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Title:: Experimental study on the proliferation and adhesion of NIH3T3 cells on graphene / gelatin nanofilm

作者:: 孙岩峰2; 刘伟1; 沈远1; 杨皛宁1; 朱惠敏1; 马金旭1; 周瑾1△; 1.军事医学科学院基础医学研究所前沿交叉学科研究室，北京 100080；2.武警总医院儿科临床肿瘤，北京 100080

Author(s):: SUN Yanfeng2; LIU Wei1; SHEN Yuan1; YANG Xiaoning1; ZHU Huimin1; MA Jinxu1; ZHOU Jin1; 1.Academy of Military Medical,Institute of Basic Medical Sciences,Department of Advanced Interdisciplinary Studies, Beijing 100080,China；2.The armed police general hospital,Pediatric, Clinical Cancer,Beijing 100080

摘要:: 本研究旨在探索氧化石墨烯/明胶（graphene oxide /gelatin）薄膜材料对小鼠胚胎成纤维细胞（ NIH3T3）活性、增殖与粘附能力的影响规律。利用扫描电镜观察材料表面微观结构。活-死细胞染色检测氧化石墨烯/明胶薄膜材料上的 NIH3T3 细胞活性，CCK-8 法进行细胞增殖评价。利用免疫荧光、Westernblot检测 Vimentin 表达，采用实时定量RT-PCR 检测粘附相关基因α-actin、 Col Ⅰ、 ColⅢ、 Vinculin、Vimentin的 mRNA 表达。添加氧化石墨烯后材料表面形成明显的褶皱结构。与对照组相比，实验组的成纤维细胞活性良好，具有更强的增殖能力，且高表达α-actin、 ColⅠ、 ColⅢ、及 Vinculin。氧化石墨烯/明胶薄膜材料促进 NIH3T3 细胞的增殖与粘附。

Abstract:: To explore the effect of Graphene oxide / gelatin（GO/gelatin） on the growth and adhesion of mouse embryonic fibroblasts (NIH3T3).The microstructure of the material was examined by scanning electron microscopy. NIH3T3 cells were inoculated on the graphene/gelatin membrane. Cell proliferation was detected by CCK-8, and the cell viability was observed by live-dead staining. The expression of Vimentin was detected by immunofluorescence and Western blotting，the mRNA expression levels of adhesion-related genes α-actin, ColⅠ, ColⅢ, and Vinculin were detected by real-time quantitative RT-PCR. The results showed that after adding graphene oxide, the surface of the material becam rough, forming a more obvious fold structure. The results of CCK-8 detection showed that the fibroblasts grew on the graphene-enriched material had good activity and had stronger ability to proliferate compared with the gelatin membrane after the third day. NIH3T3 cells on the graphene / gelatin membrane expressed adhesion genes such as α-actin, Col Ⅰ , Col Ⅲ , and Vinculin highly. Oxidized graphene / gelatin material promotes the proliferation and adhesion of NIH3T3 cells