The Development and Evaluation of In Vitro Tests by the FRAME Alternatives Laboratory

1995 ◽  
Vol 23 (1) ◽  
pp. 75-90
Author(s):  
Richard H. Clothier ◽  
Karen A. Atkinson ◽  
Michael J. Garle ◽  
Rachel K. Ward ◽  
Angela Willshaw
Keyword(s):  
Research Programme ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
In Vitro Tests ◽  
Dermal Toxicity ◽  
Mechanisms Of Toxicity ◽  
The Us ◽  
Detergent Association ◽  
Assay Methods

This review outlines the work which has been conducted in the FRAME Alternatives Laboratory during the first ten years of the FRAME Research Programme. A number of in vitro tests, including the kenacid blue, neutral red release and fluorescein leakage assay methods, have been evaluated and have subsequently been included in validation schemes organised by the US Soap and Detergent Association, the US Cosmetic, Toiletry and Fragrance Association, the European Commission and the European Cosmetic, Toiletry and Perfumery Association, as well as in the Scandinavian multicentre evaluation of in vitro cytotoxicity testing scheme. More recently, research has been undertaken in the areas of phototoxicity, immunotoxicity, dermal toxicity and intercellular communication, in addition to investigations into fundamental mechanisms of toxicity.

In Vitro Toxicity: Mechanisms, Alternatives and Validation — A Report from the 19th Annual Scientific Meeting of the Scandinavian Society for Cell Toxicology

2002 ◽  
Vol 30 (3) ◽  
pp. 307-308
Author(s):  
Anna Forsby
Keyword(s):  
Animal Experiments ◽  
Scientific Meeting ◽  
In Vitro Cytotoxicity ◽  
Annual Scientific ◽  
In Vitro Toxicity ◽  
In Vitro Tests ◽  
Mechanisms Of Toxicity ◽  
Scandinavian Society ◽  
Wide Range

The Scandinavian Society for Cell Toxicology (SSCT) has arranged annual scientific meetings since 1983. These workshops were the forum for the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) programme. Along with the MEIC programme, which was completed in 1998, a wide range of topics relating to cytotoxicity have been discussed. The meetings have also given an opportunity for graduate students and young scientists to present their work to an international audience. At the same time, experts in the fields of in vitro toxicity have been invited as speakers. The 19th SSCT scientific meeting, which was held in 2001 at Sørup Manor in Ringsted, Denmark, was no exception. The meeting consisted of four sessions: mechanisms of toxicity; environmental toxicological testing; alternatives to animal experiments; and validation of in vitro tests.

Toxicity of 20 Chemicals from the MEIC Programme Determined by Growth Inhibition of L-929 Fibroblast-like Cells

1997 ◽  
Vol 25 (1) ◽  
pp. 55-59
Author(s):  
Lena Järkelid ◽  
Per Kjellstrand ◽  
Evi Martinson ◽  
Anders Wieslander
Keyword(s):  
Cell Growth ◽  
Growth Inhibition ◽  
Exposure Period ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
Mouse Fibroblast ◽  
In Vitro Tests ◽  
Cytotoxicity Test ◽  
Uptake Method

The Multicentre Evaluation of In vitro Cytotoxicity (MEIC) programme is an international project aimed at evaluating the relevance of in vitro tests in predicting human toxicity. We have screened 20 chemicals (MEIC codes 31-50) from the programme, by using a cytotoxicity test based on growth inhibition of the mouse fibroblast-like L-929 cell line. Inhibition of cell growth was determined by the neutral red uptake method, which is well established and is used for screening the cytotoxicity of chemicals and plastics for pharmaceuticals and medical devices. The concentrations causing 50% inhibition of cell growth after a 72-hour exposure period varied from 3.1μM for hexachlorophene, to 1.4mM for caffeine. This is within the same range as results recently obtained with five other cell models. However, with some chemicals (chloroform, carbon tetrachloride and dichloromethane), no reliable results were obtained. These substances could not be dissolved in a reproducible way in any of the solvents used and, furthermore, they were highly volatile, which led to difficulties in maintaining the concentrations.

Cytotoxicity Evaluation of the First Ten MEIC Chemicals: Acute Lethal Toxicity in Man Predicted by Cytotoxicity in Five Cellular Assays and by Oral LD50 Tests in Rodents

1989 ◽  
Vol 17 (2) ◽  
pp. 83-100
Author(s):  
Björn Ekwall ◽  
Inger Bondesson ◽  
José V. Castell ◽  
Maria José Gómez-Lechón ◽  
Sven Hellberg ◽  
...  
Keyword(s):  
In Vitro Cytotoxicity ◽  
In Vitro Tests ◽  
Human Toxicity ◽  
Blood Concentrations ◽  
Cell Systems ◽  
Cellular Assays ◽  
Preliminary Validation ◽  
Partial Least Squares Model ◽  
Lethal Dosage

