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Innovation examples
HealthToxicology
Zebrafish in toxicity testing
Zebrafish are increasingly recognised as a useful model for toxicity testing of chemical substances. Testing strategies are becoming more based on mechanisms of toxicity structured in adverse outcome pathways describing the chain of events leading to toxicity or disease. Using a battery of dedicated in vitro and in silico assays, insight can be gained in how exposure leads to disease. For certain diseases it is known that toxicity relies on the interaction between different organs and cell types, which requires research on whole organisms in addition to simple in vitro models. The zebrafish is considered a valuable whole organism model in a mechanism-based testing strategy. At RIVM, the zebrafish embryo model is used for testing the effect of chemical substances on several adverse outcomes and diseases.
For more information see: https://ehp.niehs.nih.gov/doi/10.1289/EHP9888; https://doi.org/10.3390/ijerph18136717; www.linkedin.com/in/harm-heusinkveld

Innovation examples
HealthToxicologyIn silico
AI agents for safer science: How AI is Changing Chemical Risk Assessment
This video introduces a novel approach to chemical safety, where intelligent digital agents guided by large language models support scientists in making faster, more transparent decisions. By automating complex workflows and integrating tools like the OECD QSAR Toolbox, these agentic systems help prioritise research, reduce reliance on animal testing, and pave the way for safer, more sustainable innovation.

Innovation examples
HealthToxicologyIn vitro
Thyroid Hormone & Brain Development: animal-free models for human safety assessment
The environment can have a significant impact on a child's health even before birth. Brain development begins in the first trimester and continues until the age of 25, with thyroid hormone playing a critical role. During early pregnancy, the fetus depends on the mother's thyroid hormone, and a disruption in the thyroid hormone balance can lead to cognitive and motor impairments in the child. As part of the VHP4Safety project, we are developing in vitro tests to measure the developmental neurotoxic effects caused by disturbances thyroid hormone concentrations. Current testing guidelines do not always include testing for neurodevelopmental effects, highlighting the need for new non-animal methods. At the Erasmus Medical Center, human cell lines representing brain cell types are cultured to study the effect of chemicals on the thyroid hormone balance. RIVM uses human stem cells to create neuron-astrocyte networks that mimic brain development. By combining these different assays and models, we are creating a comprehensive human-based testing strategy to assess developmental neurotoxicity. These advances are a critical step toward eliminating animal testing while protecting the health and environment of future generations.

Expert interviews
Toxicology
How do we use human data in risk assessment
In this video, EFSA explains how they do risk assessment and what the role of NAMs can be in this process.

Expert interviews
Policy
Charlotte Blattner, Harvard Law School: Transition needs community efforts
Charlotte Blattner is a visiting researcher at Harvard Law School on the Animal Law & Policy Program and explains that a transition is needed to move away from animal testing. This transition needs to be a just transition, a community effort where all stakeholders are involved to replace animal testing for animal-free innovations.

Projects and initiatives
In vitroOrgan-on-Chip
SMART OoC platform: a standardized modular approach
The SMART Organ-on-Chip project aims to bring Organ-on-Chip technology to the next level, out of the pioneering labs to industrial applications. NWO awarded 4.8 million euro to a large and diverse consortium of universities, companies, research institutes and foundations, brought together by hDMT (Dutch Organ-on-Chip Consortium), that will together develop standardized Organ-on-Chip models. These models will be made to fit the scale and quality that pharmaceutical companies need to use them for development of novel drugs, with better science and less animal use as a result. The project will kick off in autumn 2021. More information on the project will follow in the course of 2021.

Innovation examples
ToxicologyIn vitroOrgan-on-Chip
Cartilage-on-a-chip for studying joint degenerative diseases
Carlo Alberto Paggi is currently a PhD candidate at the University of Twente in the research group of Prof. Marcel Karperien and Prof. Séverine Le Gac. Karperien’s lab focus on the biological aspects of osteoarthritic research while Le Gac’s specialize in organ-on-chip development. The project of Carlo Alberto is developing a joint-on-chip platform to create a reliable in vitro model to study disease progression in osteo- or rheumatoid arthritis. The model combines different organ-on-chips aimed at replicating each a tissue around the joint such as cartilage, bone and ligaments. This new technology focuses on better reproducing human models and at substituting the use of animal models for drug research. If you want to know something more about the project and the groups, you can follow the link in the video.
Carlo Paggi was nominated for the Hugo van Poelgeest prize for his research on a cartilage-on-a-chip model to study joint degenerative diseases
Karperien’s lab of Developmental Bioengineering: https://www.utwente.nl/en/tnw/dbe/
Le Gac’s lab of Applied Microfluidics for BioEngineering Research: http://www.severinelegac.com/
Linkedin: https://www.linkedin.com/in/carlo-alberto-paggi-76500b135/

