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In vitro Evaluation
Published in Raj Bawa, János Szebeni, Thomas J. Webster, Gerald F. Audette, Immune Aspects of Biopharmaceuticals and Nanomedicines, 2019
The lactate dehydrogenase assay (LDH assay) measures LDH activity in the extracellular media, which gives an indication of the rupture of the cell membrane [33, 39, 40]. It was first developed to test the cytotoxicity in neuronal cells [41]. Specifically, this test measures the production of lactate and NAD+ from pyruvate and NADH, reaction that only takes place in the presence of the LDH enzyme, by measuring the absorbance of NADH [34, 41]. Therefore, only in the presence of LDH, the absorbance should decrease due to the oxidation of NADH [41]. Since it is a measure of the extracellular LDH, the highest the LDH activity, the more damaged the cell membrane and, consequently, the lowest the viability of cells [17].
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Published in Valerio Voliani, Nanomaterials and Neoplasms, 2021
Alaaldin M. Alkilany, Catherine J. Murphy
There are many assays used to measure the cellular impact of a drug that can also be applied to measure the impact of nanoparticle exposure on cells. One common assay is the LDH assay, which is a colorimetric assay measuring the release of lactate dehydrogenase (LDH) into the culture media as an indicator of cellular membrane disruption (Marquis et al. 2009). A metabolic assay considered the “gold standard” for cytotoxicity is the MTT assay, which is a colorimetric assay that measures the enzymatic activity of cellular mitochondria. If cells properly metabolize the MTT dye, the cell culture will turn blue, allowing for simple absorbance measurements to be used to quantify cellular activity (Marquis et al. 2009).
Effects of zinc oxide nanoparticles on sludge anaerobic fermentation: phenomenon and mechanism
Published in Journal of Environmental Science and Health, Part A, 2020
Baodan Jin, Yue Yuan, Ping Zhou, Jiahui Niu, Jintao Niu, Jingwen Dai, Nuonan Li, Hongfan Tao, Zhigang Ma, Ju Zhang, Zhongfang Zhang, Yu Li
ZnO NPs could disrupt cell membranes by being adsorbed on the cell surface and reacting with the organic matter on these cell membranes. Lactate dehydrogenase (LDH) is an important biological enzyme that catalyzes the redox reaction between lactic acid and pyruvate. Therefore, LDH represents a key DH in WAS fermentation. Similar to LDH, DH can be released because of membrane damage.[35,36] DH activity declined with ZnO NPs addition in the order of ZnO1 NPs > ZnO2 NPs > ZnO3 NPs > ZnO4 NPs, however, no obvious change was noted under ZnO NPs shock (Fig. 4f). The maximum DH activity was 1.4373, 1.4523 and 1.4132 EU/mg VSS for ZnO1 NPs, which was 2.60–2.73 times higher than that for ZnO4 NPs (0.5263, 0.5324 and 0.5432 EU/mg VSS). The results indicate that ZnO NPs inhibited DH activity. Furthermore, high ZnO NPs concentrations inhibited the cellular activity, which might be related to the reduction reaction between Zn2+ and other metal ions released from WAS.[37,38] In ZnO NPs, the electron holes on ZnO surface react with H2O to generate •OH, and the exciting electron diffusing to the surface reacts with O2 to generate O2•−.[39] Thus, under anaerobic conditions, ZnO NPs produced H2O2 and the electron holes on ZnO NPs and H2O generated •OH.[37] This phenomenon probably changed the membrane morphology and activity, which declined DH activity.
Toxicoproteomic assessment of liver responses to acute pyrrolizidine alkaloid intoxication in rats
Published in Journal of Environmental Science and Health, Part C, 2018
Yan-Hong Li, William Chi-Shing Tai, Imran Khan, Cheng Lu, Yao Lu, Wing-Yan Wong, Wood-Yee Chan, Wen-Luan Wendy Hsiao, Ge Lin
Lactate dehydrogenase (LDH) assay was performed according to the protocol of the commercially available kit (Catalog No, 04744926001, Roche Diagnostics Ltd, Hong Kong). Serum total bilirubin level was tested according to a previously published protocol26 with slight modifications. Serum and hepatic GSH concentration was determined by the method of Tietze.27