Evaluation of Viability and Proliferation Profiles on Macrophages Treated with Silica Nanoparticles <i>In Vitro</i> via Plate-Based, Flow Cytometry, and Coulter Counter Assays

Bancos, Simona; Tsai, De‐Hao; Hackley, Vincent A.; Weaver, James L.; Tyner, Katherine M.

doi:10.5402/2012/454072

Cited by 27 publications

(16 citation statements)

References 44 publications

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“…In live dopaminergic neurons, lactate dehydrogenase is contained within the cell; however, upon membrane perforation, it is released into the cytoplasm. Thereby, lactate dehydrogenase levels in the cell supernatant are a reliable indicator of injury of dopaminergic neurons (22). Furthermore, reverse transcriptase polymerase chain reaction and western blot were also applied in the present study.…”

Section: Discussionmentioning

confidence: 99%

Neurotoxicity of cerebro-spinal fluid from patients with Parkinson’s disease on mesencephalic primary cultures as an in vitro model of dopaminergic neurons

Kong

Zhang

Lei

et al. 2015

Molecular Medicine Reports

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Abstract.Parkinson's disease is a degenerative disorder of the central nervous system. In spite of extensive research, neither the cause nor the mechanisms have been firmly established thus far. One assumption is that certain toxic substances may exist in the cerebro-spinal fluid (CSF) of Parkinson's disease patients. To confirm the neurotoxicity of CSF and study the potential correlation between neurotoxicity and the severity of Parkinson's disease, CSF was added to cultured cells. By observation of cell morphology, changes in the levels of lactate dehydrogenase, the ratio of tyrosine hydroxylase-positive cells, and the expression of tyrosine hydroxylase mRNA and protein, the differences between the two groups were shown. The created in vitro model of dopaminergic neurons using primary culture of mouse embryonic mesencephalic tissue is suitable for the study of neurotoxicity. The observations of the present study indicated that CSF from Parkinson's disease patients contains factors that can cause specific injury to cultured dopaminergic neurons. However, no obvious correlation was found between the neurotoxicity of CSF and the severity of Parkinson's disease.

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Section: Discussionmentioning

confidence: 99%

Neurotoxicity of cerebro-spinal fluid from patients with Parkinson’s disease on mesencephalic primary cultures as an in vitro model of dopaminergic neurons

Kong

Zhang

Lei

et al. 2015

Molecular Medicine Reports

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“…NPs were opsonized by serum proteins and agglomerated over time when exposed to cell culture media. 21,22 All NPs tested below the detection limit for endotoxin (0.25 EU/mL) for the concentrations used in this study.…”

Section: Nps Characterizationmentioning

confidence: 99%

“…In addition, a suspended culture was utilized for the majority of experiments for several reasons: 1) The suspended state tends to reduce NP assay interference. 21 2) In adherent cell cultures, the exact number of cells is not known at the initiation of the experiment. By utilizing cell suspensions, the amount of cells used in the assay may be fixed, thus more tightly controlling the ratio of cells/NP.…”

mentioning

confidence: 99%

“…28,29 Particle size and zeta potential were characterized in both water and relevant testing conditions (ie, cell culture media, Table S1). 21,22 Note that both Z-average (Z-Ave) and intensityweighted average are complicated by the presence of serum proteins in the media. With the exception of the SiO 2 NPs (10 nm), all NPs were initially dispersed as individual NPs.…”

mentioning

confidence: 99%

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Effect of silica and gold nanoparticles on macrophage proliferation, activation markers, cytokine production, and phagocytosis in vitro

Bancos¹,

Stevens²,

Tyner³

2014

IJN

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The accumulation of durable nanoparticles (NPs) in macrophages following systemic administration is well described. The ultimate biological impact of this accumulation on macrophage function, however, is not fully understood. In this study, nontoxic doses of two durable NPs, SiO 2 and Au, at particle sizes of ~10 nm and 300 nm were used to evaluate the effect of bioaccumulation on macrophage function in vitro using RAW 264.7 mouse macrophage-like cells as a model system. Cell proliferation, cell cycle, cytokine production, surface marker activation, and phagocytosis responses were evaluated through a panel of assays using flow cytometry and confocal microscopy. The most dramatic change in RAW 264.7 cell function was a reduction in phagocytosis as monitored by the uptake of Escherichia coli . Cells exposed to both 10 nm Au NPs and 10 nm SiO 2 NPs showed ~50% decrease in phagocytosis, while the larger NPs caused a less dramatic reduction. In addition to modifying phagocytosis profiles, 10 nm SiO 2 NPs caused changes in proliferation, cell cycle, and cell morphology. Au NPs had no effect on cell cycle, cytokine production, or surface markers and caused interference in phagocytosis in the form of quenching when the assay was performed via flow cytometry. Confocal microscopy analysis was used to minimize this interference and demonstrated that both sizes of Au NPs decreased the phagocytosis of E. coli . Overall, our results demonstrate that Au and SiO 2 NP uptake by macrophages can influence macrophage phagocytosis in vitro without altering surface markers and cytokine production in vitro. While the biological impact of these findings remains unclear, our results indicate that bioaccumulation of durable NPs within the macrophages may lead to a suppression of bacterial uptake and possibly impair bactericidal activity.