Innovation examples
EducationInnovation
Avatar Zoo - teaching animal anatomy using virtual reality
Animals are essential to train the next generation of scientists understand diseases and develop treatments for humans as well as animals. Therefore, animals are used for educational purposes. Technologies such as Virtual Reality and Augmented Reality can be employed to reduce the number of animals in the future. Prof. Dr. Daniela Salvatori is working on the development of 'Avatar Zoo' together with UMCU and IT. Live animals are replaced by holographic 3D in this flexible platform. With these holograms one is able to study the anatomical, physiological and pathological systems and processes of all kinds of animals.
Avatar Zoo won the Venture Challenge 2021 for the development of virtual reality models that can be used for anatomy classes and practical training.

Meetings & conferences
HealthIn vitroAdvanced
In vitro predictive models of particle-induced granulomas
Léa Hiéronimus is a PhD student at the Louvain centre for Toxicology and Applied Pharmacology (LTAP, UCLouvain, Belgium). Léa is working in François Huaux's team, where we are trying to better understand how certain inhaled particles exert their toxicity. The goal is to better diagnose and treat individuals exposed to particles, but also to identify the particle characteristics which induce, or do not induce, toxic effects. For this, Léa studies a very particular cell type which seems to be involved in particle responses. Indeed, we have found the specific accumulation of the innate subset of B-lymphocytes called “B-1 lymphocytes”, which occurred during granuloma formation/maturation induced by inhaled particles in mice. According to the literature, this accumulation can be attributed to their migration from mesothelial cavities such as the peritoneum, acting as a reservoir.
In addition to conventional particles-induced granulomas, which formation rely on macrophages responses, we developed new models relying on B-1 lymphocytes. Indeed, B-1 lymphocytes show a unique clustering property, that is not observed using macrophages or other subsets of B-lymphocytes (conventional B-2 lymphocytes) as purified B-1 lymphocytes regroup granuloma-inducing particles (carbon nanotubes CNT7, crocidolite asbestos, micrometric silica MinUSil and MSS, cobalt oxide,…) but not carbon black, a particle not-inducing granuloma in vivo. Additionally, we developed a model aiming to recapitulate the lung after B-1 lymphocytes migration and found that macrophages and epithelial cells (MHS and LA4 cell lines) where grouped to form spheroids when in coculture with B-1 and not B-2 lymphocytes.
These models will serve as tools to identify new mediators of granuloma formation, which could serve as biomarkers and/or therapeutic targets for exposed individuals. On the other hand, we aim to propose new bioassays for the prediction of granuloma-inducing materials using alternative models.
Lab website: https://uclouvain.be/en/research-institutes/irec/ltap
Contact: lea.hieronimus@uclouvain.be

Projects and initiatives
The Beyond Animal Testing Index
The Beyond Animal Testing Index (BATI) was designed after the Access to Medicine Index with the aim to be a transparent, objective and independent benchmark that provide public research organisations and their stakeholders insight in what efforts and contributions they make in the transition to animal free innovation and to provide organisations incentive to learn from and inspire each other with regard to the implementation of research practices without the use of animals for the benefit of science.

Questions
HealthInnovationIn vitro
Helpathon #8 – Can you help Germaine?
Germaine Aalderink is investigating the uptake of lipids travelling from the gut into the lymphatic system and further explore the merits of this alternative drug intake strategy. Can you help Germaine make an intestinal and lymphatic model with an alternative for Matrigel that is animal-free? She wants to know what components are essential in each phase of intestinal development and is interested in both the positive and negative experiences of other researchers with the use of alternatives for Matrigel.
Click on the link in the video to sign up and read more information on this Helpathon on the website (https://www.helpathonhotel.org/coming-up).

Innovation examples
In vitroOrgan-on-Chip
Unified organoid system for modeling heart and kidney interaction on-a-chip
Beatrice Gabbin is a PhD candidate at the Anatomy and Embryology Department of the Leiden University Medical Center. Her project is shared with the Nephrology Department and focusses on the study of the cardiorenal axis in vitro. Both heart and kidneys have vital functions in the human body and reciprocally influence each other’s behavior: pathological changes in one can damage the other. There are already multiple independent in vitro (human) models of heart and kidney, but none have so far captured their dynamic crosstalk. The aim of the project is therefore to develop a microfluidic system which can be used to study heart and kidney interaction in vitro. For this purpose, cardiac microtissues and kidney organoids derived from human induced pluripotent stem cells are generated and loaded onto a 3D perfusion chip for their dynamic co-culture. This system enables the study the cardiac and kidney interaction with a high level of control. The validation of a unified organoid system will enable the investigation of diseases involving the two organs and their potential treatments. Read more via the link in the video and https://doi.org/10.1016/j.mtbio.2023.100818.