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

该提案旨在检查N-α-苯甲酰基-N5-（2-氯-1-亚氨基乙基）-L-鸟氨酸 酰胺（Cl-脒）洗脱电纺模板调节中性粒细胞胞外 陷阱（NET），其已显示在急性相互作用的中性粒细胞中诱导。的 模板将被静电纺丝，使得所得的结构在纤维直径、孔隙 尺寸、载药量和表面化学，以创造不同的微环境， 检查每个变量在调节NET形成和随后宿主中的作用 反应最重要的假设是，NETosis的程度（或模板的水平） 预处理NET）可以通过包括Cl-脒浸渍和/或 吸附到电纺纤维上，并且洗脱的药物起到使NET最小化的作用， 形成将导致期望的、加速的边缘组织愈合和增强的组织 模板的整合。更具体地说，目标1将定义各种表面的作用- 面积体积比、降解速率、表面化学和纤维的Cl-脒负载 静电纺丝过程中的浸渍和/或模板后处理表面吸附， 表征体外释放曲线，并最终确定体外抑制功能。 NETosis。然后，目标2将确定Cl-脒负载的PDO模板的能力，以降低 调节体内NETosis的程度并确定组织愈合/整合结果 与大鼠皮下模型中氯脒下调NETosis程度相关。 预计增加模板纤维直径/孔径和改变表面化学性质 具有显著的突释药物能力将引起最小程度的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- 模板相互作用，更重要的是，提供了额外的模板衍生的证据， 在下调慢性炎症的潜在引发剂中的功能性（Cl-脒洗脱） 和受损的组织愈合。此功能可能是一个关键的缺失组件， 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， （急性模板预处理），这将导致原位再生的增强，或更多 准确地说，最大限度地减少慢性炎症和纤维化引起的失败。

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.