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Cell Biology and Toxicology

, Volume 15, Issue 1, pp 63–75 | Cite as

Use of skin cell cultures for in vitro assessment of corrosion and cutaneous irritancy

  • R. Roguet
Article

Abstract

Skin cell culture is one of the most promising tools for in vitro evaluation of both cutaneous irritancy and corrosion. New culture methodologies, including three-dimensional reconstruction of skin, allow the evaluation of a wide range of compounds and complex formulations. A number of tests have already been developed for the evaluation of cytotoxicity and many end-points are now currently used, including cell viability, alteration of cell growth or cell function. In recent years parameters more closely related to in vivo irritancy effects such as synthesis of inflammatory mediators and/or their release by keratinocytes after exposure to potential skin irritants have been evaluated. This paper reviews technological aspects and results of validation using skin cell culture for in vitro assessment of corrosion and skin irritancy. Advantages and limits of skin cell cultures are also presented. Current questions about the validation process of cutaneous irritation and corrosion are also considered.

corrosion cutaneous irritancy in vitro assessment skin cell culture 

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References

  1. Augustin C, Collombel C, Damour O. Use of dermal equivalent and skin equivalent for in vitro cutaneous irritation testing of cosmetic products: comparison with in vivo human data. J Toxicol Cutan Ocul Toxicol. 1998;17:5-17.Google Scholar
  2. Baden HP, Kubilus J, Kvedar JC, Steinberg ML, Wolman SR. Isolation and characterization of a spontaneous arising long-lived line of human keratinocytes (NMI). In Vitro Cell Dev Biol. 1987;23:205-13.Google Scholar
  3. Bartnik FG, Pitterman WF. Skin organ culture for the study of skin irritancy. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:171-81.Google Scholar
  4. Basketter DA, Whittle E. A review of human in vitro skin corrosivity test. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:151-9.Google Scholar
  5. Boelsma E, Tanojo H, Boddé HE, Ponec M. An in vivo-in vitro study of the use of a human skin equivalent for irritancy screening of fatty acids. Toxicol in Vitro. 1997;11:365-76.Google Scholar
  6. Botham PA, Hall TJ, Dennet R et al. The skin corrosive test in vitro. Results of a inter-laboratory trial. In Vitro. 1992;6:191-205.Google Scholar
  7. Botham PA, Chamberlain M, Barratt MD et al. A prevalidation study on in vitro skin corrosivity testing. The report and recommendation of ECVAM Workshop 6. ATLA. 1997;23:219-55.Google Scholar
  8. Botham PA, Earl LK, Fentem JH, Roguet R, Van de Sandt JJM Alternatives methods for skin irritation testing: the current status. ECVAM Skin Irritation Task Force Report 1. ATLA. 1998;2:195-211.Google Scholar
  9. Boukamp P, Petrussevka RT, Breitkreutz D et al. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol. 1988;106:761-71.Google Scholar
  10. Braa SS, Triglia D. Predicting ocular irritation using 3-dimensional human fibroblast cultures. Cosmetics Toiletries. 1991;106:55-60.Google Scholar
  11. Boyce ST, Ham RG. Cultivation, frozen storage and clonal growth of normal epidermal keratinocytes in serum-free media. J Tissue Culture Methods. 1985;9:83-93.Google Scholar
  12. Burleson FG, Limardi LC, Sikorski EE, Rheins LA, Donnelly TA, Gerberick GK. Cytokine mRNA expression in an in vitro human skin model SKIN2. Toxicol in Vitro. 1996;10:513-21.Google Scholar
