Learning objectives
The aim of the course is to let the students to acquire knowledge and to be able to understand in detail the biological basis of articular cartilage tissue (in terms of chondrogenesis and cell differentiation), the cellular and molecular aspects of the cartilage pathology and the cellular and molecular basis of the regenerative processes with emphasis on the in vitro models of cell therapy and tissue engineering; as a result of learning, the student will have acquired skills:
- to independently investigate on the study of regenerative medicine/ reparative articular cartilage;
- to fully understand the significance of the study of cell models and biotechnological techniques and biomaterials used for tissue engineering the repair / regeneration of the articular cartilage;
- to critically evaluate the information contained in the scientific literature;
- to analyze what the scientific research has developed in this area;
- to acquire, with the laboratory work, the skills necessary for the preparation of primary cell cultures of articular chondrocytes and for their maintenance and differentiation in three-dimensional systems for the evaluation of biocompatibility and the choindroinductive characteristics of film and scaffold of biomaterials.
The student will also demonstrate to know and understand the notions taught on the various topics during the frontal teaching and practical activity, and moreover to know how to apply them in the design, standardization and methodological validation of the experimental plans for the evaluation of biocompatibility in in vivo and in vitro mainly applied in the orthopedic and dental field. The student must know the anatomical and histological fundamentals and the basic principles of biocompatibility in the osteogenic field to understand the rationale for the choice of an experimental models in vitro and in vivo, both as models for the study and as a models for development of an innovative applications. tHe student must be able to apply the acquired knowledge to identify the most suitable experimental model for a given experimental application. Moreover, the student must be able to present orally or in writing, in a clear, concise and exhaustive manner, using appropriate technical terminology and developing his own critical thinking, the knowledge on the experimental in vitro and in vivo models and biotechnology techniques used for the repair of articular cartilage tissue and bone. The student must be able, with adequate language proficiency, to express the scientific concepts learned and to understand and interpret scientific articles relative to the field.
Another goal of the Course is also to provide the fundamental knowledge on the chemistry of natural and sysntetic polymers , on their behaviour in solution and in the biological fluids, the bases for the design of tridimensional polymeric structures for the cell growth and drug delivery. At the end of the course the student should be able to apply the acquired knowledge to the design and realization of polymeric structures, constituted of polymers of natural origin or synthetic, for the preparation of tridimensional scaffolds or films devoted to the controlled release of substance and able to act as support for cell colonisation and growth. The student will learn also the method for studying the kinetics of drug release from scaffolds/matrices in order to be able to optimize the drug release rate.
Prerequisites
- - -
Course unit content
The main content of the Integrated Course concern the biological concepts, the in vitro and in vivo models models for the sudy of biocompatibility, the techniques and the biomaterials used for the repair/regeneration of cartilage and bone with, moreover, a detailed discussion on the chemistry of synthetic and natural biomaterials also used in other biological fields. Specifically the content of the various teachings is:
Osteochondrogenesis: Models of biocompatibility in vivo and in vitro . Mod A"
The lectures deal with, in a first part, the general aspects of chondrogenesis, biology of synovial joints and articular cartilage and pathophysiology of articular cartilage such as pathogenesis of degenerative and discondroplastic joint diseases (osteoarthritis and osteochondrosis). In a second part, the biological basis of biotechnologic applications for articular cartilage regeneration/repair are treated as follows: chondrocyte differentiation and cellular models used in the innovative biotechnology techniques applied for tissue regeneration (tissue engineering, biocompatible biomaterials and systems for induction of chondrogenesis).
Osteo-chondrogenesis: Models of biocompatibility in vivo and in vitro . Mod B"
The first part of the course focuses on the deepening of knowledge about the locomotor system emphasizing on the embryological development, morphology, biodynamics aspects and considering the species peculiarities. It then deals with the classification and treatment of biomaterials used in the orthopedic and dental field, analyzing the historical path of development and refinement, focusing on the biotechnological strategies for the design of biomimetic materials, that have the ability to interact biochemically with cells and tissues. The course then focused on the features of bone tissue and the study of different in vitro and in vivo experimentals models aimed to evaluate the colonization, osteogenic response, osteointegration and resorption will be considered. The concepts of tissue engineering, replacement medicine, regenerative medicine and nanomedicine will also be described. The practical part of the course will focus on the methodologies of preparation and analysis of histological preparations containing implants. The practical part of the course will focus on the methodologies for the histological preparation and microscopic analysis of samples containing implants. The module program takes into consideration topics that are not referable to a single text. In addition to the suggested reference textbooks, the teaching materials of the lesson as well as a selection of the scientific literature provided by the teacher during the course will be made available to the student. The practicals treats the management in vitro of cultures of chondrocytes isolated from articular cartilage.
