The course aims at providing knowledge in the field of the design of in vitro and in silico 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 disease 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 2010163, 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) (lntroduction of the course). After this introduction, in vivo animal trials will be illustrated including animal welfare principles and applicative examples. Alternative preclinical testing methods currently under development will be illustrated, including: - Traditional 2D in vitro models - 3D in vitro models: spheroids and organoids (definition and main uses; examples from literature ); wider part on in vitro models by tissue engineering approaches (characteristics and main uses: examples from literature ) with focus on skin and cardiac tissue engineering; brief and general outlook of characteristics and main uses of organ-on-chips models. - Classroom practical exercises on statistical analysis of data from in vitro models. Some of the models will be described/prepared in practical lectures hold in the laboratories. Typical in vitro assays exploited for experimental 3D in vitro model characterizations will be described in the classrooms and in practical lectures in the laboratories. Additionally, students will learn how also in vitro tests can involve the use of animals, through the use of animals-derived media or materials, and the need for a paradigm shift. In silico models will be also described including molecular and computational approaches in support of 3Rs principle, involving a practical part with classroom exercises (15 hours) Seminars/webinars from Biomedical Industries working in the sector of in silico and in vitro models will be hold. The course will have practical activities in the laboratory which will also help the students to develop their exam project and to better understand the in vitro models.

Learning Outcomes include knowledge on: 1) 3R Principle and Directive 2010163; methods of refinement and reduction in the planning of animal studies; replacement methods for safety and biocompatibility assessments: potentialities and limitations. 2) Preclinical experimental models in vivo (animal trials) and their integration with in vitro cell trials with examples of mouse models of human diseases 3) Protection of animal welfare in European legislation, according to the animal species and their native social organization. 4) Experimental in vitro models and their most recent applications: 2D cultures and 3D models with practical laboratories 5) Statistical analysis of experimental data from testing in in vitro models: procedure for data analysis (basic knowledge) 6} In silico models in preclinical experimentation and their main recent applications. The ability to apply the knowledge and understanding of the topics will be gained through classroom discussions during the development of projects and the analysis of scientific literature. The teachers will give tips for excellent scientific presentations and project writing, which could be beneficial for the final thesis activity. The students will learn group working and will visit the laboratories for practical lectures to better learn the theoretical concepts.

1) Basic knowledge of general chemistry, organic chemistry and biochemistry. 2) Basic knowledge of science and technology of inorganic and polymeric materials. 3) Basic knowledge of cell biology and physiology. 4) Basic knowledge of techniques for determining the surface and massive properties of materials. 5) Basic knowledge of mechanics of fluids and solids 6) Knowledge on Tissue Engineering principles lf some knowledge is missing, the teachers will provide additional information to students requiring them. Teachers will provide complementary video lectures on basic cell biology including inner cell pathways discussed during lectures. Such video lectures will be uploaded on the portal.

1) lntroduction and European Standards for preclinical experimentation 2) Animal models, principles for the protection of animal welfare in EU 3) Brief recall on cell biology (supplemented by additional video lectures loaded on the course portal for those students who require such knowledge) 4) In vitro experimental models (2D vs. 3D models); biologicaI assays exploited in in vitro models 5) Computational fluidodynamic and molecular models in preclinical (and clinical) experimentation and their main applications 6) Statistical analysis of experimental data: software and practical methods 7) Laboratory activities on experimental in vitro models 8) Group work for the project exam

The course collects interdisciplinary topics and different teachers participate with their own block of lectures. There are 4 main blocks of subjects (animal trials, in vitro experimental models, in silico models and laboratory activities) The official language is English. lf foreign students are not attending the course, it will be possible to hold part of the most complex theoretical lectures in ltalian on request by students.

