Kozmetiklerin Deri irritasyon/Korozyon Potansiyellerinin Değerlendirilmesi için Yöntemler

Year 2016, Issue: 2, 150 - 169, 01.06.2016

Özge Köse Suna Sabuncuoğlu Pınar Erkekoğlu Belma Koçer Gümüşel

Abstract

İnsan derisi, organizmayı çevresel faktörlere karşı koruyan, farmasötik formülasyonların ve kozmetik ürünlerin içeriğinde bulunan birçok kimyasal maddeye karşı engel teşkil eden önemli bir organdır. Kimyasal maddelere maruziyet sonrasında oluşan lokal toksik etkiler arasında en sık görüleni deri irritasyonudur. Geçmişte kozmetik ürünlerinin deride irritasyon oluşturma potansiyelleri hayvan deneyleri ile değerlendirilmiştir. Etik açıdan, deri irritasyon ve deri korozyon testlerinin laboratuvar hayvanları üzerinde yapılması, hayvanlara önemli derecede rahatsızlık ve acı verme potansiyeline sahiptir. 3R ilkelerinin yayınlanmasından bu yana kimyasal maddelerin potansiyel toksik etkilerinin değerlendirilmesi için, birçok alternatif metot geliştirilmiş, valide edilmiş ve düzenleyici kuruluşlar tarafından kabul edilmiştir. 11 Mart 2013 tarihinde Avrupa Birliği’nde hayvanlar üzerinde test edilen her tür kozmetik ve kişisel bakım ürününün satışı yasaklanmasından sonra kozmetik ürünlerin güvenlilik değerlendirmeleri, alternatif in vitro toksisite testleri ile yapılmaya başlanmıştır. Alternatif yöntemler olarak; in vitro yeniden yapılandırılmış insan epidermis doku modelleri, insan derisine morfolojik olarak yakın benzerliği nedeni ile tercih edilmektedir. Bu derleme kapsamında, kozmetik ürünlerin irritasyon/korozyon potansiyellerinin değerlendirlmesinde kullanılan in vivo ve alternatif in vitro yöntemlerden bahsedilmektedir.

Keywords

Alternatif yöntemler, in vitro, deri irritasyonu, deri korozyonu, kozmetik ürünler

The Methods for Evaluation of Skin Irritation/Corrosion Potentials

Abstract

The human skin is an important organ that protects the organism against environmental factors and chemicals in the pharmaceutical formulations and cosmetic products. Skin irritation is the most common local toxic effect after exposure to cosmetic products. The potentials of the cosmetic products to induce skin irritation were evaluated by animal experiments in the past. In ethical terms, the irritation and skin corrosion tests have the potential to cause significant pain Since 3R principle was published, many alternative methods, which were validated and accepted by the regulatory authorities, have been developed for the evaluation of the potential toxic effects of the chemicals. On March 11, 2013, the commercial sale of any types of cosmetics and personal care products that were tested on animals was banned in the European Union; and since then, the safety evaluation of the cosmetics is being conducted with alternative in vitro toxicity tests. The in vitro restructured human epidermis tissue models are preferred as an alternative method because of their morphological similarity to the human skin. In this review, the traditional and alternative methods used for the evaluation of the irritation and corrosion potentials of cosmetic products are being discussed.

Keywords

Alternative methods, in vitro, skin irritation, skin corrosion, cosmetic products