  13. Chamberlain M, Earl L. Use of cell cultures in irritancy testing. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:59-67.Google Scholar
  14. Cohen C, Dossou G, Rougier A, Roguet R. Measurement of inflammatory mediators produced by human keratinocytes in vitro: a predictive assessment of cutaneous irritation. Toxic in Vitro. 1991;5:407-10.Google Scholar
  15. Cohen C, Dossou G, Rougier A, Roguet R. Episkin: an in vitro model for the evaluation of phototoxicity and sunscreen photoprotective properties Toxicol in Vitro. 1994a;8:669-71.Google Scholar
  16. Cohen C, Selvi-Bignon C; Barbier A, Rougier A, Lacherretz F, Roguet R Measurement of proinflammatory mediators production by cultured keratinocytes: a predictive assessment of cutaneous irritancy. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:83-96.Google Scholar
  17. Cotovio J, Leclaire J, Roguet R. Cytochrome P450-dependent enzyme activities in adult human keratinocytes and transformed human keratinocytes. In Vitro Toxicol. 1997;10:207-16.Google Scholar
  18. DeLéo VA, Harber LC, Kong BM, De Salva SJ. Surfactant induced alteration of arachidonic acid metabolism on mammalian cells in culture. Proc Soc Exp Med. 1987;184:477-82.Google Scholar
  19. DeLéo VA, Hondon D, Scheide S, Kong BJ, De Salva SJ. The effect of surfactant on the metabolism of choline phospholipids in human epidermal keratinocytes in culture. J Toxicol Cutan Ocul Toxicol. 1989;8:227-40.Google Scholar
  20. De Wever B, Rheins LA. Skin2: an in vitro human skin analog. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:121-31.Google Scholar
  21. Dickson FM, Lawrence JN, Benford DJ Surfactant-induced cytotoxicity in cultures of human keratinocytes and commercially available cell line (3T3). Toxicol in Vitro. 1993;7: 381-4.Google Scholar
  22. Dickson FM, Lawrence JN, Benford DJ. Cytotoxicity of 12 chemicals of known human and animal skin irritation potential in human keratinocyte cultures. Toxicol in Vitro 1994;8:661-3.Google Scholar
  23. Duffy PA. Irritancy testing — a cultured approach. Toxicol in Vitro. 1989;3:157-8.Google Scholar
  24. Dykes PJ, Edwards MJ, O'Donovan MR, Merret V, Morgan HE, Marks R. In vitro reconstruction of human skin: the use of skin equivalents as potential indicators of cutaneous toxicology. Toxicol in Vitro. 1991;5:1-8.Google Scholar
  25. Eun HC, Chung JH, Jungs Y, Cho KH, Kim KH. A comparative study of the cytotoxicity of skin irritants on cultures of human oral and skin keratinocytes. Br J Dermatol. 1994;130:24-8.Google Scholar
  26. Fentem JH, Archer GEB, Balls M 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. 1998;12:483-524.Google Scholar
  27. Flaxman BA, Harper RA, Chiarello S, Feldman AML. Effects of methotrexate on the proliferation of human keratinocytes in vitro. J Invest Dermatol. 1977;68:66-9.Google Scholar
  28. Gajjar L, Benford DJ. Irritancy testing in cultured keratinocytes. Mol Toxicol. 1987;1:513-23.Google Scholar
  29. Gallo RL, Staszewskini R, Sauder DN et al. Regulation of GMCSF and IL-3 production from murine keratinocytes cell line PAM 212 following exposure to ultraviolet radiation. J Invest Dermatol. 1991;97:203-9.Google Scholar
  30. Gay R, Swiderek M, Nelson D, Ernesi A. The living skin equivalent as a model in vitro for ranking the toxic potential of dermal irritants. Toxicol in Vitro. 1992;6:303-15Google Scholar
  31. Gery I, Gershon RK, Waxman BH. Neutralization of the lymphocyte response mitogens — the responding process. J Exp Med. 1972;136:128-35.Google Scholar
