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THE FUTURE IS ANIMAL-FREE - ACCELERATING HUMANE AND HUMAN-RELEVANT SCIENCE

The ethical and scientific crossroads

Millions of animals suffer in laboratories across Europe and globally each year, including dogs, cats, monkeys, and rodents (1). This practice is undergoing a significant re-evaluation due to ethical concerns and scientific limitations. Animal experimentation, long a staple of biomedical research and regulatory testing, is increasingly challenged by evidence of its poor applicability to human health.

A paradigm shift is underway, driven by scientific innovation, ethical pressure, and policy reforms, moving towards animal-free research methodologies (2). These Non-Animal Methodologies (NAMs) aim not just to reduce animal suffering but to provide scientifically superior tools. The language is evolving from "alternatives" to "replacements" and "superior methods," reflecting growing confidence in human-based approaches (3). Advanced technologies like artificial intelligence (AI), organoids, and organs-on-a-chip, coupled with public demand for ethical science (e.g., European Citizens' Initiatives (4)), are accelerating this transition. This article explores the failings of animal experimentation, the potential of NAMs, global policy shifts in the EU and US, and the next steps towards animal-free science. 

The failing paradigm: Why animal models fail us

Reliance on animal experimentation faces strong ethical objections and scientific deficiencies. Animal use in labs involves inherent suffering: confinement, invasive procedures, induced diseases, and premature death (5). In the US, animals can be burned, shocked, poisoned, and starved, often without adequate pain relief (6). Annually, about 10 million animals are used in EU labs (1) and 3 million in Great Britain (7). These animals are sentient beings with intrinsic value, not mere research tools. Animal experimentation is scientifically limited and an unreliable predictor of human outcomes due to poor translatability based on species differences: Significant physiological, metabolic, and genetic differences between species make extrapolating animal data to humans uncertain (5). Penicillin is toxic to guinea pigs; Paracetamol is poisonous to cats (5). Aspirin can be dangerous for some animal species (8).

This insurmountable drawback is reflected by misleading experimental results and high drug failure rates: 90% to 95% of drugs safe and effective in animal tests fail in human trials (3). This wastes resources, animal lives, and gives false hope. Thalidomide caused birth defects despite animal testing (8); Vioxx caused heart attacks in humans after showing protective effects in mice (8). Over 100 stroke drugs and 85 HIV vaccine candidates successful in animals failed in humans (8).

Another concern is that under the current testing strategy, potential human-relevant cures are ignored as animal tests might lead to abandoning effective human treatments. FK-506 (tacrolimus) was nearly shelved due to adverse animal results (8). Intravenous vitamin C treats sepsis in humans but not in mice (8).

Artificial experimental conditions and stress-induced artefacts are a major factor contributing to the poor reliability of animal experiments. Human diseases are artificially induced in animals and these animal disease models are used for biomedical research. Human tumours are grown in rodents for oncological research, genetic modification induces plaque development in animals to model Alzheimer’s disease and mice are injected with a toxin which destroys their pancreas to study diabetes. Animals do not develop complex human diseases and are not suitable for mimicking human pathology. Laboratory conditions induce chronic stress in animals, altering their biochemistry and immune function, confounding results (9). Stressed rats develop chronic inflammation (9). Enriched environments can alter disease progression in genetically predisposed mice (9). High-dose testing fallacy is a problem as exposing animals to extremely high doses of chemicals often leads to irrelevant toxic effects, not reflecting subtle, long-term human exposure (10).

The argument that animal testing is a "necessary evil" before human trials is flawed, as human trials are the ultimate test. Robust, human-relevant NAMs can make first-in-human trials safer (11). Animal experimentation is expensive and time-consuming (12). Drug development costs billions, with high failure rates contributing significantly (10). Regulatory testing for pesticides can take a decade (10). This delays beneficial products and diverts resources from developing better methods (9).

Animal-free innovation: non-animal methodologies (NAMs)

NAMs are a diverse toolkit of human-relevant methods revolutionizing research, regulatory testing, and education (2). The most relevant NAMs in the context of animal replacement include:

1) Organoids: miniature organs grown in the lab

Organoids are 3D cellular structures grown in vitro from human stem cells (pluripotent, induced pluripotent, or adult stem cells) that self-organize into miniature versions of organs like the brain, liver, and gut, mimicking their architecture and functions (13).

In fundamental research, organoids aid disease modeling. Patient-derived gut organoids are used in cystic fibrosis research for personalized drug testing (14). Brain organoids helped understand Zika virus's impact on neurodevelopment (14). Lung organoids study SARS-CoV-2 infection (15). Tumor organoids are vital in cancer research for drug screening and resistance studies (16). They also advance developmental biology by allowing in vitro study of human organ development (16).

