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

Abstract

-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.

Cell culture models of the human cornea — a comparative evaluation of their usefulness to determine ocular drug absorption in-vitro

“…In both cases, the formation of tight epithelial sheets was not observed. This cell line, which is widely used in studies of cytotoxicity, metabolism and permeation of new drug substances Goskonda et al , 2000; Tak et al 2001;Kitagaki et al 2006), was not able to express a zona occludens in our laboratory. Similar results and findings have been described by Tak et al (2001) and Becker (2006) human corneal epithelial cells and is cultured in serum-free medium, did not show TEER values more than 100 cm 2 , suggesting that the cell layers have weak barrier properties.…”

Studies with neuronal cells: From basic studies of mechanisms of neurotoxicity to the prediction of chemical toxicity

“…Such approach has been undertaken in the European-funded project ACuteTox, aimed to develop in vitro strategies for the assessment of acute human toxicity (www.acutetox.org). Previous in vitro approaches have demonstrated that general cytotoxicity assays may reasonably predict mammal acute systemic toxicity attaining a level of predicition of 70 -80 % (Halle, 2003;Ekwall et al, 1999;Kitagaki et al, 2006;Clemedson et al, 2006). However, neurotoxic events may underlie acute human toxicity that may not be predicted by using an in vitro test that exclusively relies on cell death, contributing to the observed 20 -30 % lack of predictability.…”

Prediction of Human Lethal Doses and Concentrations of MEIC Chemicals from Rodent LD50 Values: An Attempt to Make Some Reparation

“…Because of this concern, many efforts have been made to find acceptable substitutes for the rodent LD 50 test, such as in vitro cytotoxicity assays. [5][6][7][8][9][10][11] Bernson et al 12 proposed a 'Multicentre Evaluation Study of In vitro Cytotoxicity' (MEIC), based on a set of 50 very diverse organic and inorganic chemicals. The MEIC chemicals, as they came to be known, have been widely studied.…”