去细胞组织工程化神经支架的制备与形态学研究  

Scaffold of acellular tissue-engineered nerve derived from human and study of its morphology

作　　者：朱庆棠[1] 朱家恺[1] 赖英荣[2] 江长青[1] 杨光诗[1] 胡军[1] 刘小林[1] 

机构地区：[1]中山大学附属第一医院显微外科,广州市510080 [2]中山大学基础医学院病理教研室,广州市510080

出　　处：《中华显微外科杂志》2004年第1期35-37,F003,共4页Chinese Journal of Microsurgery

基　　金：国家自然科学基金 (30 1 70 962 );广州市科技攻关重大项目 (2 0 0 2Z1 E0 0 31 )

摘　　要：目的　探讨如何去除人类长段粗大神经中的细胞。为构建人类组织工程化神经提供具有仿生结构的支架。　方法　取 4段长 10cm、直径 6mm人的胫神经 ,其中 3段用 3 %三硝基甲苯和4%脱氧胆酸钠分别萃取 1、2和 3次 ,在萃取神经和未萃取神经的中段取材 ,行HE染色、砂罗铬花青染色和S 10 0免疫组化染色 ,在光镜和电镜下观察神经萃取前后形态学的变化。　结果　用三硝基甲苯和脱氧胆酸钠萃取后 ,胫神经内的细胞消失 ,而纤维性支架结构与未经萃取的神经相仿 ,电镜下可见萃取后的神经由空的神经基底膜管和管之间的胶原纤维构成。随着萃取次数的增加 ,神经内残留的S 10 0蛋白显著减少 ,但反复萃取后神经的支架结构受破坏。　结论　用三硝基甲苯和脱氧胆酸钠萃取2次可去除人体长段、粗大神经的细胞而保留完整的神经基底膜管和纤维支架结构 ,可能是制备具有仿生结构的人类组织工程化神经支架较为理想的方法。Objective To develop a procedure to remove the cells and myethin in peripheral nerve of human, so as to obtain a long and thick acellular scaffold for fabricating tissue-engineered nerve in human. Methods Four segments of tibial nerve, 10 cm in length and 6 mm in diameter, were taken from a fresh amputated lower extremity. The nerves were extracted with 3% Triton X-100 and 4% sodium deoxycholate in different times. HE, myethin and S-100 immunohistochemical staining were performed and investigated by light microscope and electron microscope. Results The cells, myethin and axons disappeared after the human tibial nerve was treated with Triton X-100 and sodium deoxycholate. The scaffold of acellular nerve was similar to the fresh nerve. Elctron microscope showed that the human acellular nerve was composed of empty basal lamina tubes and collage fibers among the tubes. There were a few S-100 proteins residue within the nerve if it was treated only one time, but they could be removed by treated once again. Conclusions The cells in long and thick human peripheral nerve can be removed by chemical extraction with Triton X-100 and sodium deoxycholate, while the basal lamina tubes and fiber scaffold can be kept intact. Maybe chemical extraction is an ideal method to prepare long and thick scaffold for fabricating tissue-engineered nerve in human.

关 键 词：去细胞技术 组织工程 神经支架 形态学 神经移植体 

分 类 号：R318.1[医药卫生—生物医学工程]