不同去细胞神经支架制备方法的对比研究  

COMPARISON OF DIFFERENT CHEMICAL METHODS FOR PREPARATION OF ACELLULAR NERVE SCAFFOLD

作　　者：高嵩涛[1] 邓展生[2] 李宝军[2] 张胜利[3] 郭晓柠[4] 

机构地区：[1]河南省肿瘤医院骨科,郑州450008 [2]中南大学湘雅医院骨科 [3]重庆市中山医院骨科 [4]中南大学湘雅二医院骨科,郑州450008

出　　处：《中国修复重建外科杂志》2008年第7期851-855,共5页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的比较不同化学方法制备去细胞神经支架的效果,提供有效的组织工程神经支架。方法2月龄雄性SD大鼠15只,体重200～250g,取双侧坐骨神经,随机分为3组,每组10条。根据处理方法不同,A组为正常未处理组;B组为传统Sondell法化学处理组;C组为采用Triton X-200、SB-10、SB-16处理的改良法组。制备后行HE染色、免疫组织化学染色及扫描电镜观察,并对去细胞程度、脱髓鞘程度及神经纤维管道完整性进行量化评价。结果HE染色:A组神经纤维横断面呈圆形,胞核呈深蓝色,髓鞘呈网格状结构;B组轴突及细胞核均消失,神经内膜结构破坏明显;C组轴突及细胞核均消失,神经内膜形成不规则圆形空腔。层粘连蛋白免疫组织化学染色:A组基底膜围绕在髓鞘外周,中间有蓝染SC核;B组SC核及髓鞘结构消失,神经基底膜结构不完整;C组SC核及髓鞘结构消失,神经基底膜形成不规则圆形空腔。S-100蛋白免疫组织化学染色:A组SC及髓鞘呈棕褐色;B组及C组未见明显髓鞘染色。扫描电镜观察:A组髓鞘、轴突等内容物形态清晰;B组髓鞘、轴突等结构均消失,基膜管壁排列欠规则;C组基膜管壁胶原纤维排列有序。去细胞程度及髓鞘染色深度评分:B、C组均优于A组(P<0.05),其中髓鞘染色深度评分C组均优于B组(P<0.05);神经纤维管道完整性评分:C组优于B组(P<0.05),与A组相似(P>0.05);综合评价去细胞处理方法以C组方法为佳。结论改良法所获得的去细胞神经的去细胞效果与传统Sondell法相似,而脱髓鞘及组织结构保留的效果优于后者,可作为一种新的神经支架材料。Objective Compare the effect of different chemical methods for preparation of acellular nerve scaffold and to provide an effective nerve scaffold for tissue engineering. Methods Fifteen male SD rats of 2 months old, weighing 200-250 g were selected; the bilateral sciatic nerves were harvested and divided into 3 groups according to preparation methods： group A （normal nerve）, group B （Sondell method） and group C （optimal method by Triton X-200, SB-10 and SB-16）. The morphology was compared by HE, immunohistochemistry and SEM after dispose; the degrees of decellularization, degrees of demyelination and integrity of the nerve fiber tube were assessed by scoring system. Results HE staining： In group A, the cross section of nerve was roundness, the cell nuclei was dark blue and the myelin sheath was reticular structure. In group B, the axon and cell nuclei disappeared and the structure of endoneurium was destroyed. In group C, the axon and cell nuclei disappeared and the endoneurium become anomalistic round cavum. The immunohistochemistry staining of Laminin： In group A, the myelin sheath was surrounded by basement membrane with dark blue SC nuclei inside. In group B, the myelin sheath and SC nuclei disappeared and the structure of basement membrane destroyed. In group C, the myelin sheath and SC nuclei disappeared and basement membrane become anomalistic round cavum. The immunohistochemistry staining of S-100： In group A, the myelin sheath and SC were brown. In groups B and C, there were no apparent stained myelin sheath. SEM： In group A, the myelin sheath and axon were clear. In group B, the axon and myelin sheath disappeared and basement membrane became anomalistic. In group C, the basement membrane was more regular than that of group B. The degrees of acellularization and demyelination of groups B and C were superior to that of group A （P 〈 0.05）, and the degrees of demyelination of group C were superior to that of group B （P 〈 0.05）. The integrity of nerve fiber tube of group C w

关 键 词：组织工程 去细胞神经 神经支架 基底膜 鼠 

分 类 号：R651.3[医药卫生—外科学] R318.08[医药卫生—临床医学]