i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; FEUP - Faculdade de Engenharia da Universidade do Porto, Rua Dr. Roberto Frias s/n, 4200-465 Porto, Portugal.
i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Rua Alfredo Allen 208, 4200-135 Porto, Portugal; ICBAS - Instituto Ciências Biomédicas Abel Salazar, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
Eur J Pharm Biopharm. 2019 May;138:111-124. doi: 10.1016/j.ejpb.2018.01.014. Epub 2018 Jan 31.
The human immunodeficiency virus (HIV) uses the brain as reservoir, which turns it as a promising target to fight this pathology. Nanoparticles (NPs) of poly(lactic-co-glycolic) acid (PLGA) are potential carriers of anti-HIV drugs to the brain, since most of these antiretrovirals, as efavirenz (EFV), cannot surpass the blood-brain barrier (BBB). Forasmuch as the conventional production methods lack precise control over the final properties of particles, microfluidics emerged as a prospective alternative. This study aimed at developing EFV-loaded PLGA NPs through a conventional and microfluidic method, targeted to the BBB, in order to treat HIV neuropathology. Compared to the conventional method, NPs produced through microfluidics presented reduced size (73 nm versus 133 nm), comparable polydispersity (around 0.090), less negative zeta-potential (-14.1 mV versus -28.0 mV), higher EFV association efficiency (80.7% versus 32.7%) and higher drug loading (10.8% versus 3.2%). The microfluidics-produced NPs also demonstrated a sustained in vitro EFV release (50% released within the first 24 h). NPs functionalization with a transferrin receptor-binding peptide, envisaging BBB targeting, proved to be effective concerning nuclear magnetic resonance analysis (δ = -0.008 ppm; δ = -0.017 ppm). NPs demonstrated to be safe to BBB endothelial and neuron cells (metabolic activity above 70%), as well as non-hemolytic (1-2% of hemolysis, no morphological alterations on erythrocytes). Finally, functionalized nanosystems were able to interact more efficiently with BBB cells, and permeability of EFV associated with NPs through a BBB in vitro model was around 1.3-fold higher than the free drug.
人类免疫缺陷病毒 (HIV) 将大脑作为储存库，使其成为对抗这种疾病的有前途的靶点。聚乳酸-共-羟基乙酸 (PLGA) 的纳米颗粒 (NPs) 是将抗 HIV 药物递送到大脑的潜在载体，因为大多数这些抗逆转录病毒药物，如依非韦伦 (EFV)，无法穿过血脑屏障 (BBB)。由于常规生产方法缺乏对颗粒最终性能的精确控制，因此微流控技术应运而生。本研究旨在通过常规和微流控方法开发靶向 BBB 的 EFV 负载 PLGA NPs，以治疗 HIV 神经病变。与常规方法相比，通过微流控技术制备的 NPs 粒径更小（73nm 对 133nm），多分散性相当（约 0.090），负 zeta 电位更低（-14.1mV 对-28.0mV），EFV 结合效率更高（80.7% 对 32.7%），载药量更高（10.8% 对 3.2%）。微流控法制备的 NPs 还表现出持续的体外 EFV 释放（前 24 小时内释放 50%）。用转铁蛋白受体结合肽对 NPs 进行功能化，预计可靶向 BBB，这在核磁共振分析中得到了证明（δ=-0.008ppm；δ=-0.017ppm）。NP 被证明对 BBB 内皮细胞和神经元细胞是安全的（代谢活性高于 70%），并且非溶血（溶血率为 1-2%，红细胞无形态改变）。最后，功能化的纳米系统能够更有效地与 BBB 细胞相互作用，并且通过体外 BBB 模型与 NPs 结合的 EFV 的通透性比游离药物高约 1.3 倍。
