Mahmoudzadeh Aziz, Mohsenifar Afshin, Rahmani-Cherati Tavoos
a Novin Medical Radiation Institute , Nanozino , Tehran , Iran ;
b Research and Development Department , South Pirozan, Shahrak Qods , Tehran , Iran.
J Immunotoxicol. 2016 Jul;13(4):526-34. doi: 10.3109/1547691X.2016.1139642. Epub 2016 Apr 4.
Macrophages are effector cells in the innate and adaptive immune systems and in situ exist within three-dimensional (3-D) microenvironments. As there has been an increase in interest in the use of 3-D scaffolds to mimic natural microenvironments in vitro, this study examined the impact on cultured mice peritoneal macrophages using standard 2-D plates as compared to 3-D collagen-chitosan scaffolds. Here, 2-D and 3-D cultured macrophages were evaluated for responses to lipopolysaccharide (LPS), dexamethasone (Dex), BSA (bovine serum albumin), safranal (herbal component isolated from safranal [Saf]) and Alyssum homolocarpum mucilage (A. muc: mixed herbal components). After treatments, cultured macrophages were evaluated for viability, phagocytic activity and release of tumor necrosis factor (TNF)-α and interleukin (IL)-1β pro-inflammatory cytokines. Comparison of 2-D vs 3-D cultures showed that use of either system - with or without any exogenous agent - had no effect on cell viability. In the case of cell function, macrophages cultured on scaffolds had increases in phagocytic activity relative to that by cells on 2-D plates. In general, the test herbal components Saf and A. muc. had more impact than any of the other exogenous agents on nanoparticle uptake. With respect to production of TNFα and IL-1β, compared to the 2-D cells, scaffold cells tended to have significantly different levels of production of each cytokine, with the effect varying (higher or lower) depending on the test agent used. However, unlike with particle uptake, here, while Saf and A. muc. led to significantly greater levels of cytokine formation by the 3-D culture cells vs that by the 2-D plate cells, there was no net effect (stimulatory) vs control cultures. These results illustrated that collagen-chitosan scaffolds could provide a suitable 3-D microenvironment for macrophage phagocytosis and could also impact on the formation of pro-inflammatory cytokines.
巨噬细胞是先天性和适应性免疫系统中的效应细胞，在体内存在于三维（3-D）微环境中。由于人们越来越关注使用3-D支架在体外模拟天然微环境，本研究考察了与3-D胶原-壳聚糖支架相比，使用标准二维平板对培养的小鼠腹腔巨噬细胞的影响。在此，评估了二维和三维培养的巨噬细胞对脂多糖（LPS）、地塞米松（Dex）、牛血清白蛋白（BSA）、藏红花醛（从藏红花中分离的草药成分[Saf]）和 Alyssum homolocarpum 黏液（A. muc：混合草药成分）的反应。处理后，评估培养的巨噬细胞的活力、吞噬活性以及肿瘤坏死因子（TNF）-α和白细胞介素（IL）-1β促炎细胞因子的释放。二维与三维培养的比较表明，使用任何一种系统（有无任何外源剂）对细胞活力均无影响。就细胞功能而言，在支架上培养的巨噬细胞相对于二维平板上的细胞，吞噬活性有所增加。一般来说，测试的草药成分Saf和A. muc. 对纳米颗粒摄取的影响比任何其他外源剂都更大。关于TNFα和IL-1β的产生，与二维细胞相比，支架细胞中每种细胞因子的产生水平往往有显著差异，其影响因所用测试剂而异（更高或更低）。然而，与颗粒摄取不同，在这里，虽然Saf和A. muc. 导致三维培养细胞产生的细胞因子水平明显高于二维平板细胞，但与对照培养相比没有净效应（刺激）。这些结果表明，胶原-壳聚糖支架可为巨噬细胞吞噬作用提供合适的三维微环境，也可能影响促炎细胞因子的形成。
