Continuous Flow Microfluidic Devices for High-Throughput Synthesis and Formulation of Multifunctional Nano-systems for Enhanced Drug Targeting and Imaging

纳米粒子用于治疗和成像在再生医学和纳米医学中具有巨大的前景。然而，它们向临床的转化一直很缓慢，因为仍然很难生产出“批次间”一致的纳米颗粒，并且数量足够用于临床研究。此外，仍然缺乏快速筛选纳米颗粒的平台。因此，需要能够产生具有均匀尺寸、几何形状和化学计量的高质量纳米颗粒的改进的制备方法。微流体和芯片实验室技术领域的进步现在为实施纳米材料生产工艺提供了独特的机会。与传统的间歇式反应器相比，它们具有一系列优势，包括改善纳米材料特性的可控性和均匀性以及高通量生产。它们还允许一步装载生物活性分子（药物、蛋白质、基因、酶等），与它们的化学性质、亲水性或疏水性无关。此外，可以利用微通道内实现的快速混合和层流条件的可预测性来研究纳米材料形成动力学。为了利用我们在纳米颗粒药物递送和成像系统开发方面的成就，以及我们在微流体（MF）设备制造方面获得的专业知识，我们在用于递送和成像的纳米颗粒方面的研究的首要目标是开发用于高度复杂和调谐的多功能纳米系统的“一步”合成和制备的微流体平台，同时在同一平台上研究细胞-纳米颗粒相互作用。在这5年的发现计划中，我们特别关注1）MF辅助的基于壳聚糖的NPs的LbL自组装，2）MF辅助的杂合和高效脂质NPs的合成，3）用于合成尺寸可调的多组分聚合物-脂质NPs的MF芯片和4）MF辅助的多特征Janus NPs的合成，所有这些都用于控制释放和体外成像。为了实现这一目标，本提案的子目标是：-）分别通过模拟和使用先进的微制造技术根据特定纳米颗粒所需的特征设计和制造高度创新的3D多层MF平台; -）MF辅助合成上述纳米颗粒; -）纳米颗粒的物理化学和生物学表征;和-）作为NP特征的函数的MF辅助的NPscell相互作用的研究和针对其预期用途的概念验证研究。我们相信，微流体使能的多功能纳米粒子可以解决多种，在疾病监测的早期阶段与高通量生物测定和治疗交付的普遍问题，如果持续努力致力于这一领域的研究。

The use of nanoparticles for therapy and imaging holds tremendous promise in regenerative medicine and nanomedicine. However, their translation into the clinic has been slow because it remains difficult to produce nanoparticles that are consistent 'batch-to-batch' and in sufficient quantities for clinical research. Moreover, platforms for rapid screening of nanoparticles are still lacking. Therefore, there is a need for improved preparation methods that are capable of yielding high quality nanoparticles of uniform size, geometry and stoichiometry. Advancements in the fields of microfluidic and lab-on-a-chip technologies now provide unique opportunities for the implementation of nanomaterial production processes. They offer a range of advantages compared to conventional batch reactors, including improved controllability and uniformity of nanomaterial characteristics and high throughput production. They also allow for one-step loading of biologically active molecules (drugs, proteins, genes, enzymes, etc.), independent of their chemical, hydrophilic or hydrophobic nature. In addition, fast mixing achieved within microchannels and the predictability of laminar flow conditions can be leveraged to investigate nanomaterial formation dynamics. To take advantage of our achievements in the development of nanoparticle drug delivery and imaging systems, and our acquired expertise in the fabrication of microfluidic (MF) devices, the overarching goal of our research in nanoparticle for delivery and imaging is to develop microfluidic platforms for “one-step” synthesis and preparation of highly complex and tuned multifunctional nanosystems while investigating cell-nanoparticle interactions on the same platform. During this 5 year discovery proposal, we particularly focus on 1) MF-assisted LbL self-assembly of chitosan-based NPs, 2) MF-assisted synthesis of hybrid and highly potent lipid NPs, 3) MF chip for synthesis of size-tunable multicomponent polymeric-lipid NPs and 4) MF-assisted synthesis of multi-featured Janus NPs, all for controlled release and in vitro imaging. To reach this goal, the sub-objectives of this proposal are: -) Design and fabrication of highly innovative 3D multilayer MF platform according to the required features for the specific NPs through simulation and using advanced microfabrication technology respectively; -) MF-assisted synthesis of aforementioned NPs; -) Physicochemical and biological characterization of NPs; and -) Investigation of MF-assisted NPscell interactions as a function of NPs characteristics and proof-of-concept study towards their intended use. We believe that microfluidic-enabled multifunctional nanoparticles could resolve multiple, prevalent issues in disease monitoring at an early stage with high-throughput bioassays and therapeutic delivery if persistent effort is devoted to this field of research.