Testes in vitro como alternativa aos testes in vivo de Draize
Resumo
Os procedimentos descritos por Draize deram origem aos testes de irritação ocular e cutânea adotados internacionalmente para avaliar produtos e substâncias. Entretanto, eles são criticados por motivos éticos, devido à crueldade com os animais, mesmo após diferentes modificações terem sido propostas nos protocolos originais. Metodologias alternativas têm sido estudadas para avaliar a toxicidade de produtos usados em seres humanos. Entre as mais citadas encontram-se as que utilizam organismos inferiores, células vivas de mamíferos, sistemas organotípicos e substratos inertes, além de bancos de dados informatizados e programas que avaliam a toxicidade pela determinação de relação estrutura-atividade. Os métodos utilizando células vivas têm sido muito utilizados para predizer com segurança a irritação, contribuindo para a redução do número de animais utilizados nos testes in vivo. Até o momento, não existem métodos validados para substituir os ensaios de irritação ocular e cutânea, mas somente para avaliar substâncias corrosivas.Referências
American Society for Testing and Materials. Standard test method for agar diffusion cell culture screening for cytotoxicity: designation: F 895-84. Philadelphia: ASTM, 1995, p.276-9.
Azevedo, J.C. Avaliação de metodologia alternativa in vitro ao teste de irritação ocular de Draize. São Paulo, 1998. 145p. (Dissertação de Mestrado - Faculdade de Ciências Farmacêuticas USP).
Babich, H.; Babich, J.P. Sodium lauryl sulfate and triclosan: in vitro cytotoxicity studies with gingival cells. Toxicol. Lett., 91: 189-96, 1997.
Babich, H.; Borenfreund, E. Cytotoxic and morphological effects of phenylpropanolamine, caffeine, nicotine, and some of their metabolites studied in vitro. Toxicol. In Vitro, 6 (6): 493-502, 1992.
Bagley, D.M. et al. Eye irritation: reference chemicals data bank. Toxicol. In Vitro, 6: 487-91, 1992b.
Bagley, D.M. et al. An evaluation of five potential alternatives in vitro to the rabbit eye irritation test in vivo. Toxicol. In Vitro, 6 (4): 275-84, 1992a.
Bagley, D.M. et al. Skin irritation: reference chemicals data bank. Toxicol. In Vitro, 10:1-6, 1996.
Balls, M. Scientific validation: a crucial and unavoidable prerequisite to the acceptability of new tests and testing strategies. ATLA, Altern. Lab. Anim., 23: 474-9, 1995.
Balls, M. et al. The EC/HO international validation study on alternatives to the Draize eye irritation test. Toxicol. In Vitro, 9 (6): 871-929, 1995.
Balls, M.; Fentem, J.H. The validation and acceptance of alternatives to animal testing. Toxicol. In Vitro, 13: 837-46, 1999.
Barratt, M.D. QSARS for the eye irritation potential of neutral organic chemicals. Toxicol. In Vitro, 11: 1-8, 1991.
Barratt, M.D. et al. The ECVAM international validation study on in vitro tests for skin corrosivity. 1. Selection and distribution of the test chemicals. Toxicol. In Vitro, 12: 471-82, 1998.
Barrela, C., Roque, J.; Silva, T. Métodos alternativos à experimentação animal na indústria de cosméticos. [http:// www.fmv.utl.pt/democ/sft/ sem0001/G23.html]. 16 janeiro 2002.
Borenfreund, E.; Babich, H.; Martin-Alguacil, N. Comparisons of two
in vitro cytotoxicity assays- the neutral red (NR) and tetrazolium
MTT tests. Toxicol. In Vitro, 2 (1): 1-6, 1988.
Borenfreund, E.; Puerner, J.A. A simple quantitative procedure using
monolayer cultures for cytotoxicity assays (HTD/NR-90). J. Tissue
Cult. Methods, 9 (1): 7-9, 1984.