References

• 1. Monterio-Riviere N. In: Riviere, J. (ed), Structure and Function of Skin. Dermal Absorption Models in Toxicology and Pharmacology, CRC Press, Taylor and Francis Group, New York, NY. 2009:1-19.
• 2. Lopez-Pajares V, Yan K, Zarnegar BJ, Jameson KL, Khavari, PA: Genetic pathways in disorders of epidermal differentiation. Trends in Genetics 2013;29(1):31-40.
• 3. Eskes C: Application of alternative methods in the regulatory assessment of chemical safety related to human skin corrosion & irritation current status and future prospects, Swiss Federal Office of Public Health – FOPH. 2010.
• 4. Lavker RM, Sun TT: Heterogeneity in epidermal basal keratinocytes: morphological and functional correlations. Science 1982;215(4537):1239-1241.
• 5. Lavker RM, Sun TT: Epidermal stem cells. J Invest Dermatol 1983;81(1 Suppl):121s-127s.
• 6. Steinhoff M, Brzoska T, Luger TA: Keratinocytes in epidermal immune responses. Current Opinions in Allergy and Clinical Immunology 2001;1(5):469-476.
• 7. Basketter DA, Whittle E, Griffiths HA, York M: The identification and classification of skin irritation hazard by a human patch test. Food and Chemical Toxicology. 1994;32(8):769-775.
• 8. Basketter D, Pease C, Kasting G, Kimber I, Casati S, Cronin M, Diembeck W, Gerberick F, Hadgraft J, Hartung T, Marty JP, Nikolaidis E, Patlewicz G, Roberts D, Roggen E, Rovida C, van de Sandt J: Skin sensitisation and epidermal disposition: the relevance of epidermal disposition for sensitisation hazard identification and risk assessment. Alternatives to laboratory animals (ALTA) 2007;35(1):137-154.
• 9. Sithamparanadarajah R. Diseases caused by dermal exposure to chemicals. In: Evans, PG. (eds), Controlling skin exposure to chemicals and Wet-working: a practical book. RMS Publishing, UK. 2008:15-24.
• 10. European Commision (EC) Cosmetics Directive: Council Directive of 27 July 1976 on the approximation of the laws of the Member States relating to cosmetic products. (76/768/EEC). 1976. Available from: http://eur-lex.europa.eu/legal-content/EN/TXT/?uri=CELEX%3A31976L0768.
• 11. Scientific Committee on Consumer Products (SCCP): THE SCCP’S Notes of Guidance For The Testing of Cosmetic İngredients And Their Safety Evaluation. 6th Revision. 2006. Available from: http://ec.europa.eu/health/ph_risk/committees/04_sccp/docs/sccp _o_03j.pdf
• 12. Stitzel KA. Tiered testing strategies--acute local toxicity. ILAR J. 2002;43Suppl:S21-6.
• 13. European Commision (EC): Regulation (EC) No 1223/2009 of The European Parliament and of The Council of 30 November 2009 on Cosmetic Products, Official Journal of the European Union. 2009. Available from: http://ec.europa.eu/health//sites/health/files/ endocrine_disruptors/docs/ cosmetic_1223_2009_regulation_en.pdf
• 14. Türkiye İlaç ve Tıbbi Cihaz Kurumu Kozmetik Ürünlerde Güvenlilik Değerlendirmesi ve Güvenlilik Değerlendiricisi Hakkında Kılavuz. Available from: https://www.titck.gov.tr /Portal Admin/Uploads/ UnitPageAttachment/tYDCLux5.pdf
• 15. 24/3/2005 tarihli ve 5324 sayili Kozmetik Kanunu. Yayimlandığı Resmi Gazete Tarihi: 30/3/2005, Sayi:25771, Tertip: 5, Cilt : 44. 2005.
• 16. İyi Laboratuvar Uygulamaları Prensipleri, Test Birimlerinin Uyumlaştırılması, İyi Laboratuvar Uygulamalarının Ve Çalışmaların Denetlenmesi Hakkında Yönetmelik, Resmi Gazete Tarihi: 09.03.2010 Resmi Gazete Sayisi: 27516. 2010.
• 17. Organization for Economic Cooperation and Development (OECD). Descriptions of selected key generic terms used in chemical hazard/risk assessment: ENV/JM/MONO(2003)15— OECD Series on Testing And Assessment Number 44, OECD, Paris. 2003. Available from: http://www.oecd.org/officialdocuments/publicdisplaydocumentpdf/?cote=ENV/JM/MONO (2003)15&docLanguage=En.
• 18. Rice RH, Mauro TM. Toxic Responses of the Skin. (Klaassen CD, ed) Casarett & Doull’s Toxicology: The Basic Science of Poisons, Eighth Edition. 2012; 839-860.
• 19. Pauwels M, Rogiers V: Human health safety evaluation of cosmetics in the EU: a legally imposed challenge to science. Toxicology and Applied Pharmacology 2010;243(2):260-274.
