两种去细胞方法制备大鼠脱细胞脊髓支架效果的-第三军医大学学报.docVIP

两种去细胞方法制备大鼠脱细胞脊髓支架效果的对比研究( 郭树章，蒋 涛，任先军 （第三军医大学新桥医院骨科，重庆 400037） 摘要：目的 比较脱氧胆酸钠+TritonX100和脱氧胆酸钠+DNA+RNA酶两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2cm，运用冻融+化学萃取的组织工程学方法（脱氧胆酸钠+TritonX100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 两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，未见细胞成份存留，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+化学萃取的两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。 关键词：脱细胞脊髓；组织工程支架；大鼠，脱氧胆酸钠；TritonX-100 Fabrication of decellularized spinal cord scaffold of rats with two kinds of method GUO Shu-zhang， REN xian-jun* ，JIANG tao， Wu Bo Department of Orthopaedics， Xinqiao Hospital of Third Military Medical University，Chongqing 400037 China Abstract： [Objective] To compare the effect of fabricating decellularized scaffold of spinal cord with two kinds of method， and to provide an ideal natural spinal cord scaffold to bridge the nerve gap. [Methods] Rat spinal cord was cut and treated using the method of freeze thawing and chemical extraction. HE staining， myelin staining and SEM observation was exploited to evaluate this spinal cord scaffold. [Results] Both Cell and myelin was completely decellularized with two methods. In cross section， network of thewas presented without axon， sheath and cells nucleus being seen in the scaffold. Typicalempty tubes were viewed in longitudinal sections. [Conclusions] An ideal spinal cord scaffold can be produced with two tissue engineering methods designed in our experiment. This scaffold has similar three dimensional structure with normal spinal cord， which 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. Key words： Acellular Spinal Cord；Tissue Engineering Scaffold；Rat; sodium deoxycholate; TritonX-100 脊髓损伤后轴突再生和功能恢复一直是骨科及神经外科的难题，虽然近年来脊髓损伤修复取得了一定的进步，但仍未取得突破性进展。在目前有限可供选择种子细胞前提下，组织工程支架的选择显得尤为重要。近年来一种新的趋势是采用天然的脱细胞基质作为神经组织的修复材料，即用同种异体来源的材料，采用脱细胞技术制备的脱细胞基质，形成生物衍生材料(bio-derived material )，用于组织工程研究。为生物衍生组织工程脊髓支架材料Trito