Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, Instituto de Investigación Sanitaria del Hospital Clínico San Carlos (IdISSC), 28040 Madrid, Spain.
Int Immunopharmacol. 2021 May;94:107457. doi: 10.1016/j.intimp.2021.107457. Epub 2021 Mar 20.
The use of nanoparticles for intracellular drug delivery could reduce the toxicity and side effects of the drug but, the uptake of these nanocarriers could induce adverse effects on cells and tissues after their incorporation. Macrophages play a central role in host defense and are responsible for in vivo nanoparticle trafficking. Assessment of their defense capacity against pathogenic micro-organisms after nanoparticle uptake, is necessary to prevent infections associated with nanoparticle therapies. In this study, the effects of hollow mesoporous SiO-CaO nanospheres labeled with fluorescein isothiocyanate (FITC-NanoMBGs) on the function of peritoneal macrophages was assessed by measuring their ability to phagocytize Candidaalbicans expressing a red fluorescent protein. Two macrophage/fungus ratios (MOI1 and MOI5) were used and two experimental strategies were carried out: a) pretreatment of macrophages with FITC-NanoMBGs and subsequent fungal infection; b) competition assays after simultaneous addition of fungus and nanospheres. Macrophage pro-inflammatory phenotype markers (CD80 expression and interleukin 6 secretion) were also evaluated. Significant decreases of CD80 macrophage percentage and interleukin 6 secretion were observed after 30 min, indicating that the simultaneous incorporation of NanoMBG and fungus favors the macrophage non-inflammatory phenotype. The present study evidences that the uptake of these nanospheres in all the studied conditions does not alter the macrophage function. Moreover, intracellular FITC-NanoMBGs induce a transitory increase of the fungal phagocytosis by macrophages at MOI 1 and after a short time of interaction. In the competition assays, as the intracellular fungus quantity increased, the intracellular FITC-NanoMBG content decreased in a MOI- and time-dependent manner. These results have confirmed that macrophages clearly distinguish between inert material and the live yeast in a dynamic intracellular incorporation. Furthermore, macrophage phagocytosis is a critical determinant to know their functional state and a valuable parameter to study the nanomaterial / macrophages / Candida albicans interface.
纳米颗粒用于细胞内药物递送可以降低药物的毒性和副作用，但这些纳米载体的摄取可能会在被细胞摄取后对细胞和组织产生不良反应。巨噬细胞在宿主防御中起核心作用，负责体内纳米颗粒的转运。评估其对摄入纳米颗粒后的致病微生物的防御能力对于预防与纳米颗粒治疗相关的感染是必要的。在这项研究中，通过测量其吞噬表达红色荧光蛋白的白色念珠菌的能力来评估用异硫氰酸荧光素（FITC-NanoMBGs）标记的中空介孔 SiO-CaO 纳米球（FITC-NanoMBGs）对腹腔巨噬细胞功能的影响。使用两种巨噬细胞/真菌比值（MOI1 和 MOI5）并进行了两种实验策略：a）用 FITC-NanoMBGs 预处理巨噬细胞，然后进行真菌感染；b）同时添加真菌和纳米球后进行竞争测定。还评估了巨噬细胞促炎表型标志物（CD80 表达和白细胞介素 6 分泌）。在 30 分钟后，观察到 CD80 巨噬细胞百分比和白细胞介素 6 分泌显著下降，这表明同时摄取 NanoMBG 和真菌有利于巨噬细胞的非炎症表型。本研究证明，在所有研究条件下，这些纳米球的摄取不会改变巨噬细胞的功能。此外，在 MOI1 下，在短时间的相互作用后，细胞内的 FITC-NanoMBGs 诱导巨噬细胞吞噬真菌的短暂增加。在竞争测定中，随着细胞内真菌数量的增加，细胞内 FITC-NanoMBG 含量以 MOI 和时间依赖性的方式减少。这些结果证实，巨噬细胞在动态的细胞内摄取过程中，能够清楚地区分惰性物质和活酵母。此外，巨噬细胞吞噬作用是了解其功能状态的关键决定因素，也是研究纳米材料/巨噬细胞/白色念珠菌界面的有价值参数。
