Nanoparticle Remodeling of Pulmonary Junctions

 描述（由申请人提供）：工程化纳米颗粒正朝着临床应用的药物输送，成像和治疗剂。研究了纳米颗粒与细胞以及偶尔与生物体相互作用的副作用（例如，毒性）但测量的输出通常是钝的（例如，活/死测定），并且对引发炎症和组织损伤的纳米颗粒化学和机制提供很少的了解。此外，许多体外试验是基于急性纳米颗粒暴露于细胞单一培养物，而对组织生理学的影响可能只出现在慢性暴露后。另一种方法是使用动物模型，但这些研究的结果并没有提供足够的机制见解来证明广泛的动物试验的费用和伦理挑战是合理的。越来越复杂的细胞培养实验允许更好地预测体内对药剂的反应。纳米颗粒最有可能进入人体的部位是肺部。除了使用原代细胞培养物之外，将机械拉伸、空气-水界面和流体流动添加到标准细胞条件更好地近似肺内的生理环境。此外，基于蛋白质的荧光报告分子的动态成像能力使得能够原位真实的实时监测信号传导和改变的组织表型，而不需要固定。这项工作的目标是联合收割机这些进步来测量肺细胞阵列的响应，一套明确的尺寸，形状和表面化学的纳米粒子后，机械拉伸和流动条件下的急性和慢性的时间尺度。将在芯片上监测疾病进展（炎症），重点关注炎症特征，包括活性氧和细胞因子产生、细胞骨架重塑和受损的细胞间屏障完整性。纳米颗粒的表面化学将被改性以抗生物污染，以泰特纳米颗粒的表面化学改性可以减轻不利影响的假设。总的来说，这种方法可能会导致更现实的器官芯片模型，而不使用动物来预测新出现的污染物的影响。

 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.