Designing Drug-eluting Regeneration Templates to Down-regulate Neutrophil Extracellular Traps

该提案旨在审查N-α-benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine的能力 电纺模板洗脱对中性粒细胞胞外水平的调节作用 陷阱(Net)，已被证明是在急性相互作用的中性粒细胞中诱导的。这个 模板将被电纺，从而得到的结构在纤维直径、孔 大小、载药量和表面化学，以创造多样化的微环境，从而 考察每个变量在调节网络形成和后续寄主中的作用 回应。压倒一切的假设是NETsis的程度(或模板的水平 预调节网络)可通过包括氯-嘧啶浸渍和/或 吸附在电纺纤维上，洗脱出的药物起到最小化净作用 形成所需的加速边缘组织愈合和强化组织 模板的集成。更具体地说，目标1将定义变化的表面的作用- 纤维的面积体积比、降解率、表面化学性质和氯化亚胺载量 在电纺和/或模板后处理表面吸附期间浸渍， 表征体外释放谱，并最终确定体外抑制功能 网纹病。然后，AIM 2将确定负载氯-酰胺的PDO模板的能力，以降低- 调节体内网织红细胞增多症的程度并决定组织愈合/整合结果 在一个大鼠皮下模型中，与氯嘧啶下调NETsis的程度有关。 预计增加模板纤维直径/孔大小和改变表面化学 具有显著的突发性药物释放能力将引发最小程度的Net，有利于组织 再生。我们预计这项研究将开始在中性粒细胞方面取得新的进展。 模板相互作用，以及更重要的是，为额外的模板派生提供证据 在下调慢性炎症的潜在起始物方面的功能性(氯-嘧啶洗脱) 以及受损的组织愈合。此功能可能是一个关键的缺失组件，它可能 为组织工程师提供了一种更有效地利用先天免疫反应的方法 (急性模板预适应)将导致增强原位再生，或更多 准确地说，将慢性炎症和纤维化导致的失败降至最低。

The proposal aims to examine the ability of N-α-benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine amide (Cl-amidine) eluting electrospun templates in regulating the level of neutrophil extracellular traps (NETs) which have been shown to be induced in acute interacting neutrophils. The templates will be electrospun such that the resulting architectures vary in fiber diameter, pore size, drug loading, and surface chemistry to create diverse microenvironments to allow examination of the role of each variable in regulating the NET formation and subsequent host response. The overriding hypothesis is that the degree of NETosis (or the level of template preconditioning NETs) can be down regulated by the inclusion of Cl-amidine impregnation and/or adsorption onto the electrospun fibers and that the eluted drug functioning to minimize NET formation will lead to a desired, accelerated marginal tissue healing and enhanced tissue integration of the templates. More specifically, Aim 1 will define the role of the varied surface- area-to-volume ratios, degradation rates, surface chemistries, and Cl-amidine loading by fiber impregnation during electrospinning and/or template post-processing surface adsorption, characterize the in vitro release profiles, and finally determine the in vitro functionality in inhibiting NETosis. Aim 2 will then determine the Cl-amidine loaded PDO templates capacity to down- regulate the degrees of NETosis in vivo and determine the tissue healing/integration outcomes associated with the Cl-amidine down-regulated degree of NETosis in a rat subcutaneous model. It is anticipated that increasing template fiber diameters/pore sizes and altering surface chemistry with a significant burst drug release capacity will invoke a minimal degree of NETs, favoring tissue regeneration. We anticipate that this study will begin to break new ground regarding neutrophil- template interaction, and more critically, provide evidence for additional template-derived functionality (Cl-amidine elution) in down-regulating a potential initiator of chronic inflammation and impaired tissue healing. This functionality is potentially a critical missing component that may provide a means for tissue engineers to more efficiently harness the innate immune response (acute template preconditioning) that will lead to the enhancement of in situ regeneration, or more precisely, minimize failure derived by chronic inflammation and fibrosis.