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首页> 外文期刊>Acta biomaterialia >Endocytotic uptake of iron oxide nanoparticles by cultured brain microglial cells
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Endocytotic uptake of iron oxide nanoparticles by cultured brain microglial cells

机译：培养的脑小胶质细胞对铁氧化物纳米颗粒的胞吞摄取

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摘要

Microglia are the phagocytotic cells of the brain that respond rapidly to alterations in brain homeostasis. Since iron oxide nanoparticles (IONPs) are used for diagnostic and therapeutic applications in the brain, the consequences of an exposure of microglial cells to IONPs are of particular interest. To address this topic we have synthesized and characterized fluorescent BODIPY?-labelled IONPs (BP-IONPs). The average hydrodynamic diameter and the ζ-potential of BP-IONPs in water were ～65 nm and -49 mV, respectively. Both values increased after dispersion of the particles in serum containing incubation medium to ～130 nm and -8 mV. Exposure of cultured rat microglial cells with BP-IONPs caused a time-, concentration- and temperature-dependent uptake of the particles, as demonstrated by strong increases in cellular iron contents and cellular fluorescence. Incubation for 3 h with 150 and 450 μM iron as BP-IONPs increased the cellular iron content from a low basal level of ～50 nmol iron mg-1 to 219 ± 52 and 481 ± 28 nmol iron (mg protein)-1, respectively. These conditions did not affect cell viability, but exposure to higher concentrations of BP-IONPs or for longer incubation periods severely compromised cell viability. The BP-IONP fluorescence in viable microglial cells was co-localized with lysosomes. In addition, BP-IONP accumulation was lowered by 60% in the presence of the endocytosis inhibitors 5-(N-ethyl-N-isopropyl)amiloride, tyrphostin 23 and chlorpromazin. These results suggest that the rapid accumulation of BP-IONPs by microglial cells is predominantly mediated by macropinocytosis and clathrin-mediated endocytosis, which direct the accumulated particles into the lysosomal compartment.
机译：小胶质细胞是大脑的吞噬细胞，对大脑动态平衡的变化迅速做出反应。由于氧化铁纳米颗粒（IONP）用于大脑中的诊断和治疗应用，因此小胶质细胞暴露于IONP的后果尤其令人关注。为了解决这个问题，我们合成并表征了荧光BODIPYα标记的IONP（BP-IONPs）。水中BP-IONP的平均流体动力学直径和ζ电位分别为〜65 nm和-49 mV。颗粒在含孵育液的血清中分散后，两个值均增加至〜130 nm和-8 mV。 BP-IONPs对培养的大鼠小胶质细胞的暴露引起时间，浓度和温度相关的颗粒摄取，这可以通过细胞铁含量和细胞荧光的强烈增加来证明。随着BP-IONPs与150和450μM铁温育3 h，使细胞铁含量从低基础水平的〜50 nmol铁mg-1分别增加到219±52和481±28 nmol铁（mg蛋白）-1 。这些条件不会影响细胞活力，但是暴露于较高浓度的BP-IONPs或较长的孵育时间会严重损害细胞活力。活的小胶质细胞中的BP-IONP荧光与溶酶体共定位。另外，在存在内吞抑制剂5-（N-乙基-N-异丙基）阿米洛利，酪氨酸磷酸化酶23和氯丙嗪的情况下，BP-IONP的积累降低了60％。这些结果表明，小胶质细胞快速积累BP-IONPs主要是由巨胞饮和网格蛋白介导的内吞作用介导的，后者将积累的颗粒导向溶酶体区室。

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