• Toxicological Research
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• v.16 no.1
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• pp.77-82
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• 2000

This study was conducted to assess a possible alternative method as replacements for in vivo test. The human fibroblasts were exposed to several photoxic chemicals (promethazine, neutral red, chlortetracyclone, amiodatone, bithional, 8-methyooxypsorale) and non-phototoxi substance, ammonium laureth sulfate and irradiatied with 5 J/$cm^2$ of UVA (3320~420nm). The cell viability was measured by NRU (neutral red uptake) assay. The photoxic potential of test chemicals in the NRU PT (phototoxicity test) was assessed by determining the PIF (photoirritancy Factor) by using a cut-off value of 5. The NRU PT responses of most chemicals showed a close agreement with in vivo response except bithinol. There was a relatively good agreement between in vitro NRU assay and in vivo data. These results suggest that NRU assay using fibroblast could be used to predict the phototoxicity.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.3
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• pp.193-202
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• 2009

This study was done to assess an alternative method as a replacement of in vivo phototoxicity test. The human fibroblasts were exposed to several phototoxic chemicals (promethazine, chlorpromazine chlortetracycline, 8-methoxypsoralen, neutral red, bithionol) and non-phototoxic materials (cinnamic aldehyde, p-aminobenzoic acid, sodium lauryl sulfate, L-cysteine). The cell viability was measured by neutral red uptake (NRU) assay. The results of the NRU phototoxicity (PT) assay showed a close agreement with in vivo test except bithionol. We also have tested the cosmetic ingredients including $Medimin^{(R)}$ A, $Medimin^{(R)}$ D, $LG^{(R)}$ 106W, $Phytoclear^{(R)}$ EL-1, Carex humilis L. extract, Canna indica L. extract, Salvia miltiorrhira Bunge extract, $Parsol^{(R)}$ MCX and $Parsol^{(R)}$ 1789. Most materials except Salvia miltiorrhira Bunge extract did not show any phototoxicity.

• Proceedings of the Korean Society of Toxicology Conference
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• 2001.05a
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• pp.109-109
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• 2001

In order to evaluate the neutral red uptake assay as an alternative method for phototoxicity test, we compared the potential of phototoxicity in vitro in cultured human fibroblasts and 3T3 fibroblast cells derived from Balb/c mice. Both fibroblasts were exposed to various known phototoxic chemicals (promethazine, neutral red, chlorpromazine, chlortetracycline, amiodarone, bithionol, 8-methoxypsoralen) and non-phototoxic chemical (ammonium laureth sulfate) and irradiated with 5 J/cm$^2$ of UVA.(omitted)

• The Korean Journal of Pesticide Science
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• v.18 no.1
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• pp.8-13
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• 2014

This study evaluated in vitro cytotoxicity of 5 pesticides, including 2 herbicides, 2 germicides, and an insecticide, as an alternative to the fish acute toxicity test. The in vitro cytotoxicity was tested using a neutral red uptake (NRU) assay with epithelioma papulosum cyprini (EPC) cells that originated from the epidermal tissue of Cyprinus carpio (common carp). An in vivo fish acute toxicity test was conducted according to OECD Test Guideline No. 203 using Aphyocypris chinensis (Chinese bleak), Oryzias latipes (Japanese medaka), and C. carpio. The results showed that the sensitivity of the cell viability assay for the pesticides was similar to the fish acute test in ranking order despite having approximately 10 times less absolute sensitivity. The $r^2$ correlation values were calculated as 0.38 (p = 0.26), 0.76 (p = 0.05) and 0.90 (p = 0.01) for A. chinensis, O. latipes, and C. carpio, respectively. These results suggested that the potential of EPC cell viability assay as an alternative to the fish acute toxicity test due to their good correlation and NRU assay is expected to serve as a useful tool for predicting acute fish lethality for pesticides if further studies with a large set of pesticides are conducted.

• Korean Journal of Pharmacognosy
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• v.33 no.1 s.128
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• pp.13-17
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• 2002

Helicobacter pylori infection is associated with type B gastritis, peptic ulcer, and gastric cancer. The vacuolation of cells induced by H. pylori is thought to be essential for the initiation and maintenance of gastric infection. The roles of H. pylori cytotoxin, urease, and ammonia in the vacuolation of HeLa cells were determined. Ammonium chloride augmented the neutral red uptake induced by H. pylori toxin. Acetohydroxamic acid (AHA) failed to block the neutral red uptake induced by H. pylori toxin. Leweifang significantly prevented the vacuolation of HeLa cells induced by H. pylori toxin or H. pylori toxin and ammonium chloride. Further investigation is required to determine the mechanisms of Leweifang for the inhibition of vacuole formation of eukaryotic cells in response to the H. pylori toxin.