In regulatory testing, organoids are used for drug discovery and toxicity assessment. Liver organoids show high predictive value for drug-induced liver injury (15). The FDA Modernization Act 2.0 supports using organoids in new drug applications (see below) (14).

2) Microphysiological systems (MPS): The body-on-a-chip

Organ-on-chip (OoC) are microfluidic devices with living human cells in perfused micro-chambers replicating functional units of human organs (17). OoC systems are able to mimic interorgan communication and a blood circuit. They offer more accurate human physiological responses to drugs than traditional methods (17). Applications include lung-on-a-chip for respiratory diseases (18), Blood-Brain Barrier (BBB)-on-a-chip for neurological therapies (17), and tumor-on-a-chip for cancer studies (17). "Body-on-a-Chip" systems integrate multiple organ models to study systemic drug effects (17). This technology is also supported by the FDA Modernization Act (19). 

3) In silico & artificial intelligence: The power of prediction

In silico (computational) modelling describes models using computer simulations (e.g., QSAR) to predict chemical properties and effects, reducing lab testing (20). They predict toxicity (like DART (20)), drug efficacy, and ADMET properties (21). The US FDA uses in silico modeling for product evaluation (22)..Artificial Intelligence (AI), especially machine learning, accelerates drug discovery. Traditional drug development is slow and costly ($2.5 billion, 13-15 years) (23). AI analyzes vast datasets from various sources (in vitro assays, 'omics') to identify patterns (2). Deep learning models predict hazards and benefits of new chemicals, sometimes outperforming animal tests in areas like liver and developmental toxicity (2).

The strength of NAMs lies in their integration: organoids in OoCs, data from these systems feeding AI models (2). This synergistic approach surpasses single methodologies. Many NAMs also allow for personalization, crucial for precision medicine (2).

4) Animal-free models in education and training

Animal-free methods are replacing harmful animal use in teaching. High-fidelity human simulators, e.g. mannequins like TraumaMan (24) and SimMan 3G Plus (24) allow practice of surgical and medical procedures. Specialized anatomical models including BoneClones (25) and veterinary simulators like the Haptic Cow (25) offer detailed training. Computer software and virtual reality (VR) offers interactive 3D anatomy software and VR dissection platforms provide immersive learning (25).

NAM tools compared to animal experiments are often more accurate, allow repetitive practice, and can simulate rare scenarios (24). A review found humane teaching methods as effective or more so than animal use in 90% of cases (26).

Table 1: Overview of key new approach methodologies (NAMs) 

NAM Type

Brief Description

Key Applications in Research/Testing

Examples

Key Advantages

Organoids

3D cell cultures derived from human stem cells that self-organize to mimic organ structure and function.

Disease modeling (genetic, infectious, cancer), developmental biology studies, drug discovery & toxicity testing, personalized medicine.

Brain organoids for Zika virus (14); Gut organoids for cystic fibrosis (14); Patient-derived tumor organoids(16).

Human-specific, patient-specific potential, 3D architecture, good for complex diseases.

Organs-on-a-Chip (OoC) / Microphysiological Systems (MPS)

Microfluidic devices with human cells that replicate organ-level functions and physiological responses in a controlled microenvironment.

Drug efficacy & toxicity prediction, disease modeling (lung, BBB (blood brain barrier), cancer, cardiovascular), ADME studies, personalized medicine.

Lung-on-a-chip (18); BBB-on-a-chip (17); "Body-on-a-Chip"(17).

Human-relevant physiology, controlled environment, potential for multi-organ systems, high-throughput screening.

In Silico Models (Computational)

Computer simulations and mathematical models (e.g., QSAR) to predict chemical properties, toxicity, efficacy, and pharmacokinetics.

ADMET prediction, toxicity prediction (DART), hazard identification, risk assessment.

QSAR models for DART (20); PBPK models for pharmacokinetics (21).

Rapid, cost-effective, high-throughput, reduces animal use, can integrate diverse data.

Artificial Intelligence (AI)

Machine learning and deep learning algorithms to analyze large datasets, predict biological responses, and accelerate drug discovery.

Drug discovery, toxicity prediction (liver, cardiac), pathway analysis, personalized medicine.

DICTrank Predictor, DILIPredictor (27); BioMorph (27); AI in neurology research (24).

Handles complex data, high predictive power, can outperform animal tests, identifies novel patterns, ethical benefits.

Advanced Educational Simulators

High-fidelity mannequins, specialized models, computer software, and VR tools for medical and veterinary training.

Surgical skills training, anatomy education, emergency procedure practice, clinical decision-making.

