Traditionally, animal testing has been considered the 'gold standard' in determining potential effects of chemicals on humans. However, this dogma is increasingly being questioned, not only due to ethical and financial i...Traditionally, animal testing has been considered the 'gold standard' in determining potential effects of chemicals on humans. However, this dogma is increasingly being questioned, not only due to ethical and financial implications, but also because of the poor translatability of the results of animal tests to humans. Therefore, there is a need to modernise the concept of the gold standard and ensure that any new approach is flexible, adaptable and as future-proof as possible. Herein, we reflect on recent suggestions for updating, or redefining, the traditional gold standard of animal testing and propose a new definition. This proposal focuses more on the process of answering a specific question, using all available tools, rather than seeking to recapitulate an animal test. New Approach Methodologies (NAMs) provide an ever-expanding array of tools that can assist product development and play a key role in chemical assessment strategies. Ten recommendations for developing fit-for-purpose NAMs, to increase their acceptance and accelerate their adoption, are presented.
Huang H, Sanders JN, Warren GL
… +16 more, Patron A, Ninobla A, Inoue K, Valk H, Sadler M, Amorelli B, Hanson M, Acquaviva E, Lear E, Creelman M, Nabua C, Pilat E, Latifzadeh D, Price G, Raabe H, Costin GE
Toxicological profiling is a critical component of the regulatory process required to inform the hazard characterisation and labelling of chemicals, thus guiding their safe handling and use. Skin irritation is a key endp...Toxicological profiling is a critical component of the regulatory process required to inform the hazard characterisation and labelling of chemicals, thus guiding their safe handling and use. Skin irritation is a key endpoint for regulatory registrations, traditionally assessed by using the Draize rabbit test, which was introduced in 1944. Advances in New Approach Methodologies (NAMs), and particularly those based on reconstructed skin models, have led to more human-relevant alternatives, as compared to the test method. Here, we report on some groundbreaking high-throughput adaptations to the validated Skin Irritation Test (SIT; OECD TG 439) with the commercially available EpiDerm™ reconstructed human epidermis (RhE) model from MatTek Corporation. These minor adaptations enabled the safety evaluation of over 3000 chemicals in just ten months. This was necessary due to the large number of chemicals that needed to be pre-screened for dermal safety, after the insect repellent or insecticidal activity was predicted and assessed. The data obtained in the SIT were then used to train a new Artificial Intelligence (AI) tool, designed to predict the skin irritation potential of large libraries of molecules within (but not limited to) the chemical space which the AI tool is based upon. Our work exemplifies the integration of AI with existing non-animal-based approaches, in order to further develop chemical safety assessment by taking advantage of this new technology.
One Welfare is an emerging concept that seeks to integrate the welfare of animals, humans and the environment. This systematic review examines the role of laboratory animal veterinarians in promoting One Welfare within t...One Welfare is an emerging concept that seeks to integrate the welfare of animals, humans and the environment. This systematic review examines the role of laboratory animal veterinarians in promoting One Welfare within the context of animal use for scientific research. By analysing 65 publications, we explored the diverse roles of laboratory animal veterinarians in advancing animal welfare, supporting human health and promoting environmental sustainability. The reviewed literature described veterinarians' involvement in areas such as animal welfare oversight, implementation of the Three Rs principles, regulatory compliance and ethical review processes. Several themes emerged that are related to the One Welfare framework, namely: veterinary oversight, welfare assessment and implementation, emotional wellbeing, professional support, and contribution to education and training. The literature highlights the potential need for specialised training in laboratory animal medicine, and outlines the challenges veterinarians encounter regarding ethical considerations and cognitive dissonance in research environments. Based on the identified gaps, we propose measures to reinforce the principles of One Welfare, including interdisciplinary collaboration and the adoption of strategies to safeguard the environment (e.g. the use of composting for minimally-infectious waste and digesters for high-impact waste). Additionally, we recommend establishing institutional sustainability committees to oversee governance and resource management across research institutions.
