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Methods and Protocols for In Vitro Animal Nanotoxicity Evaluation: A Detailed Review
Published in Vineet Kumar, Nandita Dasgupta, Shivendu Ranjan, Nanotoxicology, 2018
Venkatraman Manickam, Leema George, Amiti Tanny, Rajeeva Lochana, Ranjith Kumar Velusamy, M. Mathan Kumar, Bhavapriya Rajendran, Ramasamy Tamizhselvi
Oxidative damage specific markers analyzed during toxic response include malondialdehyde (MDA), 8-epi-PGF2α, and 8-hydroxy-2′-deoxyguanosine (8-oxo-dG) concentrations. Likewise, protein based markers heat shock protein-A1A and heme oxygenase-1 are routinely studied during toxic assays. Using these markers, oxidative changes induced by silver nanoparticles in A549 and HepG2 cells were recently reported (Xin et al. 2014). Lethal dose-50 (LD-50) value in a cellular system during toxicity evaluation is a preliminary factor to be evaluated during the viability assays. LD-50 is the lethal exposure dose or concentration of nanomaterial which leads to the death of 50% of the cells in the experiment. Acute or chronic exposure of the LD-50 value could provide us with information regarding cytotoxic response and help in choosing the relevant model system for further in vitro studies. Though MTT is the most popular and traditional method for evaluating cell proliferation and thus cytotoxicity (Figure 12.9), there are some limitations with nanomaterials like graphene-related materials and carbon nanotubes. So WST-8, an alternate tetrazolium salt based assay, is preferred in place of MTT based toxicity evaluations (Liao et al. 2011).
In Vivo Evaluation of the Hepatonephrotoxicity of Polymeric Nanoparticles in Rats
Published in Bertrand Henri Rihn, Biomedical Application of Nanoparticles, 2017
Mosaad A. Abdel-Wahhab, Olivier Joubert, Aziza A. El-Nekeety, Khaled G. Abdel-Wahhab, Carole Ronzani, Ramia Safar, Ahmed A. El-Kady, Nabila S. Hassan, Fathia A. Mannaa, Bertrand Henri Rihn
The present study also showed that MDA in the liver or kidney tissues was disturbed at all time periods of PO or IP administration but it was in the normal range by 3 weeks. However, TAC in serum, liver, or kidney tissue showed a significant increase in serum after 4 hours and 1 week, in the liver at all time points, and a significant decrease in kidney tissue at 4 hours, but was in the normal range thereafter. A previous study revealed that NPs may enter the circulation and produce adverse effects on different organs (Mohanan and Rathinam 1995), especially the liver, a major site where endogenous and exogenous substances accumulate. Free radicals are generated during the detoxification of reactive metabolites by cytochrome P-450 located in the smooth endoplasmic reticulum of hepatocytes and also by NADPH oxidase enzyme (Oesch et al. 1985) and also by NADPH oxidase enzyme (Eloisa et al. 1995) in the activated Kupffer cells (Wheeler et al. 2001). Reactive species, above a threshold, oxidize polyunsaturated fatty acids resulting in the onset of lipid peroxidation (Zacharias 2011), giving rise to peroxyl and alkoxyl radicals, the primary end products. Malondialdehyde is one of the toxic secondary products of lipid peroxidation which can diffuse and impairs cellular constituents such as lipids, nucleic acids, and proteins by their electrophilic nature, leading to cell death (Jain et al. 2008).
Phyto-Nano Interaction
Published in Ramesh Raliya, Nanoscale Engineering in Agricultural Management, 2019
Divya Vishambhar Kumbhakar, Debadrito Das, Bapi Ghosh, Ankita Pramanik, Sudha Gupta, Animesh Kumar Datta
NP-treated cells generate oxidative stress, showing an increase in concentration of malondialdehyde (MDA). MDA is the outcome of enhanced decomposition of polyunsaturated fatty acids in membranes as an indicator of lipid peroxidation conferring membrane damage (Mittler 2002, Tanou et al. 2009, Halliwell and Gutteridge 2015). H2O2 (primary indicator of stress accumulation due to its enhanced half-life) and MDA show differential elevation (Majumdar et al. 2014, Yang et al. 2017) in different plant species following NPs treatments, suggesting asynchrony in host response to NP-induced oxidative stress.
