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1 New Technologies in biomedical research should replace animal testing Jeremy J. Hansen Department of Computer Science, ECPI University ENG120: BO Advanced Composition Catherine Gorman-Sayles October 24, 2023

2 New Technologies in biomedical research should replace animal testing Jeremy Hansen, with his adopted dog from the laboratory breeding facility named Envigo, was shut down in 2022. Taken by Elizabeth Hansen. Most people do not know that animal testing on companion animals, farm animals, and primates still exists. This is because it is widely assumed that animal testing is only done on rodents. Even though $45 billion in public tax dollars goes into medical research, these pharmaceutical companies feel they do not need to tell the public how they are testing their products (National Institutes of Health, 2019). However, in 2014, Jeremy discovered that dogs, mostly beagles, are still used in harmful experiments. As a dog lover, this fact disturbed Jeremy greatly, and he decided to get involved by speaking out against what he felt was unethical and immoral. The significant change is when Jeremy was part of a protest against a dog breeding facility in Virginia named Envigo. They had over 5,000 beagles in their warehouse, and they were caught being cruel to animals in 2021 by an undercover investigator from People for the Ethical Treatment of Animals (PETA). After they were caught and reported to the authorities, the U.S. Department of Agriculture cited them for 70 federal Animal Welfare Act violations and were told to shut down. After adopting one of those beagles, Jeremy decided there had to be a better way of discovering new pharmaceutical drugs. Jeremy’s research has found that new

3 technologies can better predict human safety in developing new drugs and should replace animal testing. Recently, scientists have relied on animal testing for toxicity screening on human drugs. This is causing unnecessary harm and waste to animal and human lives. An estimated 110 million animals are used each year. Out of that number, 71,921 were primates, 44,847 dogs, and 12,595 were cats, which does not account for private companies that do not have to report this information (Cruelty Free, 2023). Testing on animals is a very inefficient process. It takes $2.6 billion to develop a new drug, and then 92% of those drugs fail in human clinical trials. With the advancement in non-animal research, scientists can better predict the effects of drugs on humans by using technology that gives human results (Gail, 2019). This is better for researchers and humans who eventually must try the drug. Scientists, research staff, and research animal caretakers experience high stress performing animal testing, causing the industry to lose possible candidates for moral and ethical reasons (Kang et al., 2018). This hinders pharmaceutical companies from finding qualified staff who want to study drugs. (peta, 2013) It is easy to see how going to work every day conducting painful experiments on animals can lead to an incredible amount of stress, especially when it is on companion animals and primates.

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4 According to a State-Trait Anxiety Phycological (STAI) test conducted by the Ewha Womans University, the primary staff involved suffer emotionally by breaching their personal moral and ethical standards. The test showed that 82% of women and 66% of men were concerned and sympathized with the animal being tested on. Stress in staff workers leads to excessive alcohol drinking, job dissatisfaction, sleep disorders, and mental depression, causing a loss in productivity (Kang et al., 2018). Is it fair to ask our most brilliant people who invested an incredible amount of time and money in their education to live in depression by breaking their morals and ethics? We also do not know how many people choose different career paths because they do not want to test on animals. Many exciting new technologies are being used for biology testing that are incredibly effective. The most commonly used today are 3D tissue bioprinting, organoids, and organ-on-a- chip (OOC). (3D Bioprinting Artificial Organs Could Become Quicker and Easier, n.d.) 3D Bioprinting uses natural or synthetic substances that contain living stem cells from any tissue from the human body. This technology can be used to print tissues from a technique called bioink to be fully functional organs for pharmaceutical science. This gives the researchers better human results, increasing the accuracy of what the drug will do to the patient, decreasing the

5 harm to humans and animals, and eventually decreasing the expense of producing the drug (Panja et al., 2022). Organoids are laboratory-grown 3D organs derived from human stem cells that are exact replicates of an actual human organ. This provides researchers with an actual human organ that responds on a life-like scale and has the genetic characteristics of the actual organ and how it will respond to pharmaceutical drugs (Yang et al., 2023). This life-like simulation provides safe, precise research that does not harm humans or animals. Organ-on-a-chip is a small hand-held device that uses microfluidic cells. These fluids are used together to represent a functional organ. One chamber has blood vessel cells, while the other has organ tissue cells. The chip allows for stretching or relaxing, mimicking human body Liver Organoid Neural Organoid Intestinal Organoid ( Organoid Research, n.d.) A lung-on-a-chip device. Image courtesy of Wyss Institute, Harvard University. (The MIT Press Reader,2022) (Organ-On-a-Chip: Microfluidic Technology That Can Revolutionize the Pharmaceutical Industry, n.d.)

