Alternative in vitro and in silico models to reduce animal use: a crucial technological advance in dermatological sciences.

Of mice and Mendel. The predicted rise in the use of knock-out and transgenic mice should cause us to reflect on our justification for the use of animals in research.

First Society for Investigative Dermatology Career Development Retreat for PhD Trainees in Dermatological Science

The recent Scientific Committee on Health, Environmental and Emerging Risks Final Opinion on "The need for nonhuman primates in biomedical research, production and testing of products and devices" (2017 SCHEER) highlights approaches that could significantly contribute to the replacement, reduction, and refinement of nonhuman primate (NHP) studies. Initiatives that have the potential to affect NHP welfare and/or their use are expected to be appropriate, fair, and objective and publicly disseminated information focused on NHPs in biomedical research, which includes toxicologic and pathologic research and testing, should be objectively evaluated by stakeholder scientists, researchers, and veterinarians. Thus, IQ Consortium member companies convened to develop an informed and objective response, focusing on identifying areas of agreement, potential gaps, or missing information in 2017 SCHEER. Overall, the authors agree that many positions in the 2017 SCHEER Opinion generally align with industry views on the use of NHPs in research and testing, including the ongoing need of NHPs in many areas of research. From the perspective of the IQ Consortium, there are several topics in the 2017 SCHEER that merit additional comment, attention, or research, as well as consideration in future opinions.

Chapter 5 - Bioprinting of skin

A World of Scientific Endeavors and Friendships

Established in 1998, the FRAME Reduction Committee (FRC) (now the FRAME Reduction Steering Committee [FRSC]) has continued to pursue its aim of reducing the number of animals used in biomedical science. Through its expertise in statistics, experimental design, animal welfare and research on alternatives, it has contributed to raising awareness of the need for reduction and the means of achieving and demonstrating it. In recognising the need for training of scientists to appreciate and understand the concept of reduction, the FRSC has organised dedicated workshops and training schools. Some of the Committee's major achievements are described, and, bearing in mind the current year-on-year increases in the number of scientific procedures on animals, its future activities are outlined.

The scholarship of integration is concerned with making connections across scientific disciplines, placing the work of individual investigators and their specialty fields into a larger context, and educating nonspecialists. The authors focus their comments on the biomedical sciences, but observe that closer integration of the biomedical and behavioral sciences will be particularly crucial to advance understanding of the human brain. They observe that as biomedical sciences become more technologically sophisticated, progress is increasingly dependent on sciences such as physics, chemistry, engineering, and related fields. However, the scholarship of integration has been slower than other forms of scholarship to gain acceptance as an integral activity of the professoriate. The isolation of disciplines from one another, particularly at large universities, and the perception of interdisciplinary work as risky and professionally unrewarding are among the forces that may discourage integrative scholarship. In addition, a troubling disconnect exists between the scientific community and the larger public in the understanding of science. Leaders in academic medicine and science must develop strategies to move interdisciplinary work from the margins into the mainstream of academia. Solutions that have been proposed include creating new research entities and funding mechanisms dedicated to interdisciplinary work; reinvigorating the integrative role of the physician-scientist; and training specialists in translational research. The scientific community must also work to develop more effective means of communicating the importance of its work to the public.

The advancement of high-throughput screening technologies facilitates the generation of massive amount of biological data, a big data phenomena in biomedical science. Yet, researchers still heavily rely on keyword search and/or literature review to navigate the databases and analyses are often done in rather small-scale. As a result, the rich information of a database has not been fully utilized, particularly for the information embedded in the interactive nature between data points that are largely ignored and buried. For the past 10 years, probabilistic topic modeling has been recognized as an effective machine learning algorithm to annotate the hidden thematic structure of massive collection of documents. The analogy between text corpus and large-scale genomic data enables the application of text mining tools, like probabilistic topic models, to explore hidden patterns of genomic data and to the extension of altered biological functions. In this paper, we developed a generalized probabilistic topic model to analyze a toxicogenomics dataset that consists of a large number of gene expression data from the rat livers treated with drugs in multiple dose and time-points. We discovered the hidden patterns in gene expression associated with the effect of doses and time-points of treatment. Finally, we illustrated the ability of our model to identify the evidence of potential reduction of animal use.

