新型纳米支架对神经干细胞生物行为及相关基因表达的影响  

Influence of novel nanoscaffolds on biological behaviors of neural stem cells and the related gene expression

作　　者：周继辉 姚猛[2] 王岩松[2] 李新志[1] 周游 黄卫[1] 陈文瑶[1] Zhou Jihui;Yao Meng;Wang Yansong;Li Xinzhi;Zhou You;Huang Wei;Chen Wenyao(Department of Orthopedics,Affiliated Renhe Hospital of China Three Gorges University,Yichang 443001,Hubei Province,China;Department of Spine Surgery,Second Affiliated Hospital of Harbin Medical University,Harbin 150086,Heilongjiang Province,China)

机构地区：[1]三峡大学附属仁和医院骨科,湖北省宜昌市443001 [2]哈尔滨医科大学附属第二医院脊柱外科,黑龙江省哈尔滨市150086

出　　处：《中国组织工程研究》2021年第4期532-536,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:利用纳米技术制备的支架可形成与天然生物体内基底膜结构相似的表面结构,对种子细胞的行为进行有效调节。目的:观察平行与交错排列纳米纤维膜对神经干细胞增殖及分化等生物学行为的影响。方法:以Ⅰ型胶原为材料,利用静电纺织技术制备平行排列纳米纤维膜与交错排列的纳米纤维膜。将新生大鼠神经干细胞接种于两种纳米纤维膜表面,以单独的细胞培养为对照,采用MTT法检测细胞增殖,流式细胞仪检测细胞增殖周期变化,免疫组化分析细胞神经元分化率,实时定量PCR技术检测Bcyclin D1及CDK2基因表达。结果与结论:①平行排列组、交错排列组培养1,3,5,7,9 d的细胞增殖吸光度值均高于对照组(P<0.05),平行排列组培养3,5,7,9 d的细胞增殖吸光度值高于交错排列组(P<0.05);②平行排列组、交错排列组增殖期细胞比率高于对照组(P<0.05),并且平行排列组高于交错排列组(P<0.05);③免疫组化显示,3组细胞的神经元分化率比较差异无显著性意义(P>0.05);④实时定量PCR技术检测显示,平行排列组、交错排列组Bcyclin D1及CDK2基因表达量高于对照组(P<0.05),并且平行排列组高于交错排列组(P<0.05);⑤结果表明,平行及交错排列纳米纤维膜可促进神经干细胞的增殖,对细胞分化无明显影响,能够从基因表达水平调节神经干细胞的生物学行为。BACKGROUND:The scaffold made by nanotechnology may have a similar surface structure to basement membrane of natural organism,which can effectively regulate the behavior of seed cells.OBJECTIVE:To observe the effects of parallel and staggered nanofiber membranes on the proliferation and differentiation of neural stem cells.METHODS:The parallel and staggered nanofiber membranes were prepared using electrostatic spinning technology with type I collagen as raw material.Neural stem cells of newborn rats were seeded on the surface of two kinds of nanofiber membranes.Cell culture alone was used as control.Cell proliferation was detected by MTT assay.Cell proliferation cycle was detected by flow cytometry.Cell differentiation rate was detected by immunohistochemistry.The gene expression of Bcyclin D1 aned CDK2 was detected by real-time quantitative polymerase chain reaction.RESULTS AND CONCLUSION:(1)The absorbance values of cell proliferation of parallel arrangement group and staggered arrangement group at 1,3,5,7 and 9 days were higher than that of the control group(P<0.05).The absorbance value of cell proliferation of parallel arrangement group was higher than that of staggered arrangement group at 3,5,7 and 9 days(P<0.05).(2)Cell differentiation rate in the parallel arrangement group and the staggered arrangement group was higher than that in the control group(P<0.05),and that in the parallel arrangement group was higher than that in the staggered arrangement group(P<0.05).(3)Immunohistochemistry showed that there was no significant difference in neuronal differentiation rate among the three groups(P>0.05).(4)Real-time quantitative polymerase chain reaction demonstrated that the expression of Bcyclin D1 and CDK2 was higher in the parallel arrangement group and staggered arrangement group was higher than in the control group(P<0.05);and the expression was higher in the parallel arrangement group than in the staggered arrangement group(P<0.05).(5)The results confirmed that parallel and staggered nano tissue engineering

关 键 词：材料 基因 支架 纳米 干细胞 细胞增殖 分化 神经 

分 类 号：R459.9[医药卫生—治疗学] R-331[医药卫生—临床医学]