The preclinical evaluation of novel imaging agents, such as contrast agents for computed tomography (CT), still largely relies on rodent models or simplified systems, each carrying limitations with regard to ethical bur...The preclinical evaluation of novel imaging agents, such as contrast agents for computed tomography (CT), still largely relies on rodent models or simplified systems, each carrying limitations with regard to ethical burden, physiological relevance or translational value. To address this gap, we propose larvae as a scalable, ethically favourable and physiologically relevant model for use in the early-stage screening of novel CT contrast agents. We established a standardised imaging protocol utilising isoflurane-based sedation and longitudinal micro-CT scanning, and used it to evaluate three commercially available contrast agents: one clinically approved molecular agent and two nanoparticulate formulations that are used for preclinical studies. All agents were systemically distributed, well-tolerated and enabled consistent visualisation of internal anatomy over eight days. While differences in clearance kinetics and route related to particle size or formulation were less pronounced compared to vertebrate models, this larval system reliably reflected biodistribution trends and allowed non-invasive tracking of agent functionality and safety. The use of in this context enables early elimination of less promising candidates and reduces reliance on vertebrates in downstream (pre)clinical development, aligning with the Three Rs principles. Our findings support the use of as a practical intermediate model, bridging cell-based assays and rodent studies in the development of next-generation CT and imaging contrast agents.
Risk decision-making has evolved from a vernacular focused on risk assessment and management to fully integrated approaches designed to inform risk acumen. Consequently, there has been a renewed interest, particularly in...Risk decision-making has evolved from a vernacular focused on risk assessment and management to fully integrated approaches designed to inform risk acumen. Consequently, there has been a renewed interest, particularly in areas with significant paradigm shifts, to understand the complex nature of the underlying intersections between data, ethics and risk. For example, building more awareness of the risk and ethical implications for applying artificial intelligence for generative (content creation) and agentic (decision-making) purposes or relying on next generation risk assessments grounded in models reflective of the Three Rs principles (i.e. the , and of animal studies). Global thinkers in risk science and analysis have also developed frameworks and models, such as the Projector Model, to show the complex nature of these intersections relevant to public health and regulatory risk decision-making. This article builds on this work by sharing real-life examples from an expert panel discussion, which occurred during the meeting. These panel members relied on the Projector Model to navigate the discussion in a session titled . The examples showed how institutional values and norms serve as foundational elements for risk decision-making, and highlighted that data-driven science, especially that based on novel approaches and technologies, needs careful consideration from an ethical and risk perspective.
The UK's 2025 strategy introduces a commitment to end polyclonal antibody reagent production by 2030. While this represents a significant milestone, focusing exclusively on polyclonal reagents raises scientific, ethical...The UK's 2025 strategy introduces a commitment to end polyclonal antibody reagent production by 2030. While this represents a significant milestone, focusing exclusively on polyclonal reagents raises scientific, ethical and implementation challenges, given that monoclonal and other recombinant antibody formats frequently remain dependent on animal immunisation. This study examines whether current UK licensing practice and reporting frameworks are positioned to deliver genuine . A structured, Three Rs-aligned framework is articulated, reflecting the principles set out in the UK Animals in Science Committee (ASC) 2022 review. This framework is applied to UK Non-Technical Summaries (NTS), to assess how antibody reagent production is justified at the project licence stage and the extent to which applications align with ASC expectations. In parallel, UK Home Office Annual Statistics of Scientific Procedures on Living Animals are analysed, to evaluate how antibody-related procedures are categorised, and to determine whether this national reporting enables progress toward the 2030 commitment to be monitored. The analysis identifies limited implementation of ASC guidance in post-2022 licences, with few applications providing evidence-based interrogation of non-animal alternatives, documented use of validated discovery platforms, or robust justification for immunisation. The national statistics lack sufficient resolution to clearly capture immunisation-dependent antibody reagent production across commercial, basic and translational research contexts. In addition, inconsistent terminology complicates the interpretation of genuine . Importantly, the findings indicate that, without complementary regulatory, reporting and infrastructural measures, the 2030 commitment risks being met through increased reliance on other immunisation-dependent antibody formats, geographical outsourcing, or reduced visibility of antibody production that is embedded within broader research categories, rather than through genuine . Achieving the intended reduction in animal use will therefore require clearer terminology, improved reporting resolution, wider access to non-animal discovery infrastructure and the integration of a Three Rs-aligned decision framework into licensing expectations.