Classroom lessons with slides project on (37.5 hours) • Lab or practical lectures (13.5 hours) • Classroom exercises: Q&A sessions and exercises (9 hours )

• Slides and articles or material uploaded by the teachers • Your own notes during lectures in the lab

Modalità di esame: Prova orale obbligatoria; Elaborato progettuale in gruppo;

Exam: Compulsory oral exam; Group project;

... The exam consists of the preparation of a group project by 2 3 students consisting of a research on the state of the art on a topic of interest by the group in the field of 3Rs-Principle. Students will decide the group composition and project topic and will communicate it to the course teachers. Concerning the project topic, students will have the possibility to discuss it with the teachers during planned meetings. Part of the lectures will be dedicated to project discussion (during the 3 hours slots of lectures). Students will write a project document (30 pages maximum) in English, following advices guidelines provided by the teachers during lectures and planned meetings. The written document should be sent to teachers no later than 7 days before the date of the exam. Then, students will prepare a ppt presentation with slides in English. The presentation should last 10 minutes per each student in the group. Students will decide how they will alternate themselves during the presentation. The language of the oral presentation is ltalian or English depending on students' choice. Examination board will consist of at least 2 teachers. The examination board will make questions on the presentation and written project. The teachers will evaluate the comprehension of the subjects, the structure of the written project and the presence of a critical part, as well as the quality of the presentation and the student's ability to answer questions. Exam is passed if the student reaches a mark equaI or superior to 18. The maximum mark 30L is possible if the student has a mark of at least 30.5.

Gli studenti e le studentesse con disabilità o con Disturbi Specifici di Apprendimento (DSA), oltre alla segnalazione tramite procedura informatizzata, sono invitati a comunicare anche direttamente al/la docente titolare dell'insegnamento, con un preavviso non inferiore ad una settimana dall'avvio della sessione d'esame, gli strumenti compensativi concordati con l'Unità Special Needs, al fine di permettere al/la docente la declinazione più idonea in riferimento alla specifica tipologia di esame.

Modalità di esame: Prova orale obbligatoria; Elaborato progettuale in gruppo;

The exam consists of the preparation of a group project by 2 3 students consisting of a research on the state of the art on a topic of interest by the group in the field of 3Rs-Principle. Students will decide the group composition and project topic and will communicate it to the course teachers. Concerning the project topic, students will have the possibility to discuss it with the teachers during planned meetings. Part of the lectures will be dedicated to project discussion (during the 3 hours slots of lectures). Students will write a project document (30 pages maximum) in English, following advices guidelines provided by the teachers during lectures and planned meetings. The written document should be sent to teachers no later than 7 days before the date of the exam. Then, students will prepare a ppt presentation with slides in English. The presentation should last 10 minutes per each student in the group. Students will decide how they will alternate themselves during the presentation. The language of the oral presentation is ltalian or English depending on students' choice. Examination board will consist of at least 2 teachers. The examination board will make questions on the presentation and written project. The teachers will evaluate the comprehension of the subjects, the structure of the written project and the presence of a critical part, as well as the quality of the presentation and the student's ability to answer questions. Exam is passed if the student reaches a mark equaI or superior to 18. The maximum mark 30L is possible if the student has a mark of at least 30.5.

Modalità di esame: Prova orale obbligatoria; Elaborato progettuale in gruppo;

The exam consists of the preparation of a group project by 2 3 students consisting of a research on the state of the art on a topic of interest by the group in the field of 3Rs-Principle. Students will decide the group composition and project topic and will communicate it to the course teachers. Concerning the project topic, students will have the possibility to discuss it with the teachers during planned meetings. Part of the lectures will be dedicated to project discussion (during the 3 hours slots of lectures). Students will write a project document (30 pages maximum) in English, following advices guidelines provided by the teachers during lectures and planned meetings. The written document should be sent to teachers no later than 7 days before the date of the exam. Then, students will prepare a ppt presentation with slides in English. The presentation should last 10 minutes per each student in the group. Students will decide how they will alternate themselves during the presentation. The language of the oral presentation is ltalian or English depending on students' choice. Examination board will consist of at least 2 teachers. The examination board will make questions on the presentation and written project. The teachers will evaluate the comprehension of the subjects, the structure of the written project and the presence of a critical part, as well as the quality of the presentation and the student's ability to answer questions. Exam is passed if the student reaches a mark equaI or superior to 18. The maximum mark 30L is possible if the student has a mark of at least 30.5.