Chiesa Enrica, Riva Federica, Dorati Rossella, Greco Antonietta, Ricci Stefania, Pisani Silvia, Patrini Maddalena, Modena Tiziana, Conti Bice, Genta Ida
Department Drug Sciences, University of Pavia, V.le Taramelli 12, 27100 Pavia, Italy.
Department of Public Health, Experimental and Forensic Medicine, Histology and Embryology Unit, University of Pavia, Via Forlanini 10, 27100 Pavia, Italy.
Pharmaceutics. 2020 Mar 13;12(3):260. doi: 10.3390/pharmaceutics12030260.
In this study, an innovative microfluidics-based method was developed for one-step synthesis of hyaluronic acid (HA)-based nanoparticles (NPs), by exploiting polyelectrolytic interactions between HA and chitosan (CS), in order to improve reliability, reproducibility and possible scale-up of the NPs preparation. The on-chip synthesis, using a staggered herringbone micromixer, allowed to produce HA/CS NPs with tailored-made size and suitable for both parenteral (117.50 ± 4.51 nm) and loco-regional (349.15 ± 38.09 nm) administration, mainly composed by HA (more than 85% wt) with high negative surface charge (< -20 mV). HA/CS NPs were successfully loaded with a challenging water-insoluble molecule, Everolimus (EVE), an FDA- and EMA-approved anticancer drug able to lead to cell cycle arrest, reduced angiogenesis and promotion of apoptosis. HA/CS NPs resulted to be massively internalized in CD44+ human mesenchymal stem cells via CD44 receptor-mediated endocytosis. HA/CS NPs selectiveness towards CD44 was highlighted by blocking CD44 receptor by anti-CD44 primary antibody and by comparison to CS-based NPs cellular uptake. Eventually, high effectiveness in inhibiting cell proliferation was demonstrated on-chip synthetized EVE loaded HA/CS NPs by tracking in vitro DNA synthesis.
在本研究中，开发了一种基于微流控技术的创新方法，通过利用透明质酸（HA）与壳聚糖（CS）之间的聚电解质相互作用，一步合成基于HA的纳米颗粒（NPs），以提高NPs制备的可靠性、可重复性和可能的放大规模。使用交错人字形微混合器的芯片上合成能够制备尺寸定制的HA/CS NPs，适用于肠胃外给药（117.50±4.51 nm）和局部区域给药（349.15±38.09 nm），主要由具有高负表面电荷（< -20 mV）的HA组成（重量超过85%）。HA/CS NPs成功负载了一种具有挑战性的水不溶性分子依维莫司（EVE），这是一种经美国食品药品监督管理局（FDA）和欧洲药品管理局（EMA）批准的抗癌药物，能够导致细胞周期停滞、减少血管生成并促进细胞凋亡。HA/CS NPs通过CD44受体介导的内吞作用大量内化于CD44+人间充质干细胞中。通过用抗CD44一抗阻断CD44受体并与基于CS的NPs细胞摄取进行比较，突出了HA/CS NPs对CD44的选择性。最终，通过跟踪体外DNA合成，证明了芯片上合成的负载EVE的HA/CS NPs在抑制细胞增殖方面具有高效性。
