Skip to main content
SpringerLink
Log in

Applications of reconstructed skin models in pharmaco-toxicological trials

  • Cellular Engineering: Bioengineering of Skin
  • Published:
Medical and Biological Engineering and Computing Aims and scope Submit manuscript

Abstract

The development of new cosmetic formulations requires precise assessment of their safety and efficacy. Today, legislation demands quality control combined with severe safety measures, as well as a limited use of animals for such testing (European Community directive 93/35/EEC). Consequently, safety assessment protocols are oriented towards in vivo tests on human volunteers and in vitro alternative methods to animal use, especially tissue engineered skin substitutes. In this paper, dermal and skin equivalents developed in the laboratory are described. The applications of reconstructed epidermis and skin substitutes for pharmaco-toxicological trials are also discussed. These tissue models have been shown to be very useful tools to assess cutaneous irritation, phototoxicity, photoprotection and to perform efficacy tests of cosmetic molecules and finished products. In conclusion, the authors are confident that these in vitro models can contribute to reduce animal use for routine toxicity testing.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Applegate, L., andFrenk, E. (1995): ‘Oxidative defense in cultured human skin fibroblasts and keratinocytes from sun-exposed and non exposed skin’,Photodermatol. Photoimmunol. Photomed.,11, pp. 95–101

    Google Scholar 

  • Auger, F. A., Lopez-Valle, C. A., Guignard, R., Tremblay, N., Noel, B., Goulet, F., andGermain, L. (1995): ‘Skin equivalents produced with human collagen’,In Vitro Cell. Dev.,31, pp. 432–439

    Google Scholar 

  • Augustin, C., Collombel, C., andDamour, O. (1995): ‘Development of a kit for predicting cutaneous toxicity in vitro using 3 D dermal equivalent: phase I Reproducibility of dermal equivalent’,J. Cell Eng.,1, pp. 58–62

    Google Scholar 

  • Augustin, C., Lepavec, A., Venet, E., Nakazawa, K., Collombel, C., andDamour, O. (1996): ‘Abstract: From monolayered culture to pigmented skin equivalent: complexification of the skin cells culture and contributions in pharmaco-toxicology’,Cell Biol. Toxicol.,12, p. 384.

    Article  Google Scholar 

  • Augustin, C., Collombel, C., andDamour, O. (1997A): ‘Use of in vitro dermal equivalent and skin equivalent kits for evaluating cutaneous toxicity of cosmetics products’,In Vitro Toxicol.,1, pp. 21–29

    Google Scholar 

  • Augustin, C., Collombel, C., andDamour, O. (1997B): ‘Use of dermal equivalent and skin equivalent models for identifying phototoxic compoundsin-vitro’,Photodermatol Photoimmunol Photomed.,13, pp. 27–36

    Google Scholar 

  • Augustin, C., Frei, V., Perrier, E., Huc, A., andDamour, O. (1997C): ‘Anin vitro selection of new cosmetic active compounds: from screening tests on fibroblast monolayered culture to efficacy study on dermal equivalent’,J Appl Cosmetol.,15, pp. 1–11

    Google Scholar 

  • Augustin, C., Frei, V., Perrier, E., Huc, A., andDamour, O. (1997D): ‘A skin equivalent model for cosmetological trials: anin vitro study of a new biopeptide’,Skin Pharmacol.,10, pp. 63–70

    Google Scholar 

  • Augustin, C., Collombel, C., andDamour, O. (1998A): ‘Use of dermal equivalent and skin equivalent models forin vitro cutaneous irritation testing of cosmetic products: comparison within vivo data’,J. of Toxicol—Cutaneous & Ocular Toxicol.,17, 1, pp. 5–17

    Google Scholar 

  • Augustin, C., Collombel, C., andDamour, O. (1998B): ‘Measurement of the protective effect of topically applied sunscreens usingin vitro determal and skin equivalents’,Photochem & Photobiol.,66, pp. 853–859

