红细胞膜包覆PEI用于基因递送

摘要

随着细胞生物学和材料科学的发展，纳米粒子能够被精确制备和优化，从而得到性能优化的材料。特别是，细胞治疗受到越来越多的关注，因为它们能够在整个循环系统中运输各种物质；它们被装载到纳米粒子的表面或内部。细胞膜伪装纳米粒子是一类新型的仿生纳米粒子，它结合了细胞膜的独特功能和合成纳米材料的多功能性，能够有效传递治疗药物。血细胞包括红细胞、白细胞和血小板已经广泛地用作药物载体，它们具有许多独特的优点，如循环时间长（尤其是红细胞），靶向释放能力（特别是血小板）和天然粘合剂性质（白细胞和血小板）。这些性能使得血细胞修饰递送系统以及基于细胞膜载体优于其他药物递送系统。 本论文利用红细胞制备了红细胞囊泡，并合成了双硫键改性聚乙烯亚胺基因载体，并将红细胞囊泡通过静电相互作用，修饰聚乙烯亚胺载体，得到仿生基因递送系统。通过马尔文粒度仪对其粒径和电位进行了表征，并通过透射电镜对超声处理的红细胞进行了形貌表征，并用荧光标记表征了该基因载体递送系统的基因递送过程。

目录

声明

Chapter 1.Introduction

1.1.Fundamental aspects of gene therapy strategy

1.1.1.Limitations associated with gene delivery system

1.1.2. Strategies to introduce therapeutic cargo

1.2.Syntheticvectors for gene delivery

1.2.1.Cationic lipid based delivery system

1.2.2.Polymeric delivery system

1.3.Cell penetrating peptides (CPPs)

1.3.1. Classificationof CPPs

1.3.2. Mechanismof uptake

1.3.3. Applications of CPPs

1.4.RBC vesicles as disguised therapeutic carriers

1.4.1. Pharmacokinetics, biodistribution, and targets ofRBC-loaded drugs

1.4.2. Drug delivery by modified RBC(mRBC)

1.4.3. Characteristics ofRBCs mediated nanoparticles

Chapter2.Experimental analysis

2.1. Introduction

2.2. Materials and methods

2.2.1. Materials

2.2.2. Synthesis of disulfide polyethylenimine(SS-PEI)

2.2.3. Characterization of polymer

2.2.4. Buffer capacity of cationic polymer

2.2.5. Formation of polymer-pDNA complexes with different N/P ratios

2.2.6. Preparation of RBCs vesicles and RBCs-polyplexes

2.2.7. Physiochemical characterization of cationic polymer, polymer-DNA complexes and RBC-polyplexes nanoparticles

2.2.8. Hemolysis assay

2.2.9. Cytocompatibilityof Complex

2.2.9.Macrophage Uptake

Chapter3. Results andDiscussions

3.1. Preparation of disulfide PEI1.8kDa

3.2. Buffer capacity

3.3. Hydrodynamic size and polydispersity index

3.4. Zeta potential or surface charge

3.5. Morphology of RBC vesicles

3.6. Preparation of the NP/pZNF580/RBCs

3.6.Cytocompatibility of Complex

3.7.Macrophage Uptake

Chapter4. Conclusion andfuture perspectives

4.1. Future perspectives

参考文献

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