Development of a high throughput in vitro toxicity screen predictive of high acute in vivo toxic potential

Evans, Sean M. H.; Casartelli, Alessandro; Herreros, Esperanza; Minnick, Dana T.; Day, Charles; George, Elisabeth; Westmoreland, Carl

doi:10.1016/s0887-2333(01)00064-9

Cited by 87 publications

(58 citation statements)

References 4 publications

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“…The fluorescence spectra of both forms of the probe show a maximum emission at 590 nm for resorufin and a negligible fluorescence for resazurin. In agreement with Maeda et al (27), we found that 570 nm is the optimal excitation wavelength of resorufin, although 530 nm is commonly used (5,15,28,29).…”

Section: Discussionsupporting

confidence: 92%

A new nondestructive cytometric assay based on resazurin metabolism and an organ culture model for the assessment of corneal viability

Perrot¹,

Dutertre-Catella

Martin³

et al. 2003

Cytometry Pt A

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Background: Three-dimensional culture or human corneal equivalents for safety testing are difficult to investigate with classic cytometric or biochemical methods. So a fluorometric method is proposed using resazurin probe. Methods: Absorbance and fluorescence spectra of the oxidized and reduced forms of resazurin were performed to determine optimal measurement conditions. More than 100 enucleated porcine eyes were used for this experiment. Twelve corneas were immersed in resazurin solution and fluorescence was measured hourly from 1 to 10 h. Ninety benzalkonium chloride-treated corneas and control corneas were used as toxicity controls, and corneal viability was compared with in vivo rabbit eye irritation. Results: After analysis of spectra, the optimal measurement condition of resazurin metabolism proved to be a fluorescence measurement using 570 nm excitation wavelength and 590 nm emission wavelength. The reduction of

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Section: Discussionsupporting

confidence: 92%

A new nondestructive cytometric assay based on resazurin metabolism and an organ culture model for the assessment of corneal viability

Perrot¹,

Dutertre-Catella

Martin³

et al. 2003

Cytometry Pt A

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“…Cell viability and cell counts were quantified using resazurin [7-Hydroxy-3H-phenoxazin-3-one 10-oxide] (Alamar Blue) indicator dye (34). A working solution of resazurin was prepared in sterile HBSS minus phenol red (0.5 mg/ml), and then added (15% v/v) to each sample.…”

Section: Hanksmentioning

confidence: 99%

HTP Nutraceutical Screening for Histone Deacetylase Inhibitors and Effects of HDACis on Tumor-suppressing miRNAs by Trichostatin A and Grapeseed (Vitis vinifera) in HeLa cells

Mazzio

Soliman

2017

CGP

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“…No animal testing was performed for this study. Following the method described by Evans et al (2001), the lowest LD 50 value of a test chemical that 2) MEIC; Multicentre Evaluation of In Vitro Cytotoxicity (Ekwall et al 1983). …”

Section: Ld 50 Valuesmentioning

confidence: 99%

“…As in a previous report (Stobbe et al, 2003), the experimentally derived IC 50 and IC 35 were compared to the in vivo data for each of the 6 parameters (accuracy, sensitivity, specificity, prevalence, positive predictability and negative predictability) ( Table 2). Evans et al (2001), the in vitro cut-off concentration corresponding to the in vivo oral cut-off point (2,000 mg/kg) was calculated from the regression equation to be 4,225 μg/mL (Fig. 1).…”

Section: Calculation Of Correlation Parametersmentioning

confidence: 99%

“…ICCVAM/ECVAM is presently validating the Neutral Red Uptake (NRU) assay with both mouse fibroblast cell line (BALB/c 3T3) and primary normal human keratinoctyes (NHK) (ICCVAM, 2001;Gennari et al, 2004). Recently, another method using Chinese hamster ovary (CHO) cells and the experimentally derived in vitro IC 35 values has been reported to be valuable in predicting the in vivo acute toxic potential of chemicals (Evans et al, 2001). The aims of these in vitro studies are to reduce, replace and refine animal use for acute toxicity determinations and to predict the starting dose for in vivo acute toxicity testing (ICCVAM, 2001;Gennari et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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Sirc-CVS Cytotoxicity Test: An Alternative for Predicting Rodent Acute Systemic Toxicity

et al. 2006

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-An in vitro crystal violet staining method using the rabbit cornea-derived cell line (SIRC-CVS) has been developed as an alternative to predict acute systemic toxicity in rodents. Seventy-nine chemicals, the in vitro cytotoxicity of which was already reported by the Multicenter Evaluation of In vitro Toxicity (MEIC) and ICCVAM/ECVAM, were selected as test compounds. The cells were incubated with the chemicals for 72 hrs and the IC 50 and IC 35 values (μg/mL) were obtained. The results were compared to the in vivo (rat or mouse) "most toxic" oral, intraperitoneal, subcutaneous and intravenous LD 50 values (mg/kg) taken from the RTECS database for each of the chemicals by using Pearson's correlation statistics. The following parameters were calculated: accuracy, sensitivity, specificity, prevalence, positive predictability, and negative predictability. Good linear correlations (Pearson's coefficient; r>0.6) were observed between either the IC 50 or the IC 35 values and all the LD 50 values. Among them, a statistically significant high correlation (r=0.8102, p<0.001) required for acute systemic toxicity prediction was obtained between the IC 50 values and the oral LD 50 values. By using the cut-off concentrations of 2,000 mg/kg (LD 50 ) and 4,225 μg/mL (IC 50 ), no false negatives were observed, and the accuracy was 84.8%. From this, it is concluded that this method could be used to predict the acute systemic toxicity potential of chemicals in rodents.

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Development of a high throughput in vitro toxicity screen predictive of high acute in vivo toxic potential

Cited by 87 publications

References 4 publications

A new nondestructive cytometric assay based on resazurin metabolism and an organ culture model for the assessment of corneal viability

A new nondestructive cytometric assay based on resazurin metabolism and an organ culture model for the assessment of corneal viability

HTP Nutraceutical Screening for Histone Deacetylase Inhibitors and Effects of HDACis on Tumor-suppressing miRNAs by Trichostatin A and Grapeseed (Vitis vinifera) in HeLa cells

Sirc-CVS Cytotoxicity Test: An Alternative for Predicting Rodent Acute Systemic Toxicity

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