• 20. Berardesca E, Distante F: The modulation of skin irritation. Contact Dermatitis 1994;31(5): 281–287.
• 21. Organization for Economic Cooperation and Development (OECD). OECD Guidelines for the Testing of Chemicals No. 404: Acute Dermal İrritation/Corrosion. Paris, France. 2002.
• 22. Robinson MK, Cohen C, de Fraissinette Ade B, Ponec M, Whittle E, Fentem JH: Non-animal testing strategies for assessment of the skin corrosion and skin irritation potential of ingredients and finished products. Food and Chemical Toxicology 2002;40(5):573-592.
• 23. Organization for Economic Cooperation and Development (OECD). OECD Guidelines for the Testing of Chemicals No. 430: İn vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER). Paris, France: Organisation for Economic Cooperation and Development. 12pp.
• 24. Lores M, Llompart M, Alvarez-Rivera G, Guerra E, Vila M, Celeiro M, Lamas JP, Garcia-Jares C: Positive lists of cosmetic ingredients: Analytical methodology for regulatory and safety controls - A review. Analytica Chimica Acta 2016;915:1-26.
• 25. Natsch A: What Makes Us Smell: The Biochemistry of Body Odour and the Design of New Deodorant Ingredients. Chimia (Aarau) 2015;69(7-8):414-420.
• 26. Chiu CH, Huang SH, Wang HM: A Review: Hair Health, Concerns of Shampoo Ingredients and Scalp Nourishing Treatments. Current Pharmaceutical Biotechnology 2015;16(12):1045-1052.
• 27. Welss T, Basketter DA, Schröder KR: İn vitro skin irritation: facts and future State of the art review of mechanisms and models, Toxicology in Vitro 2004;18(3):231-243.
• 28. Osborne R, Perkins M.A: An approach for development of alternative test methods based on mechanisms of skin irritation. Food and Chemical Toxicology 1994;32(2):133–142.
• 29. Luger TA: Epidermal cytokines, Acta Dermatologica Venereologica Suppl (Stockholm) 1989;151: 61–76.
• 30. Nickoloff BJ, Naidu Y: Pertubation of epidermal barrier function correlates with initiation of cytokine cascade in human skin, Journal of American Academy of Dermatology 1994;30(4):535–546.
• 31. Corsini E, Galli CL: Cytokines and irritant contact dermatitis. Toxicology Letters 1998;102-103, 277–282 (1998)
• 32. Dinarello CA: İnterleukin-1, interleukin-1 receptors and interleukin-1 receptor antagonist, İnternational Reviewa of Immunology 1998;16(5-6):57-99.
• 33. Stylianou E, Saklatvala J: İnterleukin-1, The International Journal of Biochemistry & Cell Biology 1998;30(10):1075–1079.
• 34. Camera E, Jensen C, Stab S, Scala G, Baadsgaard O, Picardo M: Correlation between antioxidant levels and skin reactivity to irritants, Journal of Dermatological Science 2003;16(1): 49-54.
• 35. Willis CM, Reiche L, Wilkinson JD: İmmunocytochemical demonstration of reduced Cu,Znsuperoxide dismutase levels following topical application of dithranol and sodium lauryl sulphate: an indication of the role of oxidative stress in acute irritant contact dermatitis. European Journal of Dermatology 1998;8 (1):8–12
• 36. Rogers JV, Gunasekar PG, Garrett CM, Kabbur MB, McDougal JN: Detection of oxidative species and lowmolecular-weight DNA in skin following dermal exposure with JP-8 jet fuel. Journal of Applied Toxicology 2001;21 (6):521–525.
• 37. Camhi SL, Lee P, Choi AM: The oxidative stress response. New Horizon 1995;3(2):170–182.
• 38. Allen RG, Tresini M: Oxidative stress and gene regulation, Free Radical Biology and Medicine 2000;28(3):463–499.
• 39. Draize JH, Woodard G, Calvery HO: Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes, Journal of Pharmacology and Experimental Therapeutics 1944;82, 377–390.
• 40. Basketter DA, Whittle E, Griffiths HA, York M: The identification and classification of skin irritation hazard by a human patch test. Food and Chemical Toxicology 1994;32(8):769–775.
• 41. Basketter DA, Chamberlain M, Griffiths HA, Rowson M, Whittle E, York M: The classification of irritants by human patch test. Food and Chemical Toxicology 1997;35(8):845-852.
• 42. Basketter DA, York M, McFadden JP, Robinson MK: Determination of skin irritation potential in the human 4-h patch test. Contact Dermatitis 2004;51(1): 1–4.
