二氧化硅纳米颗粒／胶原蛋白神经支架体外生物学特性研究  

A biology research of collagen/silica composite nerve scaffolds in vitro

作　　者：景尚斐[1] 温树正[1] 蒋电明[3] 郭文[1] 许鹏成[1] 王继宏[1] 冀云涛[1] 杨晨瑗[2] 

机构地区：[1]内蒙古医科大学第二附属医院手足显微Ⅱ科,呼和浩特010030 [2]内蒙古医科大学附属人民医院影像科 [3]重庆医科大学第一临床医院骨科

出　　处：《中华显微外科杂志》2016年第5期465-469,共5页Chinese Journal of Microsurgery

基　　金：内蒙古自治区卫生和计划生育委员会医疗卫生科研计划项目基金（201303079）：内蒙古医科大学第二附属医院基金（2013YJJ20）

摘　　要：目的探讨不同配比SiO2纳米颗粒/胶原蛋白复合材料作为神经支架的生物学特性。方法2013年3月至2014年5月，采用冻干技术制作浓度分别为5、10、25、50、100和200 μg/ml SiO2纳米颗粒浓度的胶原蛋白神经支架，不含SiO2纳米颗粒的胶原支架作为本研究的对照组。动态光散射仪测量SiO2纳米颗粒的直径和大小，SEM观察纳米颗粒的形态。支架内接种培养许旺细胞（SCs），检测支架的细胞相容性，光镜下观察不同样本中SCs形态，MTT检测SCs在SiO2纳米颗粒/胶原复合支架的附着和增殖效果，荧光分光光度计测定支架内细胞经H33258染色后的DNA含量。上述实验数据经多重比较检验法后进行单向方差分析，P 〈 0.05为差异有统计学意义。结果SiO2纳米颗粒呈球形，平均直径分布在150~190 nm，SCs在支架上培养了1 d和3 d后，所有培养的细胞中（除了100 μg/ml支架中生长的细胞）都处于正常状态，其他样品的所有细胞中都呈纺锤形并伴随轴突延伸，细胞没有表现出明显的形态学差异。MTT细胞相容性检测显示，SCs的活性和增殖度先升高，后随着SiO2颗粒浓度的增加而降低，SCs的活性和增殖度在25 μg/ml中表现最高，而当浓度大于50 μg/ml时明显降低，这与细胞形态变化的相符。DNA含量起初增加后随着浓度的增加而降低，25 μg/ml时DNA含量最多（P 〈 0.05），DNA含量与细胞形态变化相符。结论25 μg/ml的SiO2纳米颗粒/胶原蛋白支架有可能成为一种新的周围神经缺损修复材料。Objective To discuss the effect of collagen/silicon composite scaffolds in the peripheral nerve regeneration, and evaluate the adhesion and proliferation of Schwann cells within the scaffolds. Methods In this study, from March, 2013 to May, 2014, the collagen/silicon composite scaffolds with 5, 10, 25, 50 and 100 μg/ml were made by freeze-drying technology, and the single collagen scaffold without silica nanopartieles was made with the same method as a control group. The diameter and size of the silica nanoparticles were measured by light scattering, and the morphology of silica nanoparticles were scanned by electron microscope. The cellular compatibility of composite scaffolds was inspected by cultivating Schwann cells on the scaffolds in vitro. The morphology of the Schwann cells in different sample were observed under optical microscope. The adhesion and proliferation effect of the Schwann cells on the scaffolds were determined by MTT. The DNA content of the cells were measured by fluorescence spectrophotometer. Experimental data were expressed by mean ± standard deviation （SD） values. Data were analyzed using One-way ANOVA followed by Bonferroni＇s post hoc comparison tests. Statistics with a value of P 〈 0.05 indicated statistical significance. Results The average diameter of the nanoparticles mainly distributed from 150 to 190 nm and showed a spheric morphology. Morphologically, all the cultured cells were observed to he in healthy state in most scaffolds except for the scaffolds with 100 μg/ml silica nanoparticles. Whereas all the cells on other samples displayed spindle shape with axons spreading, and there was no obvious morphology difference for the cells on these samples. The results of cytocompatibility of the collagen/silica composite scaffolds using MTT assay showed that both the viability and proliferation of Schwann cells increased firstly and then decreased with the increasing concentration of silica nanoparticles. And the cells showed much better viability and proliferation when the co

关 键 词：二氧化硅纳米颗粒 胶原蛋白 神经支架 许旺细胞 

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