Biology Issue Report Subject: Are the results of animal testing in the cosmetics industry relevant and applicable

to humans?
The testing of cosmetics on animals has been banned in the EU since 2009; however other countries around the world (such as the USA) still use animals in these tests. In an attempt to predict safety and effectiveness of cosmetics and toiletries, regulators in countries like America require companies to prove that their products are safe and harmless towards human beings. The cosmetics companies are required to conduct these experiments - they do not simply test on animals because they want to. Many thousands of animals (such as rabbits, mice, rats and guinea pigs) are used every year in the tests, but how reliable and applicable are the results to humans?

The problem
The Draize Eye Irritancy Test is just one example of the many experiments animals are used for in countries outside of the EU. The test is often associated with rabbits; however dogs and nonhuman primates can also be used for this procedure. John H. Draize developed the test in 1944, in order to evaluate the irritation caused by a number of chemicals that are used in the cosmetics industry. In the test, the substance that the scientist is trying to evaluate is placed into one eye, whilst the other eye acts as a control. The test involves the rabbits being restrained in order to prevent them from reacting as they naturally would to the irritation. Their eyes are assessed after one hour, and then at 24-hour intervals for up to 14-21 days. The level of irritation to the eyes is recorded numerically by observation of the three major tissues of the eye - the cornea, conjunctiva, and iris. (1) Animal tests for chemicals used in the cosmetics industry (such as the Draize test) are recognised as showing poor quality results that do not relate to human beings. It is thought that the test conditions are often unrealistic, with tremendously high doses administered by abnormal routes. Furthermore, the results of the tests must be scaled up in order to make the results proportionate to humans; however the mathematical formulae used has not actually been proved accurate. An example of this can be seen in table 1(2), which demonstrates how three separate observations of injury to three ocular tissues within the eye are converted into a solitary numerical result, which is then used to estimate the overall eye injury. The total Draize score, which illustrates the possible eye irritation of a chemical that is used in a cosmetics product, is not precise.(9) Due to animal-to-animal variability, as well as the subjective nature of the unrealistic observations made when the results are scored, this makes it practically impossible to reproduce the final Draize score, which therefore means that results of Draize eye tests are not reproducible, and could be seen as producing unrealistic and inaccurate results. The Draize eye test has been criticised for a number of other reasons. For example, the structure of the cornea of the eye of a rabbit contrasts significantly from that of a human. Rabbits also

produce a smaller volume of tears than humans; this causes chemicals and other irritants that are placed into their eyes to loiter for a greater period of time and cause more problems. This makes the Draize eye test unreliable and therefore a problem requiring evaluation in counties outside of the EU so that the law might be changed. Moreover, the subjective nature of the gross observations made during the scoring of the test, plus normal animal-to-animal variability, make it virtually impossible to routinely reproduce the final Draize score, especially for midrange irritants.(3) Evidently, animals are not appropriate models for cosmetic testing. In spite of this, their use remains rooted in modern science in some countries around the world. (4) Hence, the demand for alternative methods that more precise results comes about not only from animal activists that are alarmed for the welfare of said animals, but also from scientists that are concerned with the inaccurate and imprecise results that animal tests produce.
Figure 2 shows a rabbit being experimented on as part of a Draize test (5) in order to assess whether chemicals used in the production of cosmetics have bad effects upon rabbits, the results of which are then applied to humans.

Science can provide evidence that animals and humans differ too much in order for them to provide accurate results. Nonanimal tests however, provide more reliable, predictive results and therefore lead to the production of safer
Figure 1 A rabbit undergoing the Draize test.

products which humans are able to use without harm. The

(6)

non-animal tests must undergo rigorous validation in order to ensure the results are valid, and can show matched data of the toxicity in humans. This therefore means that alternative methods of the Draize test that do not involve animals produce results that are more valid than animal tests, and are more easily applicable and relevant to humans. As well as this, these modern methods have been found to predict human reactions better than the traditional outdated animal tests. This makes them increasingly more efficient and appropriate than the Draize tests that are still in practice in countries such as the USA.
Figure 2 shows researchers at ECVAM developing alternatives to animal toxicity tests. (7)

The main solution

An example of an experiment that can replace animal
Figure 2

testing in the cosmetics industry (particularly Draize eye tests) is In-vitro techniques. This involves the study of isolated molecules, cells and tissues, which can be sourced from humans, other animals, microorganisms or even plants. An example of a non-animal test for cosmetic products can be seen in Toshiaki Takezawas latest discovery. Takezawa (along with a group of Japanese scientists) developed a form of artificial corneal tissue that can be used in place of Draize tests. Researchers from the National Institute of Agrobiological Societies, Tsukuba, carried out an experiment in which chemicals were

tested by leaving them on an ultrathin collagen sheet, and then observing the number of corneal cells that were damaged after three minutes.
Figure 3 shows the thin sheets of collagen developed by Takezawa (8). The researchers were able to cultivate human corneal cells, and create the equivalent of a human cornea.

