新型合成水凝胶神经支架材料机械性与体外生物相容性研究  

Mechanical properties and in vitro biocompatibility of novel synthetic hydrogel nerve scaffold materials

作　　者：诸葛彩英 张成业 周岩 Zhuge Caiying;Zhang Chengye;Zhou Yan(Graduate School of Hebei North University,Zhangjiakou 075031,China;Department of Neurosurgery,the Air Force Medical Center of PLA,Beijing 100048C,hina)

机构地区：[1]河北北方学院研究生院,张家口075031 [2]空军特色医学中心神经外科,北京100048

出　　处：《中华实验外科杂志》2024年第9期1980-1983,共4页Chinese Journal of Experimental Surgery

基　　金：空军军医大学军事医学“珠峰工程”(2019ZFB011)。

摘　　要：目的制备用于周围神经损伤修复的新型合成水凝胶支架材料并评估机械性能及其体外生物相容性.方法利用聚二甲基硅氧烷(PDMS)制备新型合成水凝胶,万能试验机检测其机械性能,X线能谱仪分析材料元素组成,电镜观察其微观形貌.培养PC12细胞(PC12,中国科学细胞库)并分为对照组(常规培养)和实验组(新型合成水凝胶),细胞计数试剂盒(CCK-8)、细胞免疫荧光、划痕实验、细胞凋亡实验明确新型合成水凝胶对PC12细胞增殖、迁移及凋亡的作用.蛋白质印迹法(Western blot)检测E-钙黏蛋白(E-cadherin)、Snail、波形蛋白(Vimentin)、半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-8、Caspase-3表达量,明确新型合成水凝胶对细胞的黏附作用影响.两组间比较采用非配对t检验.结果新型合成水凝胶具有良好的拉伸弹性,由初始长度2 cm最大可拉伸至16 cm;元素分析显示材料主要由碳、氧、硅组成;扫描电镜可见新型合成水凝胶具有致密均一网状结构.细胞增殖实验结果显示,PC12细胞对照组吸光度值为1.390±0.154,新型合成水凝胶组为1.093±0.152,差异无统计学意义(t=2.372,P>0.05).细胞免疫荧光实验结果显示,PC12细胞对照组成熟度为(68.712±2.877)%,新型合成水凝胶组为(74.555±2.412)%,差异无统计学意义(t=2.289,P>0.05).划痕实验结果显示PC12细胞对照组划痕愈合面积比为(66.576±3.958)%,新型合成水凝胶组为(64.091±2.635)%,差异无统计学意义(t=1.139,P>0.05).细胞凋亡实验结果显示PC12细胞的凋亡率对照组为(10.360±1.807)%,新型合成水凝胶组为(11.580±1.311)%,差异无统计学意义(t=0.947,P>0.05).Western blot结果显示,E-cadherin、Snail、Vimentin、Caspase-8、Caspase-3相对表达量与新型合成水凝胶组分别为0.698±0.040比0.785±0.036、0.646±0.050比0.752±0.053、0.759±0.051比0.844±0.051、0.772±0.059比0.836±0.045、0.822±0.024比0.822±0.024,差异无统计学意义(t=2.744、2.539、2.Objective To prepare novel synthetic hydrogel scaffold materials for peripheral nerve injury repair and evaluate mechanical properties and their in vitro biocompatibility.Methods The novel synthetic hydrogel was prepared by polydimethylsililoxane(polydimethylsililoxane,PDMS),its mechanical properties were detected by universal testing machine,the element compositions of the material was ana-lyzed by X-ray energy spectrometer,and the micromorphology was observed by electron microscope.PC 12 cells were cultured and divided into control group(conventional culture)and experimental group(novel synthetic hydrogel).The cell counting kit-8 assay,cellular immunofluorescence,scratch migration assay and apoptosis assay were used to detect the effects of the novel synthetic hydrogel on the proliferation,mi-gration and apoptosis.The relative expression levels of E-cadherin,Snail,Vimentin,cysteinyl aspartate-specific protease(Caspase)-8,and Caspase-3 were detected by Western blotting to determine the adhesion effects of the novel synthetic hydrogel on cells.The unpaired student's t-test was used to compare two groups.Results The novel synthetic hydrogel had good tensile elasticity,from the initial length 2 cm to 16 cm.Elemental analysis showed that the material was mainly composed of carbon,oxygen and silicon.It had dense uniform mesh structure.The results of cell counting kit-8 assay showed that the absorbance value of PC12 cells in the control group(1.390±0.154)was higher than the novel synthetic hydrogel group(1.093±0.152,t=2.372,P>0.05).Cellular immunofluorescence results showed that the maturity of PC12 cells in the control group[(68.712±2.877)%]was lower than the novel synthetic hydrogel group[(74.555±2.412)%,t=2.289,P>0.05].The scratch experiment results showed that the scratch healing area ratio of PC12 cells in the control group[(66.576±3.958)%]was higher than the novel syn-thetic hydrogel group[(64.091±2.635)%,t=1.139,P>0.05].The apoptosis rate of PC 12 cells in the control group[(10.360±1.807)%]was lower than

关 键 词：周围神经损伤 神经再生 神经支架 力水凝胶 

分 类 号：R318.08[医药卫生—生物医学工程]