Detection and quantification of nanoparticle-induced intracellular ROS in live cells by laser scanning confocal microscopy

Amaldoss, Maria John Newton; Pandžić, Elvis; Koshy, Pramod; Kumar, Naresh; Sorrell, Charles C.; Unnikrishnan, Ashwin

doi:10.1016/j.ymeth.2022.08.005

Cited by 7 publications

(5 citation statements)

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“…Prior studies have suggested that the mechanism by which gold, silver, and platinum NPs inhibit growth involves the generation of reactive oxygen species (ROS) [ 32 ]. To test whether the same mechanism of action is true for iron oxide NPs, we examined the role of ROS by adding the antioxidant Trolox [ 33 , 34 ] to the Toxoplasma growth inhibition assay medium.…”

Section: Resultsmentioning

confidence: 99%

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Goethite and Hematite Nanoparticles Show Promising Anti-Toxoplasma Properties

Ishii,

Akahoshi,

Adeyemi

et al. 2024

Pharmaceutics

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Toxoplasma gondii is an intracellular parasitic protozoan with a high infection rate in mammals, including humans, and birds. There is no effective vaccine, and treatment relies on antiparasitic drugs. However, existing antiprotozoal drugs have strong side effects and other problems; therefore, new treatment approaches are needed. Metal nanoparticles have attracted increased interest in the biomedical community in recent years because of their extremely high surface area to volume ratio and their unique reactivity that could be exploited for medicinal purposes. Previously, we confirmed the anti-Toxoplasma effects of gold, silver, and platinum nanoparticles, in a growth inhibition test. Here, we asked whether the anti-Toxoplasma effect could be confirmed with less expensive metal nanoparticles, specifically iron oxide nanoparticles (goethite and hematite). To improve the selective action of the nanoparticles, we modified the surface with l-tryptophan as our previous findings showed that the bio-modification of nanoparticles enhances their selectivity against T. gondii. Fourier-Transform Infrared Spectroscopy (FTIR) analysis confirmed the successful coating of the iron oxide nanoparticles with l-tryptophan. Subsequently, cytotoxicity and growth inhibition assays were performed. L-tryptophan-modified nanoparticles showed superior anti-Toxoplasma action compared to their naked nanoparticle counterparts. L-tryptophan enhanced the selective toxicity of the iron oxide nanoparticles toward T. gondii. The bio-modified nanoparticles did not exhibit detectable host cell toxicity in the effective anti-Toxoplasma doses. To elucidate whether reactive oxygen species contribute to the anti-Toxoplasma action of the bio-modified nanoparticles, we added Trolox antioxidant to the assay medium and found that Trolox appreciably reduced the nanoparticle-induced growth inhibition.

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Section: Resultsmentioning

confidence: 99%

“…Prior studies have suggested that the mechanism of action of gold, silver, and platinum NPs involves decreasing the mitochondrial membrane potential by generating ROS [ 32 ]. We confirmed the effect of adding Trolox, an antioxidant, on the relative viability of Toxoplasma in the presence of the NPs.…”

Section: Discussionmentioning

confidence: 99%

Goethite and Hematite Nanoparticles Show Promising Anti-Toxoplasma Properties

Ishii,

Akahoshi,

Adeyemi

et al. 2024

Pharmaceutics

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“…Acoustic spectroscopy AS [51][52][53] Analytical electron microscopy AEM [54,55] Asymmetric flow field-flow fractionation AF 4 [56,57] Atmospheric pressure photo-ionization mass spectrometry APPI-MS [58,59] Atomic force microscopy AFM [60,61] Capillary electrophoresis CE [62,63] Confocal laser scanning microscopy CLSM [64,65] Cross-flow filtration CFF [66,67] Table 2. Cont.…”

Section: Methods Name Abbreviation Referencesmentioning

confidence: 99%

Characterization of Nanoparticles in Antimicrobial Coatings for Medical Applications—A Review

Rezić,

Meštrović

2023

Coatings

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This review discusses relevant topics concerning the understanding of the characterization of antimicrobial coatings due to powerful antimicrobial nanoparticles in their composition. These coatings are utilized in the surface modification of yarns and materials designed for use in medical and dental applications. Various physical and chemical methods are employed to create these coatings, ensuring the development of efficient, homogeneous, and uniform layers on diverse surfaces and materials. The primary objective is to confer antimicrobial and/or antiviral properties upon these materials. For these coatings to be effective, they must incorporate active compounds that can combat a wide array of microorganisms, including those that have developed resistance to antibiotics. Examples of such active compounds include metallic nanoparticles such as silver, copper, and gold, as well as nanoparticles of metal oxides such as zinc, titanium, and aluminum. Upon the application of these coatings to medical materials, extensive testing and characterization procedures are undertaken, which will be thoroughly detailed in this review. It is crucial to emphasize that the absence of proper characterization and testing of nanoparticles in antimicrobial coatings could lead to the absence of standards, norms, or procedures necessary to safeguard human health and the environment. Despite their widespread application in the medical field, concerns have been raised regarding the potential toxicity of nanoparticles to living organisms. Consequently, this paper provides a comprehensive overview of the current state-of-the-art methodologies for characterizing nanoparticles in antimicrobial coatings, specifically focusing on materials with varying roughness and structures. Additionally, it outlines the issues associated with the potential accumulation of antimicrobial nanoparticles within the human body.

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“…Parameters such as selection of the region of interest (ROI), filtering background noise, focus and threshold intensity were set to avoid excessive or weak signal. All quantitation and analysis were performed using three images per treatment (Amaldoss et al, 2022;Deshpande et al, 2021). All treatments (n = 21) encompassing control, NPs and Cu individually and all their combinations were assessed.…”

Section: Intracellular Reactive Oxygen Speciesmentioning

confidence: 99%

“…Intracellular ROS production, plasma membrane disruption and fungal biomass were assessed by one-way factorial permutational multivariate analysis of variance (PERMANOVA, α ≤ 0.05) with treatments as a factor (Amaldoss et al, 2022;Deshpande et al, 2021). Matrices of similarity were based on Bray-Curtis similarity with 9999 permutations and Type III sums of squares (Anderson, 2017;Anderson et al, 2008).…”

Section: Statistical Analysesmentioning

confidence: 99%