Effects of Charged Oxime Reactivators on the HK-2 Cell Line in Renal Toxicity Screening

Handl, Jiří; Maliňák, Dávid; Čapek, Jan; Andrýs, Rudolf; Roušarová, Erika; Hauschke, Martina; Brůčková, Lenka; Česla, Petr; Roušar, Tomáš; Musílek, Kamil

doi:10.1021/acs.chemrestox.0c00489

Cited by 10 publications

(3 citation statements)

References 27 publications

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“…Conventional cell viability and cytotoxicity assays enable fast and cheap screening of newly synthetized compounds. During primary oxime cytotoxicity screening, the IC 50 value is usually established on several cell lines, including HepG2 cells, SH-SY-5Y, or HK-2. , The concentration of oximes used in this in vitro study (IC 50 at 24 h) exerted varying effects on cell viability and induced observable cytotoxic effects in a time-dependent manner. This allowed us to monitor the dynamics of oxidative stress induction and its associated damage after 1 and 4 h of exposure.…”

Section: Discussionmentioning

confidence: 99%

In Vitro Evaluation of Oxidative Stress Induced by Oxime Reactivators of Acetylcholinesterase in HepG2 Cells

Váňová,

Múčková,

Kalíšková

et al. 2023

Chem. Res. Toxicol.

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Oxime reactivators of acetylcholinesterase (AChE) are used as causal antidotes for intended and unintended poisoning by organophosphate nerve agents and pesticides. Despite all efforts to develop new AChE reactivators, none of these drug candidates replaced conventional clinically used oximes. In addition to the therapeutic efficacy, determining the safety profile is crucial in preclinical drug evaluation. The exact mechanism of oxime toxicity and the structure−toxicity relationship are subjects of ongoing research, with oxidative stress proposed as a possible mechanism.In the present study, we investigated four promising bispyridinium oxime AChE reactivators, K048, K074, K075, and K203, and their ability to induce oxidative stress in vitro. Cultured human hepatoma cells were exposed to oximes at concentrations corresponding to their IC 50 values determined by the MTT assay after 24 h. Their potency to generate reactive oxygen species, interfere with the thiol antioxidant system, and induce lipid peroxidation was evaluated at 1, 4, and 24 h of exposure. Reactivators without a double bond in the four-carbon linker, K048 and K074, showed a greater potential to induce oxidative stress compared with K075 and K203, which contain a double bond. Unlike oximes with a three-carbon-long linker, the number of aldoxime groups attached to the pyridinium moieties does not determine the oxidative stress induction for K048, K074, K075, and K203 oximes. In conclusion, our results emphasize that the structure of oximes plays a critical role in inducing oxidative stress, and this relationship does not correlate with their cytotoxicity expressed as the IC 50 value. However, it is important to note that oxidative stress cannot be disregarded as a potential contributor to the side effects associated with oximes.

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Section: Discussionmentioning

confidence: 99%

In Vitro Evaluation of Oxidative Stress Induced by Oxime Reactivators of Acetylcholinesterase in HepG2 Cells

Váňová,

Múčková,

Kalíšková

et al. 2023

Chem. Res. Toxicol.

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“…Since pyridoxal (vitamin B6) acts as a cofactor in different metabolic processes and therefore has important physiological roles [ 48 ], it can directly affect the dysregulation of cell processes and lead to cell decay. Thus, this additional essential information was assessed here by cytotoxicity screening studies, performed on two cell types, neuronal (as a main target site of oximes) and kidney (as a main organ that participates in the excretion of xenobiotics) [ 49 , 50 ]. As the results demonstrated, cytotoxicity was not observed for the tested compounds, indicating that they do not influence cell homeostasis or processes leading to cell death.…”