Recent global initiatives are accelerating the shift toward human-centric approaches, reducing reliance on animal models in preclinical research and other domains. In this changing landscape, objectively evaluating the s...Recent global initiatives are accelerating the shift toward human-centric approaches, reducing reliance on animal models in preclinical research and other domains. In this changing landscape, objectively evaluating the scientific relevance and merit of research involving animal models, and assessing their translational relevance is increasingly critical. Over the past decade, several tools have been developed to assess translational relevance, accuracy/appropriateness and efficacy of preclinical animal models, evaluate risk-of-bias in preclinical research, support harm-benefit analyses, and facilitate the adoption of non-animal replacement strategies. However, the uptake of such tools remains limited. To address this, a Biomedical Research for the 21st Century (BioMed21) Collaboration workshop on 'Evaluating translational value of animal models in preclinical research - Tools, challenges, and strategies', was convened by Humane World for Animals (30 June-1 July 2025). The event brought together tool developers and diverse global interest-holders to review current assessment tools, discuss their strengths, complementarity, limitations and feasibility, and explore opportunities for cross-sector collaboration. This paper summarises key outcomes of these presentations and discussions, highlighting knowledge gaps and barriers to the adoption of these tools and frameworks by researchers, funders and regulators. Strategies to raise awareness and promote the use of the tools and frameworks, to better inform funding decisions, regulatory approval and the appraisal of preclinical research, are also proposed.
Assessment of the Culture of Care (CoC) within an institution is essential in understanding the gaps in current CoC practices and highlighting what needs to be further improved. As the first responders in laboratory anim...Assessment of the Culture of Care (CoC) within an institution is essential in understanding the gaps in current CoC practices and highlighting what needs to be further improved. As the first responders in laboratory animal facilities, laboratory animal care staff (i.e. laboratory animal technicians and laboratory animal attendants) play a key role in establishing and sustaining a CoC. We employed the previously introduced Capability Maturity Model (CMM) to assess the CoC among Sri Lankan laboratory animal facility staff. A questionnaire-based survey was distributed, in order to assess the current state of a particular facility's CoC, based on the CMM. The survey also included questions to help identify the challenges faced by staff and evaluate the impact of Laboratory Animal Science education and training on improving the CoC. To identify further issues, a roundtable discussion involving a number of laboratory animal care staff was also conducted. This study revealed that most Sri Lankan laboratory animal facilities are currently at Level 1 of the CMM, with some showing early signs of transitioning to Level 2. Lack of management commitment and insufficiency of funds were identified as the key challenges. Lack of knowledge and training, and inadequate understanding and co-operation between co-workers and also with researchers, were stated as additional issues during the roundtable discussion. Thus, we urge training providers, such as the Sri Lanka Association for Laboratory Animal Science (SLALAS), to conduct more targeted training programmes, focusing on good CoC practices, laboratory animal welfare standards and ethical decision-making.
Minimally invasive catheter-based techniques for cardiac diseases, such as patent ductus arteriosus occlusion, pulmonic stenosis balloon valvuloplasty, atrial septal defect occlusion and transseptal approaches, have gain...Minimally invasive catheter-based techniques for cardiac diseases, such as patent ductus arteriosus occlusion, pulmonic stenosis balloon valvuloplasty, atrial septal defect occlusion and transseptal approaches, have gained popularity in veterinary cardiology owing to their high success and low complication rates. However, the technical complexity of these procedures and the lack of accessible training opportunities have limited their widespread adoption in clinical practice. In this study, we evaluated the effectiveness of a 3D-printed canine cardiac model as a simulator-based training tool for improving proficiency in these interventional procedures. The 3D model was developed by using computed tomography (CT) data of a dog diagnosed with type IIa patent ductus arteriosus. A virtual prototype of the model was created by segmenting the CT data. A hard-type photopolymer resin was used for 3D printing the model to ensure durability. To enhance the structural stability and applicability of the model, iterative optimisation was carried out by incorporating feedback from testers who used it for the different procedural simulations. The model accurately replicated cardiovascular structures and artificial defects, such as atrial and ventricular septal defects. Patent ductus arteriosus occlusion, pulmonic stenosis balloon valvuloplasty, atrial septal defect occlusion and transseptal approaches were performed, in order to evaluate the compatibility of the model with fluoroscopy. To assess the effectiveness of the model as a training tool, time-trial assessments and pre- and post-training evaluations involving six participants were conducted. The 3D-printed canine cardiac model improved procedural efficiency and understanding of cardiac anatomy. The 3D-printed model of canine cardiac diseases introduced here is scalable and valuable as a teaching tool in veterinary interventional cardiology.
