In a recent study beyond this thesis, Panotan Exopolysacharide, secreted from Pantoea sp. BCCS 001 GH was extracted from nectarine fruit and used as a naturally derived polymer. The polymer closely mimics the Polysaccharide structure of the native extracellular matrix (ECM) in the body. To this end, it has been shown that Methacrylated Panotan Exopolysacharide (PAMA) can be used to form a hydrogel by UV crosslinking. Specifically, PAMA-based hydrogels with different degrees of methacrylation and polymeric concentrations were synthesized and characterized in term of their chemical, degradability, physical and mechanical properties. In order to fabricate a scaffold that better resembles the ECM of skeletal muscle tissue in terms of elasticity and bioactivity to promote cellular proliferation and differentiation these PAMA-based hydrogels were modified with Gelin-s (Thiol-modified gelatin). In the present master project, we encapsulated the C2C12 myoblast cell line within the PAMA-based hydrogel. Furthermore, the cellular viability, alignment and differentiation of C2C12 myoblast into myotubes within these hydrogels were investigated by culturing the systems in different mediums, differentiation media (Dm) and growth media (Gm). According to our results, the hydrogel supports cellular alignment and differentiation of C2C12’s into myotubes, especially when exposed to differentiation media. Finally, we used a photolitography technique to gain better control over the physical properties of the cellular microenvironment. In this regard, our results demonstrated a higher myotube alignment with better morphological quality within microchannel patterned hydrogels as compared to their non-patterned counterparts.
In uno studio oltre questa tesi, Panotan Exopolisacharide, secreto da Pantoea sp. BCCS 001 GH è stato estratto dalla frutta della nettarina e usato come un polimero naturale. Il polimero simula strettamente la struttura polisaccaridica nativa della matrice extracellulare (ECM) nel corpo. A tal fine, è stato dimostrato che il Panotan Exopolisacharide metacrilato (PAMA) può essere utilizzato per formare un idrogel mediante reticolazione UV. Nello specifico, gli idrogel a base di PAMA con diversi gradi di metacrilazione e concentrazioni di polimero sono stati sintetizzati e caratterizzati in termini di proprietà chimiche, degradabilità, fisiche e meccaniche. Al fine di fabbricare uno scaffold che somigli meglio all'ECM del tessuto muscolare scheletrico in termini di elasticità e bioattività per promuovere la proliferazione e la differenziazione cellulare, questi idrogel a base di PAMA sono stati modificati con Gelin-s (gelatina Thiol-modificata). Nell'attuale progetto principale, abbiamo incapsulato la linea cellulare C2C12 myoblast all'interno dell'idrogel basato su PAMA. Inoltre, la vitalità cellulare, l'allineamento e la differenziazione del mioblasto C2C12in miotubi all'interno di questi idrogel sono stati studiati coltivando i sistemi in diversi mezzi, mezzi di differenziazione (Dm) e mezzi di crescita (Gm). Secondo i nostri risultati, l'idrogel supporta l'allineamento cellulare e la differenziazione di C2C12 in miotubi, specialmente se esposti a mezzi di differenziazione. Infine, abbiamo usato una tecnica di fotolitografia per ottenere un migliore controllo sulle proprietà fisiche del microambiente cellulare. A questo proposito, i nostri risultati hanno dimostrato un allineamento del miotubo più elevato con una migliore qualità morfologica all'interno degli idrogel a microcanali rispetto alle loro controparti non modellate.
Panotan exopolysaccharide-based hydrogel for skeletal muscle tissue engineering application
RIYAHI KHORASGANI, FARINAZ
2017/2018
Abstract
In a recent study beyond this thesis, Panotan Exopolysacharide, secreted from Pantoea sp. BCCS 001 GH was extracted from nectarine fruit and used as a naturally derived polymer. The polymer closely mimics the Polysaccharide structure of the native extracellular matrix (ECM) in the body. To this end, it has been shown that Methacrylated Panotan Exopolysacharide (PAMA) can be used to form a hydrogel by UV crosslinking. Specifically, PAMA-based hydrogels with different degrees of methacrylation and polymeric concentrations were synthesized and characterized in term of their chemical, degradability, physical and mechanical properties. In order to fabricate a scaffold that better resembles the ECM of skeletal muscle tissue in terms of elasticity and bioactivity to promote cellular proliferation and differentiation these PAMA-based hydrogels were modified with Gelin-s (Thiol-modified gelatin). In the present master project, we encapsulated the C2C12 myoblast cell line within the PAMA-based hydrogel. Furthermore, the cellular viability, alignment and differentiation of C2C12 myoblast into myotubes within these hydrogels were investigated by culturing the systems in different mediums, differentiation media (Dm) and growth media (Gm). According to our results, the hydrogel supports cellular alignment and differentiation of C2C12’s into myotubes, especially when exposed to differentiation media. Finally, we used a photolitography technique to gain better control over the physical properties of the cellular microenvironment. In this regard, our results demonstrated a higher myotube alignment with better morphological quality within microchannel patterned hydrogels as compared to their non-patterned counterparts.| File | Dimensione | Formato | |
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https://hdl.handle.net/10589/142677