TraumaMan (24); SimMan 3G Plus (24); Haptic Cow (25); VR dissection (25).

Human/animal-relevant, safe repetitive practice, ethical, can simulate rare conditions, often more effective than animal labs.

The advantages of animal-free science

Transitioning to animal-free research offers improved predictive accuracy, accelerated timelines, cost savings, and enhanced public trust. NAMs are based on human cells, tissues, and data, offering superior human relevance compared to animal experiments (3). Organoids and OoCs recapitulate human organ function with greater fidelity (28). AI models trained on human data can predict human-specific responses (27). This leads to superior predictive accuracy, potentially reducing drug attrition rates (3). Key opportunities of NAM-based research are:

1) Accelerating safety, discovery and medical progress

Animal-free methods are often quicker and more efficient (12). High-throughput screening with NAMs can test thousands of compounds rapidly (16). AI can analyze data almost instantaneously. This speeds up therapy identification and evaluation.

2) Cost effectiveness

Animal experimentation is expensive (12). NAMs, especially in silico and many in vitro assays, can be more cost-effective. By improving predictive accuracy and reducing drug failures, NAMs can save billions in R&D (29). The US FDA acknowledges NAMs' "enormous cost saving potential" (30).

3) Ethical congruence and enhanced public trust

Animal-free science eliminates animal suffering, aligning with societal values and public demand (e.g., 1.2 million signatures of ECI 'Save Cruelty-Free Cosmetics' (4); >85% of Americans support phasing out animal experiments (31), also the vast majority of European citizens (32)). Adopting humane, advanced methods enhances public trust (30). It also represents a more responsible allocation of scientific resources towards methods more likely to benefit human and environmental safety and human health (11). 

Policy shifts towards animal-free regulation

A global trend towards animal-free testing is accelerating, driven by policy shifts in the EU and US.

A. The European Drive: Leading the Charge

The catalyst: ECI 'Save Cruelty-Free Cosmetics – Commit to a Europe Without Animal Testing'

This ECI, with over 1.2 million signatures (4), demanded strengthening the cosmetics animal testing ban, transforming EU chemicals regulation towards animal-free methods, and a roadmap to phase out all animal testing (4). In response (July 2023), the European Commission pledged a roadmap towards phasing out animal testing for chemical safety assessments and actions to reduce animal testing in research and education (33).

The European Commission's Roadmap Towards Phasing Out Animal Testing for Chemical Safety Assessments aims to phase out animal testing for chemical safety, targeting finalization by Q1 2026 (33). It involves Working Groups (Human Health, Environmental Safety, Change Management) and extensive stakeholder consultation (34). A key focus is identifying NAMs for immediate implementation and addressing complex toxicological endpoints (34).

The relevant European Agencies demanding animal tests for safety assessment are ECHA (European Chemicals Agency), EMA (European Medicines Agency) and EFSA (European Food Safety Authority). ECHA operates under REACH (animal testing as a last resort) (35) and promotes NAMs via critical examination of testing proposals, data sharing, tools like QSAR Toolbox and IUCLID, and read-across (35). Its 2023 report showed increased in vitro method use (36). However, under the current system, ECHA is still requesting and relying on animal experiments, although NAMs for replacement are available. For certain testing areas such as endocrine disruptors or mixtures, even increasing animal testing is under discussion. ECHA is heavily involved in the Roadmap Development and needs to align the contents with the REACH revision which is planned for 2025. EMA (European Medicines Agency) and EFSA (European Food Safety Authority) both have strategies to reduce and replace animal tests (37). For a successful and effective Roadmap towards animal-free safety testing, it is essential that all relevant agencies align their efforts and commit to a strategy to replace animal testing.

The European Research Area (ERA) Action is another approach within the Commission’s response to the ECI (38). The aim of this ERA Action is promoting Non-Animal Approaches in Biomedical Research and aims to promote NAMs in biomedical research and pharmaceutical testing, coordinating Member States' actions (39). It focuses on NAM development, validation, education, and data sharing, with impacts expected by 2027-2028 (39). In contrast to the Roadmap, the ERA Action is not restricted to regulatory testing but aims at replacing animal experiments within the European landscape, including education and fundamental research.

B. United States Initiatives: A New Trajectory

In 2022, the US Food and Drug Administration (FDA) announced the FDA Modernization Act 2.0 which was a major step towards acceptance of NAMs for regulatory use.

The act removed the explicit requirement for animal testing for new drug approvals, allowing FDA to accept NAM data (cell-based assays, organ chips, computer models) (3).

