McKiel Laura A, Woodhouse Kimberly A, Fitzpatrick Lindsay E
Department of Chemical Engineering, Queen's University.
Department of Chemical Engineering, Queen's University;
J Vis Exp. 2020 Jan 7(155). doi: 10.3791/60317.
The persistent inflammatory host response to an implanted biomaterial, known as the foreign body reaction, is a significant challenge in the development and implementation of biomedical devices and tissue engineering constructs. Macrophages, an innate immune cell, are key players in the foreign body reaction because they remain at the implant site for the lifetime of the device, and are commonly studied to gain an understanding of this detrimental host response. Many biomaterials researchers have shown that adsorbed protein layers on implanted materials influence macrophage behavior, and subsequently impact the host response. The methods in this paper describe an in vitro model using adsorbed protein layers containing cellular damage molecules on polymer biomaterial surfaces to assess macrophage responses. An NF-кB/AP-1 reporter macrophage cell line and the associated colorimetric alkaline phosphatase assay were used as a rapid method to indirectly examine NF-кB/AP-1 transcription factor activity in response to complex adsorbed protein layers containing blood proteins and damage-associated molecular patterns, as a model of the complex adsorbed protein layers formed on biomaterial surfaces in vivo.
宿主对植入生物材料的持续炎症反应，即所谓的异物反应，是生物医学设备和组织工程构建体开发与应用中的一项重大挑战。巨噬细胞作为一种先天性免疫细胞，是异物反应的关键参与者，因为它们会在设备的整个使用周期内一直留在植入部位，并且通常会对其进行研究以了解这种有害的宿主反应。许多生物材料研究人员已经表明，植入材料上吸附的蛋白质层会影响巨噬细胞的行为，进而影响宿主反应。本文中的方法描述了一种体外模型，该模型使用聚合物生物材料表面上含有细胞损伤分子的吸附蛋白质层来评估巨噬细胞反应。使用NF-κB/AP-1报告基因巨噬细胞系和相关的比色碱性磷酸酶测定法，作为一种快速方法来间接检测NF-κB/AP-1转录因子在响应含有血液蛋白和损伤相关分子模式的复杂吸附蛋白质层时的活性，以此作为体内生物材料表面形成的复杂吸附蛋白质层的模型。