International Investigative Dermatology 2018 Meeting: Promoting Global Skin Biology and Skin Disease Research

Researchers in biomedical sciences must continually re-evaluate their investment in experiments using laboratory animals. Our group is interested in various signalling pathways that underlie physiological and pathophysiological functioning of the mammalian heart. Two important resources for this type of work are isolated cardiomyocytes and homogenized or preserved sections of whole myocardium. In order to improve our experimental approach ethically, we devised an adaptation of the Langendorff whole-heart retrograde perfusion technique that allows the isolation of adult rat ventricular cardiomyocytes and processing/storage of myocardium from the same heart. This could result in a 50% reduction in the number of animals required for certain experiments. We believe that a generalized adoption of this method would represent a meaningful reduction of animal use in our field of research and, furthermore, strengthen data sets by permitting correlation between myocyte function and various parameters of myocardial biochemistry/structure in individual hearts. This approach is of particular relevance for studies of cardiac pathology, given the cost and time involved in generating experimental disease models.

The quality of research (tests) conducted by departmental laboratories of veterinary medicine enterprises to confirm the quality of products, in particular veterinary drugs, feed additives, concerns not only the well-being of livestock and poultry farming, but also human health and ensuring environmental safety. Therefore, the regulatory provisions enshrined in the Law of Ukraine “On Veterinary Medicine”, DSTU ISO/IEC-17025 (2019) regulate specific measures to improve the work of testing laboratories, including products for veterinary medicine and animal husbandry. It should be borne in mind that the quality of preventive and therapeutic measures depends not only on the quality of the direct implementation of these measures, but primarily on the quality of medicines, feed additives, which are already used in clinical and diagnostic practice. Accordingly, this work should focus on ensuring guarantees of the reliability of research (tests) carried out by product quality control laboratories, in particular departmental laboratories of veterinary medicine enterprises, through strict compliance with the normative and regulatory requirements of their work, which should be reflected in the quality guidelines as a guiding document for the provision of testing services. And perhaps the main priority in their activities should be specifically fixed personal responsibility of both the manager and each laboratory employee for impeccable objectivity in conducting research (tests) of products on quality indicators, as well as the responsibility of employees of enterprises who select and deliver product samples from control points according to production technology or suppliers of the relevant pharmaceutical raw materials. Accordingly, the requirements for the qualification of employees of the departmental laboratory should be approved in accordance with the established procedure. As for the competence of departmental laboratories to conduct research (testing) of products for veterinary medicine and animal husbandry for quality, according to the Regulation on the Institute, confirmation of the competence of such laboratories is entrusted to the State Committee for Veterinary Medicine and Feed Additives. Confirmation of the competence of laboratories certifies its capabilities for research (testing) of products manufactured by veterinary medicine enterprises, which are their priority, namely: – official recognition of the level of quality, conformity and competence; – guarantees of the reliability of research (testing) of products; – minimizing the possibility of obtaining and issuing erroneous results; – participation in the development and implementation of a comprehensive product quality management system at the enterprise. It is characteristic that the requirements for laboratory competence should be implemented in the practice of its work not chaotically, but logically, consistently and completely, reflecting the full life cycle of conducting research (testing) of products for quality.

Biomedical endeavours can be divided into three major categories: research, education, and testing. Within the context of each of these categories, activities involving whole animals have made major contributions and will continue to do so in the future. However, with technological developments in the areas of biotechnology and computers, new methods are already reducing the use of whole animals in certain areas. This article discusses the general issues of alternatives and then focuses on the development of new approaches to toxicity testing.