Medical products for advanced therapies
Chemistry of natural and synthetic polymers used in biomedical applications, in particular for the production of tridimensional biocompatible systems. The course provides also theoretical knowledge on drug release mechanisms of biologically active substances from polymeric matrices.
Full programme
Osteochondrogenesis: Models of biocompatibility in vivo and in vitro . Mod A.
Theoretical part.
a) FIRST PART. The chondrogenesis : molecular aspects of chondrogenic differentiation in vitro and in vivo: gene expression and influence of growth factors. Molecular regulation of the chondrogenesis . - Structure and physiology of synovial joints. Biology of articular cartilage : microscopic organization, molecular structure, function and metabolism. The chondrocyte and its role in the pathology and repair. - Pathophysiology of articular cartilage : macroscopic and microscopic aspects and the pathogenesis of degenerative and discondroplasic diseases of articular cartilage ( osteoarthritis and osteochondrosis).
b) SECOND PART. The repair/regeneration of articular cartilage: models of study of biocompatibility in vitro and in vivo. Problems and prospects. Basics on the chondroprotective molecules. Models and techniques for cell regeneration / repair of cartilage: differentiated chondrocytes and mesenchymal stem cells, the prospects and limitations of the cell transplantation , the chondrocyte differentiation and de-differentiation, the induction of chondrogenesis. Innovative biotechnology application of cartilage repair: concepts of tissue engineering , biomaterials, principles of biocompatibility of the chondro-conductive and chondro-inductive systems.
Practicals: 1) Isolation and primary culture of articular chondrocytes ; 2) Models for the culturing and maintenance in vitro chondrocyte differentiation (bi- and three-dimensional systems) and for the assessment of biocompatibilty of biomaterials.
Osteochondrogenesis: Models of biocompatibility in vivo and in vitro . Mod B
Theoretical part:
Locomotor system: the organogenesis, the general features, the species peculiarities and the biodynamics aspects:
- Osteology
- Myology
- Arthrology
Components of bony tissue.
Histogenesis of bone tissue. Differentiating markers and pathways of bone deposition and remodeling. Fundamentals of morphology and bone structure, deposition dynamics and remodeling of pre and post-embryo ossification.
Endosseous biomaterials in orthopedic and dental fields: history and definitions, classifications. Biomaterials of first, second and third generation. Biological phenomena occurring at the bone-implant interface. Implant functionalization strategies to promote implantation osteointegration: physical, chemical and biochemical.
Definition of Tissue Engineering; substitutive medicine and regenerative medicine; components of a tissue engineering system, choice of cellular and animal models for experimental evaluation.
Nanomedicine; classification, therapeutic and diagnostic applications.
Experimental models for the assessment of cellular response in vitro.
In vitro development of matrices and tissues, bioreactors and physico-chemical stimuli.
In vivo experimental models for the evaluation of tissue response.
Experimental models and design, sites and implant types. Concepts of legislation.
Methods for the evaluation of osseointegration: use of vital bone markers, micro CT analysis, polarized light analysis, histoenzymatic and immunohistochemical reactions, histological stains.
Practical part:
Locomotor system: recognition and isolation of muscles, bones and joint. Histological preparations techniques; paraffin and resin embedding. Microtomy and histological staining. Observation of the histological slides with brighfield, polarized and fluorescence microscopy; concepts of computerized digital image processing.
Medical products for advanced therapies
Polymer definition and classification. Linear and crosslinked polymers, method of polymerisation and crosslinking. Polymer molecular weight and its determination. Solid state of polymers, crystalline and amorphous structure. Glassy and rubbery state, transition temperature. Polymers in solution. Natural and synthetic polymers used in biomedicine. Tridimensional structure of polymeric scaffolds. Chemical modification of polymers. Solid state manipulation. Physico-chemical characterisation of tridimesional polymeric structures, and films. Polymeric scaffolds as drug delivery systems. Drug release kinetics from matrix systems.