• 43. York M, Griffiths HA, Whittle E, Basketter DA: Evaluation of a human patch test for the identification and classification of skin irritation potential. Contact Dermatitis 1996;34:204-212.
• 44. Robinson MK, McFadden JP, Basketter DA: Validity and ethics of the human 4-h patch test as an alternative method to assess acute skin irritation potential. Contact Dermatitis 2001;45(1):1-12.
• 45. Organization for Economic Cooperation and Development (OECD). Guidance Document on the Validation and İnternational Acceptance of New or Updated Test Methods for Hazard Assessment. Environmental Health and Safety Monograph Series on Testing and Assessment No. 34.2005.
• 46. Weil CS, Scala RA: Study of intra- and interlaboratory variability in the results of rabbit eye and skin irritation tests, Toxicology and Applied Pharmacology 1971;19 (2):276-360.
• 47. Jirova D, Basketter D, Liebsch M, Bendova H, Kejlova K, Marriott M, Kandarova H: Comparison of human skin irritation patch test data with in vitro skin irritation assays and animal data. Contact Dermatitis 2010;62(2):109-116.
• 48. Nixon GA, Tyson CA, Wertz WC: Interspecies comparisons of skin irritancy. Toxicology and Applied Pharmacology 1975;31(3):481–490.
• 49. Phillips L, Steinberg M, Maibach Hİ, Akers A: A comparison of rabbit and human skin response to certain irritants. Toxicology and Applied Pharmacology 21, 369–382 (1972)
• 50. Benassi L, Bertazzoni G, Seidenari S: In vitro testing of tensides employing monolayer cultures: a comparison with results of patch tests on human volunteers. Contact Dermatitis 1999;40(1):38-44.
• 51. Zuang V, Balls M, Botham PA, Coquette A, Corsini E, Curren RD, Elliott GR, Fentem JH, Heylings JR, Liebsch M, Medina J, Roguet R, van de Sandt JJ, Wiemann C, Worth AP: Follow-up to the ECVAM prevalidation study on in vitro tests for acute skin irritation. The European Centre for the Validation of Alternative Methods Skin Irritation Task Force report 2. Alternatives to laboratory animals (ALTA) 2002;30(1):109-129.
• 52. Eskes C, Detappe V, Koëter H, Kreysa J, Liebsch M, Zuang V, Amcoff P, Barroso J, Cotovio J, Guest R, Hermann M, Hoffmann S, Masson P, Alépée N, Arce LA, Brüschweiler B, Catone T, Cihak R, Clouzeau J, D’Abrosca F, Delveaux C, Derouette JP, Engelking O, Facchini D, Fröhlicher M, Hofmann M, Hopf N, Molinari J, Oberli A, Ott M, Peter R, Sá-Rocha VM, Schenk D, Tomicic C, Vanparys P, Verdon B, Wallenhorst T, Winkler GC, Depallens O: Regulatory assessment of in vitro skin corrosion and irritation data within the European framework: Workshop recommendations. Regulatory Toxicology and Pharmacology 201;62(2):393-403.
• 53. European Comission. Communication from the commission to the European Parliament and the councilon the animal testing and marketing ban and on the state of play in relation to alternative methods in the field of cosmetics. 2013. Available from: http://ec.europa.eu/consumers/sectors/ cosmetics/files/pdf/animal_testing/com_at_2013_en.pdf
• 54. European Comission. Directive 2010/63/EU of the European Parliament and of the council of 22 September 2010 on the protection of animals used for scientific purposes. Official Journal of the European Union 2010; 276:33–79.
• 55. Kandárová H, Letašiová S: Alternative methods in toxicology: pre-validated and validated methods, İnterdisciplinary Toxicology 2011;4(3):107–113.
• 56. Tinois E, Tiollier J, Gaucherand M, Dumas H, Tardy M, Thivolet J: In vitro and post-transplantation differentiation of human keratinocytes grown on the human type IV collagen film of a bilayered dermal substitute. Experimetal Cell Research 1991;193(2):310-319.
• 57. Niwa M, Nagai K, Oike H, Kobori M: Evaluation of the skin irritation using a DNA microarray on a reconstructed human epidermal model. Biological and Pharmaceutical Bulletin 2009 ;32(2):203-208.
• 58. Hoffmann J, Heisler E, Karpinski S, Losse J, Thomas D, Siefken W, Ahr HJ, Vohr HW, Fuchs HW: Epidermal-skin-test 1,000 (EST-1,000)--a new reconstructed epidermis for in vitro skin corrosivity testing. Toxicol In Vitro 2005;19(7):925-9.