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“…As it has been shown that NMs can interact with LDH and decrease enzyme activity, [108] likely through structural deformation of the enzyme, this could result in inaccurate results regarding cell viability whereby toxicity would be masked. [84,85] NM generation of ROS The physicochemical properties of some NMs enable them to undergo oxidation and produce ROS that can potentially reduce dyes to their fluorescent forms, [92,100] resulting in false positive tests of toxicity. For example, incubation of carbon-based NMs with 2 0 ,7 0 -dichlorodihydrofluorescin diacetate (DCFH-DA) (a highly oxidisable dye that is a commonly used as a marker of oxidative stress) results in the production of fluorescent 2 0 ,7 0 -dichlorodihydrofluorescein (DCF) molecules even when cells are not present (Fig.…”

Section: Optical Interferencementioning

confidence: 99%

Aquatic toxicity of manufactured nanomaterials: challenges and recommendations for future toxicity testing

et al. 2014

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Environmental context. The increased use of nanomaterials in industrial and consumer products requires robust strategies to identify risks when they are released into the environment. Aquatic toxicologists are beginning to possess a clearer understanding of the chemical and physical properties of nanomaterials in solution, and which of the properties potentially affect the health of aquatic organisms. This review highlights the main challenges encountered in aquatic nanotoxicity testing, provides recommendations for overcoming these challenges, and discusses recent studies that have advanced our understanding of the toxicity of three important OECD nanomaterials, titanium dioxide, zinc oxide and silver nanomaterials.Abstract. Aquatic nanotoxicologists and ecotoxicologists have begun to identify the unique properties of the nanomaterials (NMs) that potentially affect the health of wildlife. In this review the scientific aims are to discuss the main challenges nanotoxicologists currently face in aquatic toxicity testing, including the transformations of NMs in aquatic test media (dissolution, aggregation and small molecule interactions), and modes of NM interference (optical interference, adsorption to assay components and generation of reactive oxygen species) on common toxicity assays. Three of the major OECD (Organisation for Economic Co-operation and Development) priority materials, titanium dioxide (TiO 2 ), zinc oxide (ZnO) and silver (Ag) NMs, studied recently by the Natural Sciences and Engineering Research Council of Canada (NSERC), National Research Council of Canada (NRC) and the Business Development Bank of Canada (BDC) Nanotechnology Initiative (NNBNI), a Canadian consortium, have been identified to cause both bulk effect, dissolution-based (i.e. free metal), or NM-specific toxicity in aquatic organisms. TiO 2 NMs are most toxic to algae, with toxicity being NM size-dependent and principally associated with binding of the materials to the organism. Conversely, dissolution of Zn and Ag NMs and the subsequent release of their ionic metal counterparts appear to represent the primary mode of toxicity to aquatic organisms for these NMs. In recent years, our understanding of the toxicological properties of these specific OECD relevant materials has increased significantly. Specifically, researchers have begun to alter their experimental design to identify the different behaviour of these materials as colloids and, by introducing appropriate controls and NM characterisation, aquatic nanotoxicologists are now beginning to possess a clearer understanding of the chemical and physical properties of these materials in solution, and how these materials may interact with organisms. Arming nanotoxicologists with this understanding, combined with knowledge of the physics, chemistry and biology of these materials is essential for maintaining the accuracy of all future toxicological assessments.

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Evaluation of Viability and Proliferation Profiles on Macrophages Treated with Silica Nanoparticles In Vitro via Plate-Based, Flow Cytometry, and Coulter Counter Assays

Cited by 27 publications

References 44 publications

Neurotoxicity of cerebro-spinal fluid from patients with Parkinson’s disease on mesencephalic primary cultures as an in vitro model of dopaminergic neurons

Neurotoxicity of cerebro-spinal fluid from patients with Parkinson’s disease on mesencephalic primary cultures as an in vitro model of dopaminergic neurons

Effect of silica and gold nanoparticles on macrophage proliferation, activation markers, cytokine production, and phagocytosis in vitro

Aquatic toxicity of manufactured nanomaterials: challenges and recommendations for future toxicity testing

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