聚乳酸电纺纤维表面粗糙度对小鼠骨髓间充质干细胞生长影响的研究

发布时间：2018-04-17 02:15

本文选题：聚乳酸 + 静电纺丝　；参考：《西南交通大学》2012年硕士论文

【摘要】：目前纳米材料表面的形态特征对细胞功能的影响,吸引了越来越多学者的关注。在再生医学和组织工程支架材料研究中的一个基本研究目的是细胞与材料之间的相互作用,其中材料表面的微观结构,尤其是微纳米结构对细胞的生物调控作用是研究的重要内容之一。材料的表面形貌和特殊纳米性质能够影响细胞的行为,如黏附、增殖、迁移和分化等。首先,本论文利用静电纺丝的方法对纳米纤维的表面粗糙度进行调节,通过调整聚合物溶液的浓度来控制纤维上珠子的形貌,其表面特征用扫描电子显微镜(SEM)来表征,利用软件计算珠子的长宽比,用原子力显微镜(AFM)来表征表面粗糙度(△h)。其次,我们在本实验室自提取出生10天左右的SD鼠胫骨骨髓原代细胞,定时更换培养基除去造血细胞,经过培养、传代时严格控制胰酶的量和消化时间,不断纯化得到骨髓间充质干细胞(rat-BMSCs)。然后,通过在体外与材料共培养骨髓间质干细胞,我们研究了纤维表面粗糙度对细胞黏附、铺展、增殖等细胞行为的影响。研究结果显示：静电纺丝纳米纤维上存在的串珠状结构对细胞的生长不会带来任何负面影响,反之,珠子在表面堆积,产生高低不同的凸起,即产生不同的纳米粗糙度,可以调节和促进细胞的行为。因此,通过实验得出具有合适的粗糙度的静电纺纤维,在一定程度上更适合作为组织工程支架。最后,为了进一步研究纤维表面粗糙度对成骨诱导的效应,探究其在骨代替物的作用,我们将它们进行了体外矿化和体内植入实验。实验结果显示：白鼠皮下植入后,炎症反应并不明显,具有合适粗糙度的纤维具有比较显著的体内成骨诱导现象,并且在第6周时,纤维能在体内部分降解。体内实验进一步说明具有合适的粗糙度的静电纺纤维具有较大的作为组织工程支架应用的潜力。
[Abstract]:At present, the effect of surface morphology on cell function of nanomaterials has attracted more and more attention.One of the basic purposes of research in regenerative medicine and tissue engineering scaffold materials is the interaction between cells and materials, in which the microstructure of the surface of materials,In particular, the biological regulation of microstructures on cells is one of the important research contents.The surface morphology and special nanometer properties of the materials can affect cell behavior, such as adhesion, proliferation, migration and differentiation.Firstly, the surface roughness of nanofibers was adjusted by electrostatic spinning. The morphology of beads on the fibers was controlled by adjusting the concentration of polymer solution, and the surface characteristics were characterized by scanning electron microscopy (SEM).The aspect ratio of beads was calculated by software and the surface roughness was characterized by atomic force microscope (AFM).Secondly, we extracted the primary cells from the bone marrow of SD rats born about 10 days in our laboratory, and periodically changed the culture medium to remove hematopoietic cells. After the culture, we strictly controlled the amount of trypsin and digestion time during passage.The bone marrow mesenchymal stem cells (BMSCs) were purified continuously.Then, by co-culture of bone marrow mesenchymal stem cells in vitro, we studied the effects of fiber surface roughness on cell adhesion, spreading and proliferation.The results show that the beaded structure on the electrospun nanofibers does not have any negative effect on cell growth. Instead, beads accumulate on the surface and produce different bulges, that is, different nano-roughness.Can regulate and promote the behavior of cells.Therefore, the electrospun fibers with suitable roughness were found to be more suitable for tissue engineering scaffold.Finally, in order to further study the effect of fiber surface roughness on osteogenesis and its role in bone substitute, we carried out in vitro mineralization and in vivo implantation experiments.The results showed that the inflammatory reaction was not obvious after subcutaneous implantation of white rats, and the fibers with suitable roughness had significant osteogenic induction in vivo, and at the 6th week, the fibers could be partially degraded in vivo.In vivo experiments further show that electrospun fibers with suitable roughness have great potential as scaffolds for tissue engineering.
【学位授予单位】：西南交通大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.08

【引证文献】