Microfluidic Devices for Eye Irritation Tests of Cosmetics and Cosmetic Ingredients

  • Tian Tian
  • Sujin Cho
  • Seog Woo RheeEmail author
Original Article


In this paper, we describe the development and application of a simple microfluidic device for in vitro irritation tests of cosmetics. The device was fabricated with a three-compartment diffusion system to mimic a hen’s egg test-chorioallantoic membrane (HET-CAM) system. Human umbilical vein endothelial cells were cultured in the three compartments, and the tested substances were injected into the compartment, and then were drawn through the small microfluidic channels. The mortalities of cells in each compartment were monitored using a fluorescent microscope. The TS (toxicity score) values were evaluated on the basis of the mortalities of cells as a function of time when the cells were exposed to the test substances. Four kinds of cosmetic materials were tested with the microfluidic system, and the results were compared with IS (irritation score) values of a HET-CAM test to validate the in vitro irritation test system. The method developed in this study could be used for evaluation of the ocular toxicity of cosmetic ingredients, and could be widely used as an alternative test for cosmetic and medical tests or for the development of new medicines.


Microfludic device HUVEC HET-CAM Eye irritation Alternative test method 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Rousseau, F., Bonaventure, J., Legeai-Mallet, L., Pelet, A., Rozet, J.M., Maroteaux, P., Le Merrer, M. & Munnich, A. Mutations in the gene encoding fibroblast growth factor receptor-3 in achondroplasia. Nature 371, 252–254 (1994).CrossRefGoogle Scholar
  2. 2.
    Directive 2003/15/EC of the European parliament and of the council of 27 February 2003 amending council directive 76/768/EEC on the approximation of the laws of the member states relating to cosmetic products. O. J. L66, 26–35 (2003).Google Scholar
  3. 3.
    Pfuhler, S., Fautz, R., Ouedraogo, G., Latil, A., Kenny, J., Moore, C., Diembeck, W., Hewitt, N.J., Reisinger, K. & Barroso, J. The cosmetics Europe strategy for animal-free genotoxicity testing: Project status update. Toxicol. In Vitro 28, 18–23 (2014).CrossRefGoogle Scholar
  4. 4.
    Almeida, A., Sarmento, B. & Rodrigues, F. Insights on in vitro models for safety and toxicity assessment of cosmetic ingredients. Int. J. Pharm. (Amsterdam, Neth.) 519, 178–185 (2017).Google Scholar
  5. 5.
    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, 377–390 (1944).Google Scholar
  6. 6.
    Burton, A.B., York, M. & Lawrence, R.S. The in vitro assessment of severe eye irritants. Food Cosmet. Toxicol. 19, 471–475 (1981).CrossRefGoogle Scholar
  7. 7.
    OECD, Test No. 437: OECD guideline for the testing of chemicals, “Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage. Section 4, OECD Publishing, Paris (2017).Google Scholar
  8. 8.
    Luepke, N.P., & Kemper, F.H. The HET-CAM test: An alternative to the draize eye test. Food Chem. Toxicol. 24, 495–496 (1986).CrossRefGoogle Scholar
  9. 9.
    Gilleron, L., Coecke, S., Sysmans, M., Hansen, E., Van Oproy, S., Marzin, D., Van Cauteren, H. & Vanparys. P. Evaluation of a modified HET-CAM assay as a screening test for eye irritancy. Toxicol. In Vitro 10, 431–446 (1996).CrossRefGoogle Scholar
  10. 10.
    Steiling, W., Bracher, M., Courtellemont, P. & de Silva, O. The HET—CAM, a useful in vitro assay for assessing the eye irritation properties of cosmetic formulations and ingredients. Toxicol. In Vitro 13, 375–384 (1999).CrossRefGoogle Scholar
