Validation ECVAM

Validated No ECVAM validation ECVAM validation validation through Consortia... 

At present there are no validated alternative methods able to completely replace the use of animals in the field of acute toxicity. Validation of an alternative model for acute oral toxicity is very complex and the time estimated to achieve complete animal replacement for acute toxicity is not clearly defined, nor can it be estimated at anything less than 10 years. However, there are some alternative methods currently available in different validation status prevalidated (MEIC, Multicenter Evaluation of in vitro cytotoxicity tests), under prevalidation (ECVAM), and validation (ECVAM, ICCVAM). 

Under validation (ECVAM, ICCVAM) we have the BALB/c 3T3 neutral red uptake (NRU) cytotoxicity assay and the normal human keratinocyte NRU cytotoxicity assay. Both methods are based on the ability of viable cells to incorporate and bind neutral red (NR), a supravital dye. NR penetrates cell membranes and accumulates in lysosomes. Alterations in the cell surface lead to decreased uptake and binding of NR (Nottingham and Spielmann, 1995 Spielmann et al, 1999). 

Mouse skin integrity function test (SIFT) Excised mouse skin Transepidermal water loss (TEWL) and electrical resistance Under validation (ECVAM)... 

Three embryotoxicity tests have been formally validated (ECVAM) in order to replace the OECD guideline 414 (developmental toxicity testing) ... 

European Centre for the Validation of Alternative Methods (ECVAM), six in vitro systems for chronic neurotoxicity testing are recommended for further consideration (Worth and Balls 2002). These are described as in vitro models that may be suitable for long-term toxicity testing. The systems are... 

In vitro tools could be used alone or in test batteries with increased potency of the description of cellular events and changes. The chapter provides a brief introduction on the components of an in vitro system, the main differences between models for research and models for testing and a list of validated alternative methods according to the European Centre for the Validation of Alternative Methods (ECVAM) (http // ecvam.jrc.it/, http //ecvam.jrc.ec.europa.eu/) evaluation. 

ECVAM as part of the JRC fulfills the task to validate alternative methods. Its advisory group ES AC advises ECVAM scientifically and gives expert judgment on the different proposed non-animal tests. 

An important point, made by the European Centre for the Validation of Alternative Methods (ECVAM), is that these 3 aspects (3Rs) should not be considered as alternatives that could replace each other, but as parts of an integrated system which should lead to progress in the development of non-animal tests and testing strategies. Another important key point recently defined by ECVAM is that the use of existing information must be maximized instead of testing methods. 

Alternative tests will have to be validated scientifically and established as acceptable by industry and the regulators before being used for registration. The EU body which undertakes this validation process is the European Centre for the Validation of Alternative Methods (ECVAM), and the US equivalent is the Interagency Co-ordinating Committee on the Validation of Alternative Methods (ICCVAM). 

ECVAM European Centre for the Validation of PEC Predicted environmental concentration... 

To test the irritancy potential of substances, two tests which can reliably distinguish between skin corrosives and noncorrosives are endorsed by the European Centre for the Validation of Alternative Methods (ECVAM). The testing procedures are based on the transcutaneous electrical resistance (TER) measurements of rat skin and on a human skin model. Both test systems [141-145] will be briefly outlined below. Nevertheless, these tests are not suited for the group of mild irritants which do not induce an acute effect on the barrier function. For those substances, new markers need to be evaluated. First results are available for heat shock protein 27 where higher levels were observed in skin models after exposure to mildly irritating chemicals [146, 147]. 

The following in vitro methods based on reconstructed human skin models are validated (by the ECVAM) for predicting skin corrosion— EPISKIN , EpiDerm , Corrositex —and irritation EPISKEM , EpiDerm , PREDISKIN , SIFT [142], Additionally, the SkinEthic was assessed and... 

ECVAM. News and Views. Statement on the scientific validity of the rat skin transcutaneous electrical resistance (TER) test (an in vitro test for skin corrosivity). ATLA 26 275-280 (1998). 

K. Meyer, C. Steinhoff, C. Tornier, B. De Wever, and M. Rosdy. Assessment of the skin irritation potential of chemicals by using the SkinEthic reconstructed human epidermal model and the common skin irritation protocol evaluated in the ECVAM skin irritation validation study. Altern. Lab. Anim. 34 393-406 (2006). 

Le Ferrec E, Chesne C, Artursson P, Brayden D, Fabre G, Gires P, Guillou F, Rousset M, Rubas W, Scarino ML (2001) In vitro models of the intestinal barrier. The report and recommendations of ECVAM Workshop 46. European Centre for the Validation of Alternative methods. Altern Lab Anim 29 649-668. 