• 59. De Wever B, Goldberg A, Eskes C, Roggen E, Vanparys P, Schröder K, Le Varlet B, Maibach H, Beken S, De Wilde B, Turchina C, Bogaert G, Bogaert J-P. Open Source’’–Based Engineered Human Tissue Models: A New Gold Standard for Nonanimal Testing Through Openness, Transparency, and Collaboration, Promoted by the ALEXANDRA Association Applıed In Vitro Toxicology. 2015;1:5-9.
• 60. Poumay Y, Dupont F, Marcoux S, Leclercq-Smekens M, Hérin M, Coquette A: A simple reconstructed human epidermis: preparation of the culture model and utilization in in vitro studies. Archieves of Dermatological Research 2004;296(5):203-211.
• 61. Kandárová H, Liebsch M, Spielmann H, Genschow E, Schmidt E, Traue D, Guest R, Whittingham A, Warren N, Gamer AO, Remmele M, Kaufmann T, Wittmer E, De Wever B, Rosdy M: Assessment of the human epidermis model SkinEthic RHE for in vitro skin corrosion testing of chemicals according to new OECD TG 431. Toxicology in Vitro.2006;20(5):547-559.
• 62. Borlon C, Godard P, Eskes C, Hartung T, Zuang V, Toussaint O: The usefulness of toxicogenomics for predicting acute skin irritation on in vitro reconstructed human epidermis. Toxicology 2007;241(3):157-166.
• 63. Dumont C, Prieto P, Asturiol D, Worth A: Review of the Availability of In Vitro and In Silico Methods for Assessing Dermal Bioavailability. Applied In Vitro Toxicology 2015;1(2):147-164
• 64. European Centre for the Validation of Alternative Methods (ECVAM) Scientific Advisory Committee (ESAC). Statement on the scientific validity of in vitro tests for skin irritation testing. 2010.
• 65. Rosdy M: Assessment of the human epidermis model SkinEthic RHE for in vitro skin corrosion testing of chemical according to new OECD TG 431. Toxicology in Vitro. 2006;20:547–559.
• 66. European Centre for the Validation of Alternative Methods (ECVAM). Performance Standards for In-Vitro Skin Irritation Test Methods 5 based on Reconstructed Human Epidermis (RhE). 2009. Available from: https://eurl-ecvam.jrc.ec.europa.eu/validation-regulatory-acceptance/docs-skinirritation-1/DOC8-updated_ECVAM_PS_2009.pdf
• 67. European Centre for the Validation of Alternative Methods (ECVAM) Scientific Advisory Committee (ESAC). Statement on the Performance under UN GHS of three in vitro assays for Skin İrritation testing and the Adaptation of the Reference chemicals and defined accuracy values of the ECVAM Skin İrritation Performance Standards. 2009.
• 68. Alépée N, Robert C, Tornier C, Cotovio J: The usefulness of the validated SkinEthic™ RHE test method to identify skin corrosive UN GHS subcategories. Toxicology in Vitro. 2014;28:616-625.
• 69. European Centre for the Validation of Alternative Methods (ECVAM) Scientific Advisory Committee (ESAC). Statement on the application of the SkinEthicTM human skin model for skin corrosivity testing, issued by the ECVAM Scientific Advisory Committee. 2006.
• 70. Ponec M, Weerheim A, Kempenaar J, Mulder A, Gooris GS, Bouwstra J, Mommaas AM: The formation of competent barrier lipids in reconstructed human epidermis requires the presence of vitamin C. Journal of Investigative Dermatology. 1997;109(3):348-355.
• 71. Ponec M, Boelsma E, Gibbs S, Mommaas M: Characterization of reconstructed skin models. Skin Pharmacology and Applied Skin Physiology 2002;15 Suppl 1:4-17.
• 72. Robson KJ, Stewart ME, Michelsen S, Lazo ND, Downing DT: 6-Hydroxy-4-sphingenine in human epidermal ceramides. Journal of Lipid Research 1994;35(11):2060-2068.
• 73. Stewart, ME, Downing DT: A new 6-hydroxy-4-sphingeninecontaining ceramide in human skin, Journal of Lipid Research 1999;40(8):1434–1439.
• 74. Ponec M, Boelsma E, Weerheim A, Mulder A, Bouwstra J, Mommaas M: Lipid and ultrastructural characterization of reconstructed skin models. International Journal of Pharmacy 2000;203(1- 2):211-225.
• 75. Boelsma E, Gibbs S, Faller C, Ponec M: Characterization and comparison of reconstructed skin models: morphological and immunohistochemical evaluation. Acta Dermatologica Venereologica 2000;80(2):82-88.