    Google Scholar 

  • Balls, M., Blaauboer, B., Brusick, D., Frazier, J., Lamb, D., Pemberton, M., Reinhardt, C., Roberfroid, M., Rosenkranz, H., Schmid, B., Spielmann, H., Stammat, A. L., andWalum, E. (1990): ‘Report and recommendations of the CAAT/ERGATT. Workshop on the validation of toxicity test procedures’,ATLA,18, pp. 313–337

    Google Scholar 

  • Bartnik, F., Pitterman, W., Mendorf, N., Tillman, U., andKunstler, K. (1990): ‘Skin organ culture for the study of skin irritancy’,Toxicol In Vitro,4, pp. 293–301

    Article  Google Scholar 

  • Bell, E., Ivarson, B., andMerrill, C. (1979): ‘Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro’,Cell Biol.,76, pp. 1274–1278

    Google Scholar 

  • Bell, E., Rosenberg, M., Kemp, P., Gay, R., Green, G., Muthukumaran, N., andNolte, C. (1991): ‘Recipes for reconstituting skin’,J. Biomed. Eng.,113, pp. 113–119

    Google Scholar 

  • Berthod, F., Damour, O., andCollombel, C. (1993): ‘Collagen synthesis by fibroblasts cultured within a collagen sponge’,Biomat.,14, pp. 749–754

    Article  Google Scholar 

  • Berthod, F., Sahuc, F., Hayek, D., Damour, O., andCollombel, C. (1996): ‘Deposition of collagen fibril bundles by long-term culture of fibroblasts in a collagen sponge’,J. Biom. Mat. Res.,32, pp. 87–94

    Article  Google Scholar 

  • Boyce, S., Christianson, D., andHansbrough, J. (1988): ‘Structure of a collagen-GAG dermal skin substitute optimized for cultured human epidermal keratinocytes’,J. Biomed. Mat.,22, pp. 939–957

    Article  Google Scholar 

  • Brinckmann, J., Bodo, M., Brey, M., Wolff, H., andMuller, P. (1993): ‘Analysis of the age-related composition of human skin collagen and collagens synthesised by fibroblast culture’,Arch. Dermatol. Res.,286, pp. 391–395

    Article  Google Scholar 

  • Bruner, L. H., Carr, G. J., Chamberlain, M., andCurren, R. D. (1996): ‘Validation of alternative methods for toxicity testing’,Toxicol. In Vitro,10, pp. 479–501

    Article  Google Scholar 

  • Cannon, C. L., Neal, P. J., Kubilus, J., andKlausner, M. (1993): ‘New epidermal model for dermal irritancy testing’. Presentation during meeting in ‘Current trends of in vitro Skin Toxicology and Eye Irritancy testing’. Ottawa,, Canada

  • Chardon, A., Moyal, D., Hourseau, C., andLang, G. (1991): ‘Skin immediate pigment darkening applied to UVA protection assesment’,Photochem. Photobiol.,53, p. 115S

    Google Scholar 

  • Cohen, C., Dossou, K., Rougier, A., andRoguet, R. (1994): ‘Episkin: an in vitro model for the evaluation of phototoxicity and sunscreen photoprotective properties’,Toxic In Vitro,8, pp. 669–671

    Article  Google Scholar 

  • Cole, C. (1993): ‘Multicenter evaluation of sunscreen UVA protectiveness with protection factor test method’,J. Am. Acad. Dermatol.,30, pp. 729–736

    Article  Google Scholar 

  • Collombel, C., Damour, O., Gagnieu, C., Marichy, C., andPoinsignon, F. ‘Biomaterials with a base of collagen, chitosan and glycosaminoglycans; process for preparing them and their application in human medicine’. French patent 8708252 (1987), European patent 884101948 (1988), US patent PCT/FR/8800303 (1989)

  • Davies, R, Harper, K., andKynoch, S. (1972): ‘Inter-species variation in dermal reactivity’,J. Soc. Cosm. Chim.,23, pp. 371–381