  32. Gueniche A, Ponec M. Use of human skin cell cultures for the estimation of potential skin irritants. Toxicol in Vitro. 1993;7:15-24.Google Scholar
  33. Harvell JD, Gay R, Maibach HI. The living skin equivalent in cutaneous irritancy assessment. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994a:117-19.Google Scholar
  34. Harvell J, Tsai Y-C, Maibach HI et al. An in vivo correlation with three in vitro assays to assess skin irritation potential. J Toxicol Cutan Ocular Toxicol. 1994b;13:171-83.Google Scholar
  35. Helman R, Hall JW, Kao JY. Acute dermal toxicity: in vivo and in vitro comparisons in mice. Fundam Appl Toxicol. 1986;7:94-100.Google Scholar
  36. Hobson DW, Blank JA. In vitro alternative methods for the assessment of dermal irritation and inflammation. In Hobson DW, ed. Dermal and ocular toxicology fundamentals and methods. CRS Press; 1990:323-68.Google Scholar
  37. James LC, Moore AM, Wheeler LA, Murphy GM, Dowd PM, Greaves MW. Transforming growth factor α — in vivo release by normal human skin following UV irradiation and abrasion. Skin Pharmacol. 1991;4:61-4.Google Scholar
  38. Kao J, Hall J, Holland JM. Quantitation of cutaneous toxicity: an in vitro approach using skin organ culture. Toxicol Appl Pharmacol. 1986;68:206-17.Google Scholar
  39. Koschier FJ, Roth RN, Wallace KA, Curren RD, Harbell JW. A comparison of three-dimensional human skin models to evaluate the dermal irritation of selected petroleum products. In Vitro Toxicol. 1997;10:391-405.Google Scholar
  40. Lawrence JN. Application of in vitro human skin models to dermal irritancy: a brief overview and future prospects. Toxicol in Vitro. 1997;11:305-12.Google Scholar
  41. Lawrence JN, Starkey S, Dickson FM, Benford DJ. Use of human and rat keratinocyte cultures to assess skin irritation potential. Toxicol in Vitro. 1996;10:331-40Google Scholar
  42. Lewis RW, Botham PA. Measurement of transcutaneous electrical resistance to assess the skin corrosivity potential of chemicals. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:161-9.Google Scholar
  43. Liebsch M, Döring B, Donelly TA, Logemann P, Rheins LA, Spielmann H. Application of the human dermal model skin 2 ZK 1350 to phototoxocity and skin corrosivity testing. Toxicol in Vitro. 1995;9:557-62.Google Scholar
  44. Luger TA, Köck A, Danner M, Colot M, Micksche M. Production of distinct cytokines by epidermal keratinocytes. Br J Dermatol. 1985;113:145-56.Google Scholar
  45. Marzulli FN, Maibach HI. The rabbit as a model for evaluating skin irritants: a comparison of results obtained on animals and man using repeated skin exposure. Food Cosmet Toxicol. 1975;13:533-40.Google Scholar
  46. Moore KG, Schofield BH, Higuchi K et al. Two sensitive in vitro monitors of chemical toxicity to human and animal skin (in short-term organ culture) I: paranuclear vacuolization in glycol methacrylate tissue sections II interference with 14C leucine incorporation. J Toxicol Cutan Ocul Toxicol. 1986;5:285-302.Google Scholar
  47. Mosman T. Rapid colorimetric assay for cell growth and survival — application to proliferation and cytototxicity assays. J Immunol Methods. 1983;7:465-73.Google Scholar
  48. Müller-Decker K, Fürstenberger G, Marks F. Development of an in vitro alternative assay to the Draize skin irritancy test using human keratinocyte-derived proinflammatory key mediators and cell viability as test parameters. In Vitro Toxicol. 1992;5:191-209.Google Scholar