Borenfreund, E.; Puerner, J.A. Toxicity determined in vitro by
morphological alterations and neutral red absorption. Toxicol. Lett.,
: 119-24, 1985.
Botham, P. et al. Cell function-based assays. Food Chem. Toxicol.,
: 67-77, 1997.
Bradlaw, J.A.; Wilcox, N.L. Workshop on eye irritation testing:
Practical applications of non-whole animal alternatives. Food Chem.
Toxicol., 35: 1-11, 1997.
Brantom, P.G. et al. A summary report of the COLIPA international
validation study on alternatives to the Draize rabbit eye irritation
test. Toxicol. In Vitro, 11: 141-79, 1997.
Bruner, L.H. et al. Validation of alternative methods for toxicity
testing. Toxicol. In Vitro, 10: 479-501, 1996.
Cade, P. Avaliação da ação antiirritante, comparação entre métodos
in vivo e in vitro. Cosmet. Toiletries, Ed. Port., 2: 13-7, 1990.
Chamberlain, M. et al. Organotypic models for the assessment/prediction
of ocular irritation. Food Chem. Toxicol., 35: 23-37, 1997.
Chamberlain, M.; Parish, W.E. Hazard and risk based on in vitro test
data. Toxicol. In Vitro, 4 (4/5): 694-7, 1990.
Chiba, K. et al. Interlaboratory validation of the in vitro eye irritation
tests for cosmetic ingredients (9): evaluation of cytotoxicity test on
Hela cells. Toxicol. In Vitro, 13: 189-98, 1999.
Chiba, K., Kawakami, K., Tohyama, K. Simultaneous evaluation of
cell viability by neutral red, MTT and crystal violet staining assays of
the same cells. Toxicol. In Vitro, 12: 251-8, 1998.
Ciapetti, G. et al. Application of a combination of neutral red and
amido black staining for rapid, reliable cytotoxicity testing of
biomaterials. Biomaterials, 17 (13): 1259-64, 1996.
Clothier, R. et al. Assessment of initial damage and recovery following
exposure of MDCK cells to an irritant. Toxicol. In Vitro, 13: 713-7, 1999.
Combrier, E., Castelli, D. The agarose overlay method as a screening
approach for ocular irritancy: application to cosmetic products. ATLA,
Altern. Lab. Anim., 20 (3): 438-44, 1992.
Cooper, K.J. et al. Prediction of ocular irritancy of prototype shampoo
formulations by the isolated rabbit eye (IRE) test, and bovine corneal
opacity and permeability (BCOP) assay. Toxicol. In Vitro, 15: 95-
, 2001.
Courtellemont, P. et al. Relevance and reliability of the PREDISAFE
assay in the COLIPA eye irritation validation program (phase 1).
Toxicol. In Vitro, 13: 305-12, 1999.
Cronin, M.T.D.; Basketter, D.A., York, M. A quantitative structureactivity
relationship (QSAR) investigation of a Draize eye irritation
database. Toxicol. In Vitro, 8 (1): 21-8, 1994.
Curren, R.D. et al. Other assays. Food Chem. Toxicol., 35: 127-58,
De Angelis, I. et al. In vitro toxicity of some cosmetic ingredients.
Food Chem. Toxicol., 24 (6/7): 477-9, 1986.
Dickson, F.M., Lawrence, J.N.; Benford, D.J. Surfactant-induced
cytotoxicity in cultures of human keratinocytes and a commercially
available cell line (3T3). Toxicol. In Vitro, 7 (4): 381-4, 1993.
Draize, J.H.; Woodard, G.; Calvery, H.O. Methods for the study of
irritation and toxicity of substances applied topically to the skin and
mucous membranes. J. Pharmacol. Exp. Ther., 82: 377-90, 1944.
Earl, L.K. et al. Comparison of five potential methods for assessing
ocular irritation in vitro. Toxicol. In Vitro, 9 (3): 245-50, 1995.