One Health initiatives are modern paradigms for research and health care practices in various fields. Concrete definitions of the One Health framework, however, remain heterogeneous, leading to conceptual problems and un...One Health initiatives are modern paradigms for research and health care practices in various fields. Concrete definitions of the One Health framework, however, remain heterogeneous, leading to conceptual problems and uncertainties in the application of the framework. This article discusses several approaches to the One Health concept, and their associated consequences, with special focus on animal experimentation. The first issue addressed is how One Health should be defined, as well as what (and who) should be considered within a One Health approach. In order to shed further light on this, we explore the history of animals in biomedical science, highlighting historical milestones in the use of animal models, as well as the development and current state of ethical considerations in the field of animal experimentation. The second issue comes with the inclusion of animal experimentation as part of the One Health concept. Therefore, particular attention is paid to bioethical principles and the resulting problems that can arise when applying them to the One Health concept. Arguments such as the idea of inequality between humans and non-human animals, and the premise that all actions are done for the benefit of humans, are raised and then used to explore the question of whether the One Health concept is compatible with existing bioethical principles. Based on the bioethical principles of protecting the environment, the biodiversity and biosphere, this paper seeks an inclusive perspective of the One Health concept. Successful solutions will be based on this concept, which embraces all living beings. The authors conclude that a multispecies ethics approach could help create a more ethical ecosystem that is aligned with the wellbeing of all life on a shared planet.
This paper critically examines the legal coherence of the EU's chemical safety regulations, REACH and CLP, in light of the EU's commitment to the phasing out of animal testing. While both instruments express normative su...This paper critically examines the legal coherence of the EU's chemical safety regulations, REACH and CLP, in light of the EU's commitment to the phasing out of animal testing. While both instruments express normative support for non-animal approaches, their operational provisions remain structurally biased toward animal-based evidence. This bias persists despite scientific concern about the predictive value of animal data, and the demonstrable inefficiency of animal testing for chemical safety assessment, as well as the growing availability of scientifically sound non-animal methodologies. Alongside cultural and institutional barriers, such as regulatory conservatism and a rigid validation system, legal obstacles embedded in the legislative design of REACH and CLP also impede a paradigm shift away from animal testing. Applying the principle of proportionality, the article argues that the animal-centric operational provisions of these regulations are: (i) neither suitable nor necessary to ensure a high level of human health protection; and (ii) that they disproportionately encroach upon animal welfare, in relation to their benefits. The article therefore calls for a legislative recalibration, to realign REACH and CLP with primary Union law. Without such changes, the EU Roadmap toward the phasing out of animal testing in chemical safety assessment, risks having limited practical effect.
Gabapentin is a structural analogue of gamma-aminobutyric acid. Initially introduced as an anti-epileptic drug, it has also been shown to be effective in the treatment of pain in humans and non-human animals. The main pu...Gabapentin is a structural analogue of gamma-aminobutyric acid. Initially introduced as an anti-epileptic drug, it has also been shown to be effective in the treatment of pain in humans and non-human animals. The main purpose of this study was to test the suitability of the cockroach (Blattodea: Blaberidae) as a non-vertebrate model for evaluation of the antinociceptive properties of gabapentin. After the intersegmental abdominal membrane administration of different doses of gabapentin (50, 150 and 250 mg/kg), two acute thermal pain tests were performed: the hot plate test and the hot box escape test. In the hot plate test, 50 mg/kg gabapentin was the most potent dose in terms of reducing thermal stimulus-associated movement of the cockroaches; in the hot box escape test, 150 mg/kg gabapentin was the most efficacious dose. Thus, when comparing the two different tests, gabapentin exerted effective antinociception effects in cockroaches at different doses. This is similar to the effects observed in mice and rats exposed to thermal stimuli. Our results therefore demonstrate that could be a suitable non-vertebrate model for the study of pain, and suggest that it could be effectively used for the evaluation and screening of other analgesic compounds.
The acquisition of surgical skills is an essential component of veterinary training. The use of live animals or cadavers for early surgical skills development is limited by ethical, logistical and financial constraints,...The acquisition of surgical skills is an essential component of veterinary training. The use of live animals or cadavers for early surgical skills development is limited by ethical, logistical and financial constraints, highlighting the need for accessible and animal welfare-oriented training alternatives. The objective of this study was to evaluate the effectiveness of a low-cost, balloon-based simulator for teaching basic surgical knot techniques to undergraduate veterinary students with no prior surgical experience. A cohort of 20 students practised slip knots, Miller's knots and transfixation knots, using a low-fidelity balloon simulator under a structured training protocol. Performance was assessed by using standardised rubrics, i.e. execution time and a binary success/failure classification based on air leakage. From the 13th repetition onward, all students successfully executed slip knots and Miller's knots, while competency in the transfixation knot was achieved by the 14th repetition. Execution time decreased significantly from the 11th repetition for all three techniques, indicating progressive improvement in technical efficiency. It was evident that the balloon-based simulator facilitated the acquisition of basic surgical knot-tying skills, and represents an accessible, ethical and effective tool for early-stage surgical training in veterinary education.
