The course aims at providing knowledge in the field of the design of models for the preclinical testing of biomedical implants, drugs and advanced therapies. Innovations in preclinical models, stimulated by EU regulation, will prospect new work opportunities for Biomedical Engineers. Currently, preclinical investigations on biomedical implants, drugs and advanced therapies proceed through initial in vitro trails, using 2D human cell cultures, followed by in vivo animal tests. However, 2D human cell culture models are not able to closely reproduce cell-cell and cell-extracellular matrix interactions present in human tissues, which indeed possess a 3D complex structure. Animal experiments have greatly contributed to the understanding of the mechanisms of diseases and to the study of new treatments for diseases. However, main weak points of animal experimentation include: (1) the influence of laboratory environment and other variables on animal study outcomes; (2) recognized differences between animal models of diseases and human diseases; (3} specie-specific physiology and genetics; (4) the large use of experimental animals in basic and applied research, associated with animal suffering. Based on EU Directive 2010/63, alternative methods for preclinical validation are under development or - in a few cases - have been already approved , to Replace, Reduce and Refine animal testing according to the 3Rs Principle (Replacement, Reduction and Refinement) (as analysed in the lntroduction of the course). Alternative preclinical testing methods currently under development will be then illustrated in the course as described below. During 2023-24, some modifications will be performed, expanding the in vitro modelling part. PART 1 - IN VITRO MODELS: - Traditional 2D in vitro models and use of animals-derived components in cultures, underlining the need for a paradigm shift. - 3D in vitro models: spheroids, organoids and organs-on-chip (definition and main uses) with focus on in vitro models by tissue engineering approaches (particularly skin tissue engineering) and applications of organoids. - Understanding the effect of 3D versus 2D cultures on cell behaviour. - Some of the studied in vitro models and their validation will be analysed during practical lectures in laboratories. Typical in vitro assays exploited for 3D in vitro model characterizations will be described in the classrooms and in practical lectures in the laboratories. - Case studies, e.g. cardiac tissue and skeletal muscle tissue modelling in preparation to the project exam. PART 2 - STUDENTS' PROJECTS After this first part dedicated to in vitro experimental models and lab exercises, the rules for Exam Project will be the subject of a lecture, followed by specific lectures aimed at providing a general knowledge on the project subjects before project assignments to students' groups. In the following part of the course, some of the lectures will be dedicated to project activity in the classroom under teacher's supervision. PART 3 - IN VIVO ANIMAL MODELS The remaining theoretical lectures will be dedicated to provide knowledge on in vivo models, including principles of animal welfare and main animal models used for preclinical validation of regenerative medicine solutions. PART 4 - IN SILICO MODELS Finally, in silico models will be described including molecular and computational approaches in support of 3Rs principle, involving a practical part with classroom exercises in groups. The in silico model part could be integrated into the project work. EXTERNAL ACTIVITIES During the course, exernal activity in the form of seminars/webinars from researchers and Biomedical Industries working in the sector of in silico and in vitro models will be held. PRACTICAL ACTIVITIES The course will have practical activities in classrooms (in silico part) and in laboratory (in vitro part) which will also help the students to better understand the in vitro models and to carry out their project.