The use of in vitro assays for the assessment of cytotoxicity on the example of MTT test

Twarużek, Magdalena; Zastempowska, Ewa; Soszczyńska, Ewelina; Ałtyn, Iwona

doi:10.1515/fobio-2017-0006

Cited by 15 publications

(18 citation statements)

References 5 publications

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“…The tetrazolium salt (MTT) (Life Technologies Corporation, Eugene, OR, USA) assay, which is based on the cellular conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide into formazan, was performed to determine the cell viability after the cell exposure to FB1 and AFB1, while the sulforhodamine B (Sigma-Aldrich, Co., St. Louis, MO, USA) colorimetric assay (SRB) was conducted to measure the protein content in the Caco-2 and HepG2 cells [71,72]. Briefly, Caco-2 and HepG2 cells were seeded in 96-well plates at a density of 20,000 per well for 24 h before the treatment at 37 °C in a sterilized incubator with a humidified atmosphere of 5% CO 2 to allow the cells' adhesion.…”

Section: Cell Viability and Protein Content Assaysmentioning

confidence: 99%

Bioenergetic Status of the Intestinal and Hepatic Cells after Short Term Exposure to Fumonisin B1 and Aflatoxin B1

Chen

Abdallah

Grootaert

et al. 2022

IJMS

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Fumonisin B1 (FB1) and aflatoxin B1 (AFB1) are frequent contaminants of staple foods such as maize. Oral exposure to these toxins poses health hazards by disrupting cellular signaling. However, little is known regarding the multifaced mitochondrial dysfunction-linked toxicity of FB1 and AFB1. Here, we show that after exposure to FB1 and AFB1, mitochondrial respiration significantly decreased by measuring the oxygen consumption rate (OCR), mitochondrial membrane potential (MMP) and reactive oxygen species (ROS). The current work shows that the integrity of mitochondria (MMP and ROS), that is the central component of cell apoptosis, is disrupted by FB1 and AFB1 in undifferentiated Caco-2 and HepG2 cells as in vitro models for human intestine and liver, respectively. It hypothesizes that FB1 and AFB1 could disrupt the mitochondrial electron transport chain (ETC) to induce mitochondrial dysfunction and break the balance of transferring H+ between the mitochondrial inner membrane and mitochondrial matrix, however, the proton leak is not increasing and, as a result, ATP synthesis is blocked. At the sub-toxic exposure of 1.0 µg/mL for 24h, i.e., a viability of 95% in Caco-2 and HepG2 cells, the mitochondrial respiration was, however, stimulated. This suggests that the treated cells could reserve energy for mitochondrial respiration with the exposure of FB1 and AFB1, which could be a survival advantage.

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Section: Cell Viability and Protein Content Assaysmentioning

confidence: 99%

Bioenergetic Status of the Intestinal and Hepatic Cells after Short Term Exposure to Fumonisin B1 and Aflatoxin B1

Chen

Abdallah

Grootaert

et al. 2022

IJMS

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“…The colorimetric MTT assay is characterized by high sensitivity and therefore it can be used to study a large number of different samples 21 . The amount of MTT assay is proportional to the number of metabolically active cell cultures.…”

Section: Resultsmentioning

confidence: 99%

“…They can divide about 50 times with a culture medium that is frequently changed. Samples with absorption values less than 50% of division activity are considered toxic 21 . In our studies, the biocompatible effect of the samples was studied by indirect contact 19 .…”

Section: Resultsmentioning

confidence: 99%

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Evaluation of the biocompability and corrosion activity of resorbable CaMgZnYbBAu alloys

