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纸质出版:2010
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郭树章,蒋涛,任先军.2种去细胞方法制备大鼠脱细胞脊髓支架效果的对比研究[J].陆军军医大学学报(原第三军医大学学报),2010,32(01):19-22.
Guo Shuzhang,Jiang Tao,Ren Xianjun.Comparison of 2 decellularizing methods to prepare rat acelluar allograft spinal cord scaffold[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(01):19-22.
目的 比较脱氧胆酸钠+Triton X-100和脱氧胆酸钠+DNA+RNA酶2种脱细胞方法制备脱细胞的同种异体脊髓支架的效果。 方法 取成年SD大鼠胸段脊髓2 cm,运用冻融+化学萃取的组织工程学方法(脱氧胆酸钠+Triton X-100和脱氧胆酸钠+DNA+RNA酶)处理大鼠脊髓组织,对处理后的脊髓支架分别进行HE染色,髓鞘染色和扫描电镜观察。 结果 2种方法均较为完全地脱去了细胞和髓鞘成分,脊髓横断面上呈网状结构,未见细胞成分存留,纵切面上呈互相交错的管状通路;未见轴突、髓鞘和细胞核。 结论 采用冻融+化学萃取的2种组织工程学方法均可制备出理想的天然脊髓支架,该支架与脊髓三维组织结构具有高度相似性,有望作为脊髓损伤后桥接物和神经组织工程种子细胞的支架。
Objective To compare the effect of 2 decellularizing methods
sodium deoxycholate plus Triton X-100 or plus DNase and Rnase
in the preparation of acelluar allograft spinal cord scaffold in order to provide an ideal natural spinal cord scaffold to bridge the nerve gap. Methods Spinal cord was removed from health rats
and then decellularized by the method of freeze thawing (immersing in 3% sodium deoxycholate followed by the mixture of 1×10
3
U/ml DNase and RNase)
or by chemical extraction (immersing in 1% Triton X-100 and then 1% sodium deoxycholate). HE staining
myelin staining and scanning electron microscopy (SEM) were employed to evaluate the spinal cord scaffold after the 2 methods of decellularization. R
esults Both cells and myelin were completely decellularized with the 2 methods. In cross section
network of the extracellular matrix was presented without axon
sheath and cells nucleus being seen in the scaffold. Typical network of empty tubes were viewed in longitudinal sections. Conclusion An ideal spinal cord scaffold can be produced with these 2 decellularizing methods in tissue engineering. The scaffold made by the 2 methods have similar three dimensional structure with normal spinal cord
so can be used as a graft to bridge the nerve gap directly or as a scaffold to implant the seeding cells in spinal cord tissue engineering.
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