1. PREAMBLE

One of the main objectives of Council Directive 76/768/CE of 27 July 1976 on the approximation of the laws of the member states relating to cosmetic products, as stated in its recitals is "the safeguarding of public health". Moreover "cosmetic products must not be harmful under normal or foreseeable conditions of use... taking into account the possibility of danger to zones of the body which are contiguous to the area of application".

These positions have been maintained by means of a set of Commission Directives, and reiterated in a series of Amendments to the Council Directive.

In 1977 the Commission Decision 78/45/EEC of 19 December established a Scientific Committee on Cosmetology (SCC) to assist the Commission in the process of drafting and amending the Community cosmetic laws. The SCC is formed by scientists highly qualified in the fields of medicine, toxicology, biology, chemistry and other similar disciplines.

In 1997 Commission Decision 97/18/EEC of 23 July 1997 reorganized such precious technical assistance to the Commission by establishing a "Scientific Committee on Cosmetic and Non-Food Products intended for Consumers" (SCCNFP) to be consulted in the case laid down by Community legislation, and on questions of particular relevance to consumers' health. The SCCNFP has been requested to produce "scientific advice concerning matters relating to consumers' health in its strict sense".

Art. 4 of Council Directive 76/768/ECC, amended by Council Directive 93/35/EEC, affirms a prohibition of the marketing of cosmetic products containing "ingredients or combinations of ingredients tested on animals after 1 January 1998". The date was later postponed to 30 June 2000 by Commission Directive 97/18/EEC of 17 April 1997.

The Commission shall submit draft measures to postpone the date of implementation of this provision by 1 January 2000. A justification of a possible postponement of the deadline of 30 June 2000, could be an insufficient progress in developing alternative methods to replace animal testing or, also, other cases where alternative methods would not have been scientifically validated as offering an equivalent level of protection to consumers. Before submitting such measures, the Commission shall consult the SCCNFP.

In particular, the Commission has requested the SCCNFP as regards the status of alternative methods for the safety assessment of cosmetic ingredients according to the current state-of-the-art. Specifically, the Commission has requested the SCCNFP to assess the possibility to replace the data obtained on the basis of animal tests with data obtained making use of alternative methods in the safety evaluation of cosmetic ingredients, and to indicate those end-points for which alternative methods to animal testing are not available yet (Doc. no. 16831 of 11 August 1998).

2. SAFETY EVALUATION OF COSMETIC INGREDIENTS

The SCCNFP and previously to it, the SCC, has illustrated in a set of documents the concepts and the criteria of the present procedure of safety evaluation of cosmetic ingredients, based on the experience developed during more than 20 years, by regulating ca. 800 individual cosmetic ingredients of which over 400 have been proposed for a ban, due to their toxic and harmful effects to consumers' health (Report EUR 8794; SPC/803-5/90; XXIV/1878/97; SCCNFP/0119/99 Rev.1 draft).

As stressed several times by the SCCNFP, the primary goal of the safety testing of all cosmetic ingredients presented to the European Commission for their possible inclusion in the technical Annexes of the Cosmetic Directive, is to determine the potential of these ingredients for their harmful effects in an experimental model, so that it makes possible, by extrapolation, to predict the same effect or the absence of harmful effects for consumers.

According to medical science, the safety studies should permit a quantitative determination of the potential for a cosmetic ingredient, or a mixture of cosmetic ingredients to produce local and systemic adverse effects and allow a determination of factors which may influence the nature, severity and possible reversibility of effects (Ref. 1).

Information necessary to the above purposes can be obtained only from carefully designed and well conducted studies. Toxicology testing programmes generally begin with single exposure in vivo or in vitro studies and progress to evaluate the effects of long-term repeated exposures.

The most used and recognized adequate models for safety testing are those represented by living laboratory animals (mice, rats, rabbits, guinea pigs etc.) which have been the object of millions of experimental studies developed by the toxicological research. In the last twenty years new toxicological systems no longer based on animal models, have been employed by scientists and accepted by national, continental (EU, Ref. 2, 3) and international (OECD, Ref. 4) regulations.

