Tissue engineering constitutes an important support to regenerative medicine to repair damaged tissues towards the use of polymeric biomaterials. Synthetic and natural polymers are employed to produce three-dimensional highly porous fibers-made scaffolds in order to guide cells organization and tissue reconstruction. In the initial part of this thesis project an overview about the state of the literature and the different available methods used in the field of tissue engineering was obtained at SUPSI (Manno, TI). Therefore, the attention was focused on the aims and main treatment options of cartilage repair, in particular the use of biomaterials to produce hollow microfibers that allow the cells used in this kind of applications, mesenchymal stem cells, to adhere to the surface and grew inside the cavity of the fibers. In order to create fiber-based scaffolds for cartilage tissue repair, two artificial biomaterials were investigated: poly-L-lactic acid (PLLA) and poly-caprolactone (PCL). The apparatus used in this study was realized by Gimac (Castronno, Italy) and is based on the dry-wet spinning technique to produce different types of porous fibers through the control of process parameters. This production process requires the determination of the appropriate operative conditions to allow phase inversion, in order to obtain fibers with the suitable porosity for their final use. Therefore, on the basis of the theory behind polymeric mixtures, the experiments were conduced using different polymeric solutions and non-solvents. Then, through a purposeful procedure, two arrangements of the hollow fibers (random and ordered) were used to produce different types of scaffolds. For regenerative medicine aims, it is necessary that these scaffolds allow cells adhesion on the target tissue to ensure significant clinical results. For this reason, mesenchymal stem cells were seeded on the matrices produced to investigate their adhesion on the surface of biomaterial used, their vitality and growth. Biological characterization of the samples was conduced, through in vitro tests, at University of Pisa, to evaluate the effective functioning of scaffolds realized during this project.
L’ingegneria dei tessuti costituisce un valido supporto per la medicina rigenerativa per riparare i tessuti danneggiati attraverso l’uso di biomateriali polimerici. I polimeri sintetici e naturali sono impiegati per produrre scaffolds tridimensionali costituiti da fibre porose al fine di guidare l’organizzazione cellulare e la ricostruzione del tessuto. Nella parte iniziale di questo progetto di tesi, svolta presso la SUPSI (Manno, TI), è stata fatta una panoramica riguardante la letteratura e i differenti metodi disponibili nell’ambito dell’ingegneria dei tessuti. Sono stati quindi analizzati gli scopi e le principali opzioni di trattamento della riparazione della cartilagine, in particolare l’uso di biomateriali per produrre fibre cave che consentano alle cellule usate per questo tipo di applicazioni, cellule staminali mesenchimali, di aderire alla superficie e crescere all’interno della cavità delle fibre. Sono stati considerati due biomateriali artificiali per la produzione di fiber-based scaffolds per riparare i tessuti cartilaginei: acido polilattico (PLLA) e policaprolattone (PCL). Il macchinario usato in questo lavoro è stato realizzato dalla ditta Gimac (Castronno, Italia) e il suo funzionamento è basato sulla tecnica della filatura secco-umido per produrre differenti tipi di fibre porose tramite il controllo dei parametri di processo. Il processo di produzione richiede la determinazione di appropriate condizioni operative per permettere l’inversione di fase, in modo da ottenere fibre con porosità adeguata per il loro uso finale. Quindi, sulla base della teoria sulle miscele polimeriche, sono state condotte delle prove sperimentali utilizzando diverse soluzioni polimeriche e non solventi. Utilizzando quindi una procedura specifica, sono state realizzate due disposizioni delle fibre cave (disordinata e ordinata) per produrre differenti tipologie di scaffolds. Per gli obiettivi della medicina rigenerativa è necessario che gli scaffolds consentano l’adesione cellulare sui tessuti bersaglio al fine di ottenere risultati clinici significativi. Per questo motivo, le matrici prodotte sono state caricate con cellule staminali mesenchimali per verificarne l’adesione sulla superficie del biomateriale utilizzato, la loro vitalità e crescita. La caratterizzazione biologica dei campioni è stata condotta, attraverso test in vitro, presso l’Università di Pisa, per valutare l’effettivo funzionamento degli scaffolds realizzati durante questo progetto.
Development and fine tuning of a GMP production process for the manufacturing of 2D-3D scaffolds for regenerative medicine
SORIGA, ILARIA;PIREDDA, FABIO
2013/2014
Abstract
Tissue engineering constitutes an important support to regenerative medicine to repair damaged tissues towards the use of polymeric biomaterials. Synthetic and natural polymers are employed to produce three-dimensional highly porous fibers-made scaffolds in order to guide cells organization and tissue reconstruction. In the initial part of this thesis project an overview about the state of the literature and the different available methods used in the field of tissue engineering was obtained at SUPSI (Manno, TI). Therefore, the attention was focused on the aims and main treatment options of cartilage repair, in particular the use of biomaterials to produce hollow microfibers that allow the cells used in this kind of applications, mesenchymal stem cells, to adhere to the surface and grew inside the cavity of the fibers. In order to create fiber-based scaffolds for cartilage tissue repair, two artificial biomaterials were investigated: poly-L-lactic acid (PLLA) and poly-caprolactone (PCL). The apparatus used in this study was realized by Gimac (Castronno, Italy) and is based on the dry-wet spinning technique to produce different types of porous fibers through the control of process parameters. This production process requires the determination of the appropriate operative conditions to allow phase inversion, in order to obtain fibers with the suitable porosity for their final use. Therefore, on the basis of the theory behind polymeric mixtures, the experiments were conduced using different polymeric solutions and non-solvents. Then, through a purposeful procedure, two arrangements of the hollow fibers (random and ordered) were used to produce different types of scaffolds. For regenerative medicine aims, it is necessary that these scaffolds allow cells adhesion on the target tissue to ensure significant clinical results. For this reason, mesenchymal stem cells were seeded on the matrices produced to investigate their adhesion on the surface of biomaterial used, their vitality and growth. Biological characterization of the samples was conduced, through in vitro tests, at University of Pisa, to evaluate the effective functioning of scaffolds realized during this project.| File | Dimensione | Formato | |
|---|---|---|---|
|
2014_12_Soriga_Piredda.pdf
accessibile in internet per tutti
Descrizione: Testo della tesi
Dimensione
11 MB
Formato
Adobe PDF
|
11 MB | Adobe PDF | Visualizza/Apri |
I documenti in POLITesi sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
https://hdl.handle.net/10589/102753