005 to October 2007. MATERIALS： A total of 30 healthy male Sprague Dawley rats were equally and randomly divided into 3 groups： control group, OB-OEC transplantation group and OM-OEC transplantation group. METHODS： Human nasal septum mucosa and human embryo olfactory bulb were obtained for OEC incubation. Following 10 days of incubation, cells were used for cell transplantation. Sprague Dawley rat models of caudate nucleus hemorrhage were established. 10 μL of cell suspension was separately collected from the OB-OEC transplantation group and OM-OEC transplantation group, injected into rat brain tissues （1 μL/min） using a microinjector under the stereotaxis. The same volume of medium was infused into the right caudate nucleus from rats of the control group. MAIN OUTCOME MEASURES： In vitro culture of OB-OECs and OM-OECs was observed. Following OEC transplantation, anima behavior was assessed. The severe injury indicated a high score. Immunocytochemistry analysis results were observed at the target area of tissue slices. RESULTS： No significant differences were detected in morphology and phenotype of both cells. Longa 5 point method and forelimb place test were significantly lower in the OB-OEC transplantation group and OM-OEC transplantation group compared with the control group at 14, 28 and 56 days （P 〈 0.05）. No significant differences were found between the two transplantation groups （P 〉 0.05）. No significant differences were seen in the transplantation groups at various time points （P 〉 0.05）. CONCLUSION： There are no significant differences in cell characteristics and transplantation outcome using OB-OEC and OM-OEC transplantation for repairing neurological function.