Eisenbrand, G. et al. Methods of in vitro toxicology. Food Chem.
Toxicol., 40: 193-236, 2002.
FDA.U.S. Food and Drug Administration. Animal testing. [http://
vm.cfsan.fda.gov/~dms/cos-205html]. 8 março 2001.
Fentem, J.H. et al. The ECVAM international validation study on in
vitro tests for skin corrosivity. 2. Results and evaluation by the
management team. Toxicol. In Vitro, 12: 483-524, 1998.
Fentem, J.H. et al. A prevalidation study on in vitro tests for acute skin
irritation: results and evaluation by the management team. Toxicol.
In Vitro, 15: 57-93, 2001.
Gautheron, P. et al. Interlaboratory assessment of the bovine corneal
opacity and permeability (BCOP) assay. Toxicol. In Vitro, 8 (3):
-92, 1994.
Gettings, S.D. et al. The CTFA evaluation of alternatives program: an
evaluation of in vitro alternatives to the Draize primary eye irritation
test. (Phase II) oil/water emultions. Food Chem. Toxicol., 32 (10):
-76, 1994.
Gettings, S.D. et al. Comparison of low-volume, Draize and in vitro
eye irritation test data. I. Hydroalcoholic formulations. Food Chem.
Toxicol., 34: 737-49, 1996b
Gettings, S.D. et al. The CFTA evaluation of alternatives program: an
evaluation of in vitro alternatives to the Draize primary eye irritation
test. (Phase III) Surfactant-based formulations. Food Chem. Toxicol.,
(1): 79-117, 1996a.
Gettings, S.D. et al. A comparison of low volume, Draize and in vitro
eye irritation test data. II. Oil/water emulsions. Food Chem. Toxicol.,
: 47-59, 1998.
Gordon, V.C. The scientific basis of the EYTEX system. ATLA,
Altern. Lab. Anim., 20: 537-48, 1992.
Guess, W.L. et al. Agar diffusion method for toxicity screening of
plastics on cultured cell monolayers. J. Pharm. Sci., 54: 1545-7,
Harbell, J.W. et al. Cell cytotoxicity assays. Food Chem. Toxicol.,
: 79-126, 1997.
Hockley, K.; Baxter, D. Use of the 3T3 cell-neutral red uptake assay
for irritants as an alternative to the rabbit (Draize) test. Food Chem.
Toxicol., 24 (6/7): 473-5, 1986.
Husoy, T.; Syversen, T.; Jenssen, J. Comparisons of four in vitro
cytotoxicity tests: the MTT assay, NR assay, uridine incorporation
and protein measurements. Toxicol. In Vitro, 7 (2): 149-54, 1993.
International Organization for Standartization. ISO 10.993-5:
Biological evaluation of medical devices. Part 5. Test for cytotoxicity:
in vitro methods. Geneva: ISO, 1992. 7p.
Itagaki, H. et al. An in vitro alternative to the Draize eye-irritation
test: evaluation of the crystal violet staining method. Toxicol. In
Vitro, 5 (2): 139-43, 1991.
Jester, J.V. et al. Extent of initial corneal injury as a basis for alternative
eye irritation tests. Toxicol. In Vitro, 15: 115-30, 2001.
Kay, J.H.; Calandra, J.C. Interpretation of eye irritation tests. J. Soc.
Cosmet. Chem., 13: 281-9, 1962.
Kojima, H. et al. Evaluation of seven alternative assays on the main
ingredients in cosmetics as predictors of Draize eye irritation scores.
Toxicology in Vitro, 9 (3): 333-40, 1995.
Lawrence, J.N. Application of in vitro human skin models to dermal
irritancy: a brief overview and future prospects. Toxicol. In Vitro,
: 305-12, 1997.
LEE, J.K. et al. In vitro cytotoxicity tests on cultured human skin
fibroblasts to predict skin irritation potential of surfactants. Toxicol.
In Vitro, 14: 345-9, 2000.
