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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

TPI.tv videos
InnovationPolicyBeginner
TPI.tv: improving science through animal-free innovations and research
Introducing TPI.tv : a video platform by experts striving to improve science through animal-free innovations and research.

Expert interviews
HealthEducation
Daniela Salvatori, TPI Utrecht: We aim for better science with less animals
Prof. dr. Daniela Salvatori, chair of TPI Utrecht, presents the aims of the local TPI group and invites all who want to share their ideas or questions on the transition towards animal-free innovations to get in touch via uu.nl/tpi.

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.

Projects and initiatives
HealthIn vitroOrgan-on-Chip
SCREENED: developing 3D thyroid models
The European collaborative project SCREENED aims to develop three-dimensional (3D) cell-based in vitro tests to better characterize the effects of endocrine disruptors (EDs) on thyroid gland function. This method will overcome the limitations of existing tests, being more sensitive at low doses of exposure to chemicals, and enabling the prediction of their toxicity on human health in a sex-specific manner. The ambition of the SCREENED project is that these new 3D in vitro tests, as well as the increased knowledge about adverse reactions after exposure to EDs, will be used for regulatory purposes, ultimately to improve human health.

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.

TPI.tv videos
Beginner
Pro tips for making a video about your research
Need some pro-tips to make your next video on animal-free innovations? Aniek and Victoria got some for you! In this video, they share why you should want to make a video about your research, tips about the content and format of an attractive video, and how to best share your video.

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.

Meetings & conferences
HealthIn vitroIn silicoAdvanced
A hybrid in silico-in vitro cardiorespiratory simulator for medical device testing
Cardiovascular medical devices (CMDs) (e.g. artificial hearts, ventricular assist devices, ECMO, heart valves) support the cardiac and/or the respiratory function of patients. Large challenges are encountered when assessing CMDs interaction with the human body and the effects on the heart and vessels. Especially CMDs with new designs require an extensive evaluation concerning their effectiveness and safety under different pathophysiological conditions. We propose a high fidelity cardiorespiratory simulator for the testing of the hemodynamic performance of CMDs. The proposed simulator merges the flexibility of the in silico system with a hydraulic interface to test CMDs. As such, the simulator embeds a high fidelity cardiorespiratory model, allowing the reproduction of pathologies at both cardiac and respiratory level. The simulator works as a test bench for the assessment of CMDs, from prototype stage to pre-clinical stage. Thanks to its flexibility and high-fidelity, the simulator helps reducing animal testing and provides insights on how to improve CMD design to better suit different patient’s needs.
Contact: https://www.kuleuven.be/wieiswie/en/person/00098489
RE-place database: https://www.re-place.be/method/cardiovascular-modelling-medical-device-testing

Meetings & conferences
HealthIn vitroAdvanced
Liquid marbles for cardiac organoids development
Advances in three-dimensional (3D) culture techniques have shown several advantages over 2D cultures, especially by more accurately mimicking the in vivo environment. This has led to improved reproducibility and reliability of experimental results, which are important criteria in disease modelling and toxicity testing. Induced pluripotent stem cells (iPSC) provide an unlimited source for the derivation of all cell types of the adult body, including cardiomyocytes. To improve the current culture methods for multicellular cardiac spheroids, such as the hanging drop method, we explored the use of hydrophobic powders. Fumed silica nanoparticles can be used to encapsulate liquid drops, which could serve as a microenvironment for cell cultures. This microbioreactor stimulates cell coalescence and 3D aggregation while providing optimal gas exchange between the interior and the surrounding environment. Moreover, the properties of liquid marble microbioreactors render them ideal for co-culture experiments. This liquid marble technique has been previously explored and optimized for other cell types. Here we describe a protocol that allows for the derivation of functional cardiac mini organoids, consisting of co-cultured cardiomyocytes and cardiac fibroblasts. These cardiospheres can be valuable for modelling cardiac diseases in vitro and assessing cell interactions to decipher disease mechanisms.
Lab website: https://www.medicalcellbiologylab.com/
Contact: https://www.researchgate.net/profile/Jeffrey-Aalders
RE-place database: https://www.re-place.be/method/liquid-marbles-cost-effective-platform-generate-cardiospheres-co-cultured-cardiomyocytes-and

Innovation examples
HealthToxicologyIn vitro
Assessing respiratory toxicity using in vitro models
The airways form a barrier for inhaled compounds, however, such compounds may cause local effects in the airways or may lead to lung diseases, such as fibrosis or COPD. Cell models of the respiratory tract, cultured at the air-liquid-interface (ALI) are a relevant model to assess the effects of inhaled compounds on the airways. Such models allow human relevant exposure, which is via the air, and assessment of effects on the epithelial cell layer. At RIVM we use air-liquid-interface cultured cell models and expose these to airborne compounds to assess the effects of agents such as nanomaterials, air pollutants or compounds from cigarette smoke. By using a mechanism-based approach to assess the effects of these compounds we invest in animal-free alternatives that better predict adverse effects in humans.

Questions
Helpathon #7 - Can you help Jesmond and Duco?
Can you help Jesmond Dalli, Professor at Barts, the London School of Medicine and Dentistry and Queen Mary University of London and Duco Koenis, Post-Doctoral Fellow in his team, to identify animal free research methods to discover novel drug targets for resolving inflammation in rheumatoid arthritis and bacterial infections? Join Helpathon #7 – first of its kind as it will take place in the UK, on 10-11th of October 2022.