• Journal of Pharmaceutical Investigation
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• v.25 no.4
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• pp.365-369
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• 1995

The acute cytotoxicities of chloroquine sulfate, propranolol, ascorbic acid, acetylsalicylic acid and acrylamide on cultured adult rat hepatocytes were evaluated by the use of LDH leakage and NR uptake test. On the basis of $IC_{50}$ values, the rank order of cytotoxicities of these drugs in both tests was chloroquine sulfate > propranolol > acetylsalicylic acid > ascorbic acid. The $IC_{50}$ of LDH test was very similar to that of NR uptake test. Thus, we concluded that both tests are reliable and sensitive methods in detecting toxicity in adult cultured rat hepatocytes.

• Journal of the Korean Society of Tobacco Science
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• v.29 no.2
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• pp.110-117
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• 2007

In vitro toxicity tests such as cytotoxicity, mutagenicity and genotoxicity assay are useful for evaluating the relative toxicity of smoke or smoke condensates obtained from different cigarette configurations. A major disadvantage of these tests is relatively time-consuming, complicated and expensive. Recently, a cell-free glutathione consumption assay (GCA) as a rapid and simple screening method for the toxicity assessment of smoke has been reported by Cahours et al. (CORESTA, 2006). This study was carried out to assess the GCA application capable of predicting the toxicity of gas/vapor phase (GVP) of cigarette smoke and to identify individual compounds responsible for the glutathione (GSH) consumption in smoke. Each GVPs from 2R4F, standard cigarette, carbon filter cigarette (ExC) and new carbon filter cigarette (ExN), test cigarettes were collected by automatic smoking machine and evaluated the relative toxicity by GCA and neutral red uptake (NRU) assay. Toxic compounds existed in smoke were also chosen, relative toxicities of these compounds were screened by using two methods and compared individually. The overall order of toxicity by GCA was 2R4F > ExC > ExN, which was consistent with the result of Neutral Red Uptake assay. The levels of carbonyl compounds of ExN were lower than those of 2R4F and ExC, indicating that GSH consumption was associated with carbonyl compound yields. A major toxicant under current study is acrolein, which contributed to more than half of the GSH consumption. Collectively, the toxicity of GVP determined by GCA method may be mainly attributed to acrolein.

• Toxicological Research
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• v.18 no.3
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• pp.227-232
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• 2002

In order to find out the appropriate in vitro method for high correlation with in vivo, we com-pared the sensitivities of phototoxicity (PT) in vitro method between in human keratinocytes, HaCaT cells and in 3T3 fibroblast cells derived from Balb/c mice. Both cells were exposed to six known phototoxic chemicals : promethazine, neutral red, chlortetracycline, amiodarone, bithionol, 8-methoxypsoralen, or non-phototoxic chemical, ALS (ammonium laureth sulfate) and then irradiated with 5 J/$cm^2$ of UVA. Cell viability ($IC_{50}$ ) was measured by neutral red uptake (NRU) assay. The ratio of $IC_{50}$ value of chemicals in the presence and absence of UVA was determined by the cut-off value. The phototoxic potential of test chemicals in NRU assay was determined by measuring the photoirriation factor (PIF) with a cut-off value of 5. In both 3T3 and HaCaT cells, all known phototoxic chemicals were positive (over 5 of PIF value), except that bithionol was found to be non-phototoxic to HaCaT cells, and ALS, non-phototoxic chemical was negative. These results suggest that Balb/c 3T3 cell was more sensitive than HaCaT cell to predict phototoxicity potential.

• Toxicological Research
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• v.7 no.1
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• pp.13-20
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• 1991

To investigate the cytotoxicity and genotoxicity of the DNA alkylating agnet, mitomycin C and the antimetabolite, 5-Fluorouracil (5-FU) in cultured rat fibroblasts, the colorimetric assay of netural red (NR) for cytotoxicity and for genotoxicity, sister chromatid exchange (SCE) assay and the measurement of the rate of DNA synthesis were performed in cells cultured in media containing various concentrations of mitomycin C and 5-FU. The uptake ability of neutral red decreased does-dependently. NR90 and NR50 values of mitomycin C were 1.49 nM and 6.87mM and 5-FU were 38.4mM AND 284.4Mm respectively.

• International Journal of Oral Biology
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• v.34 no.3
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• pp.143-150
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• 2009

Amino acid transporters play important roles in supplying nutrients to cells. In our current study, we investigated the expression of LAT1 and measured the amino acid uptake in ameloblast cultures to further elucidate the roles of this transporter during the differentiation of these cells. RT-PCR, observations of cell morphology, Alizaline red-S staining, and uptake analyses were performed following the experimental induction of differentiation in the cultures. LAT1 mRNA was detectable and found to gradually increase over time whereas LAT2 mRNA was not evident in the ameloblast cultures. Transcripts of 4F2hc, a cofactor of LAT1 and LAT2, were also found to be expressed in ameloblast cultures and increase with time. Amelogenin mRNA was expressed in the early stage ameloblast cultures. L-leucine uptake was observed to increase over 14 days of growth in culture. Our data suggest that LAT1 has a key role in the differentiation of ameloblasts and in providing these cells with neutral amino acids, including several essential amino acids.