The Human Organotypic Skin Explant Culture (hOSEC) model utilises skin fragments, mainly obtained from plastic surgery procedures, as the primary source of material. The model is used for testing that requires human tis...The Human Organotypic Skin Explant Culture (hOSEC) model utilises skin fragments, mainly obtained from plastic surgery procedures, as the primary source of material. The model is used for testing that requires human tissue, and as the basis for various research models. The objectives of the current study were to review, analyse and summarise the published literature on the use of human skin explants in experimental studies in Brazil. The literature search was conducted within three databases, using terms related to explants and study location combined with defined eligibility criteria. Sixteen studies were considered eligible for further detailed analysis in the review. The geographical location of these studies was concentrated in the South-Eastern region of Brazil, and demonstrated a prevalence of explants obtained during plastic surgery procedures. Five main focus areas were identified in the studies, namely: disease pathology, cellular ageing, pharmacological testing of cosmetic ingredients, wound healing, and cell culture optimisation - with pathology-focused models being the most common. Despite the search strategy being capable of identifying the diverse characteristics of the studies, the research protocols used in the studies were heterogeneous. This is an intrinsic limitation of the published literature in general, which prevents direct comparisons and hinders reproducibility. Human skin explants were shown to represent versatile tools, with potential for expansion into other areas. The creation and dissemination of standardised methodologies and guidelines for human skin explant-based research are essential. This information, as well as the promotion of the use of such models, will contribute to a reduction in the use of animals in experiments.
has been used as a model organism in biomedical research for over a century, facilitating a number of fundamental breakthroughs and Nobel Prize-winning discoveries in genetics, developmental biology and disease mechanism...has been used as a model organism in biomedical research for over a century, facilitating a number of fundamental breakthroughs and Nobel Prize-winning discoveries in genetics, developmental biology and disease mechanisms. This bibliometric analysis assessed 140,962 -related publications indexed in the Web of Science Core Collection from 1984 to 2024. Publication output increased rapidly until the late 1990s, with an average annual growth rate of 13%, before stabilising. Co-authorship analysis revealed extensive international collaborative networks spanning multiple continents, while keyword co-occurrence analysis identified seven major research clusters: developmental biology, cell biology, circadian biology, ageing, evolutionary biology, molecular biology and immunology. Temporal trend analyses demonstrated sustained growth in fields such as neuroscience, neurodegenerative diseases, ageing, circadian biology, immunology, pharmacology, toxicology and environmental sciences. These findings provide a comprehensive overview of contemporary -based research, highlighting its ongoing global relevance, evolving applications and emerging research frontiers. The unique advantages of for investigating complex biological processes establish it as a more ethical and efficient alternative to vertebrate models in modern biomedical science.
We established a 3D model of human NT2/D1-derived early neural progenitor cells immobilised in alginate microfibres as a system for testing the neurotoxicity of energy drinks and their components, either alone, together,...We established a 3D model of human NT2/D1-derived early neural progenitor cells immobilised in alginate microfibres as a system for testing the neurotoxicity of energy drinks and their components, either alone, together, or in combination with alcohol. The system supports the retinoic acid-induced neurogenesis of NT2/D1 cells, and the proliferative capacity of the NT2/D1-derived early neural progenitor cells was maintained in the 3D environment. Cell cycle distribution and the expression of pluripotency markers (, and ), early neural markers (, and ), and (a marker of neural commitment), showed profiles characteristic of early neural progenitors. Treatments with an energy drink and its major components (caffeine and taurine) - either alone, together, or in combination with alcohol - had different effects on the proliferative capacity of the NT2/D1-derived early neural progenitor cells in the 2D and 3D models. In the 2D-cultured cells, all treatments except for caffeine led to a significant decrease, while cells within the 3D model exhibited a significant increase after treatment with caffeine, or after combined treatment with energy drink and alcohol. Preliminary findings suggesting that there were no treatment effects on OCT4 and PAX6 protein expression in either model, should be further confirmed. This human cell-based 3D model could potentially represent a rapid and cost-effective system for assessing the acute and long-term neurotoxicity of various compounds.
The aim of this integrative review was to investigate the use of non-animal derived substrates and models as alternatives to bovine enamel in restorative dental materials research. It included studies on the use of vario...The aim of this integrative review was to investigate the use of non-animal derived substrates and models as alternatives to bovine enamel in restorative dental materials research. It included studies on the use of various synthetic alternatives to animal-derived enamel in such experimental research. Excluded from the review were: 1) reviews, clinical cases, letters, chapters, conference abstracts and editorials; and 2) studies assessing datasets, alternatives to other types of tissues such as dentin or bone, biomaterials in the context of enamel repair, and laboratory studies using substrates of animal and human origin without any comparison with enamel-like synthetic materials. It was demonstrated that calcium phosphate-based ceramics, multiscale materials, 3D-printed and hydroxyapatite-based materials have the capability to emulate the mechanical and chemical properties of dental enamel. However, these materials have inherent limitations, with some needing further investigation in order to understand their responses in certain laboratory tests. Even so, some materials appear promising and have already performed well with regard to the most relevant aspects that are commonly evaluated in testing. It is thus recommended that new studies prioritise the use of such synthetic substitutes as standard in laboratory protocols whenever possible, in order to gradually abandon the use of animal-derived tissues and transition to a more ethical science without contributing to animal exploitation by acquiring bovine teeth from abattoirs.