6 movements. All of these sequences represent a human organ, such as a heart, lung, or any organ in the human body. Multiple organ chips can be linked together to represent a human body-on-a- chip. This allows scientists to understand how drugs move through the body, affecting human organs (Hansen, 2020). This is a complicated, new, and expensive technology, but so is animal testing. However, so is all technology when it first comes out. How much does it cost society when people die from inaccurate pharmaceutical drugs or $2.6 billion tax dollars are spent on drugs that do not work? 92% fail rate is unacceptable in any business or government entity; why is this acceptable in pharmaceuticals when there is a more accurate way? The obvious answer is that we must invest in the proper methods, or the actual costs of improper testing will keep increasing. Scientists want to check a drug's toxicity in every way possible before it gets to human trials. Even though non-animal research methods are 90% accurate, there is still a 10% risk. Non- animal research technologies are not considered complex human beings, and some scientists consider them inferior to testing on animals. Testing on animals should be done as a precaution. This reduces the risk of people getting hurt or dying. New pharmaceutical drugs are unpredictable until researchers properly analyze the drug. An animal should suffer and die before a human (Knight, 2019). Animal testing does not give the same results as testing in humans. They are two different complex models. There will always be risk in pharmaceutical research; maybe setting a standard below 10% is unreasonable. Now, researchers can combine methods to make non-animal testing methods more complex. Combining multiple technologies such as organoids, 3D bioprinting, and organ-on-a-chip labs can create a whole human representation, making non-animal testing

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7 methods far superior to animals because we are testing the drugs on the proper species (Knight, 2019). Life-saving drugs are an international government issue, and some countries require researchers to conduct animal testing before it is allowed on the market. This is considered in the scientific research industry as a lack of harmonization across countries. If a drug were tested in France and the researchers decided to use non-animal experiments, that drug may not meet the requirements to be sold in the United States unless proper animal testing was involved. So, the pharmaceutical companies in France would have to use animal testing in order to make their product more accessible. A pharmaceutical company would then have to invest in animal testing and the non-animal testing process to sell their drug internationally. This increases the expense and the investment in drug testing with the same potential profit that can be made (Knight, 2019). Creating life-saving drugs is not just a profitable business for investors but also a humanitarian issue. People deserve the benefit of life-saving drugs created safely and efficiently. These investments are not just coming from investors. In 2018, the National Institute of Health (NIH), a federal agency funded by the American people's tax dollars (USAGov, n.d.), contributed over $100 billion in research for 210 new drugs approved by the FDA and gave pharmaceutical companies $76 billion in tax breaks (Meller & Ahmed, 2019). American people are giving pharmaceutical companies money to produce life-saving drugs, the priority needs to be utilizing the best methods to produce the drugs and not the bottom line. The European Medicines (EU) is responsible for harmonizing drugs and modern testing method acceptance across countries, which complies with the Organization for Economic Co- operation (OECD). If the drug is approved by both of these entities, then it must be published by