    Google Scholar 

  • Draize, J., Woodard, G., andCalvery, H. (1944): ‘Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membrane’,J. Pharmacol. Exp. Ther.,82, pp. 377–390

    Google Scholar 

  • Duffy, P., Bennett, A., andRoberts, M. (1987): ‘Prediction of phototoxic potential using human A431 cells and mouse 3T3 cells’,Molecular Toxicol.,1, pp. 579–587

    Google Scholar 

  • Dugard, P., andWalker, M. (1983): ‘Comparison of the permeability of human and laboratory animal skin’,J. Invest. Dermatol.,80, p. 347

    Google Scholar 

  • Edward, S., Donnelly, T., Sayre, R., andRheins, L. (1994): ‘Quantitative in vitro assessment of phototoxicity using a human skin model Skin2’,Photodermatol. Photoimmunol. Photomed.,10, pp. 111–117

    Google Scholar 

  • Gabriel, K., Mark, R., Jackson, E., Davies, R., andForbes, P. (1987): ‘Sun protection factor testing: comparison of FDA and DIN methods’,J. Cut. Ocul. Toxicol.,6, pp. 357–370

    Google Scholar 

  • Gordon, V. (1992): ‘SOLATEXTM Presentation at SFC/ISIPCA congres’ Versailles, France

  • Gueniche, A., andPonec, M. (1993): ‘Use of human skin cell cultures for the estimation of potential skin irritants,’J. Toxicol. Cut. Ocular. Toxicol.,12, pp. 49–57

    Google Scholar 

  • Guinter, K., andFleischaker, B. (1971A): ‘Evaluation of phototoxicity of salicylanides and similar compounds by photohemolysis’,J. Invest. Dermatol.,56, pp. 91–98

    Article  Google Scholar 

  • Guinter, K., andFleischaker, B. (1971B): ‘Red blood cell hemolysis by photosensitizing compounds’,J. Invest. Dermatol.,56, pp. 85–91

    Article  Google Scholar 

  • Kaidbey, K., andKligmann, A. (1978A): ‘Identification of systemic phototixic drugs by human intradermal assay’,J. Invest. Dermatol.,70, pp. 272–274

    Article  Google Scholar 

  • Kaidbey, K., andKligmann, A. (1978B): ‘Identification of topical photosensitizing agents in human.’J. Invest. Dermatol.,70, pp. 149–151

    Article  Google Scholar 

  • Kao, J., Hall, J., andHolland, J. (1983): ‘Quantitation of cutaneous toxicity: an in vitro approach using skin organ culture’,Toxicol. Appl. Pharmacol.,68, pp. 206–217

    Article  Google Scholar 

  • Koeter, H. (1995): ‘Validation: a highly charged concept’,Toxicol. In Vitro,9, pp. 851–856

    Article  Google Scholar 

  • Lapiere, C. (1988): ‘Aging of fibroblasts, collagen and the dermis’,Cutaneous aging pp. 47–60

  • Laska, D., Poulsen, R., Horn, J., Meador, V., andHoover, D. (1992): ‘An evaluation of Testskin: an alternative dermal irritation model’,In Vitro Toxicol.,5, pp. 177–189

    Google Scholar 

  • Lehmann, P., Holzle, E., Melnik, B., andPlewig, G. (1991): ‘Effects of ultraviolet A and B on the skin barrier: a functional, electron microscopic and lipid biochemistry study’,Photodermatol. Photoimmunol. Photomed.,8, pp. 129–134

    Google Scholar 

  • Lock, S., andFriend, V. (1986): ‘Phototoxicity testing in vitro: evaluation of mammalian cell culture techniques’,Food Chem. Toxic.,6, pp. 789–793

    Article  Google Scholar 

  • Loprieno, N., andLoprieno, G. (1995A): ‘Cosmetic regulation in the European union’,In vitro Toxicol.,8, pp. 419–426

