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Effect of silica nanoparticles on endothelial permeability and inflammation in human brain microvessel endothelial cells

机译：二氧化硅纳米颗粒对人脑微血管内皮细胞内皮通透性和炎症的影响

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Introduction: Silica nanoparticles (SiO_2-NPs) have certain unique characteristics which have been exploited in many biomedical applications. However, these unique features such as small size also enable NPs to negotiate various biological barriers such as blood-brain barrier (BBB) in the body and cause unexpected effects. The core element of the BBB is the cerebral blood vessel endothelial cells (ECs), which have continuous intercellular tight junctions (TJs) and greatly limit the movement of molecules through the BBB. However, to date, far less effort has been put forth concerning the adverse effects of SiO_2NPs on brain microvessel endothelial cells (BMECs). Thus, a fundament understanding of the interaction of SiO_2-NPs with human BMECs (HBMECs) was investigated in this study. Materials and Methods: SiO_2-NPs was prepared by a sol-gel method according to the previous study. HBMECs was purchased from ScienCell Research Laboratories (Carlsbad, CA, USA). Firstly, changes in endothelial permeability were measured by transendothelial electrical resistance (TER), and then F-actin organization was observed by rhodamine-phalloidin staining under laser scanning confocal microscope (LSCM). Moreover, flow cytometry and enzyme-linked immunosorbent assay(ELISA) were used to detect inflammatory cytokines and cells adhesion molecules (CAMs) (CD54.CD106 and CD62E). Result and Discussion: The synthesized SiO_2-NPs were near-spherical and the size was approximately 20 nm. Measurements of TER reflecting endothelial monolayer permeability changes showed that administration of 100 μg/mL of SiO_2-NPs resulted in a time-dependent decrease in TER (Figure 1 A), indicating that SiO_2-NPs could mediate the breakdown of barrier integrity in HBMECs. Before stimulation, HBMECs did not display stress fibers, but after 24 h of SiO_2-NPs exposure, cells exhibited prominent stress fiber formation and paracellular gaps (Figure 1B-C). Pro-inflammatory stimuli may induce actin stress fibres, cell elongation and contractility in endothelial cells. To investigate whether SiO_2-NPs induce inflammatory response in MBMECs, the hallmark of ECs activation, cell adhesion molecules such as VCAM-1 (CD106), ICAM-1 (CD54) and E-selectin (CD62E) was detected. Herein, we observed a significant increase of CD54 and CD62E, indicating an inflammatory effect of SiO_2-NPs on HBMECs(Figure 2). It is likely that SiO_2-NPs induce stress fibers formation and increase endothelial permeability by pro-inflammatory stimuli. Conclusion: This study showed that SiO_2-NPs could induce a proinflammatory effect on HBMECs and increase the endothelial permeability, suggesting SiO_2-NPs may disrupt the BBB, which may further induce brain inflammation and neurotoxicity.
机译：简介：二氧化硅纳米粒子（SiO_2-NPs）具有某些独特的特性，已在许多生物医学应用中得到利用。但是，这些独特的功能（例如体积小）也使NP能够协商各种生物屏障，例如体内的血脑屏障（BBB），并产生意想不到的效果。 BBB的核心元素是脑血管内皮细胞（EC），它们具有连续的细胞间紧密连接（TJ），并极大地限制了分子通过BBB的运动。但是，迄今为止，关于SiO_2NPs对脑微血管内皮细胞（BMECs）的不利影响的研究工作已经很少。因此，本研究研究了对SiO_2-NP与人BMEC（HBMEC）相互作用的基本理解。材料与方法：SiO_2-NPs是根据以前的研究通过溶胶-凝胶法制备的。 HBMEC是从ScienCell研究实验室（美国加利福尼亚州卡尔斯巴德）购买的。首先，通过跨内皮电阻（TER）测量内皮通透性的变化，然后在激光扫描共聚焦显微镜（LSCM）下通过若丹明-鬼笔环肽染色观察F-肌动蛋白的组织。此外，流式细胞仪和酶联免疫吸附试验（ELISA）用于检测炎症细胞因子和细胞粘附分子（CAMs）（CD54.CD106和CD62E）。结果与讨论：合成的SiO_2-NPs为近球形，粒径约为20 nm。反映内皮单层通透性变化的TER的测量结果表明，施用100μg/ mL的SiO_2-NPs会导致TER的时间依赖性下降（图1 A），表明SiO_2-NPs可以介导HBMECs中屏障完整性的破坏。在刺激之前，HBMECs没有显示应力纤维，但是在SiO_2-NPs暴露24小时后，细胞表现出显着的应力纤维形成和细胞旁间隙（图1B-C）。促炎性刺激可诱导内皮细胞中肌动蛋白应激纤维，细胞伸长和收缩。为了研究SiO_2-NPs是否在MBMECs中引起炎症反应，检测了ECs活化的标志，检测了细胞粘附分子，例如VCAM-1（CD106），ICAM-1（CD54）和E-选择素（CD62E）。在这里，我们观察到CD54和CD62E的显着增加，表明SiO_2-NPs对HBMECs有炎症作用（图2）。 SiO_2-NPs可能通过促炎性刺激诱导应激纤维形成并增加内皮通透性。结论：这项研究表明，SiO_2-NPs可以诱导对HBMECs的促炎作用并增加内皮通透性，表明SiO_2-NPs可以破坏血脑屏障，从而进一步诱发脑部炎症和神经毒性。

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