Section: Discussionmentioning

confidence: 99%

Potential of Vitamin B6 Dioxime Analogues to Act as Cholinesterase Ligands

Gašo-Sokač

Zandona

Roca

et al. 2022

IJMS

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Seven pyridoxal dioxime quaternary salts (1–7) were synthesized with the aim of studying their interactions with human acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The synthesis was achieved by the quaternization of pyridoxal monooxime with substituted 2-bromoacetophenone oximes (phenacyl bromide oximes). All compounds, prepared in good yields (43–76%) and characterized by 1D and 2D NMR spectroscopy, were evaluated as reversible inhibitors of cholinesterase and/or reactivators of enzymes inhibited by toxic organophosphorus compounds. Their potency was compared with that of their monooxime analogues and medically approved oxime HI-6. The obtained pyridoxal dioximes were relatively weak inhibitors for both enzymes (Ki = 100–400 µM). The second oxime group in the structure did not improve the binding compared to the monooxime analogues. The same was observed for reactivation of VX-, tabun-, and paraoxon-inhibited AChE and BChE, where no significant efficiency burst was noted. In silico analysis and molecular docking studies connected the kinetic data to the structural features of the tested compound, showing that the low binding affinity and reactivation efficacy may be a consequence of a bulk structure hindering important reactive groups. The tested dioximes were non-toxic to human neuroblastoma cells (SH-SY5Y) and human embryonal kidney cells (HEK293).

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“…The WST-1 test measures the activity of mitochondrial dehydrogenases. 46 After incubation with nanomaterials, 10 µL of WST-1 reagent was added to each well containing cells in 100 µL of culture medium according to the manufacturer’s instructions. After 1 h, the change of absorbance was measured at the wavelength of 440 nm using SPARK microplate reader (Tecan, Austria) or at 450 nm with 650 nm reference wavelength using SPEKTRAFluor Plus (Tecan, Austria) while incubated at 37 ℃.…”

Section: Methodsmentioning

confidence: 99%

Evaluating the Use of TiO2 Nanoparticles for Toxicity Testing in Pulmonary A549 Cells

Báčová¹,

Knotek²,

Kopecká³

et al. 2022

IJN

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Purpose Titanium dioxide nanoparticles, 25 nm in size of crystallites (TiO 2 P25), are among the most produced nanomaterials worldwide. The broad use of TiO 2 P25 in material science has implied a request to evaluate their biological effects, especially in the lungs. Hence, the pulmonary A549 cell line has been used to estimate the effects of TiO 2 P25. However, the reports have provided dissimilar results on caused toxicity. Surprisingly, the physicochemical factors influencing TiO 2 P25 action in biological models have not been evaluated in most reports. Thus, the objective of the present study is to characterize the preparation of TiO 2 P25 for biological testing in A549 cells and to evaluate their biological effects. Methods We determined the size and crystallinity of TiO 2 P25. We used four techniques for TiO 2 P25 dispersion. We estimated the colloid stability of TiO 2 P25 in distilled water, isotonic NaCl solution, and cell culture medium. We applied the optimal dispersion conditions for testing the biological effects of TiO 2 P25 (0–100 µg.mL −1 ) in A549 cells using biochemical assays (dehydrogenase activity, glutathione levels) and microscopy. Results We found that the use of fetal bovine serum in culture medium is essential to maintain sufficient colloid stability of dispersed TiO 2 P25. Under these conditions, TiO 2 P25 were unable to induce a significant impairment of A549 cells according to the results of biochemical and microscopy evaluations. When the defined parameters for the use of TiO 2 P25 in A549 cells were met, similar results on the biological effects of TiO 2 P25 were obtained in two independent cell laboratories. Conclusion We optimized the experimental conditions of TiO 2 P25 preparation for toxicity testing in A549 cells. The results presented here on TiO 2 P25-induced cellular effects are reproducible. Therefore, our results can be helpful for other researchers using TiO 2 P25 as a reference material.

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Effects of Charged Oxime Reactivators on the HK-2 Cell Line in Renal Toxicity Screening

Cited by 10 publications

References 27 publications

In Vitro Evaluation of Oxidative Stress Induced by Oxime Reactivators of Acetylcholinesterase in HepG2 Cells

In Vitro Evaluation of Oxidative Stress Induced by Oxime Reactivators of Acetylcholinesterase in HepG2 Cells

Potential of Vitamin B6 Dioxime Analogues to Act as Cholinesterase Ligands

Evaluating the Use of TiO2 Nanoparticles for Toxicity Testing in Pulmonary A549 Cells

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