    Google Scholar 

  • Loprieno, N., Bruner, L., Carr, G., Chamberlain, M., Cottin, M., De Silva, O., andKato, S. (1995B): ‘Alternatives in cosmetics testing’,In vitro Toxicol.,9, pp. 827–838

    Article  Google Scholar 

  • Maier, S., Schmitt-Landgraf, R., andSiegemund, B. (1991): ‘Development of an in vitro test system with human skin cells for evaluation of phototoxicity’,Toxicol In Vitro,5, pp. 457–461

    Article  Google Scholar 

  • Michel, M., L'Heureux, N., Auger, F. A., andGermain, L. (1997): ‘from newborn to adult: Phenotypic and functional properties of skin equivalent and human skin as a function of donor age’,J. Cell. Phys.,171, pp. 171–179

    Article  Google Scholar 

  • Morison, W., Daniel, M., McAullife, J., Parrish, J., andBloch, K. (1978): ‘In vitro assay for phototoxic chemicals’,J. Invest. Dermatol.,70, pp. 149–151

    Article  Google Scholar 

  • Mosmann, T. (1983): ‘Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assay’.J. Immunol. Meth.,65, pp. 55–63

    Article  Google Scholar 

  • Moysan, A., Clement-Lacroix, P., Michel, L., Dubertret, J., andMorliere, P. (1995): ‘Effects of UVA and antioxydant defense in cultured fibroblasts and keratinocytes’,Photodermatol. Photoimmunol. Photomed.,11, pp. 192–197

    Google Scholar 

  • Naughton, G., Jacob, L., andNaughton, B. (1989): ‘A Physiological skin model for in vitro toxicity studies’. in:Goldberg A. (Ed.): ‘In vitro toxicology: mechanisms and new technology’ (Mary A. Liebert inc. New York) pp. 183–189

    Google Scholar 

  • Nelson, D., andGay, R. (1992): ‘Effects of UV irradiation on a living skin equivalent’,Photochem. Photobiol.,57, pp. 830–837

    Google Scholar 

  • Nixon, G., Tyson, C., andWertz, W. (1975): ‘Interspecies comparisons of skin irritancy’,Toxol. Appl. Pharmacol.,31, pp. 480–490

    Google Scholar 

  • Noel-Hudson, M. S., Brant-Boucher, F., Robert, M., Aubry, M., andWepierre, J. (1997): ‘Comparison of six different methods to assess UVA cytotoxicity on reconstructed epidermis. Relevance of a fluorimetric assay (the calcium-AM) to evaluate the photoprotective effects of α-tocopherol’,Toxicol. In Vitro,11, pp. 645–651

    Article  Google Scholar 

  • Oliver, G. J., Pemberton, M. A., andRhodes, C. (1986): ‘An in-vitro corrosivity test: modifications and validation’,Food Chem. toxicol.,24 (6–7), pp. 507–512

    Article  Google Scholar 

  • Osborne, R., andPerkins, M. (1991): ‘In vitro skin irritation testing with human skin cell cultures’,Toxicol. In Vitro,5, pp. 563–567

    Article  Google Scholar 

  • Paquet, I., Chouinard, N., andRouabhia, M. (1996): ‘Cutaneous cell and extracellular matrix responses to ultraviolet-B irradiation’,J. Cell. Physiol.,166, pp. 296–304

    Article  Google Scholar 

  • Pearse, A., andEdwards, C. (1993): ‘Human stratum corneum as a substrate for in vitro sunscreen testing’,Int. J. Cosm. Sci.,15, pp. 234–244

    Google Scholar 

  • Prunieras, M., Regnier M., andWoodley, D. (1983): ‘Methods for cultivation of keratinocytes with an air-liquid interface.’J. Invest. Dermatol.,81, pp. 28s-33s.