Advances in multicellular two-dimensional (2D) and three-dimensional (3D) cell culture systems are providing parasitologists with new tools to investigate host-parasite interactions in vitro. These models offer tissue-specific and, increasingly, host-specific alternatives to traditional 2D monoculture and animal systems, with applications across protozoan and helminth biology. Spheroids, organoids, and emerging assembloid platforms capture key aspects of tissue architecture and function, enabling co-culture of parasites and their products, including dynamic analysis of interactions at defined host interfaces. In recent years, these systems have been adapted to model infection processes, parasite development, immune modulation and tissue remodelling across a range of parasite taxa and tissue types, particularly of the gastrointestinal tract. This review outlines applications of multicellular 2D and 3D cell culture systems in parasitology, drawing on examples from both human and veterinary research. We highlight lessons learned from published works to date that have accelerated the uptake and refinement of these approaches. We also examine technical challenges, including issues of standardisation, scalability, model accessibility, and species representation, particularly for livestock hosts. Looking ahead, the integration of immune, stromal, and microbial components into these models, as well as advances in imaging and omics technologies, and CRISPR-Cas9-mediated engineering of host organoids, promise increasingly sophisticated platforms for studying parasite biology, host tissue responses and pathogenesis. With continued investment and cross-disciplinary collaboration, multicellular culture systems are poised to play a central role in reducing animal use, improving model predictiveness, and supporting the development of next-generation antiparasitic therapies and interventions, including drugs and vaccines.

BackgroundIn drug development, liver failure is the cause of approximately 30% of post marketing withdrawals of pharmaceuticals. Drug-induced liver injury (DILI) remains the leading cause of acute liver failure (ALF), accounting for approximately 15% of the cases.Materials and methodsIn this study, we developed a novel human three-dimensional (3D) liver tissue model by seeding adult primary human hepatocytes onto cell culture inserts under Air-Liquid Interface (ALI) condition for extended culture periods. The engineered tissues were thoroughly characterized for barrier integrity using transepithelial electrical resistance (TEER) measurements and assessed for tissue morphology and structure via hematoxylin and eosin (H&E) staining and immunohistochemistry. Expression levels of drug transporters and drug-metabolizing enzymes were evaluated by quantitative PCR (qPCR). The functionality of the tissue model for drug toxicity assessment was demonstrated by comparison with conventional two-dimensional (2D) monolayer hepatocyte cultures and liver spheroids. To evaluate the model's relevance for DILI studies, we exposed the 3D liver tissues to compounds with well-documented hepatotoxic profiles in humans. Liver function was monitored by quantifying biomarkers such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) released into the culture medium.ResultsThe engineered 3D liver tissue model exhibited distinct apical and basolateral surfaces, reflecting a polarized and stratified architecture that closely mimics native liver tissue. Morphological and phenotypic analyses confirmed the tissue's organotypic features. Gene expression profiling revealed elevated levels of liver-specific genes involved in drug transport, metabolism, and clearance. Functionally, the tissue metabolized midazolam--a substrate of the cytochrome P450 3A4 (CYP3A4) enzyme--into its primary metabolite, 1-hydroxymidazolam. Upon repeated exposure to fialuridine, a discontinued anti-hepatitis B drug known for causing severe liver toxicity in humans, the tissue model exhibited barrier compromise, reduced albumin production, and increased levels of ALT and AST in a time- and concentration-dependent manner.DiscussionThe results strongly suggest the model's physiological relevance and functionality in predicting drug responses in humans. Thus, the engineered 3D organotypic human liver tissue model which can be cultured for weeks and produced in a semi-high throughput format creates an opportunity to study drug-induced liver toxicity in an in vitro microenvironment. The reconstructed 3D liver tissue model can serve as a tool for alternative methods intended to reduce animal use in experimentation.

In this perspective, we identify emerging frontiers in clinical and basic research of melanocyte biology and its associated biomedical disciplines. We describe challenges and opportunities in clinical and basic research of normal and diseased melanocytes that impact current approaches to research in melanoma and the dermatological sciences. We focus on four themes: (1) clinical melanoma research, (2) basic melanoma research, (3) clinical dermatology, and (4) basic pigment cell research, with the goal of outlining current highlights, challenges, and frontiers associated with pigmentation and melanocyte biology. Significantly, this document encapsulates important advances in melanocyte and melanoma research including emerging frontiers in melanoma immunotherapy, medical and surgical oncology, dermatology, vitiligo, albinism, genomics and systems biology, epidemiology, pigment biophysics and chemistry, and evolution.