摘要
目的:观察并比较兔坐骨神经组织细胞微囊化处理与单纯兔坐骨神经组织细胞移植至大鼠损伤脊髓及大鼠单纯脊髓损伤3组神经生长因子表达的变化。方法:实验于2003-05/2005-05在九江学院医学科研中心完成。①制备不同移植材料:取10只成年家兔双侧完整坐骨神经制备细胞悬液;兔坐骨神经组织细胞悬液微囊化处理,将细胞浓度调整为(2.0～2.5)×106L-1后与15g/L海藻酸钠生理盐水溶液混合,经双腔喷头将海藻酸钠-坐骨神经组织细胞悬液混合液喷入20mmol/L的氯化钡生理盐水溶液中,加入生理盐水洗涤2次。②脊髓损伤大鼠模型制作:取健康SD大鼠90只,随机分为3组,微囊化兔坐骨神经组织细胞悬液组(微囊化组);单纯兔坐骨神经组织细胞悬液组(单纯悬液组)和单纯损伤组,每组30只。健康SD大鼠腹腔麻醉(30mg/kg)成功后,以T10为中心,暴露T10棘突及椎板,作脊髓右半侧横向切开,并去除约2mm脊髓组织。立即在损伤腔内植入约2mm×2mm×2mm的明胶海绵作为填充支架,微囊化组在明胶海绵上吸附10μL微囊化兔坐骨神经组织细胞悬液;单纯悬液组在明胶海绵上吸附10μL单纯兔坐骨神经组织细胞悬液;单纯损伤组不做任何移植。③神经生长因子检测:于移植后1,3,7,14,28d,取出大鼠脊髓损伤平面为中心的2cm脊髓标本,每组6个标本共18张切片,进行免疫组织化学染色(SABC法),神经生长因子染色以细胞浆呈明显的棕黄色为阳性染色,分别计算每平方毫米阳性神经细胞数。结果:90只大鼠全部进入结果分析。①各组神经生长因子阳性神经元测定结果:微囊组在1,3,7,14,28d均高于单纯损伤组和单纯悬液组;在第7,14,28天明显高于单纯损伤组和单纯悬液组犤7d:(28.55±2.26),(7.65±3.35),(19.35±2.39)个/mm2;14d:(30.52±3.51),(8.10±2.45)(20.45±3.45)个/mm2;28d:(27.50±4.50),(6.59±3.85),(17.50±4.65)个/mm2,P<0.01或0.05犦;第14天达到顶峰(P<0.01)。②神�
AIM： To observe and compare the impacts on nerve growth factor （NGF） expression in rats of three groups： microencapsulated rabbit sciatic nerve （SN） cells transplantations on rats with spinal cord injury （SCI）, rabbit SN cells transplantations on rats with SCI and rats only with SCI groups.
METHODS： The experiment was finished in the Centre of Medical Science Research, Jiujiang University from May 2003 to May 2005.①Preparations of different transplanted materials： The complete SN of 10 rabbits were used for making cell suspension; With the adjusted cell concentration of （2.0-2.5） ×10^6 L^-1, the microencapsulated suspension was mixed with the alginate saline solution, and ejaculated to 20 mmol/L barium chloride saline solution by double-cavity ejaculator, then rinsed twice by saline. ②SCI rats models establishment： Totally 90 healthy SD rats were randomly divided into 3 groups with 30 rats in each： Microencapsulated rabbit SN cells suspension group （Microencapsulated group）; Rabbit SN cells suspension group （Only suspension group）; Only injured group. After anesthetized with 30 mg/kg peritoneal injection, T10 spinous process and vertebra lamina of rats were exposed as T10 center. The spinal cord tissues of 90 rats were removed from rats by spinal cord right hemisection. The gelatin sponges with the size of 2 mm×2 mm×2 mm were grafted as filling cage, and adsorpted 10μL microencapsulated rabbit SN cells （Microencapsulated group） and 10 μL rabbit SN cells （Only suspension group） respectively; There was no graft in only injured group. ③NGF detection： Six rats were selected on each time point （1^st, 3^rd, 7^th, 14^th, 28^th days） to take out the 2 cm spinal cord samples, and totally 18 sections of 6 samples in each group were detected with the immunohistochemistry staining. The plasm presented obvious brown-yellow meant the positive staining of NGF. The number of positive nerve cells in per square millimeter was counted.
RESULTS： All the 90 rats were i
出处
《中国临床康复》
CAS
CSCD
北大核心
2006年第17期7-9,共3页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(30060034)
江西省自然科学基金(0140126)
江西省卫生厅(20052039)
九江市指导性计划项目(200447)
九江学院重点项目(05kj25)~~