8 the official EU regulations. This is a two-year delay before researchers are allowed to use the new technology they invested in, making non-animal testing a more expensive process than it should be. There is no need to have two entities monitoring the testing methods, causing delays and added expense in producing new technologies. Two years to get a permit is unacceptable in any industry, and the EU needs to make this process more streamlined (Knight, 2019). Lives are at stake. Humans have always used animals for labor, pets, and food. Over five billion animals are killed annually in the United States for food alone. Animals such as mice share 98% of the same biological DNA as humans. They also share the same diseases, such as cancer and polio. The polio vaccine is still being tested for safety in monkeys. Animals are easy to grow and examine. Many have less life span than humans, showing the drug tests through the complete life cycle. Eighty-five percent of animals being tested are mice and rats (“Read ‘Science, Medicine, and Animals’ at NAP.edu,” 1991). The Animal Welfare Act (AWA) requires scientists only to use animal testing when necessary and defines the 3Rs: replace, reduce, and refine animal testing. This requires the scientist to look at the experiment to see if they can replace animal testing with non-animal testing methods, reduce the number of animals conducted in the research to the minimum necessary, and refine the test to reduce stress and pain or enhance the well-being of the animal (Alternatives to Animal Testing, n.d.). This federal law is enforced by the Investigative and Enforcement Services (IES). The big Envigo scandal shows that these inspections are sometimes not conducted. Envigo, a pharmaceutical beagle dog breeding facility in Virginia, was cited for more than 70 violations of the AWA act and did not get inspected until PETA’s undercover

9 investigation went viral and forced the IES to conduct an investigation and eventually shut the breeding facility down (PETA Investigates Mill Breeding Beagles for Experiments at NIH, n.d.). Even if animals share 98% of the same biological DNA as humans, the scientific results do not support that animals are a great study of drugs. Ninety-two percent of drugs that pass animal trials fail in human clinical trials. That means 92% of people getting paid for clinical trials are experiencing harmful, toxic side effects. It is also a waste of funding when the pharmaceutical company cannot use a drug that costs an average of $2.6 billion to develop (Gail, 2019) (Hansen, 2020). This shows that not only are animals being tested, but humans are as well. The statistical documentation is unavailable after attempting to find any statistics on how many people die or get injured in human clinical trials. It is probably not public knowledge for privacy reasons and is unavailable on the FDA website. If people are being tested in human clinical trials, they should know the risks, all possible outcomes, and statistics of people getting harmed in past clinical trials. It is illegal not to inform the patient of all risks (Research, 2019). The ranching industry is not a good comparison to the medical industry. They are two separate things and two completely different issues. All deaths of animals, whether from hunting, ranching, roadside accidents, or natural causes, should not be compared to whether researchers should use animal testing. This oversimplifies the issue and makes the topic illogical in the scientific community (cite the McGraw Hill handbook). This scientific argument should be based on evidence on whether researchers should be using animals for their toxicity studies or non-animal technology. The reasons that should be explored are the better, more accurate, and ethical ways to test drugs. Not if there is a difference between eating animals and testing on them. This does not support a logical scientific argument based on evidence (The Logic of Scientific Arguments - Understanding Science, 2022).

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10 Since the development of non-animal bio-testing methods, animal testing is no longer the most effective drug testing method. This is an excellent win for the scientific community, animal activists, research animals, and humans. Producing safe, effective drugs with little to no side effects is vital for society. Unfortunately, non-animal testing technologies are a new development that will take time and money to implement. Animal testing dates back to 384 BC, and the first effective non-animal testing method was developed in 2005 (Hajar, 2011) (Simian & Bissell, 2016). It will take time for researchers and pharmaceutical companies to get comfortable using alternative methods. The scientific community needs to keep moving forward, prioritizing the success of healthcare and ethics, not profits and ignorance. Looking at the statistical evidence, past dangerous drugs, and the side effects each drug produces, we can see that the past animal testing methods are ineffective, showing a need to invest in non-animal testing technologies. Pharmaceutical researchers should focus their time and resources on the best methods to produce effective and safe, life-saving cures. Society can help make drugs safer and end animal testing. Instead of listening to pharmaceutical drug advertisements and doctors, we should ask questions and get the facts before buying and taking medications. You should ask the statistics on how many people experienced ill side effects or even died from this drug. How was this drug tested? What kinds of animals were used? Is this against your ethics? Do you find this drug to be unsafe? Are the side effects worse than the actual problem? Educate yourself about the medications before you take them. Voice your concerns to your doctor and pharmaceutical companies about their drugs not meeting your safety or ethical requirements. You are the customer, and your tax dollars go into pharmaceutical research. You should speak out and have a say in how these drugs are developed.

