Neutral red uptake assay for the estimation of cell viability/cytotoxicity

Repetto, Guillermo; Peso, Ana del; Zurita, Jorge L.

doi:10.1038/nprot.2008.75

Cited by 1,815 publications

(1,270 citation statements)

References 14 publications

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“…This is supported by a previous study where Co not affected the viability of THP‐1 cells in concentrations up to 200 µM 31. As cytotoxicity test, NRU has previously been validated as a good technique for quantification of viable cells 44. Our results showed that IL‐1β released in the protein detection analysis was measured in noncytotoxic conditions.…”

Section: Discussionsupporting

confidence: 88%

Effect of cobalt ions on the interaction between macrophages and titanium

Pettersson

Thorén

et al. 2018

J Biomedical Materials Res

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Inflammation and bone reduction around dental implants are described as peri‐implantitis and can be caused by an inflammatory response against bacterial products and toxins. Titanium (Ti) forms aggregates with serum proteins, which activate and cause release of the cytokine interleukin (IL‐1β) from human macrophages. It was hypothesized that cobalt (Co) ions can interact in the formation of pro‐inflammatory aggregates, formed by titanium. To test this hypothesis, we differentiated THP‐1 cells into macrophages and exposed them to Ti ions alone or in combination with Co ions to investigate if IL‐1β release and cytotoxicity were affected. We also investigated aggregate formation, cell uptake and human biopsies with inductively coupled plasma atomic emission spectroscopy and electron microscopy. Co at a concentration of 100 µM neutralized the IL‐1β release from human macrophages and affected the aggregate formation. The aggregates formed by Ti could be detected in the cytosol of macrophages. In the presence of Co, the Ti‐induced aggregates were located in the cytosol of the cultured macrophages, but outside the lysosomal structures. It is concluded that Co can neutralize the Ti‐induced activation and release of active IL‐1β from human macrophages in vitro. Also, serum proteins are needed for the formation of metal‐protein aggregates in cell medium. Furthermore, the structures of the aggregates as well as the localisation after cellular uptake differ if Co is present in a Ti solution. Phagocytized aggregates with a similar appearance seen in vitro with Ti present, were also visible in a sample from human peri‐implant tissue. © 2018 The Authors Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A:2518–2530, 2018.

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Section: Discussionsupporting

confidence: 88%

Effect of cobalt ions on the interaction between macrophages and titanium

Pettersson

Thorén

et al. 2018

J Biomedical Materials Res

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“…This assay is based on the ability of viable cells to incorporate and bind the supravital dye neutral red in their lysosomes [29]. Differentiated LUHMES were seeded in 96 well culture plates and cultivated under differentiating conditions for 4 days as previously described.…”

Section: Lysosomal Integritymentioning

confidence: 99%

Toxicity of organic and inorganic mercury species in differentiated human neurons and human astrocytes

Lohren

Blagojevic

Fitkau

et al. 2015

Journal of Trace Elements in Medicine and Biology

106

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a b s t r a c tOrganic mercury (Hg) species exert their toxicity primarily in the central nervous system. The food relevant Hg species methylmercury (MeHg) has been frequently studied regarding its neurotoxic effects in vitro and in vivo. Neurotoxicity of thiomersal, which is used as a preservative in medical preparations, is to date less characterised. Due to dealkylation of organic Hg or oxidation of elemental Hg, inorganic Hg is present in the brain albeit these species are not able to readily cross the blood brain barrier. This study compared for the first time toxic effects of organic MeHg chloride (MeHgCl) and thiomersal as well as inorganic mercury chloride (HgCl 2 ) in differentiated human neurons (LUHMES) and human astrocytes (CCF-STTG1). The three Hg species differ in their degree and mechanism of toxicity in those two types of brain cells. Generally, neurons are more susceptible to Hg species induced cytotoxicity as compared to astrocytes. This might be due to the massive cellular mercury uptake in the differentiated neurons. The organic compounds exerted stronger cytotoxic effects as compared to inorganic HgCl 2 . In contrast to HgCl 2 exposure, organic Hg compounds seem to induce the apoptotic cascade in neurons following low-level exposure. No indicators for apoptosis were identified for both inorganic and organic mercury species in astrocytes. Our studies clearly demonstrate species-specific toxic mechanisms. A mixed exposure towards all Hg species in the brain can be assumed. Thus, prospectively coexposure studies as well as cocultures of neurons and astrocytes could provide additional information in the investigation of Hg induced neurotoxicity.

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“…The cytotoxicity of rosuvastatine, doxazosin, repaglinide and oxcarbazepin was tested against MCF-7, HeLa and HepG2 cells by the neutral red assay as previously described (Repetto et al, 2008). Exponentially growing cells were collected using 0.25% Trypsin-EDTA and plated in 96-well plates at 1000-2000 cells/well.…”

Section: Cytotoxicity Assaysmentioning

confidence: 99%