The FDA's Roadmap to reduce animal testing was published in April 2025. It aims to make animal studies the exception for preclinical safety testing within 3-5 years, starting with monoclonal antibodies (mAbs) (40). It encourages NAMs like AI models, organoids, OoCs, microdosing, and use of existing international human safety data (30) and includes a pilot program for mAb developers and potential regulatory incentives (30).

Beyond regulatory testing, the US National Institutes of Health (NIH) announced at the same time (April 2025) its new strategy focused on prioritizing human-based research. The aim of this initiative is to expand human-based science and reduce animal use in NIH-funded research (41). A new Office of Research Innovation, Validation, and Application (ORIVA) will coordinate efforts, expand funding and training in NAMs, and address bias in grant reviews (41).

The US Environmental Protection Agency (EPA) has committed to a NAMs work plan that aims to reduce reliance on animal testing for chemical safety (42). Initial goals included reducing mammal study requests by 30% by 2025 and eliminating them by 2035 (43). The work plan focuses on evaluating regulatory flexibility, developing metrics, establishing scientific confidence in NAMs, and stakeholder engagement (44). 

C. International harmonization efforts: a global symphony for change

The OECD (Organisation for Economic Co-operation and Development) Test Guidelines Programme develops internationally accepted Test Guidelines (TGs) for chemical safety, crucial for Mutual Acceptance of Data (MAD) to prevent duplicative testing (45). NAM-based TGs are slowly but increasingly adopted (e.g., for skin sensitization, eye irritation) (35). The OECD also publishes guidance like GIVIMP for in vitro methods (46) and identifies insufficient funding for NAM validation as a key challenge (47).

ICATM (International Cooperation on Alternative Test Methods) is a global collaboration of validation organizations (EURL ECVAM, ICCVAM, JaCVAM, etc.) to foster international cooperation on NAM development, validation, and regulatory use (48).

ICCVAM (Interagency Coordinating Committee on the Validation of Alternative Methods - US) comprises 17 US federal agencies, promoting NAM development, validation, and regulatory acceptance (49). ICCVAM develops strategic roadmaps and engages with international partners to implement NAMs and replace animal testing (50). 

Accelerating the transition: the path forward

A successful and sustainable transition towards animal-free science requires strategic action from all stakeholders. Consensus recommendations for action include (51):

  1. Strengthen Legislation & Harmonize Regulations: Revise laws to promote NAMs, remove animal testing mandates, and harmonize across sectors (51). Prioritize and incentivize NAM use.
  2. Accelerate Validation & Regulatory Acceptance: Create efficient, harmonized validation pathways focusing on human relevance (52). Increase funding for validation (47).
  3. Secure Sustained Funding & Prioritization: Invest significantly in NAM R&D, validation, and infrastructure (41). Prioritize NAMs in grant evaluations (41).
  4. Enhance Education & Training: Develop comprehensive programs for scientists, regulators, and technicians in NAMs (41).
  5. Address animal studies bias: Address bias in grant reviews (41), increase involvement of NAM-researchers in decision making processes and committees, develop approaches to support the paradigm shift for fundamental researchers focused on animal experiments.
  6. Establish Clear Timelines, Metrics & Oversight: Roadmaps need ambitious targets, milestones, and transparent monitoring (41).
  7. Global Alignment and Harmonization: Coordinate NAM promotion internationally (OECD, ICATM) (51).

On the way towards phasing-out animal experiments, critical hurdles and major challenges need to be addressed. NAM validation is clearly a bottleneck with the urgent need for new validation paradigms focused on human relevance, not just correlation with flawed animal tests. Regulatory conservatism requires proactive leadership in agencies to champion NAMs and implement pilot programs (30). For complex toxicological endpoints such as DART (developmental and reproductive toxicity) targeted research is needed including sufficient funding (34).

Last but not least, a successful paradigm shift requires a mindset shift by all parties including overcoming ingrained biases and transforming institutional practices. 

Conclusion and Future Outlook

The global scientific community is decisively moving from animal experimentation to humane, human-relevant NAMs. This is driven by ethical imperatives, scientific advancements, and supportive policy frameworks in the EU and US. The question is no longer if, but how effectively and quickly this transition will be realized.

Animal-free science promises more reliable and predictive insights into human health, leading to faster development of safer medicines and products. Increased efficiency and reduced failure rates offer economic advantages. This shift aligns science with ethics, fostering public trust.

Sustained effort is crucial. NGOs, policymakers, scientists, industry, regulators, and the public must work collaboratively while avoiding animal studies bias. With shared purpose and commitment to scientific excellence and ethical principles, we can accelerate the transition to a future where science no longer relies on animal suffering. The future of science is innovative, human-relevant, and must be animal-free.

23/06/2025
Dr. Tamara Zietek

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