How Fake Skin Will End Animal Testing (as Soon as We Can Mass Produce It)
A rabbit undergoes a Draize test, a toxicity test approved by the FDA in which chemicals are applied to the eyes of restrained animals to record the effects. (Image: PETA/Wikipedia)

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How Fake Skin Will End Animal Testing (as Soon as We Can Mass Produce It)

A new L'Oreal partnership with a bioprinting startup could finally end the practice.

If you took a shower today, the products you used were required to be tested for safety under many laws around the world. Depending on the brand, your shampoo could have been cleared using a variety of methods, but traditionally, cosmetics have been tested by rubbing the product or its chemical components onto the shaved skin of animal subjects, which were likely killed shortly after the test, according to animal rights advocates.

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With animal testing bans being enacted, and more on the horizon, many companies have been racing to come up with alternatives. Methods range from experiments on human volunteers and simulations using computer programs, to testing on high-tech lab-grown skin.

The most recent development in this realm comes in the form of a partnership between cosmetic company L'Oréal and 3D bioprinting startup Organovo, which was announced earlier this month.

Products are rubbed into the skin and eyes of animals, often causing damage

The project will allow the beauty manufacturer to 3D print human skin to test its products, something it says no other company has done before, with the potential to automate the skin manufacturing process for mass production. So how close are we to getting away from animal testing completely?

The State of Animal Testing

Testing laws vary by country, but beauty companies have been required to prove their products are safe before they hit the US market since 1938, when Congress passed the the Federal Food, Drug, and Cosmetic (FDC) Act. The law was passed in response to several high-profile tragedies involving untested products, including an eyelash dye that caused many women to go blind from chemical burns.

Although the act does not specifically call for animal testing, it requires manufacturers to substantiate the safety of products and their components in some way, which many companies still do on living subjects.

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According to figures from the Humane Society, hundreds of thousands of mice, guinea pigs, rats, and rabbits undergo testing around the world each year to determine the safety of products including lipstick, mascara, shampoo, and cologne.

Products are rubbed into the skin and eyes of animals, often causing damage including irritation, ulcers, and blindness. Animals are not given pain relief and are usually killed after the tests, according to the Humane Society, by asphyxiation, neck-breaking, or decapitation.

In recent years there has been a major move away from testing cosmetics and other products on animals, at least in the European Union and the United States, according to Kim Paschen, a program manager at cruelty-free certification program Leaping Bunny.

Leaping Bunny is run by a coalition of eight animal protection organizations in the US and Canada, and certifies companies for being free of animal testing at all stages of development.

"Even the market's established international companies cannot produce more than 2,000 tiny skin pieces a month."

Leaping Bunny has certified more than 600 companies in the US and Canada and even more in Europe, Paschen said, but the patchwork of federal testing requirements around the world makes it difficult to shut down animal testing completely.

"Animal testing is decreasing overall, but there are new products all the time that are tested on animals," she said. "In the US it's not as common anymore, but in certain countries like China, regulation and requirements are more strict. China is a big problem because they require animal testing in a lot of cases."

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China's huge and quickly-growing cosmetics market is a big contributor to the prevalence of animal testing, she said. The Humane Society has previously reported up to 300,000 animals are killed in China alone each year due to testing.

Last year, China slightly relaxed its regulations, which previously required animal testing for all cosmetic products sold in the country, saying domestic companies now have the option to use alternative testing methods to prove safety before marketing cosmetics in the country.

However, many goods are still required to be tested on animals, including imported products and "special use" products, such as hair dyes, hair growth products, deodorants, and sunscreens, according to The Humane Society.

Paschen said the global push away from animal testing has been fueled not only by animal rights groups, but also by companies' desire for more accurate testing.

"Animal testing is ineffective because you're comparing an organism that isn't a human to humans, so the results vary," she said. "More companies are simulating human skin and developing other non-animal methods for testing."