Lewis, R.W.; Mccall, J.C.; Botham, P.A. A comparison of two
cytotoxicity tests for predicting the ocular irritancy of surfactants.
Toxicol. In Vitro, 7 (2): 155-8, 1993.
Lordo, R.A.; Feder, P.I.; Gettings, S.D. Comparing and evaluating
alternative (in vitro) tests on their ability to predict the Draize
maximum average score. Toxicol. In Vitro, 13: 45-72, 1999.
Louekari, K. In vitro toxicology and the test guidelines. ATLA, Altern.
Lab. Anim., 24: 435-8, 1996.
Lovell, D.P. Principal component analysis of tissue scores from
substances used in the COLIPA eye irritation validation study. Toxicol.
In Vitro, 13: 491-503, 1999.
Majmudar, G., Smith, M. Técnicas de screening in vitro em
dermatologia. Cosmet. Toiletries, Ed. Port., 10: 44-9, 1998.
Mosmann, T. Rapid colorimetric assay for cellular growth and survival:
application to proliferation and cytotoxicity assays. J. Immunol.
Methods, 65: 55-63, 1983.
Noser, F. Cultura de células a serviço da cosmetologia. Cosmet.
Toiletries, Ed. Port., 3: 46-7, 1991.
O’Brien, K.A.F. et al. An in vitro study of the eye irritation potential
of new shampoo formulations. Toxicol. In Vitro, 8 (2): 257-61,
Ohno, Y. et al. Interlaboratory validation of the in vitro eye irritation
tests for cosmetic ingredients. (1) overview of the validation study
and Draize scores for the evaluation of the tests. Toxicol. In Vitro,
: 73-98, 1999.
Okumura, H. et al. Interlaboratory validation of the in vitro eye
irritation tests for cosmetic ingredients. (10) evaluation of cytotoxicity
test on CHL cells. Toxicol. In Vitro, 13: 199-208, 1999.
Olivier, P. et al. Effect of high polyol concentrations on the neutral
red absorption assay and tetrazolium- MTT test of rat hepatocytes in
primary culture. Toxicol. In Vitro, 9 (2): 133-8, 1995.
Pinto, T.J.A.; Azevedo, J.C.; Cruz, A.S. Comparative study of epithelial
and fibroblastic cell lines as an alternative cytotoxicity test to the
Draize method. J. AOAC Int., 83 (3): 665-8, 2000.
Pinto, T.J.A.; Kaneko, T.M.; Ohara, M.T. Controle biológico de
qualidade de produtos farmacêuticos, correlatos e cosméticos.
São Paulo: Atheneu, 2000. 309p.
Riddell, R.J.; Clothier, R.H.; Balls, M. An evaluation of three in vitro
cytotoxicity assays. Food Chem. Toxicol., 24 (6/7): 469-71, 1986.
Robinson, M.K. et al. Non-animal testing strategies for assessment of
the skin corrosion and skin irritation potential of ingredients and
finished products. Food Chem. Toxicol., 40: 573-92, 2002.
Robinson, M.K.; Osborne, R.; Perkins, M.A. Strategies for the
assessment of acute skin irritation potential. J. Pharmacol. Toxicol.
Methods, 42: 1-9, 1999.
Roggeband, R. et al. Eye irritation responses in rabbit and man after
single applications of equal volumes of undiluted model liquid detergent
products. Food Chem. Toxicol., 38: 727-34, 2000.
Roguet, R. et al. An interlaboratory study of the reproducibility and
relevance of Episkin, a reconstructed human epidermis, in the
assessment of cosmetics irritancy. Toxicol. In Vitro, 12: 295-304,
Roguet, R.; Schaefer, H. Overview of in vitro cell culture technologies
and pharmaco-toxicological applications. Toxicol. In Vitro, 11: 591-
, 1997.
Rosenbluth, S.A. et al. Tissue culture method for screening toxicity of
plastic materials to be used in medical practice. J. Pharm. Sci., 54:
-9, 1965.
