Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Meibergdreef 15, Amsterdam, 1105AZ, The Netherlands.
MIRA Institute for Biomedical Technology and Technical Medicine, Department of Biomaterials Science and Technology, University of Twente, Enschede, AE, 7500, The Netherlands.
J Biomed Mater Res A. 2017 Sep;105(9):2522-2532. doi: 10.1002/jbm.a.36110. Epub 2017 Jun 6.
To rapidly assess early inflammatory cell responses provoked by biomaterials in the full complexity of the living organism, we developed a zebrafish embryo model which allows real time analysis of these responses to biomaterial microspheres. Fluorescently labeled microspheres with different properties were injected into embryos of selected transgenic zebrafish lines expressing distinct fluorescent proteins in their neutrophils and macrophages. Recruitment of leukocytes and their interactions with microspheres were monitored using fluorescence microscopy. We developed a novel method using ImageJ and the plugin ObjectJ project file "Zebrafish-Immunotest" for rapid and semi-automated fluorescence quantification of the cellular responses. In the embryo model we observed an ordered inflammatory cell response to polystyrene and poly (ε-caprolactone) microspheres, similar to that described for mammalian animal models. The responses were characterized by an early infiltration of neutrophils followed by macrophages, and subsequent differentially timed migration of these cells away from the microspheres. The size of microspheres (10 and 15 µm) did not influence the cellular responses. Poly (ε-caprolactone) microspheres provoked a stronger infiltration of neutrophils and macrophages than polystyrene microspheres did. Our study shows the potential usefulness of zebrafish embryos for in vivo evaluation of biomaterial-associated inflammatory cell responses. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2522-2532, 2017.
为了快速评估生物材料在活体生物的全部复杂性中引发的早期炎症细胞反应，我们开发了一种斑马鱼胚胎模型，该模型允许实时分析对生物材料微球的这些反应。用不同性质的荧光标记微球注射到在其中性粒细胞和巨噬细胞中表达不同荧光蛋白的特定转基因斑马鱼系的胚胎中。使用荧光显微镜监测白细胞的募集及其与微球的相互作用。我们开发了一种使用 ImageJ 和插件 ObjectJ 项目文件“Zebrafish-Immunotest”的新方法，用于快速和半自动荧光定量细胞反应。在胚胎模型中，我们观察到对聚苯乙烯和聚（ε-己内酯）微球的有序炎症细胞反应，类似于描述的哺乳动物动物模型。反应的特征是早期中性粒细胞浸润，随后是巨噬细胞，随后这些细胞以不同的时间从微球迁移。微球的大小（10 和 15 µm）不会影响细胞反应。聚（ε-己内酯）微球比聚苯乙烯微球引起更强的中性粒细胞和巨噬细胞浸润。我们的研究表明，斑马鱼胚胎具有用于体内评估与生物材料相关的炎症细胞反应的潜在用途。© 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A：105A：2522-2532，2017。
