摘要
目的制备不同浓度的氧化石墨烯(GO)/聚己内酯(PCL)复合基质纳米材料,观察其在体外对大鼠棕色脂肪干细胞(BASC)成心肌分化的影响。方法利用静电纺丝技术制备GO/PCL复合纳米纤维材料。通过将BASC接种于纯PCL和GO/PCL复合纳米纤维上培养后进行CCK-8法测试、对纳米材料的扫描电镜(SEM)检测及电导率测定,表征GO/PCL复合纳米材料的生物相容性及性状特征。采用细胞学、免疫荧光染色等方法,检测心肌细胞相关蛋白的表达情况,系统评价GO/PCL复合材料对BASC活力、增殖及成心肌分化效率的影响。结果该GO/PCL复合基质纳米材料在GO浓度为0.1 mg/ml时无明显细胞毒性。蛋白荧光统计结果表明,与PCL组相比,GO/PCL组细胞的心肌细胞特异蛋白α-辅肌动蛋白(α-actinin)及闰盘相关蛋白缝隙连接蛋白-43(connexin-43,CX-43)表达水平显著提高(P<0.05)。细胞骨架染色结果表明,与PCL组相比,GO/PCL组的细胞表现出类似天然心肌细胞束的长梭样伸展,且生长方向具有一定的极性。结论本研究成功制备了GO/PCL复合纳米材料。该GO/PCL复合纳米材料可促进BASC向心肌细胞分化,并促进闰盘相关蛋白的表达。
Objective To prepare graphene oxide(GO)/polycaprolactone(PCL)composite matrix nanomaterials with different concentrations,and investigate effects of the nanomaterials on the myocardial differentiation of rat brown adipose stem cells(BASC)in vitro.Methods The GO/PCL composite nanofiber materials were prepared by electrospinning technology.The biocompatibility of the nanomaterials was tested by the CCK-8 assay after cultivation of the BASC on the pure PCL and GO/PCL composite nanofibers,and the characterization of the nanofiber materials was performed by the scanning electron microscopy(SEM)and electrical conductivity measurement.The expression of cardiomyocyte-related proteins was examined by the cytological method and immunofluorescence staining.The GO/PCL composite nanofiber materials were systematically evaluated for their effect on the viability,proliferation and myocardial differentiation of BASC.Results The GO/PCL composite nanofiber materials showed no obvious cytotoxicity at the 0.1 mg/ml GO concentration.The statistical results for the protein fluorescence intensities showed that the expression of the cardiomyocyte specific protein,α-actinin,and the intercalated disc-related protein,connexin-43(CX-43),was significantly increased in the GO/PCL group than in the PCL group(P<0.05).The cytoskeletal staining results showed that,compared with the PCL group,the cells in the GO/PCL group showed a long spindle-like stretch similar to the natural myocardial cell bundle,and the growth direction had a certain polarity.Conclusion This study successfully prepared GO/PCL composite nanomaterials,which could promote the differentiation of BASCs into cardiomyocytes and the expression of intercalated disc-related proteins.
作者
张晓静
李屹
师晓丽
朱琳
张蛟
刘惠亮
ZHANG Xiao-jing;LI Yi;SHI Xiao-li;ZHU Lin;ZHANG Jiao;LIU Hui-liang(Clinical College of the Third Medical Center,PLA General Hospital,Anhui Medical University,Beijing 100039,China;Department of Cardiology,the Third Medical Center,PLA General Hospital,Beijing 100039,China;National Nano Science Center,Beijing 100190,China;Department of Cardiology,Beijing Electric Power Hospital,State Grid Corporation of China,Beijing 100073,China)
出处
《国际药学研究杂志》
CAS
北大核心
2020年第4期283-289,共7页
Journal of International Pharmaceutical Research
基金
国家自然科学基金资助项目(81700276)。
关键词
氧化石墨烯
聚己内酯
纳米纺丝纤维
棕色脂肪干细胞
心肌分化
graphene oxide
polycaprolactone
nanospun fibers
brown adipose stem cells
myocardial differentiation