The experiment (identified as the Vitrigel-EIT method) was made up of thirty toxicity experiments
Figure 3 Thin sheets of collagen

overall, the results of which showed that the new substance offered the same results as toxicity tests

developed previously in 90% of cases. The other 10% were seen to be even more accurate than tests on rabbits eyes. The Japanese researchers said the results of their experiments showed the widespread eye irritancy of chemicals can be predicted without false-negatives by the Vitrigel-EIT method.(9) Takezawa says apart from the cruelty involved, animal testing is unsustainable in the long term, because of the physiological differences between animals and humans. (10) Takezawa also believes that his research could be applied to all cells within the human body; from skin cells to internal organs. His research into this technique of testing cosmetics could be the solution to leaving animals out of the picture completely, thus delivering more reliable results that can be applied to humans more accurately.

Implications of the main solution

A social implication of the solution addressed previously, is that non-animal experiments may pose a risk to humans. If scientists were to perform tests on computer-based models, test-tube grown cells, or lower organisms (such as invertebrates or eggs, as opposed to warm-blooded animals) the results may not show a full picture of the risks that may pose a threat towards living organisms, as they would when testing on live animals. This is due to the fact that animals (including humans) exhibit variables that may not be fully understood through methods of experimentation not involving the use of animals. It may be argued, that in order to fully comprehend a live organism's system, scientists may have to implement animal testing at some point. This argument suggests that alternative methods to animal experimentation produce results that do not reveal the full extent of the potential risks, and so pose a threat towards humans. This is therefore a major social issue, and should be taken into consideration if cosmetic companies decide not to test their products on animals. An ethical implication of the solution is the idea that the overall benefit to human beings outweighs the harm done to animals. (11) It is possible that by not using animals when testing chemicals used in the cosmetics industry, we are limiting our knowledge on the risks that may affect humans. However, by using animals in these experiments, we are able to better understand the potential risks that the non-animal tests may otherwise not identify. As the solution of using alternative methods for testing chemicals does not involve the use of animals, regulators may suggest that the results would not mirror the effects fully, and so could pose risks upon humans. This is therefore a major ethical issue, and so should be taken into great consideration if animals are not used in the testing of cosmetics.

Benefits of the solution

Alternative methods of testing chemicals in the cosmetics industry that do not involve the use of animals not only reduce the harm done to animals, but they are also very cost effective; in fact, they cost only a portion of what animal experiments cost. For example, Corrositex costs approximately $200, whereas an equivalent animal test would cost anything ranging from $1,200 - $1,800. A further benefit from this would be that users of Corrositex can also save up to $50,000 annually on shipping charges. (12) Furthermore, another benefit of non-animal experiments is that these methods of experimentation are more environmentally friendly than animal-based tests. Once the millions of animals used in experiments have fulfilled their purpose, they are disposed of, creating much hazardous waste. (13) Conversely, the method of producing cruelty-free products through in-vitro or computer based tests is not damaging and does not produce unnecessary waste, as the nonuse of animals does not create environmental damage.

Alternative solutions
An alternative solution to the use of in vitro methods of experimentation, rather than the use of animals is in silico techniques. This form of testing means that biological experiments can be carried out entirely on a computer. As you can see from the table in figure 4,
(14)

the QSAR computer model has

shown evidence that they produce results which are strong at predicting allergic reactions and irritation due to certain chemicals that come into contact with the skin. In fact, the CAESAR model was seen to correctly predict 90% of the irritancy levels of 42
Figure 4

chemicals and the effects they might have on humans; whereas The animal tests involving mice or guinea pigs [as well as rabbits] only predict human reactions 72% of the time.(15) The results of computer models show the evidence of validity to be of a high standard, which would therefore suggest that they are a more reliable and valid method of testing chemicals used in the cosmetics industry, as opposed to animal experiments.

