Nanoparticle Remodeling of Pulmonary Junctions

 描述(申请人提供)：设计的纳米颗粒正在走向临床，用作药物输送、成像和治疗剂。研究了纳米颗粒与细胞相互作用的副作用(例如，毒性)，但测量的结果通常是迟钝的(例如，活/死分析)，对纳米颗粒化学和引发炎症和组织损伤的机制几乎没有洞察力。此外，许多体外测试是基于急性纳米颗粒暴露于细胞单一培养，而对组织生理学的影响可能只有在长期暴露后才会出现。另一种选择是使用动物模型，但这类研究的结果并没有提供足够的机械洞察力来证明广泛的动物试验的费用和伦理挑战是合理的。日益复杂的细胞培养实验可以更好地预测体内对试剂的反应。纳米颗粒最有可能进入人体的地方是肺部。除了使用原代细胞培养，在标准细胞条件下增加机械拉伸、空气-水界面和液体流动更能更好地接近肺内的生理环境。此外，基于蛋白质的荧光记者的动态成像功能使人们能够实时监测信号和原位改变的组织表型，而不需要固定。这项工作的目标是结合这些进展来测量肺细胞阵列对一套定义明确的大小、形状和表面化学的纳米颗粒在急性和慢性时间尺度上的机械拉伸和流动条件下的反应。疾病进展(炎症)将在芯片上监测，重点是炎症信号，包括活性氧和细胞因子的产生，细胞骨架重构，以及受损的细胞间屏障完整性。纳米粒子的表面化学将被修饰为抗生物污垢，以验证纳米粒子的表面化学修饰可以减轻不利影响的假设。总体而言，这种方法可能会导致更现实的芯片上器官模型，而不需要使用动物来预测新出现的污染物影响。

 DESCRIPTION (provided by applicant): Engineered nanoparticles are moving toward clinical use as drug delivery, imaging and therapeutic agents. Side effects of nanoparticle interactions with cells, and occasionally organisms, are studied (e.g., toxicity) but the measured outputs are usually blunt (e.g., live/dead assays) and provide little insight into the nanoparticle chemistry and mechanisms that trigger inflammation and tissue damage. Moreover, many in vitro tests are based on acute nanoparticle exposure to cell monocultures, whereas effects on tissue physiology may only appear after chronic exposure. An alternative is to use animal models, but the outcomes of such studies have not provided sufficient mechanistic insight to justify the expense and ethical challenges of extensive animal testing. Increasingly sophisticated cell culture experiments allow for better prediction of in vivo responses to agents. The most likely point of entry of nanoparticulates to humans is the lung. Beyond the use of primary cell cultures, adding mechanical stretch, an air-water interface, and fluid flow to standard cell conditions far better approximates the physiological environment within the lung. In addition, dynamic imaging capabilities with protein-based fluorescent reporters enable the real time monitoring of signaling and altered tissue phenotype in situ, without the need for fixation. The goal of this work is to combine these advances to measure the response of pulmonary cell arrays to a suite of nanoparticles of well-defined size, shape, and surface chemistry upon mechanical stretch and flow conditions for both acute and chronic time scales. Disease progression (inflammation) will be monitored on chip, by focusing on inflammatory signatures including reactive oxygen species and cytokine production, cytoskeletal remodeling, and compromised intercellular barrier integrity. The surface chemistry of the nanoparticles will be modified to be anti-biofouling, to tet the hypothesis that surface chemical modification of nanoparticles can mitigate adverse impacts. Overall, this methodology might lead to more realistic organ-on-a-chip models without the use of animals to predict emerging contaminant impact.