羧化纳米金刚石与小鼠巨噬细胞系和原代细胞的相互作用

Christopher J. Osgood, Maisoun Bani-Hani, Stephen J Beebe1, Michael W Stacey1, Christopher Osgood2
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摘要

纳米金刚石(ND)因其在几种生物医学领域的应用而引起了人们的极大兴趣。如果ND的安全性和兼容性得到证实,这些应用将非常有用。我们评估了ND (100 nm,羧化)对原代巨噬细胞和巨噬细胞样细胞系的影响,发现这些颗粒在较低浓度下对这些细胞无毒,但可能干扰细胞功能和分化。这些细胞以时间和剂量依赖的方式内化ND,主要通过吞噬和网格蛋白依赖的内吞作用,并定位于细胞质而不进入细胞核。没有明显的炎症细胞因子诱导或降低这些细胞对脂多糖(LPS)的反应能力。然而,这些细胞的内吞活性明显降低。此外,ND暴露降低了分化骨髓细胞表达巨噬细胞表面标记物的能力。对nd处理细胞的荧光和吸光度的测量清楚地显示了这些颗粒产生不同波长信号的能力。因此,在测试ND对细胞的相互作用或影响时,考虑ND在不同比色和荧光测定中的干扰是很重要的。我们的研究结果表明,在测试浓度下,ND对巨噬细胞没有细胞毒性,但它可以干扰巨噬细胞的功能和分化,并可能通过产生不同波长的信号干扰检测结果。
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Interactions of Carboxylated Nanodiamonds With Mouse Macrophages Cell Line and Primary Cells
Nanodiamonds (ND) have attracted significant interest for their use in several biomedical applications. These applications can be very useful if the safety and compatibility of ND are proven. We assessed the effects of ND (100 nm, Carboxylated) on primary macrophages and a macrophage-like cell line and found that these particles are not toxic to these cells at lower concentrations but may interfere with cell functions and differentiation. Internalization of ND by these cells in a time- and dose-dependent manner was mostly via phagocytosis and clathrin-dependent endocytosis and localized to the cytoplasm but not into the nucleus. No significant induction of inflammatory cytokines or reduction in the ability of these cells to respond to lipopolysaccharides (LPS) was noted. However, the endocytic activity of these cells is significantly reduced. In addition, ND exposure reduced the ability of differentiating bone marrow cells to express macrophage surface markers. Measurement of the fluorescence and absorbance of ND-treated cells clearly showed the ability of these particles to produce a signal at different wavelengths. Therefore, it is important to consider interference of ND in different colorimetric and fluorometric assays when testing interactions or effects of ND on cells. Our findings suggest that ND are not cytotoxic to macrophages at the tested concentrations, but it can interfere with macrophage functions and differentiation and may interfere with assays’ result through the production of a signal at different wavelengths.
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