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首页> 外文期刊>Journal of biomedical materials research. Part B, Applied biomaterials. >Decontaminating particles exposed to bacterial endotoxin (LPS).
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Decontaminating particles exposed to bacterial endotoxin (LPS).

机译：净化暴露于细菌内毒素（LPS）的颗粒。

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摘要

Lipopolysaccharide (LPS), which comes from the cell wall of gram-negative bacteria, can stimulate murine macrophage cells to produce nitric oxide (NO), cytokines, such as tumor necrosis factor-alpha, and interleukins, such as IL-6. When examining the biological effects of particles on macrophages, it is important to have no contaminating LPS associated with the particles and none with any cell culture media or supplies since even very low levels of LPS are stimulatory. The presence or absence of LPS was observed in two ways: (1) the amount of NO produced by RAW 264.7 murine macrophage cells, and (2) the Limulus amebocyte lysate (LAL) test. Treating particles with 70% ethanol at room temperature for 48 h, followed by washing the polymethylmethacrylate (PMMA) particles with endotoxin-free phosphate-buffered saline three times, decontaminated LPS and LPS-treated PMMA particles. When given LPS that had been treated with 70% ethanol for 48 h at room temperature or at 37 degrees C, cells did not produce NO above control levels. Negative LAL tests indicated the presence of extremely low levels or the complete absence of LPS in 70% ethanol-treated LPS. Copyright 1999 John Wiley & Sons, Inc.
机译：来自革兰氏阴性细菌细胞壁的脂多糖（LPS）可以刺激鼠巨噬细胞产生一氧化氮（NO），细胞因子（例如肿瘤坏死因子-α）和白介素（例如IL-6）。当检查颗粒对巨噬细胞的生物学作用时，重要的是不要与颗粒相关的LPS污染，也不要与任何细胞培养基或补给品一起污染，因为即使LPS含量极低也具有刺激性。有两种方法可以观察到LPS的存在与否：（1）RAW 264.7鼠巨噬细胞产生的NO量，以及（2）am变形细胞裂解液（LAL）测试。在室温下用70％乙醇处理颗粒48小时，然后用无内毒素的磷酸盐缓冲盐水洗涤聚甲基丙烯酸甲酯（PMMA）颗粒3次，对LPS和经LPS处理的PMMA颗粒进行净化处理。当给予在室温或37摄氏度下用70％乙醇处理过48小时的LPS时，细胞不会产生高于对照水平的NO。阴性LAL测试表明在70％乙醇处理的LPS中存在极低的水平或完全不存在LPS。版权所有1999 John Wiley＆Sons，Inc.

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