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，它们向临床的转化一直很缓慢，因为仍然很难生产出“批对批”一致的纳米颗粒，并且用于临床研究的数量足够。此外，快速筛选纳米颗粒的平台仍然缺乏。因此，有必要改进制备方法，使其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。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过程的实施提供了独特的机会。与传统间歇式反应器相比，它们具有一系列优势，包括改进的可控性和纳米材料特性的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），而不依赖于它们的化学性质、亲水性或疏水性。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用于研究纳米材料的形成动力学。为了利用我们在纳米颗粒药物传递和成像系统的发展方面取得的成就，以及我们在微流体（MF）设备制造方面获得的专业知识，我们在纳米颗粒传递和成像方面的研究的总体目标是开发微流体平台，用于“一步”合成和制备高度复杂和调谐的多功能纳米系统，同时研究细胞-纳米颗粒在同一平台上的相互作用。在这个为期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辅助合成杂交和高效脂质NPs， 3) MF芯片合成可调节大小的多组分聚合物-脂质NPs， 4) MF辅助合成多特征Janus NPs，所有这些都是为了控释放和体外成像。为了实现这一目标，本提案的子目标是：-)分别通过仿真和采用先进的微加工技术，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特性；nf辅助NPs -细胞相互作用作为NPs特性的功能的研究和对其预期用途的概念验证研究。我们相信，如果在这一研究领域持续努力，微流控多功能纳米颗粒可以在高通量生物测定和治疗递送的早期阶段解决疾病监测中的多个普遍问题。

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.