The MEIC (multicentre evaluation of in vitro cytotoxicity) programme is a five-year programme to validate in vitro tests for general toxicity, and is organised by the Scandinavian Society for Cell Toxicology. Interested laboratories are invited, on an international basis, to test 50 published reference chemicals in their respective assays. Submitted results will then be evaluated yearly by the MEIC Committee for their relevance to various types of human toxicity, including an evaluation for the same chemicals of the prediction by animal tests of human toxicity. To establish the validation methods, a preliminary validation cycle is being performed in 1989/90 with submitted results for the first ten MEIC chemicals. The present paper is the very first step of this preliminary validation process. The prediction of human toxicity by five cytotoxicity assays (altogether 14 different cell systems/endpoints) has been evaluated, and also compared with the predictive value of rodent LD50 tests. Mouse LD50 prediction of human lethal dosage for these substances was good, while rat LD50 prediction was less satisfactory. The collective predictions by all 14 cell systems/endpoints of human toxicity in the form of a multivariate PLS (partial least squares) model of human acute lethal blood concentrations, as well as the corresponding prediction by a HeLa cell assay, were comparable to the efficiency of mouse LD50 prediction of human lethal dosage. When combined with simple toxicokinetic data (absorption of chemicals in the intestine and distribution volumes), the PLS model and the HeLa assay were able to predict human lethal dosage of the ten chemicals as accurately as the mouse LD50 value. The small number of chemicals studied to date means that general conclusions cannot be drawn from these results. Further validation of more chemicals with the in vitro methods is essential and promises to be worthwhile.

Analogy Models for Prediction of Human Toxicity

1990 ◽  
Vol 18 (1_part_1) ◽  
pp. 103-116
Author(s):  
Sven Hellberg ◽  
Lennart Eriksson ◽  
Jörgen Jonsson ◽  
Fredrik Lindgren ◽  
Michael Sjöström ◽  
...  
Keyword(s):  
Human Subjects ◽  
Toxicity Testing ◽  
In Vitro Cytotoxicity ◽  
Alternative Methods ◽  
Laboratory Animals ◽  
In Vitro Tests ◽  
Human Toxicity ◽  
Efficient Data ◽  
Basal Cytotoxicity

Estimating the toxicity to humans of chemicals by testing on human subjects is not considered to be ethically acceptable, and toxicity testing on laboratory animals is also questionable. Therefore, there is a need for alternative methods that will give estimates of various aspects of human toxicity. Batteries of in vitro tests, together with physicochemical and toxicokinetic data, analysed by efficient data analytical methods, may enable analogy models to be constructed that can predict human toxicity. It may be possible to model non-specific toxicity relating to lipophilicity, or basal cytotoxicity, for a series of diverse compounds with large variation in chemical structure and physicochemical properties. However, local models for a series of similar compounds are generally expected to be more accurate, as well as being capable of modelling more-specific interactions. Analogy models for the prediction of human toxicity are discussed and exemplified with physicochemical and cytotoxicity data from the first ten chemicals in the multicenter evaluation of in vitro cytotoxicity (MEIC) project.

An Animal Protection Sponsor's View of MEIC

1997 ◽  
Vol 25 (3) ◽  
pp. 343-345
Author(s):  
Ethel Thurston
Keyword(s):  
Gold Standard ◽  
Animal Cell ◽  
Scientific Evidence ◽  
In Vitro Cytotoxicity ◽  
In Vitro Tests ◽  
Blood Concentrations ◽  
Animal Protection ◽  
Animal Tests ◽  
Lethal Blood

The Multicenter Evaluation of In Vitro Cytotoxicity programme is most important to animal protection, since it has validated 64 in vitro tests using advanced human data for 50 chemicals as the “gold standard”. Therefore, it has been able to compare animal cell tests, human cell tests and whole-animal tests fairly with unbiased scientific evidence. Added bonuses have included the identification and development of missing in vitro information (“missing tests”), publication of time-related lethal blood concentrations for all 50 chemicals, and some preliminary plans to resolve the 50,000 untested (or poorly tested) chemicals in the chemical mountain.

scholarly journals The Addition of Cocoa, Glycerol, and Saccharose to the Tobacco of Cigarettes: Implications for Smoke Chemistry, In Vitro Cytotoxicity, Mutagenicity and Further Endpoints

2010 ◽  
Vol 24 (3) ◽  
pp. 117-138 ◽  
Author(s):  
E Roemer ◽  
S Wittke ◽  
E TrellesSticken ◽  
JJ Piade ◽  
T Bonk ◽  
...  
Keyword(s):  
Metabolic Activation ◽  
Potential Effect ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
Total Evidence ◽  
Mainstream Smoke ◽  
Particulate Phase ◽  
Cocoa Powder ◽  
Uptake Assay