Rougier, A. et al. The use of in vitro methods in the ocular irritation
assessment of cosmetic products. Toxicol. In Vitro, 8 (4): 893-905,
Rutten, A.A.J.J.L., Van de Sandt, J.J.M. In vitro dermal toxicology
using skin organ cultures. Toxicol. In Vitro, 8 (4): 703-5, 1994.
Saotome, K.; Morita, H.; Umeda, M. Cytotoxicity test with simplified
crystal violet staining method using microtitre plates and its application
to injection drugs. Toxicol. In Vitro, 3 (4): 317-21, 1989.
Sasaki, T. et al. Detergent cytotoxicity simplified assay of cytolysis
by measuring LDH activity. Toxicol. In Vitro, 6 (5): 451-7, 1992.
Seabaugh, V.M. et al. Use of ophthalmic topical anesthetics. Food
Chem. Toxicol., 31: 95-8, 1993.
Southee, J.A. et al. The performance of the tissue equivalent assay
using the skin ZK 1200 model in the COLIPA international validation
study on alternatives to the Draize eye irritation test. Toxicol. In
Vitro, 13: 355-73, 1999.
Spielmann, H.; Liebsch, M. Lessons learned from validation of in
vitro toxicity test: from failure to acceptance into regulatory practice.
Toxicol. In Vitro, 15: 585-90, 2001.
Spielmann, H. et al. CAM-based assays. Food Chem. Toxicol., 35:
-66, 1997.
Springer, J.A. et al. Number of animals for sequential testing. Food
Chem. Toxicol., 31: 105-9, 1993.
Tani, N. et al. Interlaboratory validation of the in vitro eye irritation
tests for cosmetic ingredients. (8): evaluation of cytotoxicity tests on
SIRC cells. Toxicol. In Vitro, 13: 175-87, 1999.
Tsutsui, T. et al. Quantitative comparison of cytotoxicity of dental
antiseptics to normal human oral keratinocytes in vitro. Toxicol. In
Vitro, 8 (6): 1253-8, 1994.
Uchiyama, T. et al. Interlaboratory validation of the in vitro eye
irritation tests for cosmetic ingredients (7): evaluation of cytotoxicity
test by CornePack. Toxicol. In Vitro, 13: 163-73, 1999.
United States Pharmacopeia. 22.ed. Rockville: United States
Pharmacopeial Convention, 1990. p.1495-7.
United States Pharmacopeia. 23.ed. Rockville: United States
Pharmacopeial Convention, 1995. p.1697-9.
United States Pharmacopeia. 24.ed. Rockville: United States
Pharmacopeial Convention, 1999. p.1831-2.
United States Pharmacopeia. 25.ed. Rockville: United States
Pharmacopeial Convention, 2002. p.1893-5.
United States Pharmacopeia. 26.ed. Rockville: United States
Pharmacopeial Convention, 2003. p.2026-8.
Van de Sandt, J.J.M.; Rutten, A.A.J.J.L. Differential effects of chemical
irritants in rabbit and human skin organ cultures. Toxicol. In Vitro, 9
(2): 157-68, 1995.
Van de Sandt, J.J.M.; Rutten, A.A.J.J.L.; Koeter, H.B.W.M. Cutaneous
toxicity testing in organ culture: neutral red uptake and reduction of
tetrazolium salt (MTT). Toxicol. In Vitro, 7 (1): 81-6, 1993.
Van Zutphen, L.F.M.; Van der Valk, J.B.F. Developments on the
implementation of the three Rs in research and education. Toxicol.
In Vitro, 15: 591-5, 2001.
Vian, L. et al. Comparison of three in vitro cytotoxicity assays for
estimating surfactant ocular irritation. Toxicol. In Vitro, 9 (2): 185-
, 1995.
Wilhelmus, K.R. The Draize eye test: therapeutic reviews. Surv.
Ophthalmol., 45 (6): 493-515, 2001.
