论文总字数：23139字

摘 要

Abstract 4

第一章：绪论 6

1引言 6

1.1选题背景及意义 6

1.2磁性纳米材料 7

1.2.1磁性纳米材料概述 7

1.2.2磁性纳米材料的界面特性 7

1.2.3磁性纳米材料的制备方法 8

1.2.4磁性纳米材料自组装 8

1.3纳米材料对细胞生长分化的作用 9

1.3.1纳米材料对干细胞生长的作用 9

1.3.2纳米材料对成骨细胞生长的作用 9

1.4成骨细胞培养及其微环境 9

1.5检测细胞粘附特性的方法概述 10

1.5.1电镜检测 10

1.5.2 Western Blotting 10

1.5.3 实时荧光QPCR即实时荧光定量核酸扩增检测系统 10

1.5.4 细胞粘附相关基因研究进展 10

1.6本实验的主要内容及目标 11

第二章 磁性纳米裸γ-Fe2O3材料的制备和表征 11

2．1实验原理 11

2.2实验试剂以及器材 11

2.3实验前的准备 12

2.4 裸γ-Fe2O3 纳米颗粒的纯化 12

2.5磁性纳米颗粒组装体的构建与表征 12

2.5.1自然干燥 12

2.5.2磁场下组装 12

2.5.3磁性纳米颗粒组装体的表征 13

第三章 磁性材料纳米颗粒组装体对小鼠成骨细胞分化影响研究 14

3.1小鼠成骨细胞的提取及分化时接种数目的确定 14

3.1.1实验试剂与材料 14

3.1.2成骨细胞的提取 15

3.2 磁性纳米材料上分组培养成骨细胞 16

3.2.1 24孔细胞培养板上分组情况 16

3.2.2 细胞在磁性纳米材料上分化培养 17

3.3磁性微纳基底对小鼠成骨细胞的影响 17

3.3.1 光镜和电子显微镜下观察 17

3.3.2 碱性磷酸酶染色鉴定成骨细胞分化 17

3.3.3 CCK8法检测细胞活力 18

3.3.4定量PCR检测细胞分化活力................................................................................18

第四章 小结与展望 19

参考文献： 19

致谢 20

磁性纳米基底对成骨细胞粘附作用调控机制研究

41112107 李婵娟 指导老师：刘璇

摘要

据统计数据表明，全球每年用于骨科方面的医疗费用高达 170 亿美元。许多骨科类的缺损不能自行修复，需要填充各种骨修复材料作为细胞的支撑。因此骨修复材料在全球范围内成为再生与修复医学研究的热点。所以去了解骨髓细胞的生长究竟受哪些因素影响，还有哪些因素可以促进骨细胞的生长，都有助于我们探索出利于骨修复的材料。

磁性纳米材料由于本身比表面积大，易于表面修饰等特性，在生物学领域备受关注。前人研究表明，磁场的存在下骨能够生长得更好，这说明磁对骨生长有促进作用。磁性纳米材料对生物体的作用在临床治疗疾病上已经成为热点，特别是在骨缺损，骨增生等方面的研究有极大的研究价值。

本次实验主要通过控制变量方法研究磁性纳米材料对成骨细胞的影响。主要是控制浓度不变，磁场增强来研究这种机制到底如何。并采取检测以下检测指标来探究这种作用机制，包括：光镜和扫描电镜检测细胞生长形态的变化，碱性磷酸酶染色，cck8检测细胞活力以及qPCR检测与细胞粘附相关的基因等方法。提取小鼠骨髓细胞并向成骨细胞诱导分化，通过比较培养于纳米材料基底的细胞的分化情况，发现磁性材料组细胞集落更多，而且碱性磷酸酶染色也更明显，说明组装材料组分化的更好，cck8检测细胞活力也显示磁性材料组活力更好，说明磁性作用的材料有更好的生物相容性。进一步检测细胞粘附相关基因情况，证明磁性纳米材料对骨细胞的分化的促进作用。主要从细胞粘附的角度探究磁性纳米基底对细胞分化的影响。从而更好地解释磁性纳米材料对细胞分化的作用机制，加快人类早日研究出骨修复材料的进程。

关键词：磁性纳米材料；成骨细胞；细胞粘附

Study on regulatory mechanism of magnetic nano substrate on osteoblast adhesion

41112107 Chanjuan Li Tutor:Xuan Liu

Abstract

According to the statistics show that each year for the global orthopedic medical expenses up to $ 17 billion.Many orthopedic defect class can not repair itself, we need to fill a variety of bone repair material as a cell support.Therefore, bone repair material on a global scale to become a regeneration and repair of medical research hotspot.Therefore,make a thorough inquiry of What factors affect the growth of osteoblasts,and What factors promote the growth of osteoblasts.bone repair.

Magnetic nano-materials because of their large specific surface area. Ease of surface modification.Thus, in the field of biology it is of concern.Previous studies have shown that the presence of a magnetic field under the bone can grow better, indicating that the magnetic bone growth can promote.The magnetic effect of nanomaterials on organisms in the clinical treatment of the disease has become a hot spot, especially with regard to bone defects, osteoarthritis and other studies have great research value.

In the course of this biological experiments， mainly studying the magnetic properties of nanomaterials on osteoblasts by controlling the variable method.Mainly to control the concentration of the same, but the enhanced magnetic field strength.Check the following and take detect indicators to explore the mechanism of action,Including: changes in light microscopy and scanning electron microscopy to detect cell growth morphology, alkaline phosphatase staining, cck8 , and qPCR detection of cell viability and cell adhesion genes associated methods.Extracting of bone marrow cells to osteogenic differentiation. By comparing cultured in nanomaterials base differentiation of cells, Group found that magnetic colony more and more obvious alkaline phosphatase staining, indicating assembly materials group better differentiation,cck8 cell viability magnetic group also showed better vitality, described the role of magnetic materials have better biocompatibility.Further testing of cell adhesion situation.Proof magnetic nanomaterials role in promoting the differentiation of bone cells, Explore the impact of the main magnetic nanoparticles basal cell differentiation from the perspective of cell adhesion. In order to better explain the mechanism of action of magnetic nanomaterials on cell differentiation, early humans developed to accelerate bone repair process.

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