When Testing on Humans Is the Cheap Option

To say animal testing has fallen out of favor would be an understatement. According to the Humane Society, 81 percent of Canadians, 73 percent of Americans, and 81 percent of Australians support a animal cosmetics testing ban. The European Union banned the testing of finished cosmetic products on animals in 2004 and eliminated the testing of individual ingredients on animals by 2013. The ban came after an aggressive campaign by PETA that included public protests, phone calls, and more than 20,000 emails.

In preparation for the EU ban on animal testing, major cosmetic companies worked quickly to find alternatives, pouring hundreds of thousands of dollars into research and in some cases being forced to collaborate with competitors. Companies are continuing to develop alternative methods like computer modeling and the production of artificial skin.

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"I personally really didn't like that we tested on humans."

L'Oréal claims to be one of the first entities to develop skin farming technology for this purpose, tracing its current innovation all the way back to the 70s.

"Forty years ago there was very little known about skin's physiology. Was it possible to grow skin in the lab that looked and behaved like the real skin?" a spokesperson from L'Oréal told Motherboard. "Two L'Oréal scientists responded to this need and, in 1975, were able to demonstrate the first true skin grown in vitro, before any other living organ had been reconstructed."

This story has become part of company lore, but Motherboard was unable to corroborate this alleged advancement and L'Oreal declined to provide more information on it. However, according to the book Case Studies in Innovation for Researchers, Teachers and Students, a L'Oréal researcher named Marcelle Régnier became the first person to successfully reconstruct human skin in 1983.

Around the same time, another French researcher Estelle Tinois developed a kit of artificial skin while working at the Lyon research laboratory Imedex, according to the book. She developed the method for growing skin from donated keratinocyte cells, the predominant cell type the skin's outermost layer, the epidermis, and called it Episkin. L'Oreal acquired the Episkin technology in 1997, which became a company others could purchase Episkin kits from in 2001.

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The keratinocyte cells come from tissue donated by plastic surgery patients. Skin cells harvested from the samples are grown into an artificial epidermis using a collagen matrix. The reconstructed skin grows in layers, like actual skin, and takes about a week to form. Episkin offers several models of reconstructed epidermis, including various skin colors and ages of skin.

L'Oreal integrated this technology into a kit called the Episkin model, a set that consists of 12 test tubes, each of them containing constructed epidermis. In 1998 the European Centre for the Validation of Alternative Methods (ECVAM) validated the Episkin model for alternative testing in the EU. L'Oreal's goal was to make the Episkin kits available commercially for safety purposes, and now more than 100 cosmetic, pharmaceutical, and chemical companies can purchase the technology to test products for safety.

Nadia Shakoor, a researcher who worked at L'Oreal from 2005 to 2009, says the company had already eliminated animal testing of its finished products at that time. Instead, they used a combination of human subjects and Episkin.

She said Episkin could assess traits like absorption, spreadability, feel, color change, and to some extent, irritation, but because of its steep price they would often opt for human testing instead at certain stages.

L'Oreal declined to comment on the cost of Episkin, but it was previously reported to be $70 per sample. For human testing, subjects would be sent a 48 hour patch test in exchange for monetary compensation.

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"I personally really didn't like that we tested on humans," Shakoor said. "Like all cosmetic products, sometimes people have skin allergies or sensitivities to certain chemicals, even if it's 1 to 2 percent of the population. I would much rather test on animals or tissue substitutes like Episkin than people who need money and are willing to take risks with their bodies and health to acquire it."

L'Oreal has continued to develop skin models and alternatives to animal and human testing. In addition to Episkin, L'Oreal also bought 81 percent of the share capital of SkinEthic, a former competitor that uses another method to reconstruct skin, in 2006.

Other researchers have also been working to find more efficient ways to produce more life-like artificial skin. In 2009, German research center Fraunhofer-Gesellschaft announced it was developing the first fully automatic production system for two-layer skin models.

Jörg Saxler, a scientist at the center, explained the demand for mass skin production at the time.

"The production is complex and involves a great deal of manual work," he said in a press release. "Even the market's established international companies cannot produce more than 2,000 tiny skin pieces a month. With annual requirements of more than 6.5 million units in the EU area alone, however, the industrial demand far exceeds all currently available production capacities."