Bibliography
1) Teaching material
At teacher’s discretion the slides of the lectures, schematic summary handouts on the topics treated, descriptive experimental protocols, scientific reviews for deepening of arguments , can be provided and made available on line in electronic format through a special web link.
2) Reference textbooks:
- Kyriacos A. Athanasiou, Eric M. Darling, Jerry C. Hu, Grayson D. DuRaine, A. Hari Reddi. Articular Cartilage. March 21, 2013 by CRC Press.
- Martin's Physical Pharmacy and Pharmaceutical Sciences, Fifth Edition P. J. Sinko Editor, Lippincot Williams & Wilkins, Phildelpia, 2006
- DE FELICI M., e coll.: Embriologia Umana - Morfogenesi - Processi molecolari - Aspetti Clinici, Piccin, Padova, 2014.
- PELAGALLI G.V., e coll.: Embriologia, Morfogenesi e Anomalie dello Sviluppo, Idelson - Gnocchi, Napoli, 2009.
- ADAMO S., CAMOGLIO P., DOLFI A., et al.: Istologia di V. Monesi, VI edizione, Piccin, Padova.
- ROSS M.H., PAWLINA W.: Istologia –Testo e Atlante con cenni di Biologia cellulare e molecolare, Ambrosiana, Milano.
- BARONE R.: Anatomia Comparata dei Mammiferi Domestici, Edagricole, Bologna.
- COZZI B., BALLARIN C., PERUFFO A., CARÙ F.: Anatomia degli animali da laboratorio (Roditori e Lagomorfi), Ambrosiana, Milano.
- KÖNIG H. E., LIEBICH H. G.: Anatomia dei mammiferi domestici (Testo-atlante a colori), Piccin, Padova.
- DI BELLO C.: biomateriali – Introduzione allo studio dei materiali per uso biomedico, Pàtron, Bologna.
- MANTERO S., REMUZZI A., RAIMONDI M.T., AHLUWALIA A.: Fondamenti di ingegneria dei tessuti per la medicina rigenerativa, Pàtron, Bologna.
- HOLLINGER J.O., EINHORN T.A., DOLL B.A., SFEIR C.: Bone tissue engineering , CRC PRESS, Boca Raton, Florida.
Teaching methods
The theoretical lectures, for all courses, are organized through the presentation of topics by Power Point slides. The practicals, in the course of “Osteochondrogenesis:models of biocompatibility in vivo and in vitro (Mod.A) is carried out in the biological multi-seat laboratory with the availability of microbiological hoods, thermostatically baths for tissue enzymatic isolations and inverted optical microscopes for the counting and observations of the cells and cultivations systems applied. Students will operate directly following laboratory protocols defined in the preparations of primary crops and specific cultivations systems . In the course of Osteochondrogenesis: models of biocompatibility in vivo and in vitro (Mod.B), the practice will be taken in the classroom, in the veterinary anatomical museum, in the histology laboratory and in the microscopy lab. The practical lessons foresee the subdivision into groups whose number depends exclusively on the possibility of allowing each student to operate autonomously in the proposed activity.
Assessment methods and criteria
The assessment of the achievement of the objectives of the course has based on an oral examination with one or more questions on the topics covered in the theoretical part and on the practicals in order to assess whether the student has acquired the knowledge and understanding of the content of the course and the ability to apply acquired knowledge. The student should demonstrate an adequate language proficiency and a proper reasoning skills by linking together different aspect of the topic.
The result of the examination is based on the evaluation of the response of the student to both of the questions with an overall assessment.
It 's optional to sustain an ongoing testing (written test multiple choice or Power Point presentation of specific subject with one or more oral questions) on the first part of the program; the examination will be completed, in the date of the official exams, on the topics of the second part and of the practical part with the definition of the final vote.
Other information
The teacher of Osteochondrogenesis: models of biocompatibility in vitro and in vivo (Module A) is available for educational interviews in order to further elucidation and clarification, both during the course and for the preparation of the exam, from Monday to Friday by agreement by email. Without warning, the best days, compatibly with other institutional commitments, are: Wednesday 9.30-11.30, Thursday 10.30-12.30.
The professor of medicinal products for advanced therapies is available on Wednesday 10-12.
The teacher of "Osteochondrogenesis: biocompatibility models in vitro and in vivo (Module B) is available from Monday to Friday by agreement by e-mail or phone.