Real progress has been made towards replacing animals for risk assessment purposes, particularly given the recent announcement of comprehensive roadmaps by regulatory agencies. However, given the vast number of animals t...Real progress has been made towards replacing animals for risk assessment purposes, particularly given the recent announcement of comprehensive roadmaps by regulatory agencies. However, given the vast number of animals that continue to be used for biomedical purposes, perhaps now is the time to place greater emphasis on replacing animals in biomedical research. Scientific journals have a key role to play in encouraging the uptake of new approach methodologies and providing a supportive environment for a cultural shift to humane, sustainable Three Rs-based science.
Recent advances in neural differentiation have unveiled new possibilities that could potentially be applied to the development of human-relevant non-animal models, for use in fields such as biomedical research and drug s...Recent advances in neural differentiation have unveiled new possibilities that could potentially be applied to the development of human-relevant non-animal models, for use in fields such as biomedical research and drug screening. Thus, the directed differentiation of tissue stem cells toward neural progenitor cells or neural stem cells, by small molecules and growth factors without the need for genetic manipulation, has attracted great attention. The generation of neural progenitor cells, and their proliferation and lineage commitment are regulated by signaling pathways activated by small molecules and growth factor families, including various fibroblast growth factors (FGFs). FGF-9 regulates the differentiation of neural stem cells, not only during embryonic development, but also contributes to adult neurogenesis and the protection of degenerating neurons. Here, we investigate an improved protocol for neural stem cell proliferation, differentiation and lineage commitment of adipose tissue-derived mesenchymal stem cells (AD-MSCs). We evaluated a cocktail of valproic acid (VPA), CHIR99021 and FGF-9, both alone and in combination, for the potential to induce cell differentiation. The AD-MSCs were isolated from human omentum fat and characterised immunologically by the presence of specific mesenchymal markers and multi-lineage differentiation potential. To assess the potential toxic effect of each cocktail, cell proliferation and viability were determined, followed by confirmation of neural differentiation via quantitative reverse transcriptase real-time PCR (qRT-PCR) and immunocytochemistry. The combined administration of VPA and FGF-9 promoted neurogenesis and neuronal fate commitment of AD-MSCs cultures, leading to significantly increased expression of the neural stem cell markers Nestin and Sox-2, as well as the neuronal cell marker MAP-2. This study contributes to our understanding of the role of FGF-9 in neural differentiation from human AD-MSCs. Our simplified protocol uses VPA and FGF-9 to efficiently generate neural progenitor cells, avoiding complex and prolonged induction methods, making it ideal for use in, for example, human-relevant drug screening for neurotoxicity.
