脑靶向纳米载药复合物调控血脑屏障细胞功能的有机电化学晶体管阵列体外分析

The brain-targeting nanocarrier-drug hybrids for the therapy of brain disease, especially the brain tumors, require not only good efficacy but also high biocompatibility. To address the effects of brain-targeting nanocarrier-drug hybrids on the function of blood-brain barrier cells, in this project, by utilizing the advantages of organic electrochemical transistors (OECTs) including sensitive signal transforming/amplifying and good biocompatibility, the OECT arrays with high performances are fabricated by screen-printing technique and optimization of device preparation parameters, and used to measure the in vitro weak signals of blood-brain barrier cells. The brain-targeting nanocarrier-drug hybrids are synthesized by super-molecular assembly, and etc. The micromorphologies, structure, size, and drug loading amount and etc. of the nanohybrids are characterized by kinds of analytical methods. Through the studies on the principle of OECT arrays for measuring the in vitro signals of blood-brain barrier cells, the effects and mechanism of the brain-targeting nanocarrier-drug hybrids on the function of blood-brain barrier cells, it hopes to obtain new understanding and points at the in vitro signal measuring of blood-brain barrier cells by OECT arrays and the adjusting of the cell function by the brain-targeting nanocarrier-drug hybrids, which will provide the theory basis and technique supporting for preparation of the brain-targeting nanocarrier-drug system with high efficacy and biocompatibility. At the same time, it is also hopeful to construct and develop OECT array devices with high performances and the potential for commercial applications.

用于脑部疾病尤其是脑肿瘤治疗的脑靶向纳米载药复合物除了要求高疗效，另一决定因素即生物安全性。针对脑靶向纳米载药复合物如何影响血脑屏障细胞功能的问题，本项目拟利用有机电化学晶体管（OECTs）灵敏的信号转换/放大、生物兼容性好的特点，采用丝网印刷技术、通过器件参数优化获得可用于血脑屏障细胞体外微弱信号检测的高性能OECTs阵列器件；采用超分子组装等方法制备脑靶向纳米载药复合物，利用多种分析测试方法对所制备复合物的微观形貌/结构、尺寸、载药量等进行表征。通过研究OECTs阵列检测血脑屏障细胞信号原理、脑靶向纳米载药复合物对血脑屏障细胞功能的影响及其作用机制，以期在OECTs阵列体外检测血脑屏障细胞信号、脑靶向纳米载药复合物调控血脑屏障细胞功能等方面获得新的理解和观点，为高效、安全的脑靶向纳米载药系统制备提供理论依据和技术支持；同时开发高性能、具有商业应用前景的OECTs阵列器件。

本研究利用丝网印刷、氧等离子体等技术，制备了有机电化学晶体管（OECTs）阵列器件；优化了丝网印刷模板设计/印刷参数、氧等离子体参数、导电聚合物合成/成膜工艺等条件。利用去溶剂法、超分子组装、溶胶-凝胶法等合成了基于人血清白蛋白纳米粒子、聚酰胺-胺、介孔二氧化硅纳米粒子、儿茶素纳米粒子和嵌段聚合物胶束的脑靶向纳米载药复合物。构建了血脑屏障细胞模型，详细研究了OECTs阵列器件检测血脑屏障细胞ROS、酶活性原理。探讨了栅极纳米材料改性、传感界面构建策略等OECTs阵列器件响应调控及传感检测性能影响因素；分析了载药复合物成分、微观形貌/尺寸、表面功能团等对血脑屏障细胞模型完整性/通透性、应激响应及酶活性的影响。结果表明：OECTs阵列器件具有调控灵敏、稳定性好的特点；且制备过程简便、成本较低、适于批量生产。所制备脑靶向纳米载药复合物形貌规则、尺寸可控、载药量高。通过引入聚酰胺-胺或其交联物等可有效减缓药物释放，提高纳米载药复合物生物相容性，减小对血脑屏障细胞模型通透性的影响；在纳米载药复合物中引入脑靶向功能团或活性小分子、调控复合物表面电荷/亲疏水性，可有效提高载药复合物穿过血脑屏障模型效率。利用金属-有机凝胶或有机框架材料、碳纳米管、贵金属纳米粒子、氧化铈纳米粒子等对OECTs阵列栅极进行修饰，在有效提高OECTs传输调控特性的同时，可极大提高栅极的电化学性能和生物亲和性。以此为基础构建的OECTs-Transwell传感检测平台具有高灵敏度、低检测下限、选择性好等特点；实现了纳米材料与活细胞相互作用后ROS释放的实时监测；血脑屏障模型细胞蛋白激酶A活性、血管内皮生长因子释放检测。项目的研究结果对于理解脑靶向纳米载药复合物调控血脑屏障细胞功能、OECTs阵列器件体外检测血脑屏障细胞信号等方面具有重要理论意义；在开发高效、安全脑靶向纳米载药系统，低成本、高性能OECTs阵列传感器方面具有实际应用前景。