  49. Müller-Decker K, Fürstenberger G, Marks F. Keratinocyte-derived proinflammatory key mediators and cell viability as in vitro parameters of irritancy: a possible alternative to the Draize skin irritation test. Toxicol Appl Pharmacol. 1994;127:99-108.Google Scholar
  50. Nakamura M, Rikimaru T, Yano T et al. Full thickness human skin explants for testing the toxicity of topically applied chemicals. J Invest Dermatol. 1990;95:325-33.Google Scholar
  51. Nelson D, Gay RJ. Effects of UV irradiation on a living skin equivalent. Photochem Photobiol. 1993;57:830-7.Google Scholar
  52. Oliver GJA, Pemberton MA. An in vitro skin corrosivity test. Modifications and validation. Fed Toxicol. 1996;24:507-17.Google Scholar
  53. Oliver GJA, Pemberton, Rhodes C An in vitro model for identifying skin corrosive chemicals. I. Initial validation. Toxicol in Vitro 1988;2:7-17.Google Scholar
  54. Osborne R, Perkins MA. In vitro skin irritation testing with human skin cell cultures. Toxicol in Vitro. 1991;5:563-7.Google Scholar
  55. Pemberton MA, Oliver GJA. The measurement of skin irritation in vitro using ex vivo rabbit skin. Br J Dermatol. 1986;(Suppl. 31):45-6.Google Scholar
  56. Ponec M. In vitro cultured human skin cells as alternatives to animal for irritancy screening. Int J Cosmetics 1992;14:245-64.Google Scholar
  57. Ponec M. The use of in vitro skin recombinants to evaluate cutaneous toxicity. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:107-116.Google Scholar
  58. Ponec M, Kempenaar J. Use of human skin recombinants as an in vitro model for testing the irritation potential of cutaneous irritant. Skin Pharmacol. 1995:49-59.Google Scholar
  59. Ponec M, Haverkort M, Soei YL et al. Toxicity screening of N-alkylazacycloheptan-2-one drivatives in cultured human skin cells — structure-toxicity relationship. J Pharm Sci. 1989;78:738-41.Google Scholar
  60. Prunieras M. Skin and epidermal equivalents: a review. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:97-105.Google Scholar
  61. Reichert U, Jacques Y, Grangeret M, Schmidt R. Antirespiratory and antiproliferative activity of anthralin in cultured human keratinocytes. J Invest Dermatol. 1995;84:130-4.Google Scholar
  62. Rheinwald JG, Green H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell. 1975;6:331-44.Google Scholar
  63. Rikimaru T, Nakamura M, Yano T et al. Mediators, initiating the inflammatory response released in organ culture by full-thickness human skin explants exposed to the irritant sulfur mustard. J Invest Dermatol. 1991;96:888-97.Google Scholar
  64. Roguet R. Cutaneous pharmacotoxicology in vitro. In Castell JM, Gomez-Lechon MJ, eds. In vitro methods in pharmaceutical research. Academic Press; 1997:242-64Google Scholar
  65. Roguet R, Schaefer H. Overview of in vitro cell culture technologies and pharmacotoxicological applications. Toxicol in Vitro. 1997;11:591-9.Google Scholar
  66. Roguet R, Dossou G, Rougier A. Use of in vitro skin recombinants to evaluate cutaneous toxicity: a preliminary study. J Toxicol Cutan Ocul Toxicol. 1992;11:305-15.Google Scholar
  67. Roguet R, Cohen C, Dossou G, Rougier A. Episkin, a reconstructed human epidermis assessing in vitro the irritancy of topically applied compounds. Toxicol in Vitro. 1994a; 8:283-91.Google Scholar
  68. Roguet R, Régnier M, Cohen C, Dossou G, Rougier A. The use of in vitro reconstructed human skin in dermotoxicity testing. Toxicol in Vitro. 1994b;8:635-9.Google Scholar
  69. Roguet R, Cohen C, Rougier A, Leclaire J. Measurement of proinflammatory mediators production by cultured keratinocytes. In Elsner P, Maibach HI, eds. Irritant dermatitis, new clinical and experimental aspects. Basel: Karger; 1995:230-40. (Current problems in dermatology, vol. 23).Google Scholar