These new test systems are represented by mutagenicity/genotoxicity in vitro tests which make use of individual cellular organisms (bacteria, yeast, mammalian cells) or insects. This development has enabled, in many cases, to avoid the use of a large number of animals, as requested by the very expensive and laborious long-term carcinogenicity bioassays (Ref. 1).

After the approval of Sixth Amendment, the SCC and later the SCCNFP have been monitoring the several actions developed by scientific groups, including academics, industrial research and public institutions, to stimulate the progress in the development and validation of alternative methods to the use of animal models in the safety testing of cosmetics. In particular, the SCCNFP has been discussing and evaluating together with ECVAM (European Centre for the Validation of Alternative Methods) and COLIPA (European Cosmetic, Toiletry and Perfumery Association) scientists, the results of the pre-validation and validation studies and the applicability of these results to the safety evaluation of cosmetic ingredients and cosmetic products. An opinion has been adopted by the SCCNFP on 20 January 1999 during its Plenary Meeting on the use of some alternative methods to animal testing in the safety evaluation of cosmetic ingredients (Ref. 5).

3. PRESENTLY VALIDATED ALTERNATIVE METHODS

The following notes are representing the opinion on some alternative methods which could be of some relevant use in the safety evaluation of cosmetic ingredients, and on the state-of-the-art of some in vitro methods which could be validated in the near future.

3.1. SKIN IRRITATION

The present scientific knowledge on the mechanistic basis of skin irritation in vivo is still limited, due to the complex set of reactions involved , and in the impossibility to define the key specific and relevant end points which could be evaluated by an in vitro system (Ref. 6).

However, a pre-validation study is in progress by using human skin models and a pig ear test, under the sponsorship of ECVAM.

In the evaluation of a potential skin irritant effect by a cosmetic ingredient, it is still possible by using a combination of different criteria of evaluation, to identify the corrosivity/non-corrosivity potential of chemical ingredients.

Recently, two alternative in vitro methods for skin corrosivity have been validated and demonstrated to be applicable to the procedure for safety testing also in the sector of cosmetic ingredients. A draft new guideline on skin corrosivity testing has been submitted to OECD and to the European Commission (EC) for its inclusion in the Annex V of Council Directive 67/548/EEC. The new in vitro methods are represented by the Transcutaneous Electrical Resistance (TER) Test and by the Episkin Test. The SCCNFP has proposed the use of these two methods when corrosivity of cosmetic ingredients must be tested on animals, or when humans cannot be excluded. (Ref. 5)

3.2. PHOTOTOXICITY

OECD guidelines or EC guidelines on phototoxicity testing have not been adopted yet for the testing of UV light absorbing substances on animal models. Pre-validation, validation and applicability on cosmetic ingredients, such as the UV filters have been the object of a series of studies and different projects. An in vitro model, the 3T3 NEUTRAL RED UPTAKE Phototoxicity Test has been developed and demonstrated to be valid for the identification of phototoxic UV absorbing chemicals, including cosmetic ingredients (Ref. 7). This method is based on a cell phototoxicity process, observed in a mammalian cell population in vitro.

A draft protocol for testing phototoxicity, by employing this new alternative method has been presented to the OECD and to the European Commission for its inclusion in Annex V of Council Directive 67/548/EEC.

The SCCNFP in its Plenary Meeting of 20 January 1999 has adopted an opinion which proposes to the European Commission the use of the "3T3 NRU Phototoxicity Test" as the standard method for testing the UV light absorbing cosmetic ingredients or mixtures of ingredients for phototoxic potential.(Ref. 5)

3.3. PERCUTANEOUS ABSORPTION

OECD guidelines or EC guidelines on safety testing for percutaneous absorption have not been adopted yet. However, some draft measures have been presented to OECD.

