Carbon nanodots interference with lactate dehydrogenase assay in human monocyte THP-1 cells.
Carbon nanodots interference with lactate dehydrogenase assay in human monocyte THP-1 cells.
复制标题
碳纳米点干扰人单核细胞 THP-1 细胞中的乳酸脱氢酶测定。
作者：
Wright PC;Qin H;Choi MM;Chiu NH;Jia Z
碳纳米点（CD）是一种尺寸小于10 nm的新型碳纳米材料，由于其水溶性、化学惰性和抗光漂白等优点，近年来引起了人们的广泛关注。因此，环糊精在能源、催化、生物标记、生物成像和药物输送等方面有着重要而广泛的应用。另一方面，由于缺乏可用的毒性数据，人们越来越担心CD的潜在风险。因此，准确评估CD的细胞毒性变得比以往任何时候都更加重要。乳酸脱氢酶（LDH）测定广泛用于检测各种纳米颗粒（包括CD）的细胞毒性。近来许多研究使用LDH测定来研究CD在各种细胞中的毒性。然而，这些研究未能进一步检查CD是否干扰LDH测定，从而改变其结果。本研究调查了碳纳米点对人单核细胞THP-1细胞中LDH测定的可能干扰。已知单核细胞参与易燃性血管疾病，并已被建议为CD暴露的靶点。在本研究中，使用LDH试验测定了浓度范围为0.075 - 0.60 mg/mL的CD的细胞毒性。为了验证LDH测定的结果，使用台盼蓝染色的细胞计数法。根据台盼蓝染色法，随着24小时孵育时间的延长，THP-1的细胞活力显著降低。而在LDH测定中，发现CD以剂量依赖性方式干扰340 nm处的NADH吸光度测量。本研究是首次报道CD对LDH测定的阴性干扰，在评价CD的细胞毒性时应谨慎。
Carbon nanodots (CD), a new class of carbon nanomaterials with sizes below 10 nm, have recently attracted wide attention due to their superiority in water solubility, chemical inertness, and resistance to photobleaching. As a result, CD has found important and wide applications in energy, catalysis, biological labeling, bioimaging and drug delivery. On the other hand, due to the lack of available toxicity data, there is a growing concern regarding the potential risks of CD. Hence, accurate assessment of the cytotoxicity of CD has become more important than ever before. The lactate dehydrogenase (LDH) assay is widely used to detect cytotoxicity of various nanoparticles including CD. Many recent studies used LDH assay to study the CD toxicity in various cells. However, these studies failed to further examine whether the CD were interfering with the LDH assay which would alter their findings. This study investigated the possible interference of carbon nanodots on the LDH assay in human monocyte THP-1 cells. Monocytes are known to be involved in inflammable vascular diseases, and have been suggested to be the targets for CD exposure. In this study, the cytotoxicity of CD in concentrations ranging from 0.075 to 0.60 mg/mL, was determined by using the LDH assay. To validate the results of LDH assay, the cell counting method with trypan blue staining was used. With 24 hours incubation time, the cell viability of THP-1 was significantly decreased according to the trypan blue staining method. Whereas, in the LDH assay, the CD was found to interfere in a dose-dependent manner with the NADH absorbance measurements at 340 nm. This study represents the first report on the negative interference of CD on LDH assay, and caution should be observed when evaluating the cytotoxicity of CD.
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