1. Ingegneria Tessutale applicata ai modelli in vitro
1.4 Scopo della tesi
Questo lavoro di tesi ha avuto come obiettivo lo sviluppo di scaffold a base di gelMA fabbricati attraverso tecniche di micro e nano fabbricazione.
Il connubio delle proprietà biochimiche del gelMA e la sua fabbricazione mediante tecniche di Elettrospinning, PAM2 e Soft Lithography può consentire la
realizzazione di matrici 3D nano- e micro-strutturate in grado di mimare le proprietà topologiche dell’ambiente extracellulare e di fornire alle cellule gli stimoli di natura biochimica necessari al loro sviluppo. Una valutazione più accurata dell’influenza degli scaffold in gelMA, ottenuti tramite tecniche di fabbricazione PAM2 e Soft Litography, sulla differenziazione di cellule staminali
mesenchimali estratte da midollo osseo umano verranno investigate in collaborazione con l’Università di Southampton.
Questo lavoro di tesi si compone di cinque capitoli e un’appendice.
Capitolo 1: dopo una breve introduzione sull’importanza di utilizzare dei modelli in vitro nel settore dell’ingegneria tessutale, viene presentata: una descrizione delle caratteristiche principali degli scaffold e dei biomateriali impiegati per la realizzazione di modelli in vitro, una descrizione del gelMA, in termini di processo di sintesi e proprietà del materiale, e infine una descrizione delle attuali tecniche di biofabbricazione per la preparazione degli scaffold in TE, con una maggiore attenzione alle tecniche impiegate (Elettrospinning, PAM2 e Soft Lithography) in
questo lavoro di tesi.
Capitolo 2: dopo una prima descrizione dei metodi di sintesi impiegati per la produzione del gelMA, vengono descritti i processi di realizzazione e caratterizzazione dei diversi scaffold ottenuti attraverso le tecniche di nano (Elettrospinning) e micro (PAM2 e Soft Lithography) fabbricazione. Durante
questa prima fase diversi esperimenti sono stati svolti per ottimizzare le caratteristiche morfologiche degli scaffold a base di gelMA. A tal proposito, sono stati condotti studi morfologici attraverso tecniche di microscopia ottica e test meccanici e di swelling, al fine di comprendere le proprietà meccaniche e valutare
secondo capitolo è inoltre descritto il design e la fabbricazione del nuovo setup di stampa della PAM2 proposto per l’ottimizzazione del processo di reticolazione
delle matrici in gelMA.
Capitolo 3: vengono descritti i risultati ottenuti per ogni tipo di scaffold in funzione della tecnica di fabbricazione impiegata e le relative discussioni.
Capitolo 4: vengono definiti i metodi e i risultati ottenuti dalle colture cellulari eseguite su PAM2 e Soft Lithography di cellule staminali mesenchimali estratte da
midollo osseo umano al fine di valutare l’effetto del pattern sulla differenziazione cellulare.
Capitolo 5: si definiscono le conclusioni e i possibili sviluppi futuri degli scaffold in gelMA ottenuti in questo lavoro di tesi.
In appendice è riportato il codice MATLAB realizzato per definire la geometria dello scaffold fabbricato tramite la tecnica PAM2, e le messe in tavola dei
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