The European Center for the Validation of Alternative Methods (ECVAM) uses and evaluates toxicogenomics, among other methods, e.g., in its carcinogenicity program (ECVAM 2007). 

ECVAM was created in response to a requirement in Directive 86/609/EEC on the protection of animals used for experimental and other scientific purposes, which requires that the EU Commission and the EU Member States should actively support the development, validation, and acceptance of methods, which could reduce, refine, or replace the use of laboratory animals. 

The duties of ECVAM are to coordinate the validation of alternative test methods at the EU level, to act as a focal point for the exchange of information on the development of alternative test methods, to set up, maintain, and manage a database on alternative procedures, and to promote dialogue between legislators, industries, biomedical scientists, consumer organizations, and animal welfare groups, with a view to the development, validation, and international recognition of alternative test methods. 

ECVAM has validated alternative test methods for acute oral toxicity, biologies, immunotoxicity, dermal corrosion and irritation, developmental toxicity, and pyrogenicity (ECVAM 2007). 

The following in vitro tests, replacing in vivo animal tests, have been endorsed as scientifically valid by ECVAM and have been adopted as EU test methods (EU 2006) ... 

ECETOC. 2007b. European Centre for Ecotoxicology and Toxicology of Chemicals (publications) website http /www.ecetoc.org/Content/Default.asp PagelD = 21 ECVAM. 2007. European Center for Validation of Alternative Methods website. http /ecvam.jrc.it/index.htm EU. 2006. The DG Environment REACH website. http /ec.europa.eu/environment/chemicals/reach EU. 2007. European Commision, Research Science and Society website The role of ethics in EU research. 

Currently, there are no validated and regulatory accepted in vitro methods for assessing repeated dose toxicity. Numerous in vitro systems have been developed over the last decades and have been discussed and summarized in recent ECVAM reports on repeated dose toxicity testing (Worth and Balls 2002, Prieto et al. 2005, Prieto et al. 2006). Human in vitro data, particularly on kinetics and metabolism, and in vitro test data from well-characterized target organ and target system models on, e.g., mode of action(s)/mechanism(s) of toxicity may be useful in the interpretation of observed repeated dose toxicity. 

ECVAM European Center for the Validation of Alternative Methods... 

Toda, E., Tong, W., van Delft, J.H., Weis, B. and Schechtman, L.M. (2006) Validation of toxicogenomics-based test systems ECVAM-ICCVAM/NICEATM considerations for regulatory use. Environmental Health Perspectives, 114, 420-429. 

The 35 mm dish assay detects compounds that are cytotoxic to or alter proliferation of the multipotent stem cells, but the main limitation of the 35 mm dish CFU assay is the need to microscopically identify and count the colonies to determine the compound concentration that decreased the number of colonies by 50% (IC50) or 90% (IC90). The European Community for Validation of Alternative Methods (ECVAM) supported the recommendation of using the mouse and human CFU-GM IC90 with... 

C O LI P A international in vitro phototoxicity test validation study and the ECVAM/... 

Right now, in Europe, there are three validated in vitro assays aimed at evaluating embryotoxicity (teratogenicity) potential. Three of these assays were submitted to extensive intra- and interlaboratory validations lead by the European Center for Validation of Alternative Methods (ECVAM) (3) ... 

At the 17th meeting of the ECVAM Scientific Advisory Committee in 2001, the EST, the MM assay, and the WEC assay were endorsed as scientifically validated assays and ready for consideration for regulatory acceptance. Detailed reports on the performance of these validated assays have been published (5-7). 

Three in vitro alternative assays are endorsed as scientifically validated by ECVAM in 2001 the EST, the MM, and WEC assays. The best results were obtained by combining the EST and WEC assays. The EST assay has the advantage of not using animals at all. As human cell culture technology improves, particularly regarding stem cells (8, 9), new methods will undoubtedly evolve that will enable a closer in vitro detection of in vivo human teratogenesis. 

ECVAM validation study on three in vitro embryotoxicity tests. Altern Lab Arum 32 209-244... 

Spielmann H et al (2004) Validation of the rat Kmb bud micromass test in the international ECVAM validation study on three in vitro embryotoxicity tests. Altern Lab Anim 32 245-274... 

Genschow E, Spiehnann H, Scholz G et al (2002) The ECVAM international validation study on in vitro embryotoxicity tests results of the definitive phase and evaluation of prediction models. European Centre for the Validation of Alternative Methods. Altern Lab Anim 30(2) 151-176... 

Hartung T, Bremer S, Casati S et al (2004) A modular approach to the ECVAM principles on test validity. Altern Lab Anim 32(5) 467- 72... 

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