The assessment of percutaneous absorption of cosmetic ingredients has primary relevance in the procedure for evaluating the safety of cosmetics for consumers. The SCCNFP has recently reviewed the available scientific literature and data developed by cosmetic industry in this sector of testing and has agreed with the rationale for using in vitro methods to evaluate the percutaneous absorption. An opinion has been adopted by SCCNFP during its Plenary Meeting of 20 January 1999 proposing the use of in vitro methodologies for the safety testing of cosmetic ingredients. Moreover, due to the lack of a guideline approved by OECD or by the European Commission, the SCCNFP has defined a set of basic criteria for the in vitro assessment of percutaneous absorption of cosmetic ingredients, which have been adopted during the Plenary Meeting of 23 June 1999. These basic criteria represent the recommendation put forward by the SCCNFP for all cosmetic industries, in their assessment of the percutaneous absorption (Doc. SCCNFP/0167/99 Final)

3.4. OCULAR IRRITATION

Ocular irritation testing is needed for many cosmetic ingredients applied in particular zones of the consumers' body, especially those which may come into contact with the eye.

Since the approval of the Sixth Amendment, several collaborative studies have been developed within the European Union on chemicals of different use or cosmetic ingredients; similar studies have been developed in the USA (finished cosmetic products) and Japan (cosmetic ingredients) (Ref. 8.1-8.6).

The results of such extensive studies have revealed that no single test can fully replace the Draize rabbit test; that some in vitro tests have a certain level of predictivity and some are promising; that the level of predictivity is improved when combining several and different test systems (batteries) (Ref. 9). Some of these in vitro tests combined with Structure Activity Relationships and Physicochemical data could be used to identify potentially non-ocular irritant chemicals, but the need to use in vivo tests is still requested.

A document prepared by ECVAM and COLIPA on the current status of the alternatives to eye irritation (Doc. SCCNFP/0174/99) indicates the utility of a short-time approach optimising the current strategies and methods, and a long-term approach allowing gaps in knowledge to be filled, so as to increase the current predictivity of the alternative methods, and as a basis for the development of new methods, are being developed and conducted.

Attempts are currently being carried out by COLIPA: (1) to review the validation studies concluded so far, as to extract the maximum bulk of information to help refine the currently available prediction models; (2) to optimise the tier testing strategies as a "reduction alternative" proposed by ECVAM; (3) to propose a research programme to increase the knowledge on the mechanisms of chemically induced eye irritation so as to develop complementary test methods to the current alternative or to modify these in view of improving their predictive capacity (Ref. 9).

3.5. SKIN SENSITIZATION

Skin sensitization is a complex phenomenon which implies a series of biological reactions; skin permeation by the allergen; reaction of the hapten with a skin protein; processing haptenated proteins by epidermal Langerhans cells; migration of Langerhans cells to draining lymph nodes and interaction with T cells; recognition of hapten by specific T cells; etc (Ref. 10).

It should be possible by combining computerized expert systems with appropriate biological in vitro systems to identify chemicals able to perform the initial reactions. At present the elucidation of the critical stages of the phenomenon is still under study and considerable research is being undertaken. Recently, a substantial opportunity to refine and reduce animal use in the hazard identification of skin sensitizing cosmetic ingredients has been achieved with the Murine Local Lymph Node Assay (LLNA) (Ref. 11). This aspect is being considered by the SCCNFP.

3.6. OTHER TOXICOLOGICAL END-POINTS

Besides, apart carcinogenicity, the other fields of toxicology do not seem at present to offer the possibility to substitute animal models with alternative methods not using animals. Due to the same essential basic mechanisms between certain types of carcinogenic substances (genotoxic carcinogens) and mutagenic substances, chemicals that induce mutations in somatic cells in vitro should be regarded to as potential carcinogens and hence mutagenicity screens are of value in identifying potential "genotoxic" carcinogens. This is not the case for other fields of toxicology, such as reproductive toxicology, neurotoxicity, teratogenicity, sub-chronic toxicology, etc. The scientific knowledge of the basic mechanisms of the different types of toxic events still requires development of long-term planning or research into basic and cellular events underlying toxicity.