Antibodies are indispensable tools in biomedical research, yet the widespread use of animal-derived antibodies contributes to poor reproducibility and raises serious ethical concerns. Many commercial antibodies fail to b...Antibodies are indispensable tools in biomedical research, yet the widespread use of animal-derived antibodies contributes to poor reproducibility and raises serious ethical concerns. Many commercial antibodies fail to bind their intended targets, leading to irreproducible results, wasted resources and stalled progress. In Europe alone, over a million animals are used annually for antibody production. Non-animal-derived recombinant antibodies and antibody mimetics offer a scalable, scientifically valid alternative. Endorsed by the European Union Reference Laboratory for alternatives to animal testing (EURL ECVAM), these reagents deliver greater consistency, transparency and ethical alignment. Despite their advantages, they are currently estimated to make up less than 5% of research reagents, held back by limited awareness, access barriers and persistent misconceptions about performance. To address these challenges, the Centre for Human Specific Research developed the open-access Recombinant Antibodies & Mimetics Database. This platform links researchers to ethically sourced, sequence-defined affinity reagents and features a unique colour-coded classification system to clearly indicate the extent of animal involvement. By improving transparency, the database empowers informed decision-making and supports alignment with best practices. More than a sourcing tool, the platform drives advocacy for systemic change, encouraging funders, institutions, publishers and regulators to prioritise recombinant antibodies and antibody mimetics. Its ultimate aim is to make high-quality animal-free recombinant and mimetic antibodies the global standard, supporting a more reproducible, ethical and forward-looking future for antibody science.
Angiogenesis is an essential part of bone regeneration, as neovascularisation supports the supply of necessary oxygen, nutrients, and cellular transport. Bioactive glasses (BGs) have shown promise in enhancing both angio...Angiogenesis is an essential part of bone regeneration, as neovascularisation supports the supply of necessary oxygen, nutrients, and cellular transport. Bioactive glasses (BGs) have shown promise in enhancing both angiogenesis and bone regeneration. For the evaluation of the angiogenic potential of BGs, the chorioallantoic membrane (CAM) assay constitutes an attractive experimental model and has already gained increasing attention in BG-focused research. However, there is conflicting evidence as to whether the addition of cells, such as bone-marrow-derived mesenchymal stromal cells (BMSCs) to the CAM is necessary to facilitate the evaluation of the angiogenic potency of BGs. Therefore, in this study, the angiogenic potential of mesoporous bioactive glass nanoparticles (MBGNs; molar composition = 70% silica (SiO) and 30% calcium oxide (CaO)) was assessed by using the CAM assay, both in the presence and the absence of exogenous BMSCs. Compared to the BMSC-free and MBGN-free control groups, both BMSCs alone and MBGNs without the addition of BMSCs were able to induce an equally strong, significantly enhanced angiogenic response on the CAM. The combination of MBGNs with BMSCs did not yield a more pronounced angiogenic response, as compared to MBGNs without the addition of cells. Thus, MBGNs exhibit a strong intrinsic angiogenic potential that is not dependent on the presence of exogenous BMSCs. Therefore, the CAM assay without added cells can be used as a simplified, reproducible and cost-effective method for accurate preclinical testing of the angiogenic potential of BGs.
Assays to predict the skin sensitisation potential of substances commonly address the molecular initiating event (MIE) of the adverse outcome pathway (AOP), which is the covalent binding of chemicals to proteins. This re...Assays to predict the skin sensitisation potential of substances commonly address the molecular initiating event (MIE) of the adverse outcome pathway (AOP), which is the covalent binding of chemicals to proteins. This represents the first key event in the pathway. approaches have emerged as validated alternatives to conventional animal testing methods to determine this process. This review explores the integration of biosensor-based approaches to supplement other methods, for use as alternatives to the 'gold standard' Local Lymph Node Assay (LLNA) in skin sensitisation testing. While surface plasmon resonance-based biosensors have shown promise, they exhibit limitations such as poor sensitivity. To overcome such limitations, this review highlights the use of impedance-based biosensors in the study of hapten-skin protein interactions, which are associated with the MIE in skin sensitisation. Impedance-based biosensors have exhibited impressive performance, with a specificity of 97%, a sensitivity of 83% and an accuracy of 92% being obtained in one study ( = 49 substances). The advantages of impedance-based biosensors include label-free methodologies, high sensitivity, low cost and operational simplicity. This review also covers recent advancements in the use of biosensors in cosmetic studies and skin research, as well as comparisons of the limitations of the various methods and future perspectives for skin sensitisation assessment. The use of impedance-based biosensors as part of integrated testing approaches alongside other testing methods, can represent a reliable approach for skin sensitisation assessment, while supporting the reduction and replacement of animal use in toxicity testing.