Another alternative solution that could be used in place of animal experiments is the development of organs-on-a-chip, including heart-on-a-chip and lung-on-a-chip. These miniscule devices were created by scientists and engineers at Harvards Wyss Institute (16) and contain human cells in a 3-dimensional system that mimics human organs. The microchips (which are created from a translucent, flexible plastic polymer) are the size of a computer memory stick and contain channels that are lined with living human cells (as seen in figure 5). Due to the microdevices being of a transparent nature, they provide a
Figure 5 organs-on-a-chip

window into the inner workings of human organs.

(17)

This revolutionary chip has many potential

applications; for example, it is thought that it will provide an accurate alternative solution of testing cosmetics, rather than the traditional animal tests that frequently fail to show results that will relate to humans. As well as this, it is believed that the organs-on-a-chip will be able to test both the safety of cosmetics and the toxicity of chemicals, and will supposedly offer results that are much more applicable than previous animal experiments, such as the Draize test. It is believed by the scientists and engineers at Harvards Wyss Institute that the microchips could one day form an accurate alternative to traditional animal testing.

Validity of sources
I chose the research of organs-on-a-chip as my main source of information for an alternative solution to animal experiments, as the well-respected Harvards Wyss Institute conducted it. Harvard University, located in Cambridge, Massachusetts, is a well-known prestigious and devoted Institute for providing extraordinary education, and is one of the best institutes for using this technology in the world. The Wyss Institute has the best researchers and engineers in the bioengineering sector, which leads me to believe that the information they provide is both accurate and valid, and I am able to give it my trust. I decided to include their work in my report for this reason. I chose the article In Vitro Alternatives for Ocular Irritation, by Rodger D. Curren and John W. Harbell to use as my non-internet based source, as I felt that the information within it was both useful and very relevant to my report, especially the information on the Draize Scoring Scale. I also felt that the images and tables that accompanied the article were very useful at providing a visual explanation for the alternative in-vitro tests that could be used instead of the traditional animal experiments. Dr. Rodger Curren has received many qualifications in the context of biology, for example his Bachelor of Science in biology from Purdue University, followed by a Master of Science from Ohio University, and a Doctorate from the Institute of Microbiology at Rutgers University. For this reason, I believe that the information within the article is both valid and accurate, hence my reason to use it in my report.

Bibliography
1) Curren, Rodger D., & Harbell John W. (1998). In vitro alternatives for ocular irritation. Environmental Health Perspectives, p.486. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1533382/pdf/envhper00537-0088.pdf Table 1 showing the scoring of Draize eye tests. In Vitro Alternatives for Ocular Irritation by Rodger D. Curren and John W. Harbell, April 1998. Environmental Health Perspectives, Vol 106. Supplement 2, Pages 486-487. This section can be found at the following address: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1533382/pdf/envhper00537-0088.pdf Curren, Rodger D., & Harbell John W. (1998). In vitro alternatives for ocular irritation. Environmental Health Perspectives, p.487. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1533382/pdf/envhper00537-0088.pdf Limitations and dangers of using animals in research. New England Anti-Vivisection Society (Neavs) http://www.neavs.org/research/limitations Image courtesy of PETA http://www.peta.org/issues/animals-used-for-experimentation/rabbits-laboratories/ More than a cosmetic change. NATURE, Vol 438, 10 November 2005. This article can be found at: http://ethics.ucsd.edu/journal/2006/readings/Animal_Testing_More_than_a_cosmetic_change.pdf

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7) 8)

See note 6 Screenshot of video showing Japanese scientist Takezawa see end to animal testing with the development of artificial cornea http://www.reuters.com/video/2099/01/01/reuters-tv-video?videoId=263962506 Oxford online journal Toxicological Science. Vitrigel-Eye Irritancy Test Method Using HCE-T Cells. Toshiaki Takezawa http://toxsci.oxfordjournals.org/content/early/2013/07/20/toxsci.kft159.abstract

9)

10) See note 7 11) BBC ethics Animal ethics: experimenting on animals. http://www.bbc.co.uk/ethics/animals/using/experiments_1.shtml#top 12) In Vitro international. Why Use Corrositex? http://www.invitrointl.com/products/corrosit.htm 13) Healing Cancer Naturally. Limitations of animal tests. http://www.healingcancernaturally.com/why_alternatives15.html 14) Table showing alternatives for skin sensitization. BUAV, campaigning to end animal experiments. A scientific review of non-animal tests for cosmetics. Page 7. This review can be found online at: http://www.crueltyfreeinternational.org/_assets/userfiles/files/BUAV_ScienceReport-SCREEN.pdf 15) See source 13 16) Organs-on-chips. Wyss Institute at Harvard University. http://wyss.harvard.edu/viewpage/461/ 17) See note 15