Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 20 N. Pine Street, MD, 21201, Baltimore, USA.
Department of Microbiology and Immunology, University of Maryland School of Medicine, 685 W. Baltimore Street, MD, 21201, Baltimore, USA.
AAPS J. 2021 Dec 2;24(1):6. doi: 10.1208/s12248-021-00645-2.
Nanoparticles (NPs) have emerged as a highly useful and clinically translatable drug delivery platform for vast therapeutic payloads. Through the precise tuning of their physicochemical properties, NPs can be engineered to exhibit controlled drug release properties, enhanced circulation times, improved cellular uptake and targeting, and reduced toxicity profiles. Conventional bulk methods for the production of polymeric NPs suffer from the ability to control their size and polydispersity, batch-to-batch variability, significant preparation times, and low recovery. Here, we describe the development and optimization of a high-throughput microfluidic method to produce cargo-less immunomodulatory nanoparticles (iNPs) and their formulation-dependent anti-inflammatory properties for the modulation of lipopolysaccharide (LPS)-induced macrophage responses. Using poly(lactic acid) (PLA) as the core-forming polymer, a rapid and tunable microfluidic hydrodynamic flow-focusing method was developed and optimized to systematically evaluate the role of polymer and surfactant concentration, surfactant chemistry, and flow rate ratio (FRR) on the formation of iNPs. A set of iNPs with 6 different surface chemistries and 2 FRRs was then prepared to evaluate their inherent anti-inflammatory effects using bone marrow-derived macrophages stimulated with the Toll-like receptor 4 agonist, LPS. Finally, a lyophilization study was performed using various cryoprotectants and combinations to identify preferable conditions for iNP storage. Overall, we demonstrate a highly controlled and reproducible method for the formulation of iNPs using microfluidics and their formulation-dependent inherent anti-inflammatory immunomodulatory properties, which represents a potentially promising strategy for the management of inflammation.
纳米颗粒（NPs）已成为一种非常有用且可临床转化的药物输送平台，可用于输送各种治疗有效载荷。通过精确调整其物理化学性质，可以设计 NPs 以表现出可控的药物释放特性、增强的循环时间、改善的细胞摄取和靶向性以及降低的毒性特征。传统的用于生产聚合物 NPs 的批量方法存在控制其尺寸和多分散性、批间变异性、显著的制备时间和低回收率的能力有限。在这里，我们描述了开发和优化高通量微流控方法来生产无载药免疫调节纳米颗粒（iNPs）及其制剂依赖性抗炎特性，以调节脂多糖（LPS）诱导的巨噬细胞反应。使用聚乳酸（PLA）作为核心形成聚合物，开发并优化了一种快速且可调的微流控流体聚焦方法，以系统地评估聚合物和表面活性剂浓度、表面活性剂化学和流速比（FRR）对 iNPs 形成的作用。然后制备了一组具有 6 种不同表面化学性质和 2 种 FRR 的 iNPs，以使用骨髓来源的巨噬细胞评价它们在 TLR4 激动剂 LPS 刺激下的固有抗炎作用。最后，进行了冻干研究，使用各种冷冻保护剂及其组合来确定 iNP 存储的优选条件。总体而言，我们使用微流控技术展示了一种高度可控且可重复的 iNP 制剂方法及其制剂依赖性固有抗炎免疫调节特性，这代表了一种有希望的炎症管理策略。