In 2011, L'Oreal opened a facility in France that produces 100,000 units of reconstructed tissue every year using both of these technologies, each sample a 0.5 square centimeter in size. According to Bloomberg, L'Oréal uses about half of the skin it produces for its own work and sells the rest to other cosmetic companies.

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"L'Oreal's focus right now is to continue to build on the accuracy and consistent replication of the skin engineering process."

In March 2013, L'Oreal officially stated it would completely eliminate the testing of its products and ingredients on animals. The announcement came just one year after launching its Technology Incubator based out of the company's Research & Innovation labs, meant to develop new technology in this area and others.

In addition to reconstructed skin models, the company uses predictive analysis as well, including computer programs and computer-aided design of chemical molecules to assess how ingredients will interact and affect humans.

"L'Oréal believes that the future of safety evaluation lies in the development of alternative, 'predictive' strategies," the company's website says of animal testing. "This approach is built on the substantial scientific progress being made in the fields of reconstructed skin models, molecular modeling and also high-performance data processing."

The Road to Mass Skin Production

Organovo is the latest company L'Oreal has turned to in that approach. The regenerative medicine company was founded on bioprinting technology developed at University of Missouri–Columbia in 2005 and officially incorporated in 2007.

Organovo has worked to develop a variety of human tissues using bioprinting technology in the past, including liver tissue for toxicology testing. Organovo does much of its work through partnerships with companies, but this is the first time its technology will be used in the beauty industry.

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"All of our tissues are made from human cells," Organovo told Motherboard. "It's better to think of it as 3D bioprinted human tissue."

The Organovo bioprinter is 18 by 24 inches and fits within a standard biosafety cabinet by design to keep all of the cells and tissues sterile. It works by taking living human cells and placing them in a 3D matrix where they can grow into functional tissue.

"In the case of skin that means you can have different cell types in layers, for example, and different amounts of certain types of cells in each layer," a representative from Organovo said.

A photo of a bioprinted breast cancer tumor model which looks similar to 3D bioprinted skin under the microscope. (Image: Organovo)

Recreating skin uses a similar process to how Organovo has bioprinted tissues in the past, the company representative said, but every new tissue has a learning curve.

"Skin has specific characteristics that make it interesting, including of course multiple layers, hair follicles, immune cells, and the fact that it naturally is exposed to air," a representative from Organovo said.

The representative told Motherboard that to Organovo's knowledge, it is the only company making fully cellular 3D bioprinted tissues. L'Oreal will have exclusive rights to the Organovo 3D printed skin technology.

"Some of the biggest potential advantages of 3D bioprinting are the speed of production as well as the level of precision that 3-D printing can achieve," Guive Balooch, global vice president of L'Oreal's Technology Incubator told Motherboard. "We are still in the preliminary research phase of the partnership, but L'Oreal's focus right now is to continue to build on the accuracy and consistent replication of the skin engineering process."

Organovo's 3D bioprinter (Image: Organovo)

Meanwhile, other cosmetic companies are developing their own testing technology. In January 2014, Procter & Gamble developed its own approved non-animal alternative for skin allergy testing. The method involves combining product ingredients with peptides that mimic skin tissues in small test tubes and measuring the reaction.

Unilever, the company behind cosmetic brands like Dove and Axe, invests 3 million Euros, or 3.3 million dollars each year into developing animal testing alternatives, including mathematical modeling to predict skin allergy and and other predictive methods.

Companies are also approved to use the Bovine Corneal Opacity and Permeability Test and Isolated Chicken Eye Test, which recycle eyes from animals killed for meat instead of using live animals to test for eye irritation.

Other cosmetic providers use only ingredients that have been established as safe in their products, eliminating the need to test on animals.

Although the Organovo partnership is still in early stages, the partnership is the latest step in efforts to find high-tech, accessible alternatives to animal testing. As Shakoor noted, the current alternatives to animal testing come at a cost.

"It's easy to cry foul when it comes to animals, but animal rights organizations forcing the industry to exclusively test on humans isn't without consequence," she said. "I'll be very glad when all testing is validated on artificial surfaces like the 3D skin."