    Article  Google Scholar 

  • Punnonen, K., Puntala, A., andAhotupa, M. (1991): ‘Effects of ultraviolet A and B irradiation on lipid peroxidation and activity of the antioxydant enzymes in keratinocytes in culture’,Photodermatol. Photoimmunol. Photomed.,8, pp. 3–6

    Google Scholar 

  • Regnier, M., Prunieras, M., andWoodley, D. T. (1981): ‘Growth and differenciation of adult human epidermal cells on dermal substrate’.Front Matrix.,9, pp. 4–35

    Google Scholar 

  • Regnier, M., Asselineau, D., andLenoir, M. (1990): ‘Human epidermis reconstructed on dermal substrates in vitro: an alternatives to animals in skin pharmacology’,Skin Pharmacol.,3, pp. 70–85

    Google Scholar 

  • Roguet, R., Cohen, C., andLeclaire, J. (1995): ‘The use of a reconstructed human epidermis Episkin for the assessment of safety and efficacy of cosmetic products’,In Vitro Toxicol.,8, pp. 427–429

    Google Scholar 

  • Roguet, R., andSchaefer, H. (1997): ‘Overview of in vitro cell culture technologies and pharmaco-toxicological applications’,Toxicol. In Vitro,11, pp. 591–599

    Article  Google Scholar 

  • Rosdy, M., andClauss, L. (1990): ‘Terminal epidermal differentiation of human keratinocytes grown chemically defined medium on inert filter substrates at the air-liquide interface’,J. Invest. Dermatol.,96, pp. 409–414

    Article  Google Scholar 

  • Saintigny, G., Bonnard, M., Damour, O., andCollombel, C. (1993): ‘Reconstruction of epidermis on a chitosan cross-linked collagen-GAG lattice: effects of fibroblasts,’Acta Derm. Venereol.,73, pp. 175–180

    Google Scholar 

  • Shahabeddin, L., Berthod, F., Damour, O., andCollombel, C. (1989): ‘Characterisation of skin reconstructed on a chitosan-cross-linked collagen glycosaminoglycan matrix’,Skin Pharmacol.,3, pp. 107–114

    Article  Google Scholar 

  • Steinberg, R., Akers, W., Weeks, M., McCreeh, A., andMaibach, H. (1975): ‘A comparison of test techniques based on rabbit and human skin responses with recommendations, regarding the evaluation of mildly or moderatly irritating compounds’, In Animals in Dermatology, (Editions Churchill Livingstone) pp. 1–11

  • Swisher, D., Cormier, M., Johnson, J., andLedger, P. (1988): ‘A cytotoxicity assay using normal keratinocytes: characterisation and application’,Mod. Dermatol.,4, pp. 131–137

    Google Scholar 

  • Tinois, E., Tillier, J., Gaucherand, M., Dumas, H., Tardy, M., andThivolet, J. (1991): ‘In vitro and post-transplantation differentiation of human keratinocytes growth on the human type IV collagen film of a bilayered dermal substitut’Exp. Cell. Res.,193, pp. 310–319

    Article  Google Scholar 

  • Wallace, J., Harbell, A., Triana, A., Accomando, N., andValone, S. (1992): ‘Evaluation of the human epidermal keratinocyte neutral red release and neutral red uptake assay using the first 10 MIEC materials’,Toxicol. In Vitro,6, pp. 367–371

    Article  Google Scholar 

  • Weil, C., andScala, R. (1971): ‘Study of intra- and inter-laboratory variability in the results of rabbit eye and skin irritation tests’,Toxicol. Appl. Pharmacol.,19, pp. 276–360

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire des Substituts Cutanés (CNRS UPR 412, Hôpital Edouard Herriot, 3 Place d'Arsonval, Lyon, France

    O. Damour, C. Augustin & A. F. Black

Authors
  1. O. Damour
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. Augustin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. F. Black
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to O. Damour.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Damour, O., Augustin, C. & Black, A.F. Applications of reconstructed skin models in pharmaco-toxicological trials. Med. Biol. Eng. Comput. 36, 825–832 (1998). https://doi.org/10.1007/BF02518890

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02518890

Keywords

Search

Navigation