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Local Organising Committee Alessia Di Giancamillo Laura Mangiavini Laura de Girolamo Antongiulio Marmotti Celeste Scotti BRIDGING THE GAP BETWEEN SURGERY AND BIOLOGY: HANDS-ON CARTILAGE LABORATORY COURSE Honorary President : Matteo Denti, Italy Program Chair: Giuseppe M. Peretti, Italy Helder Pereira, Portugal Congress Chairs: Giuseppe M. Peretti, Italy EVENT CO-ORGANIZED BY ESSKA CARTILAGE COMMITTEE AND RESEARCH COMMITTEE April 21 - 22 2016 DIBIT Fondazione Centro San Raffaele del Monte Tabor

Local Organising BRIDGING THE GAP BETWEEN SURGERY … · BRIDGING THE GAP BETWEEN SURGERY AND BIOLOGY: HANDS-ON CARTILAGE LABORATORY COURSE ... diversi aspetti della ricerca in campo

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Local Organising Committee

Alessia Di Giancamillo Laura Mangiavini

Laura de Girolamo Antongiulio Marmotti

Celeste Scotti

BRIDGING THE GAP

BETWEEN SURGERY AND BIOLOGY:

HANDS-ON CARTILAGE

LABORATORY COURSE

Honorary President : Matteo Denti, Italy

Program Chair: Giuseppe M. Peretti, Italy

Helder Pereira, Portugal

Congress Chairs: Giuseppe M. Peretti, Italy

EVENT CO-ORGANIZED BY ESSKA

CARTILAGE COMMITTEE AND

RESEARCH COMMITTEE

April 21 - 22 2016

DIBIT

Fondazione Centro San Raffaele del Monte Tabor

Honorary President

Matteo Denti

Program Chair Giuseppe M. Peretti

Congress Chairs

Giuseppe M. Peretti Helder Pereira

Per migliorare la pratica clinica ci serve più ricerca traslazionale! Lo scopo di questo corso è quello di permettere ai chirurghi ortopedici di prendersi una pausa dalla pratica clinica per studiare più approfonditamente la riparazione e rigenerazione cartilaginea. Il programma sarà focalizzato sui diversi aspetti della ricerca in campo cartilagineo, dal prelievo di tessuto alla coltura cellulare e alle più basilari tecniche di biologia molecolare e di istologia. Durante il corso, i partecipanti proveranno una vera esperienza di laboratorio conducendo loro direttamente diversi esperimenti; inoltre, i partecipanti acquisiranno una base teorica sulla riparazione cartilaginea fondamentale sia per intraprendere un eventuale cammino di ricerca di base nel campo della rigenerazione articolare, sia nel comprendere più a fondo i principi che stanno alla base delle moderne metodiche di ingegneria dei tessuti, che al giorno d’oggi sono utilizzati quotidianamente nella pratica clinica. Questo corso rappresenta quindi un’opportunità unica per ricercatori e chirurghi che desiderano avere un approccio diretto alla ricerca in campo cartilagineo.

For a better clinical practice, we need more translational research! The aim of this course is to allow the orthopedic surgeons to “take a short break” from patients practice in order to “go deeper” inside cartilage repair and regeneration. The program will be focused on the different aspects of cartilage research, from tissue harvesting to cell culture and basic techniques of molecular biology and histology. The attendees will experience a real laboratory practice by directly conducting several lab experiments. Additionally, the attendees will acquire a theoretical background on cartilage repair, fundamental both for the research career in joint regeneration and for a greater comprehension of tissue engineering basic principles, which are nowadays routinely applied in clinical practice. Thus, this course represents a unique opportunity for scientists and surgeons, who desire a more direct approach on cartilage research.

Laura de Girolamo (Milan, Italy)

Matteo Denti (Milan, Italy)

Alessia Di Giancamillo (Milan, Italy)

Neil Fisher (Milan, Italy)

Laura Mangiavini (Milan, Italy)

AntonGiulio Marmotti (Turin, Italy)

Silvia Mattia (Turin, Italy)

Carlotta Perucca Orfei (Milan, Italy)

Helder Pereira (Portugal)

Giuseppe M. Peretti (Milan, Italy)

Celeste Scotti (Milan, Italy)

Irene Tessaro (Milan, Italy)

Marco Viganò (Milan, Italy)

FACULTY

TUTORS

Registration Introduction and presentation of the course

THEORY

Articular Cartilage morphology, biochemistry and biomechanics. Growth plate cartilage

The articular environment Stem cells (MSC, ASC and mesenchymal cells

from umbilical cord) Growth factors and Platelet Rich Plasma (PRP) Gene therapy Culture of chondrocytes: from monolayer to

tridimensional (3D) culture

Coffee break

Molecular biology for gene expression analysis: RNA extraction, purification, quantification and measure of RNA integrity

RT PCR PCR; semi-quantitative PCR and real time PCR Fluorocitometric analysis: Fluorescence

Activated Cell Sorter (FACS) Analysis of protein expression and cell

proliferation of chondrocytes both in monolayer and 3D cultures

Use of different cell markers to determine cell phenotype and proliferation

Immunohistochemistry an immunofluorescence of mesenchymal stem cells (differentiated versus undifferentiated)

Immunohistochemistry and immunofluorescence of umbilical cord mesenchymal stem cells

Lunch break

First day (April 21st 2016)

8.30-9 am 9-9:30 am 9:30 am-1 pm

1 - 2 pm

THEORY

Engineered cartilage: animal models. Transgenic mouse models

Biological materials for engineered cartilage Introduction to lab activity

PRACTICE

Articular explants from pig knee, shoulders and elbows

Preparation of articular explants Preparation of the sterile field Collection of articular cartilage Preparation of culture medium, PBS with

antibiotics/antimycotic and collagenase solution Basics on sterility in the use of the laminar flow

cabinet Enzymatic digestion of articular cartilage

First day (April 21st 2016)

2-2:40 pm

2.45 - 6 pm

1 - 2 pm

PRACTICE

Collection of chondrocytes Evaluation of cell number and viability In vitro cell culture: cell seeding and feeding;

passaging, cell counts and viability 3D culture system: use of fibrin glue and

collagen scaffold

Lunch break

THEORY

Histology evaluation score, problems in the biopsy, preservation and storage

Histology and staining on tissue sections: 1. Hematoxylin and Eosin (H&E); different Hematoxylins 2. Safranin-O 3. Alcian Blue 4. Von Kossa 5. Masson’s Trichrome 6. TRAP 7. Immunohistochemistry and immunofluorescence 8. In situ hybridization

PRACTICE

H&E Safranin-O staining and sample analysis under

microscope Biochemistry assays: quantification of DNA and

GAGs in 3D cultures versus native cartilage

End of the Course

Second day (April 22nd 2016)

9am - 1 pm

1 – 2 pm

2 -3 pm

3 - 5 pm

DIBIT Fondazione Centro San Raffaele del Monte Tabor San Raffaele Congress Centre Via Olgettina, 58 - 20132 Milano

Registration

€ 480,00 (taxes included)

Registrations are limited to maximum 20

attendees and will be accepted in the order in

which they are received. Applications will have

to the administrative office along with a copy of

the money tranfer receipt. In case of cancellation,

there will be no refund.

Course fee includes:

Attending to the entire theorical and

practical sessions

Lunch during the I and II course days

Abstract book

Keep International Via Giuseppe Vigoni, 11 - 20122 Milano Tel. +39 0254122579 – Fax +39 0254124871 E.mail: [email protected]

www.keywordeuropa.com

GENERAL INFORMATION

Venue

Registration

Organising

Secretariat

1 - 2 pm