Dietary supplementation of garlic, propolis, and wakame improves recuperation in cadmium exposed Japanese medaka fish (Oryzias latipes)
Published in Journal of Environmental Science and Health, Part A, 2020
Henry Okechukwu Ujeh, Masaaki Kurasaki
The reactive carbonyl compound malondialdehyde (MDA) is one of the major compounds formed during lipid peroxidation. The reaction of MDA and TBA under high temperature (90–100 °C) and acidic conditions forms a MDA-TBA adduct, which is measured colorimetrically at 530–540 nm. Assay was performed with the TBARS (TCA Method) Assay Kit (Caymen Chemical, Ann Arbor, MI) following manufacturer’s protocol. Briefly, 100 µL of sample or standard was added to appropriately labeled vial. 100 µL of TCA assay reagent (10%) was added to vial and swirled to mix. Then, 800 µL of provided color reagent was added to each vial and vortexed. Vials were caped and heated in an upright position in vigorously boiling water for about one hour. After one hour, vials were immediately removed and placed in ice bath to stop reaction, with further 10 min incubation on ice. Vials were then centrifuged for 10 min at 1,600 x g at 4 °C. Two hundred µL of supernatants were carefully removed from each vial in duplicate and transferred to clear 96 well plate, and absorbance read at 540 nm. MDA values for each sample were calculated from standard curve using the equation:
Exposure of tomato (Lycopersicon esculentum) to silver nanoparticles and silver nitrate: physiological and molecular response
Published in International Journal of Phytoremediation, 2020
Azam Noori, Trevor Donnelly, Joseph Colbert, Wenjun Cai, Lee A. Newman, Jason C. White
Malondialdehyde (MDA) is the product of lipid peroxidation and is an indicator of oxidative stress. The method described by Vos et al. (1991) was used to determine the amount of MDA. Briefly, 1 g leaves were ground in liquid nitrogen and were transferred into 15 mL polypropylene tubes amended with 3 mL of TCA (20%) and 1 mL of 0.5% thiobarbituric acid. The tubes were incubated in a water bath at 100 °C for 15 min and were then cooled quickly using tap water. All samples were centrifuged at 12,000 g for 15 min. The supernatant was transferred into a new polypropylene tube and the final volume was adjusted to 10 mL by adding dH2O. The absorbance was recorded at 532 and 600 nm using the aforementioned spectrophotometer. The amount of MDA was calculated based on the dimethyl acetal (malonaldehyde bis) standard and reported as µM/g FW.
Phytoremediation of Heavy Metal- Contaminated Tailings Soil by Symbiotic Interaction of Cymbopogon Citratus and Solanum Torvum with Bacillus Cereus T1B3
Published in Soil and Sediment Contamination: An International Journal, 2019
A.K. Nayak, A. Basu, S.S. Panda, N.K. Dhal, R. K. Lal
All plants produce different antioxidant enzymes like catalase (CAT) and superoxide dismutase (SOD) for protection from different induced oxidative injury under metal stress condition (Guo, Zhang, and Zhang 2006). By inoculation of T1B3 strain, CAT and SOD activities increased significantly in both C. citratus and S. torvum as (77 and 35%) and (36 and 35%) respectively compared to uninoculated controls (Tables 3 and 4). Malondialdehyde (MDA) is a product of lipid peroxidation and an oxidative stress indicator (Yang et al. 2010). In present study, MDA contents in both inoculated C. citratus and S. torvum plants decreased 27 and 45% respectively, as compared to uninoculated controls when both grown in contaminated tailings (Tables 3 and 4). The above results indicate that T1B3 strain can play an important role in the antioxidative defense mechanism for both the plants in HMs stress conditions. Different functional bacteria have the capacity to reduce phytotoxicity, increase HM uptake and subsequently enhance plant growth in HM-contaminated soil (Rajkumar et al. 2009). In this study, by inoculating the T1B3 strain, both (C. citratus and S. torvum) plants accumulated appreciable amount of HMs. In comparison to uninoculated plants, inoculated plants accumulated more HMs in roots and shoots (Table 5).