  70. Roguet R, Cotovio J, Leclaire J. Reconstructed skin for in vitro cutaneous pharmacotoxicology: example of metabolic studies. In Vitro Toxicol. 1997;10:3-9.Google Scholar
  71. Roguet R, Cohen C, Robles C 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. 1998;12:295-304.Google Scholar
  72. Rosdy M, Clauss LC. Terminal epidermal differentiation of human keratinocytes growth in chemically defined medium on inert filter substrates at air-liquid interface. J Invest Dermatol. 1990a;95:409-14.Google Scholar
  73. Rosdy M, Clauss LC. Cytotoxicity testing of wound dressing using normal human keratinocytes in culture. J Biomed Mater Res. 1990b;24:363-77.Google Scholar
  74. Shibata M, Tsuda T, Itagaki H et al. Interleukin 1α and interleukin 8 release by human keratinocyte cell culture treated with surfactants. ATLA. 1997;25:161-71.Google Scholar
  75. Slivka SR, Landeen LK, Zeigler F, Zimber MP, Bartel RL. Characterization, barrier function and drug metabolism of an in vitro skin model. J Invest Dermatol. 1993;100:40-6.Google Scholar
  76. Shivji GM, Segal L, McKenzie RC, Sauder D. Cutaneous toxicity of surfactants in normal human keratinocytes assessed by cytotoxicity, arachidonic acid release and regulation of interleukin 1α mRNA. Toxicol Methods. 1994;4: 193-203.Google Scholar
  77. Smoot EC, Kucan JO, Roth A, Mody N, Debs N. In vitro testing for antibacterials against human keratinocytes. Plastic Reconstruct Surg. 1992;87:917-24.Google Scholar
  78. Tinois E, Gaetani Q, Gayraud B, Dupont D, Rougier A, Pouradier Duteil X. The Episkin model: successful reconstruction of human epidermis in vitro. In Rougier A, Golberg A, Maibach HI, eds. In vitro skin toxicology irritation, phototoxicity, sensibilization. New York: Mary Ann Liebert; 1994:133-40.Google Scholar
  79. Triglia D, Kidd I, de Wever B, Roonan R. Interlaboratory validation study of the advanced tissue sciences Skin 2TM dermal model and MTT cytotoxicity assay kits. AATEX. 1992;1:142-7.Google Scholar
  80. Van de Sandt JJM, Rutten AAJJL. Differential effects of chemical irritants in rabbit and human skin organ cultures. Toxicol in Vitro. 1995;9:157-68.Google Scholar
  81. Van de Sandt JJM, Rutten AAJJL, Koëter HBWM. Cutaneous toxicity testing in organ culture: neutral red uptake and reduction of tetrazolium salt (MTT). Toxicol in Vitro. 1993;7:81-6.Google Scholar
  82. Ward RK, Agrawalla S, Clothier RH. Investigation of an in vitro cytotoxicity assay for prediction of skin irritation. Toxicol in Vitro. 1994;8:659-60.Google Scholar
  83. Ward RK, Hubbard AW, Sulley H, Garle MJ, Clothier RH. Human keratinocyte culture in an in vitro approach for the assessment of surfactant-induced irritation. Toxicol in Vitro. 1998;12:163-73.Google Scholar
  84. Whittle E, Basketter DA. In vitro skin corrosivity test using human skin. Toxicol in Vitro. 1994;8:861-3.Google Scholar
  85. Whittle E, Basketter DA. The in vitro skin corrosivity test. Development of method using human skin. Toxicol in Vitro. 1993;7:265-8.Google Scholar
  86. Wilhelm KP, Samblebe MA, Siegers CP. Quantitative in vitro assesment of N-alkyl sulphate-induced cytotoxicity in human keratinocytes (HaCaT): comparison with in vivo human irritation tests. Br J Dermatol. 1994;130:18-23.Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • R. Roguet
    • 1
  1. 1.Life Sciences Research, L'Oreal RechercheClichyFrance

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