• 76. Motta S, Monti M, Sesana S, Caputo R, Carelli S, Ghidoni R: Ceramide composition of the psoriatic scale. Biochimica et Biophysica Acta. 1993;1182(2):147-151.
• 77. Deshmukh GR, Kumar KH, Reddy PV, Rao BS: İn vitro skin corrosion: Human skin model test - A validation study, Toxicology in Vitro 2012;26(6):1072-1074.
• 78. Faller C, Bracher M, Dami N, Roguet R. Predictive ability of reconstructed human epidermis equivalents for the assessment of skin irritation of cosmetics. Toxicology in Vitro 2002;16(5):557- 572.
• 79. Organization for Economic Cooperation and Development (OECD). İn vitro Skin İrritation: Reconstructed Human Epidermis Test Method. OECD Guideline for the Testing of Chemicals No. 439. Paris, France. 2010.
• 80. Organization for Economic Cooperation and Development (OECD). Test No. 431: In vitro skin corrosion: reconstructed human epidermis (RHE) test method, OECD Publishing, Paris. Available from: http://www.oecd-ilibrary.org/docserver/download/9716161e.pdf?expires= 1481878451&id= id&accname= guest&checksum= 7D973C344F4B0598966A14F512F519D7.
• 81. European Centre for the Validation of Alternative Methods (ECVAM)Validation & regulatory acceptance. Available from: https://eurl-ecvam.jrc.ec.europa.eu/validation-regulatory-acceptance
• 82. Japanese Center for the Validation of Alternative Methods (JACVAM) Homepage. Available from: http://www.jacvam.jp/en/index.html
• 83. European Union Reference Laboratory for alternatives to animal testing (EURL-ECVAM) Homepage. Available from: https://eurl-ecvam.jrc.ec.europa.eu
• 84. National Toxicology Program. About ICCVAM. Available from: http://ntp.niehs.nih.gov/pubhealth/ evalatm/iccvam/index.html
• 85. German Centre for the Protection of Laboratory Animals (Bf3R) ZEBET Homepage. Available from: http://www.bfr.bund.de/en/zebet-58194.html
• 86. Johns Hopkins University Center for Alternatives to Animal Testing Homepage. Available from: http://caat.jhsph.edu/
• 87. FRAME Homepage. Available from: http://www.frame.org.uk/
• 88. Wilhelmus KR: The Draize eye test. Surveys in Ophthalmology 2001;45(6):493-515.
• 89. Fentem JH, Briggs D, Chesné C, Elliott GR, Harbell JW, Heylings JR, Portes P, Roguet R, van de Sandt JJ, Botham PA: A prevalidation study on in vitro tests for acute skin irritation. results and evaluation by the Management Team. Toxicology in Vitro 2001;15(1):57-93.
• 90. Heylings JR, Diot S, Esdaile DJ, Fasano WJ, Manning LA, Owen HM: A prevalidation study on the in vitro skin irritation function test (SIFT) for prediction of acute skin irritation in vivo: results and evaluation of ECVAM Phase III. Toxicology in Vitro. 2003;17(2):123-38.
• 91. Kandárová H, Liebsch M, Genschow E, Gerner I, Traue D, Slawik B, Spielmann H: Optimisation of the EpiDerm test protocol for the upcoming ECVAM validation study on in vitro skin irritation tests. Alternatives to animal experimentation (ALTEX) 2004;21(3):107-14.
• 92. Portes P, Grandidier MH, Cohen C, Roguet R. Refinement of the Episkin protocol for the assessment of acute skin irritation of chemicals: follow-up to the ECVAM prevalidation study. Toxicology in Vitro 2002;16(6):765-770.
• 93. Worth AP, Cronin MTD: The use of bootstrap resampling to assess the variability of Draize tissue scores. Alternatives to laboratory animals (ALTA) 2001;29(5):557-573.
• 94. Kozmetik Sektörü. TC Ekonomi Bakanlığı. http://www.ekonomi.gov.tr/portal/content/ conn/UCM/ uuid/dDocName:EK-051185
• 95. Türkiye İlaç ve Tıbbi Cihaz Kurumu Kozmetik Ürün Bilgi Dosyasının İçermesi Gerekenlere İlişkin Kılavuz. Available from: https://www.titck.gov.tr/PortalAdmin/ Uploads/UnitPageAttachment/ ynVWPlD3.pdf
• 96. Türkiye İlaç ve Tıbbi Cihaz Kurumu Kozmetik Ürünlerde Güvenlilik Değerlendirmesi ve Güvenlilik Değerlendiricisi Hakkında Kılavuz. Available from: https://www.titck.gov.tr/ PortalAdmin/Uploads/ UnitPageAttachment/tYDCLux5.pdf.
• 97. Worth AP, Balls M. The principles of validation and the ECVAM validation process. Alternatives to laboratory animals (ALTA) 2004;32 Suppl 1B:623-629.