Neutrophil Response to Tissue Templates in Vitro: Implant Implications

该提案旨在检查中性粒细胞的细胞外陷阱方面的中性粒细胞的反应 (NET)形成（过程称为NETosis）的三维再生模板， 变化但受控的架构和组成。模板将通过空运制作- 阻抗静电纺丝使得所得结构在纤维直径，孔径， 和组成，以创建不同的微环境，以允许每个角色的检查 在调节NET形成和随后的细胞相互作用和对 这个预处理事件。最重要的假设是， 和组合物将调节NETosis的程度， 事件将促进/增强再生并减少植入物失效（即急性血栓形成和 增强的炎症/组织破坏）。更具体地说，目标1将界定以下方面的作用： 调节模板相互作用中性粒细胞NETosis中模板结构和组成，以及 确定该预处理事件的后续体外作用。目标2将确定 体内模板引起中性粒细胞NETosis的程度，从而测试体外结果是否 是相关的，以及与NETosis模板程度相关的结果 在大鼠皮下模型中预处理。预计增加模板纤维 直径/孔径和特定组成将引起最小程度的NETosis， 导致募集分泌细胞因子和MMPs的嗜中性粒细胞和巨噬细胞表型 有利于组织再生最终目标是能够定义模板架构 和能够支持先天免疫细胞系指导的组合物， 通过模板相互作用引发和/或引导的植入后增强的再生， 微环境中的中性粒细胞，以取代或修复受损或患病的组织和器官。

The proposal aims to examine the neutrophil response in terms of neutrophil extracellular trap (NET) formation (process known as NETosis) to three-dimensional regeneration templates with varying yet controlled architecture and composition. Templates will be fabricated by air- impedance electrospinning such that the resulting architectures vary in fiber diameter, pore size, and composition to create diverse microenvironments to allow examination of the role of each variable in regulating the NET formation and subsequent cellular interactions and responses to this preconditioning event. The overriding hypothesis is that an electrospun template architecture and composition will modulate the degree of NETosis such that this neutrophil preconditioning event would promote/enhance regeneration and reduce implant failure (i.e. acute thrombosis and enhanced inflammation/tissue destruction). More specifically, Aim 1 will define the role of template architecture and composition in modulating template-interacting neutrophil NETosis, and determine the subsequent in vitro role of this preconditioning event. Aim 2 will then determine the in vivo template invoked degree of neutrophil NETosis and thus test whether the in vitro results are correlative as well as the outcomes associated with the degree of NETosis-template preconditioning in a rat subcutaneous model. It is anticipated that increasing template fiber diameters/pore sizes and particular compositions will invoke a minimal degree of NETosis, leading to recruited neutrophil and macrophage phenotypes that secrete cytokines and MMPs that favor tissue regeneration. The ultimate goal is to be able to define a template architecture and composition that is capable of supporting an innate immunity cell line-instructed and enhanced regeneration upon implantation as initiated and/or directed by the template-interacting, microenvironment neutrophils to replace or repair damaged or diseased tissues and organs.