  11. 11.
    Wu, J., Wu, X. & Lin, F. Recent developments in microfluidics-based chemotaxis studies. Lab Chip 13, 2484–2499 (2013).CrossRefGoogle Scholar
  12. 12.
    Englert, D.L., Manson, M.D. & Jayaraman, A. Investigation of bacterial chemotaxis in flow-based microfluidic devices. Nat. Protoc. 5, 864–872 (2010).CrossRefGoogle Scholar
  13. 13.
    Rhee, S.W. Compartmented microfluidic device for positioning and chemotactic migration of cells. Bio-Chip J. 5, 129–136 (2011).Google Scholar
  14. 14.
    El-Ali, J., Sorger, P.K. & Jensen, K.F. Cells on chips. Nature 442, 403–411 (2006).CrossRefGoogle Scholar
  15. 15.
    Jusoh, N., Oh, S., Kim, S., Kim, J. & Jeon, N.L. Microfluidic vascularized bone tissue model with hydroxyapatite-incorporated extracellular matrix. Lab Chip 15, 3984–3988 (2015).CrossRefGoogle Scholar
  16. 16.
    Lancaster, M.A., Renner, M., Martin, C.A., Wenzel, D., Bicknell, L.S., Hurles, M.E., Homfray, T., Penninger, J.M., Jackson, A.P. & Knoblich, J.A. Cerebral organoids model human brain development and microcephaly. Nature 501, 373–379 (2013).CrossRefGoogle Scholar
  17. 17.
    Huh, D., Torisawa, Y.S., Hamilton, G.A., Kim, H.J. & Ingber, D.E. Microengineered physiological biomimicry: Organs-on-Chips. Lab Chip 12, 2156–2164 (2012).CrossRefGoogle Scholar
  18. 18.
    Whitesides, G.M., Ostuni, E., Takayama, S., Jiang, X. & Ingber, D.E. Soft lithography in biology and biochemistry. Annu. Rev. Biomed. Eng. 3, 335–373 (2001).CrossRefGoogle Scholar
  19. 19.
    Taylor, A.M., Rhee, S.W., Tu, C.H., Cribbs, D.H., Cotman, C.W. & Jeon, N.L. Microfluidic multicompartment device for neuroscience research. Langmuir 19, 1551–1556 (2003).CrossRefGoogle Scholar
  20. 20.
    Xia, Y. & Whitesides, G.M. Extending microcontact printing as a microlithographic technique. Langmuir 13, 2059–2067 (1997).CrossRefGoogle Scholar
  21. 21.
    Xia, Y. & Whitesides, G.M. Soft lithography. Angew. Chem. Int. Ed. 28, 550–75 (1998).CrossRefGoogle Scholar
  22. 22.
    NIH, ICCVAM In Vitro Ocular Evaluation Report, ICCVAM-Recommended test method protocol: Hen’s egg test-chorioallantoic membrane (HET- CAM) test method. NIH Publication No 10-7553 (2010).Google Scholar
  23. 23.
    McKenzie, B., Kay, G., Matthews, K.H., Knott, R.M. & Cairns, D. The hen’s egg chorioallantoic membrane (HET-CAM) test to predict the ophthalmic irritation potential of a cysteamine-containing gel Quantification using Photoshop® and ImageJ. Int. J. Pharm. (Amsterdam, Neth.) 490, 1–8 (2015).Google Scholar
  24. 24.
    Gillerona, L., Coecke, S., Sysmans, M., Hansen, E., van Oproy, S., Marzin, D., van Cauteren, H. & Vanparys, P. Evaluation of the HET-CAM-TSA method as an alternative to the draize eye irritation test. Toxicol. In Vitro 11, 641–644 (1997).CrossRefGoogle Scholar
  25. 25.
    Wilson, T.D. & Steck, W.F. A modified HET-CAM assay approach to the assessment of anti-irritant properties of plant extracts. Food Chem. Toxicol. 38, 867–872 (2000).CrossRefGoogle Scholar
  26. 26.
    Wilson, S. L., Ahearne, M. & Hopkinson, A. An overview of current techniques for ocular toxicity testing. Toxicology 327, 32–46 (2015).CrossRefGoogle Scholar

Copyright information

© The Korean BioChip Society and Springer 2018

Authors and Affiliations

  1. 1.Department of ChemistryKongju National UniversityKongjuKorea

Personalised recommendations