11 Reference Akhtar, A. (2015). The Flaws and Human Harms of Animal Experimentation: Cambridge Quarterly of Healthcare Ethics, (4), 407–419. https://doi.org/10.1017/s0963180115000079 3D bioprinting artificial organs could become quicker and easier. (2023). Drug Target Review. https://www.drugtargetreview.com/news/110266/3d-bioprinting-artificial-organs-could- become-quicker-and-easier/ Alternatives to Animal Testing. (n.d.). National Institute of Environmental Health Sciences . https://www.niehs.nih.gov/health/topics/science/sya-iccvam/index.cfm#:~:text=Under %20U.S.%20law%20and%20policies Berkeley University of California. (2022). Understanding Science . https://undsci.berkeley.edu/understanding-science-101/how-science-works/the-logic-of- scientific-arguments/ Hajar, R. (2011). Animal testing and medicine. Heart Views , 12(1), 42. https://doi.org/10.4103/1995-705x.81548 Hansen, E. (2022). Laboratory Dogs Rescued: From Test Subjects to Beloved Companions. McFarland & Company Inc. Kang, M., Han, A., Kim, D., Seidle, T., Lim, K.-M., & Bae, S. (2018). Mental Stress from Animal Experiments: a Survey with Korean Researchers. Toxicological Research . 34(1), 75–81. https://doi.org/10.5487/tr.2018.34.1.075 Meller, A., & Ahmed, H. (2019). How Big Pharma Reaps Profits While Hurting Everyday Americans. Center for American Progress. https://www.americanprogress.org/article/big- pharma-reaps-profits-hurting-everyday-americans/ National Institutes of Health. (2019) . Budget of National Institutes of Health (NIH). https://www.nih.gov/about-nih/what-we-do/budget Organoid Research. (n.d.). Stemcell Technologies . https://www.stemcell.com/technical- resources/area-of-interest/organoid-research.html uFluidix. (n.d.). Organ-on-a-Chip: microfluidic technology that can revolutionize the pharmaceutical industry . https://www.ufluidix.com/microfluidics-applications/organ-on- a-chip/ Panja, N., Maji, S., Choudhuri, S., Ali, K. A., & Hossain, C. M. (2022). 3D Bioprinting of Human Hollow Organs. AAPS PharmSciTech , 23(139), 1-18. https://doi.org/10.1208/s12249-022-02279-9 PETA Investigates Mill Breeding Beagles for Experiments at NIH. (n.d.). PETA Exposés and Undercover Investigations. https://investigations.peta.org/dog-beagle-breeding-mill- envigo/

12 Science, Medicine, and Animals (1991). Why Are Animals Used in Research ? https://nap.nationalacademies.org/read/10089/chapter/3 Seyler, D. & Brizee, A. (2022). Read, reason, write: An argument text and reader. Ch. 6. McGraw-Hill Education. Simian, M., & Bissell, M. J. (2016). Organoids: A historical perspective of thinking in three dimensions. The Journal of Cell Biology , 216(1), 31–40. https://doi.org/10.1083/jcb.201610056 United States Department of Agriculture (USDA). (2022 ). Animal Welfare Act. https://www.aphis.usda.gov/aphis/ourfocus/animalwelfare/sa_awa United States Food & Drug Administration (FDA). (2004). Vioxx (rofecoxib) Questions and Answers. https://www.fda.gov/drugs/postmarket-drug-safety-information-patients-and- providers/vioxx-rofecoxib-questions-and-answers United States Food & Drug Administration (FDA). (2014). Inside Clinical Trials: Testing Medical Products in People. https://www.fda.gov/drugs/information-consumers-and- patients-drugs/inside-clinical-trials-testing-medical-products-people United States Government. (n.d.). National Institutes of Health (NIH) . https://www.usa.gov/agencies/national-institutes-of-health#:~:text=The%20National %20Institutes%20of%20Health The MIT Press Reader. (2022). The Organ-on-a-Chip Revolution Is Here . https://thereader.mitpress.mit.edu/the-organ-on-a-chip-revolution-is-here/ Yang, S., Hu, H., Kung, H., Zou, R., Dai, Y., Hu, Y., Wang, T., Lv, T., Yu, J., & Li, F. (2023). Organoids: The current status and biomedical applications. MedComm , 4(3), 1-32. https://doi.org/10.1002/mco2.274

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