AbstractThe cigarette ingredients cocoa powder, glycerol, and saccharose were investigated regarding their potential effect on the resulting mainstream smoke, i.e., smoke chemistry (Hoffmann analytes), mammalian cell cytotoxicity (Neutral Red Uptake assay), and bacterial mutagenicity (Ames assay). Each ingredient was added at three concentrations to the tobacco of a 6 mg and 10 mg ‘tar’ yield experimental American blend filter cigarette (obtained under ISO/FTC smoking regime). The lowest application concentration was equivalent to the normal approximate use level of the ingredients; the highest application level was up to 5-fold higher. The resulting data were compared with the respective control cigarettes without addition of the ingredients. The addition of cocoa powder did not lead to any consistent effects on the measured mainstream smoke analytes. Neither the in vitro cytotoxicity nor the in vitro mutagenicity was affected by cocoa addition. The addition of glycerol resulted in a decrease in the delivery of several smoke constituents (generally around 20%), e.g. aldehydes, phenolics, and N-nitrosamines. Water in the particulate phase (TPM) was distinctly increased (up to +150%). The cytotoxicity of the TPM was decreased (approx. !15%). Mutagenicity was not affected. Saccharose addition consistently increased formaldehyde delivery in smoke by up to 40% and decreased tobacco-specific N-nitrosamines by up to approximately 20%. The increase in formaldehyde is discussed in the context of the human smoker. The cytotoxicity was not affected by the addition of saccharose, while the mutagenicity of the TPM was decreased in tester strain TA98 with metabolic activation (!15%). The results are in agreement with currently available literature. Some investigations summarized in this publication are novel and have not yet been reported in the literature. Based on the total evidence, it can be concluded that the three ingredients added at their current use levels do not increase the inherent toxicity of the cigarette smoke.

The Importance of Exposure Period and Cell Type in In Vitro Cytotoxicity Tests

1986 ◽  
Vol 14 (2) ◽  
pp. 86-92
Author(s):  
R.J. Riddell ◽  
D.S. Panacer ◽  
S.M. Wilde ◽  
R.H. Clothier ◽  
M. Balls
Keyword(s):  
Exposure Period ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
Test Period ◽  
Assay Method ◽  
Cell Type ◽  
Long Term Effects ◽  
3T3 Fibroblasts ◽  
Affect Cell Viability

The cytotoxicities of 50 coded chemicals from the FRAME blind trial set were estimated by the FRAME kenacid blue (KB) and Rockefeller neutral red (NR) assay methods after test periods of 24 hours and 72 hours with murine 3T3 fibroblasts. Results obtained after the two test periods were considered to be similar for 25 of the chemicals, but different for 25 others. Our results indicate that a test period of 72 hours is more suitable for the assessment of potential cytotoxicity by the KB and NR methods than the 24 hour test period, which is less likely to allow sufficient time for certain chemicals to exert their toxic effects, particularly those that inhibit cell division or affect cell viability through other long-term effects. A comparison of results obtained by the 72 hour KB method with 3T3 cells and with human BCL-D1 cells indicated that substantially different ID50 values were found for 8 of 48 chemicals. These results are discussed in terms of the importance of choice of test period, assay method and cell type used in in vitro cytotoxicity tests.

scholarly journals L929 cell response to root perforation repair cements: an in vitro cytotoxicity assay

2009 ◽  
Vol 20 (1) ◽  
pp. 22-26 ◽  
Author(s):  
Rosana Belchior Miranda ◽  
Sandra Rivera Fidel ◽  
Maria Aparecida Affonso Boller
Keyword(s):  
Morphological Changes ◽  
Mineral Trioxide Aggregate ◽  
L929 Cell ◽  
Neutral Red ◽  
In Vitro Cytotoxicity ◽  
Mean Values ◽  
Nonviable Cell ◽  
Red Dye ◽  
Positive Controls

This study compared the cytotoxicity of an experimental epoxy-resin and calcium hydroxide-based cement (MBPc), gray mineral trioxide aggregate (MTA) and white mineral trioxide aggregate (WMTA) using the agar overlay method with neutral red dye. L929 cells were seeded into 6-well culture plates where 48-h set test materials were placed on the agar overlay, in triplicate. Teflon and natural rubber served as negative and positive controls. After an incubation period of 24 h at 37ºC in a humidified atmosphere of 5% CO2 in air, a discolored area around the samples and the positive controls could be observed and measured per quadrant. The mean values were compared and converted into grades to classify the results according to the table of cytotoxicity grades according to the Standard Operating Procedures (SOP) of the Oswaldo Cruz Foundation, Brazil. The nonviable cell areas and the morphological changes in the cells were observed with an inverted microscope. The results showed grade 1 (slight) for the two types of MTA (p>0.05) and grade 2 (mild) for the MBPc (p<0.001). All samples met the requirements of the test as none of the cultures showed reactivity higher than grade 2.

Sign in / Sign up

Export Citation Format

Share Document