Laboratory of Pneumology, Unit for Lung Toxicology, KU Leuven, Leuven, Belgium.
Part Fibre Toxicol. 2012 Nov 9;9:41. doi: 10.1186/1743-8977-9-41.
Nanomaterials can be contaminated with endotoxin (lipopolysaccharides, LPS) during production or handling. In this study, we searched for a convenient in vitro method to evaluate endotoxin contamination in nanoparticle samples. We assessed the reliability of the commonly used limulus amebocyte lysate (LAL) assay and an alternative method based on toll-like receptor (TLR) 4 reporter cells when applied with particles (TiO(2), Ag, CaCO(3) and SiO(2)), or after extraction of the endotoxin as described in the ISO norm 29701.
Our results indicate that the gel clot LAL assay is easily disturbed in the presence of nanoparticles; and that the endotoxin extraction protocol is not suitable at high particle concentrations. The chromogenic-based LAL endotoxin detection systems (chromogenic LAL assay and Endosafe-PTS), and the TLR4 reporter cells were not significantly perturbed.
We demonstrated that nanoparticles can interfere with endotoxin detection systems indicating that a convenient test method must be chosen before assessing endotoxin contamination in nanoparticle samples.
纳米材料在生产或处理过程中可能会被内毒素(脂多糖,LPS)污染。在这项研究中,我们寻找了一种方便的体外方法来评估纳米粒子样品中的内毒素污染。我们评估了常用的鲎变形细胞溶解物(LAL)检测法和另一种基于 Toll 样受体(TLR)4 报告细胞的方法的可靠性,当应用于颗粒(TiO2、Ag、CaCO3 和 SiO2)时,或按照 ISO 标准 29701 中所述提取内毒素后。
我们的结果表明,凝胶凝结 LAL 检测法在存在纳米颗粒时很容易受到干扰;并且内毒素提取方案不适用于高颗粒浓度。基于显色的 LAL 内毒素检测系统(显色 LAL 检测法和 Endosafe-PTS)和 TLR4 报告细胞没有受到明显干扰。
我们证明纳米颗粒会干扰内毒素检测系统,这表明在评估纳米粒子样品中的内毒素污染之前,必须选择一种方便的测试方法。