Cellular Models for In Vitro Toxicity Testing

Zucco, Flavia; Angelis, Isabella De; Stammati, Annalaura

doi:10.1007/978-3-642-80412-0_21

Cited by 14 publications

(4 citation statements)

References 24 publications

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“…Cell culture V79 Chinese hamster lung fibroblast cells (MZ subline (60)) were used for the cytotoxicity assays (61). They were routinely cultured in 175 cm 2 tissue flasks in Ham's F-10 medium, supplemented with 10% newborn calf serum and 1% penicillin/streptomycin, in a humidified incubator at 37 C with 5% CO 2 .…”

Section: Cytotoxicity Assaysmentioning

confidence: 99%

Synergistic Effects of the Membrane Actions of Cecropin-Melittin Antimicrobial Hybrid Peptide BP100

Ferré

Melo

Correia

et al. 2009

Biophysical Journal

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BP100 (KKLFKKILKYL-NH(2)) is a short cecropin A-melittin hybrid peptide, obtained through a combinatorial chemistry approach, which is highly effective in inhibiting both the in vitro and in vivo growth of economically important plant pathogenic Gram-negatives. The intrinsic Tyr fluorescence of BP100 was taken advantage of to study the peptide's binding affinity and damaging effect on phospholipid bilayers modeling the bacterial and mammalian cytoplasmic membranes. In vitro cytotoxic effects of this peptide were also studied on mammalian fibroblast cells. Results show a stronger selectivity of BP100 toward anionic bacterial membrane models as indicated by the high obtained partition constants, one order of magnitude greater than for the neutral mammalian membrane models. For the anionic systems, membrane saturation was observed at high peptide/lipid ratios and found to be related with BP100-induced vesicle permeabilization, membrane electroneutrality, and vesicle aggregation. Occurrence of BP100 translocation was unequivocally detected at both high and low peptide/lipid ratios using a novel and extremely simple method. Moreover, cytotoxicity against mammalian models was reached at a concentration considerably higher than the minimum inhibitory concentration. Our findings unravel the relationships among the closely coupled processes of charge neutralization, permeabilization, and translocation in the mechanism of action of antimicrobial peptides.

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Section: Cytotoxicity Assaysmentioning

confidence: 99%

Synergistic Effects of the Membrane Actions of Cecropin-Melittin Antimicrobial Hybrid Peptide BP100

Ferré

Melo

Correia

et al. 2009

Biophysical Journal

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“…Established cell lines are generally used for preliminary toxicity studies, because they are a more standardised model than animal cells and other types of cell cultures, such as primary cultures or freshly isolated cells, and they allow the investigation of possible effects on basic cell functions and structures. Basic toxicity endpoints deal with cell viability, cell proliferation, cell energy consumption, and cellular membrane integrity (7,8).…”

mentioning

confidence: 99%

Cytostatic Properties of a Novel Compound Derived from Penicillium pinophilum: An In Vitro Study

Stammati

Nicoletti²,

Stefano³

et al. 2002

Altern Lab Anim

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3-O-Methylfunicone (OMF), a secondary metabolite produced by Penicillium pinophilum, inhibits the in vitro growth of plant pathogenic fungi. This specific property suggested that the compound could be used against other fungal pathogenic activities, including dermatological ones. However, for such applications, toxicological side-effects should be taken into account, in order to prevent other types of risk to mammalian cells. Therefore, investigations were made of the basic toxicity of OMF toward a human tumour cell line. The compound was found to have a cytostatic effect, which represents a counter-indication to its use as a therapeutic agent in dermatology, but suggests that it may have potential as an antitumour agent. This study confirmed the validity of in vitro systems for preliminary assays on new compounds, in order to avoid the use of animals in toxicological studies.

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“…1 Despite the importance of VOCs, in vitro toxicity testing of these compounds has encountered significant technical challenges due to their physicochemical properties. [3][4][5][6] Low water solubility and high vapour pressure are two major physico-chemical characteristics of many VOCs that may create technical challenges during the course of in vitro experimentation. Conventionally, insoluble test chemicals are solubilised in culture media using a range of organic solvents such as dimethyl sulfoxide (DMSO) or alcohols.…”

Section: Introductionmentioning

confidence: 99%

A novel in vitro exposure technique for toxicity testing of selected volatile organic compounds

et al. 2006

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Exposure to vapours of volatile chemicals is a major occupational and environmental health concern. Toxicity testing of volatile organic compounds (VOCs) has always faced significant technological problems due to their high volatility and/or low solubility. The aim of this study was to develop a practical and reproducible in vitro exposure technique for toxicity testing of VOCs. Standard test atmospheres of xylene and toluene were generated in glass chambers using a static method. Human cells including: A549-lung derived cell lines, HepG2-liver derived cell lines and skin fibroblasts, were grown in porous membranes and exposed to various airborne concentrations of selected VOCs directly at the air/liquid interface for 1 h at 37 °C. Cytotoxicity of test chemicals was investigated using the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) and NRU (neutral red uptake) assays following 24 h incubation. Airborne IC 50 (50% inhibitory concentration) values were determined using dose response curves for xylene (IC 50 = 5350 ± 328 ppm, NRU; IC 50 = 5750 ± 433 ppm, MTS in skin fibroblast) and toluene (IC 50 = 10500 ± 527 ppm, NRU; IC 50 = 11200 ± 1044 ppm, MTS in skin fibroblast). Our findings suggest that static direct exposure at the air/liquid interface is a practical and reproducible technique for toxicity testing of VOCs. Further, this technique can be used for inhalational and dermal toxicity studies of volatile chemicals in vitro as the exposure pattern in vivo is closely simulated by this method. Exposure to vapours of volatile chemicals is a major occupational and environmental health concern. Toxicity testing of volatile organic compounds (VOCs) has always faced significant technological problems due to their high volatility and/or low solubility. The aim of this study was to develop a practical and reproducible in vitro exposure technique for toxicity testing of VOCs. Standard test atmospheres of xylene and toluene were generated in glass chambers using a static method. Human cells including: A549-lung derived cell lines, HepG2-liver derived cell lines and skin fibroblasts, were grown in porous membranes and exposed to various airborne concentrations of selected VOCs directly at the air/liquid interface for 1 h at 37 1C. Cytotoxicity of test chemicals was investigated using the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) and NRU (neutral red uptake) assays following 24 h incubation. Airborne IC 50 (50% inhibitory concentration) values were determined using dose response curves for xylene (IC 50 = 5350 AE 328 ppm, NRU; IC 50 = 5750 AE 433 ppm, MTS in skin fibroblast) and toluene (IC 50 = 10 500 AE 527 ppm, NRU; IC 50 = 11 200 AE 1044 ppm, MTS in skin fibroblast). Our findings suggest that static direct exposure at the air/liquid interface is a practical and reproducible technique for toxicity testing of VOCs. Further, this technique can be used for inhalational and dermal toxicity studies of volatile chemica...

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Cellular Models for In Vitro Toxicity Testing

Cited by 14 publications

References 24 publications

Synergistic Effects of the Membrane Actions of Cecropin-Melittin Antimicrobial Hybrid Peptide BP100

Synergistic Effects of the Membrane Actions of Cecropin-Melittin Antimicrobial Hybrid Peptide BP100

Cytostatic Properties of a Novel Compound Derived from Penicillium pinophilum: An In Vitro Study

A novel in vitro exposure technique for toxicity testing of selected volatile organic compounds

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