3.7. THE USE OF HUMAN VOLUNTEERS IN THE SAFETY EVALUATION OF COSMETIC INGREDIENTS AND FINISHED COSMETIC PRODUCTS

In a recent opinion, the SCCNFP has stressed the concept that the tests on animals for skin irritation or (not yet) validated alternative methods may be limited regarding their predictive value for exposure of human population. The SCCNFP states that confirmatory tests on humans may be needed scientifically and ethically, provided that the toxicological profile of an ingredient, or a mixture of ingredients, or a finished cosmetic product, based on animal or alternative methods is available and that a degree of safety is to be expected (SCCNFP/0003/98). This opinion also stresses the concept that confirmatory skin tolerance tests of cosmetics in human should not be preferred to animal testing; that the safety testing of cosmetics on humans may not be considered an alternative method to the use of animals; and that the use of human volunteers in the safety evaluation of cosmetics is subjected to ethical concern.

The SCCNFP has recently approved Guidelines on the use of human volunteers in the testing of potentially cutaneous irritant cosmetic ingredients or mixtures of ingredients (SCCNFP/0003/98) and Guidelines on the use of human volunteers in compatibility testing of finished cosmetic products (SCCNFP/0068/98 Final).

4. OPINION OF THE SCCNFP

On the basis of the scientific literature, after the evaluation of different research programmes conducted by cosmetic industries, the European Commission (ECVAM) and other Institutions, and considering the results obtained during the period 1993-1999 on the development and validation of alternative methodologies to the use of animals in the safety testing of cosmetic ingredients, the SCCNFP expresses the following opinion to the European Commission.

1. There has been a considerable effort in the technical and scientific fields of the safety testing of chemicals in general and cosmetics in particular, to develop and validate alternative methodologies to the use of animal models which could offer to the consumers an adequate and acceptable level of protection;

2. Due to the complexity of biological mechanisms that represent the basis for the occurrence of toxic events in human organism, a significant effort of scientific research is needed to understand all different steps of the aforementioned mechanisms and their molecular events. A set of research programs have been planned in the sectors of ocular irritation, skin irritation, skin sensitization, neurotoxicity, etc. The results of such researches will considerably influence scientific knowledge on several toxic events which, on turn, will allow the identification of more rigorous and rational criteria and systems to be applied in the safety evaluation of cosmetics, by possibly reducing the need of laboratory animals (Ref. 12);

3. At present, the SCCNFP has identified with the help of the contribution made by ECVAM in this field of activity, the following alternative methods that can be used for the safety testing of cosmetics:

a. In Vitro Methods to assess skin corrosivity in the safety evaluation of cosmetic ingredients or mixtures of ingredients (SCCNFP/0070/98 Final);

b. In Vitro Methods to assess phototoxicity in the safety evaluation of cosmetic ingredients or mixtures of ingredients (SCCNFP/0069/98 Final);

c. In Vitro Methods to assess percutaneous absorption of cosmetic ingredients (SCCNFP/0088/98 Final).

4. Moreover, the SCCNFP has defined the "Basic criteria for the in vitro assessment of percutaneous absorption of cosmetic ingredients" (SCCNFP/0167/99 Final) in order to provide the cosmetic industry with a set of recommendations for an adequate protocol for applying the in vitro methods in the studies of percutaneous absorption.

5. The SCCNFP has also produced a set of guidelines on the use of human volunteers in the testing of potentially cutaneous irritant cosmetic ingredients or mixtures of ingredients (SCCNFP/0003/98 Final) and in skin compatibility testing of finished products (SCCNFP/0068/98 Final) in order to provide recommendations on the use of human volunteers in the safety evaluation of cosmetics, taking into account scientific and ethical aspects of the problem;

6. The SCCNFP will be monitoring on a regular basis, scientific progress in the development and validation of alternative methods, and it will also evaluate their applicability to the safety testing of cosmetics, as well as immediately report its opinion to the Commission.

5. REFERENCES

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12. Council Directive 86/609/EEC of 24 November 1986. Official Journal of the European Community, No. L358 of 18-12-1986.