Department of Physical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-348 Krakow, Poland.
Jerzy Haber Institute of Catalysis and Surface Chemistry PAS, 30-239 Krakow, Poland,.
Colloids Surf B Biointerfaces. 2017 Nov 1;159:200-210. doi: 10.1016/j.colsurfb.2017.07.053. Epub 2017 Jul 25.
Despite progress in the development of novel pharmacological compounds, their efficacy in the treatment of neuropathologies is not satisfactory. One strategy to achieve safe and efficient brain targeting therapy is to design nanocarriers capable of transporting antipsychotic drugs through the BBB (without affecting the normal functions of the barrier) in a defined part of the brain. Here we investigate the interaction of clozapine-loaded polymeric Nano capsules (CLO-NCs) with hCMEC/D3 (human cerebral microvascular endothelial cells, D3 clone) cells that constitutes an in vitro model of the blood brain barrier (BBB). CLO-NCs (average size of 100nm) were constructed by the technique of sequential adsorption of polyelectrolytes (LbL), using biocompatible polyanion PGA (Poly-l-glutamic acid sodium salt) and polycation PLL (poly L-lysine) on clozapine-loaded nano-emulsion cores. Pegylated external layers were prepared using PGA-g(39)-PEG (PGA grafted by PEG poly(ethylene glycol)). The influence of the physicochemical properties of the CLO-NCs (charge, size, surface modification) on cell viability was determined. Advanced studies of CLO-NCs internalization (including endocytosis and transcytosis experiments) using confocal microscopy, flow cytometry and fluorescence spectroscopy are presented. Our results indicate that among the studied NCs, the pegylated clozapine-loaded NCs were the most protected from their uptake by macrophages, and they were the least toxic to hCMEC/D3 cells. They were also the most efficient in the transcytosis experiment, which serves as an indicator of their ability to cross a model BBB.
尽管新型药理学化合物的发展取得了进展，但它们在治疗神经病理学方面的疗效并不令人满意。实现安全有效的脑靶向治疗的一种策略是设计纳米载体，使抗精神病药物能够通过血脑屏障（不影响屏障的正常功能）在大脑的特定部位进行运输。在这里，我们研究了载氯氮平的聚合物纳米囊（CLO-NC）与 hCMEC/D3（人脑血管内皮细胞，D3 克隆）细胞的相互作用，hCMEC/D3 细胞构成了血脑屏障（BBB）的体外模型。CLO-NC（平均粒径为 100nm）是通过顺序吸附聚电解质（LbL）技术构建的，使用生物相容性聚阴离子 PGA（聚谷氨酸钠盐）和聚阳离子 PLL（聚 L-赖氨酸）作为载氯氮平纳米乳液的核心。使用 PGA-g(39)-PEG（聚乙二醇接枝的 PGA）制备了 PEG 化的外部层。测定了 CLO-NC 的物理化学性质（电荷、大小、表面修饰）对细胞活力的影响。通过共聚焦显微镜、流式细胞术和荧光光谱学对 CLO-NC 的内化（包括内吞和转胞吞作用实验）进行了深入研究。我们的结果表明，在所研究的 NC 中，载 PEG 氯氮平的 NC 最能防止被巨噬细胞摄取，对 hCMEC/D3 细胞的毒性也最小。它们在转胞吞实验中也是最有效的，这是它们穿越模型 BBB 的能力的指标。
