Electrospun Template Architecture and Composition Regulate Neutrophil NETosis In Vitro and In Vivo
Electrospun Template Architecture and Composition Regulate Neutrophil NETosis In Vitro and In Vivo
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DOI：
10.1089/ten.tea.2016.0452
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发表时间：
2017-10-01
影响因子：
4.1
通讯作者：
Bowlin, Gary L.
中科院分区：
文献类型：
--
作者：
Fetz, Allison E.;Neeli, Indira;Bowlin, Gary L.
越来越多的证据表明，中性粒细胞，植入生物材料的第一反应者，通过分泌因子和通过NETosis释放中性粒细胞胞外陷阱（NET）来为招募的免疫细胞准备微环境。在这项研究中，我们研究了电纺模板结构和组成在调节NETosis中的作用。用小直径（0.25-0.35 μ m）和大直径（1.0-2.00 μ m）纤维制备静电纺丝聚对二氧环己酮（PDO）、I型胶原（COL）和混合PDO-COL模板（PC）。中性粒细胞-模板相互作用进行了评价，在体外3和24小时与人中性粒细胞，PDO模板进行了研究，在体内（大鼠皮下模型）1和7天。通过荧光显微镜和On-cell Western分析定量模板结合的NET。体外结果表明，较大的纤维直径减少了PDO模板上的NETosis，而COL的掺入减弱了NETosis，与纤维直径无关。体内结果类似地揭示了在1天时在大直径PDO模板上的较低程度的NET，导致在7天时模板的边缘组织整合。相比之下，小直径PDO模板，这是在体内24小时包被在大量的NET，被包围的胶囊状组织在7天。这些初步的体内结果验证了体外模型，并表明NETosis是一种潜在的重要生理反应和对模板的先天免疫反应的关键预处理事件。总之，这些结果表明了表征中性粒细胞与生物材料的急性对抗以工程化能够促进原位再生的模板的重要性。
Mounting evidence indicates that neutrophils, first responders to an implanted biomaterial, prime the microenvironment for recruited immune cells by secreting factors and releasing neutrophil extracellular traps (NETs) through NETosis. In this study, we investigated the role of electrospun template architecture and composition in regulating NETosis. Electrospun polydioxanone (PDO), collagen type I (COL), and blended PDO-COL templates (PC) were fabricated with small-diameter (0.25-0.35 mu m) and large-diameter (1.0-2.00 mu m) fibers. Neutrophil-template interactions were evaluated in vitro for 3 and 24 h with human neutrophils, and the PDO templates were studied in vivo (rat subcutaneous model) for 1 and 7 days. Template-bound NETs were quantified by fluorescent microscopy and an On-cell Western assay. The in vitro results indicate that larger fiber diameters reduced NETosis on PDO templates, whereas the incorporation of COL attenuated NETosis independent of fiber diameter. The in vivo results similarly revealed a lower degree of NETs on large-diameter PDO templates at 1 day, resulting in marginal tissue integration of the templates at 7 days. In contrast, the small-diameter PDO templates, which were coated in a large amount of NETs at 24 h in vivo, were surrounded by capsule-like tissue at 7 days. These preliminary in vivo results validate the in vitro model and signify NETosis as a potentially significant physiological response and a critical preconditioning event for the innate immune response to templates. In conclusion, these results demonstrate the importance of characterizing the neutrophil's acute confrontation with biomaterials to engineer templates capable of promoting in situ regeneration.