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有着巨大的前景。然而，它们进入临床的速度一直很慢，因为仍然很难生产出一致的、批量的、用于临床研究的纳米颗粒。此外，还缺乏快速筛选纳米颗粒的平台。因此，需要改进的制备方法，能够产生尺寸、几何形状和化学计量比均一的高质量纳米颗粒。微流控和芯片实验室技术领域的进步现在为纳米材料生产工艺的实施提供了独特的机会。与传统的间歇反应器相比，它们提供了一系列优势，包括提高了纳米材料特性的可控性和一致性，以及高通量生产。它们还允许一步加载生物活性分子(药物、蛋白质、基因、酶等)，而不受其化学、亲水或疏水性质的影响。此外，微通道内实现的快速混合和层流条件的可预测性可以被用来研究纳米材料的形成动力学。为了利用我们在纳米药物输送和成像系统开发方面的成就，以及我们在制造微流控(MF)设备方面获得的专业知识，我们在用于输送和成像的纳米颗粒方面的研究的首要目标是开发微流体平台，用于“一步”合成和制备高度复杂和可调的多功能纳米系统，同时在同一平台上研究细胞与纳米颗粒的相互作用。在这五年的发现计划中，我们特别关注1)微球辅助壳聚糖基纳米粒的LBL自组装，2)微球辅助合成杂化和高效的脂类纳米粒，3)微球芯片用于合成尺寸可调的多组分聚合物脂质纳米粒和4)微球辅助合成多功能Janus纳米粒，所有这些都用于控制释放和体外成像。为了达到这一目标，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NPs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NPs；-)NPs的物理化学和生物学表征；以及-)作为NPs特性的函数的MF辅助NPs-细胞相互作用的研究以及针对其预期用途的概念验证研究。我们相信，如果继续致力于这一领域的研究，微流控多功能纳米颗粒可以通过高通量生物检测和治疗传递在早期阶段解决疾病监测中的多种普遍问题。

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 NPs-cell 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.