摘要
目的:应用化学萃取法制备的同种异体神经移植物在小动物及低等哺乳类动物实验中取得良好的效果,但在萃取猕猴粗大、长段周围神经方面尚未形成标准化程序。目的:研究猕猴化学去细胞同种异体神经制备方法,以期获得低免疫反应的同种异体神经移植物。设计、时间及地点:观察实验,于2003-01/09在中山大学动物实验中心完成。材料:取2.5岁,3.5kg左右,雄性健康猕猴2只,由广州九佛动物研究中心提供。方法:取4段长5cm,直径3.5～4.0mm的坐骨神经,以4%triton-100和4%脱氧胆酸钠溶液按一定浓度和程序进行萃取,萃取神经及未萃取神经于中段取材,行苏木精-伊红染色,纤维素染色,砂罗铬花青染色,S-100免疫组织化学染色,在扫描电镜及透射电镜下观察超微结构。主要观察指标:神经的组织学及形态学观察。结果:萃取2次后的去细胞神经具有良好的黏性和弹性,细胞和髓鞘被彻底清除,神经纤维支架被保留,成为一种没有细胞髓鞘及其碎片的空的基底膜管架结构。而萃取1次不能完全去除抗原成分,萃取3次则破坏神经纤维支架。结论:应用4%triton-100及4%脱氧胆酸钠萃取2次,可去除猕猴长段、粗大周围神经中的细胞和髓鞘而保留神经的基底膜管和纤维支架结构。
BACKGROUND： The acellular nerve autograft prepared by chemical extraction method has received good effect on mammals, however, it is poorly understood how to obtain thick and segmental peripheral nerves from rhesus.
OBJECTIVE： To investigate the method of chemical extraction to remove immunogenicity of peripheral nerve in rhesus, in order to obtain nerve atlograft with lower immunogenicity.
DESIGN, TIME AND SETTING： The observational experiment was performed at the Experimental Animal Center of Sun Yat-sen University from January to September 2003.
MATERIALS： Two healthy, male rhesus monkeys, 2.5 years old, weighing 3.5 kg were supplied by Jiufo Animal Research Center. METHODS： Four segments of sciatic nerve of rhesus, 5 cm in length and 3.5-4.0 mm in diameter, were extracted with 4% Triton X-100 and 4% sodium deoxycholate in different times. Hematoxylin-eosin, fibrin, myethin and S-100 immunohistochemical staining were performed, and ultrastructure was checked by light microscope and transmission electron microscope.
MAIN OUTCOME MEASURES： The histological and morphological observation of nerves. RESULTS： The acellular nerve allograft was elasticity and ductility; the cells, axons and myelin sheath were removed and basal membrane was preserved after two extraction procedures, and become empty basal lamina tubes in the endoneurium; but it could not remove the immunogenicity by one extraction procedures or in fresh nerves; the nervous basal membrane almost all ruined after three extraction procedures.
CONCLUSION： The cells in long and thick rhesus peripheral nerve can be removed by two chemical extractions with 4% Triton X-100 and 4% sodium deoxycholate, while the basal lamina tubes and fiber scaffold can be kept intact.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第21期4133-4136,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