Szyba

Kubina

Młynarek-Żak

et al. 2022

Sci Rep

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Calcium-based alloys can be promising candidates for use as biodegradable implants because of attractive properties as mechanical, corrosive, and biocompatible. In the work, the biocompatibility authors discussed the results of the Ca32Mg12Zn38Yb18−xBx (x = 0, 1, 2, 3 at.%) and Ca32Mg12Zn38Yb18−2xBxAux (x = 1, 2 at.%) alloys. The tests were performed using a MTT assay. The corrosion behavior of such Ca-based alloys in PWE fluid at 37 °C was studied and compared with the results in Ringer’s solution from previous works. Electrochemical tests were presented by open circuit potential and potentiodynamic curves. Different concentrations of boron and gold in the alloys caused changes in the corrosion results. The best corrosion resistance in PWE solution was observed for the Ca-based alloy with 2 at.% Au due to the lowest value of the corrosion current density (jcorr), equal to 10.6 µA·cm−2. A slightly higher value of jcorr was obtained for the Ca32Mg12Zn38Yb15B3 alloy with the lowest roughness values. The results of the cytotoxicity tests also showed that the alloy with 3 at.% boron was characterized by the highest cell viability. The investigation results discussed in the work allow us to suggest that the presented calcium alloys with 3 at.% of B, and 2 at.% of Au addition may be promising materials for the use in implantology.

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“…In the present studies, the cytotoxic activity of the synthesized carriers was investigated using two healthy skin cell lines, i.e., normal human dermal fibroblasts (NHDF), human epidermal keratinocytes (HaCaT), and two skin melanoma cell lines, i.e., malignant melanoma cells (Me45) and human metastatic melanoma variant of WM164 cell line (451-Lu). The viability of cells evaluated by MTT assay, which is based on the reduction of the bromide salt of tetrazolium dye to insoluble formazan with a purple color [36,37], was performed at various solution concentrations of carriers with active substances (100-1.5 µg/mL). The MTT test was accomplished after 72 h of cells incubation with carrier solutions and allowed a toxicity assessment of the carriers even after prolonged contact with skin cells.…”

Section: Cytotoxicitymentioning

confidence: 99%

PEG Graft Polymer Carriers of Antioxidants: In Vitro Evaluation for Transdermal Delivery

2020

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The in vitro biochemical evaluation of the applicability of polymers carrying active substances (micelles and conjugates) was carried out. Previously designed amphiphilic graft copolymers with retinol or 4-n-butylresorcinol functionalized polymethacrylate backbone and poly(ethylene glycol) (PEG) side chains that included Janus-type heterografted copolymers containing both PEG and poly(ε-caprolactone) (PCL) side chains were applied as micellar carriers. The polymer self-assemblies were convenient to encapsulate arbutin (ARB) as the selected active substances. Moreover, the conjugates of PEG graft copolymers with ferulic acid (FA) or lipoic acid (LA) were also investigated. The permeability of released active substances through a membrane mimicking skin was evaluated by conducting transdermal tests in Franz diffusion cells. The biological response to new carriers with active substances was tested across cell lines, including normal human dermal fibroblasts (NHDF), human epidermal keratinocyte (HaCaT), as well as cancer melanoma (Me45) and metastatic human melanoma (451-Lu), for comparison. These polymer systems were safe and non-cytotoxic at the tested concentrations for healthy skin cell lines according to the MTT test. Cytometric evaluation of cell cycles as well as cell death defined by Annexin-V apoptosis assays and senescence tests showed no significant changes under action of the delivery systems, as compared to the control cells. In vitro tests confirmed the biochemical potential of these antioxidant carriers as beneficial components in cosmetic products, especially applied in the form of masks and eye pads.

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The use of in vitro assays for the assessment of cytotoxicity on the example of MTT test

Cited by 15 publications

References 5 publications

Bioenergetic Status of the Intestinal and Hepatic Cells after Short Term Exposure to Fumonisin B1 and Aflatoxin B1

Bioenergetic Status of the Intestinal and Hepatic Cells after Short Term Exposure to Fumonisin B1 and Aflatoxin B1

Evaluation of the biocompability and corrosion activity of resorbable CaMgZnYbBAu alloys

PEG Graft Polymer Carriers of Antioxidants: In Vitro Evaluation for Transdermal Delivery

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