微流控技术合成与筛选多层纳米颗粒用于肿瘤治疗

基于纳米材料的肿瘤治疗方法近些年正在走向临床，对治疗性纳米材料的可控合成和临床前评估筛选方法提出了更高的要求。微流控技术是生产临床批准的纳米颗粒（NPs）的最先进方法之一，可以实现NPs 的精准可控和规模化制备。微流控细胞/器官芯片为药物的快速筛选提供了高效的疾病模型。基于此，本论文设计了多级微流控芯片实现高度可控的多层脂质纳米颗粒的一步法合成，并高效包封小干扰RNA（siRNA）和小分子药物索拉菲尼（SFN）用于肝细胞癌（HCC）的治疗；基于微流控技术构建了人工肿瘤-血管模型，实现模拟血流动态环境对纳米颗粒进行筛选。

所设计的微流控芯片有三个单元，可以容纳快过程和慢过程的组装方式。第一单元用于正电荷SFN 组装体形成的快速过程（~70 cm/s 流速）；第二单元用于siRNA 电荷吸附的慢速过程（~5 cm/s 流速）；第三阶段将脂质分子对纳米复合物进行封装，产生多层LNP（mLNPs）。mLNPs 具有明确的三层壳核结构，对 siRNA/SFN 的包封效率接近100%。通过优化得到了一种具有低毒性、良好细胞摄取行为和较高体内肿瘤靶向和转染效率的siRNA/SFN_mLNPs，并负载缺氧诱导因子1α siRNA（HIF1α-siRNA）用于HCC 的治疗。HIF1α-siRNA/SFN_mLNPs（HS_mLNPs）具有优异的体外抗肿瘤效果和基因沉默效率。体内实验结果表明，HS_mLNPs 对Hep3B-HCC皮下瘤和原位瘤小鼠具有很好的体内抑瘤效果。

构建的人工肿瘤-血管模型是以仿生纳米纤维膜为支架，由单层内皮细胞和多层肿瘤细胞形成三维管状结构，模拟有血液供应的肿瘤组织。我们将该模型整合至由蠕动泵和硅胶管组成的体外循环装置中。借助微流控芯片合成了不同粒径和电位的脂质-聚合物壳核纳米颗粒，使用该模型对这些颗粒进行了细胞摄取行为的评估。

本文从芯片设计、多层纳米颗粒的可控制备和筛选模型在肿瘤治疗方面的应用做了系列研究。本文的研究成果在未来或许为纳米颗粒的设计及其微流控合成和筛选方法提供重要依据，最终促进纳米颗粒的临床转化。

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