The neuromuscular junction (NMJ) serves as a synapse between the nervous and muscle systems, which is essential for proper muscle contraction and motor control. The NMJ dysfunction leads to severe neuromuscular diseases, and thus, an advanced platform is needed to model the NMJ for understanding disease mechanisms and developing effective treatments. The emergence of 3D printing, digital light processing (DLP) in particular, enables cost-effectiveness, the ability to produce complex designs and architecture, and patient-specificity. The thesis work presents the design of a compartmentalized in vitro platform for NMJ modeling. The platform was fabricated using the DLP manufacturing method. The device design consisted of two parts, i.e., the top part as the culture chamber and the bottom part as support. The culture chamber consists of two compartments, each for a different cell type, connected by microchannels in between, allowing physical separation and connection at the same time. The DLP method was carried out using the LC OPUS printer and the commercially available resins. Prior to device fabrication, preliminary validation tests were performed on the printer to assess resolution capabilities and on the resins to evaluate biocompatibility according to ISO 10993 standard. Subsequently, the functionality of the fabricated was evaluated through the development and optimization of culture protocol for the neuronal stem cells. Results demonstrate that the selected commercially available resins provide enough printing resolution for this application while exhibiting no cytotoxic effect on cultured cells. The final design successfully achieved effective compartmentalization during the cell seeding procedure and maintained stable performance under operating conditions. Overall, this Thesis work study establishes a robust foundation for NMJ modelling applications and related endeavors.
La giunzione neuromuscolare (NMJ) funge da sinapsi tra i sistemi nervoso e muscolare, risultando essenziale per la corretta contrazione muscolare e per il controllo motorio. La disfunzione della NMJ conduce a gravi patologie neuromuscolari e, pertanto, è necessaria una piattaforma avanzata per modellare la NMJ, al fine di comprendere i meccanismi patologici e sviluppare trattamenti efficaci. L’emergere della stampa 3D, in particolare della digital light processing (DLP), consente economicità, possibilità di produrre design e architetture complesse e specificità per il paziente. Il lavoro di tesi presenta la progettazione di una piattaforma compartimentata in vitro per il modellamento della NMJ. La piattaforma è stata fabbricata mediante il metodo di produzione DLP. Il design del dispositivo consisteva di due parti, ossia la parte superiore come camera di coltura e la parte inferiore come supporto. La camera di coltura è composta da due compartimenti, ciascuno per un diverso tipo cellulare, connessi da microcanali intermedi, che consentono allo stesso tempo separazione fisica e connessione. Il metodo DLP è stato eseguito utilizzando la stampante LC OPUS e resine disponibili in commercio. Prima della fabbricazione del dispositivo, sono stati eseguiti test preliminari di validazione sulla stampante, per valutarne le capacità di risoluzione, e sulle resine, per valutarne la biocompatibilità secondo lo standard ISO 10993. Successivamente, la funzionalità del dispositivo fabbricato è stata valutata attraverso lo sviluppo e l’ottimizzazione di un protocollo di coltura per cellule staminali neuronali. I risultati dimostrano che le resine selezionate, disponibili in commercio, forniscono una risoluzione di stampa sufficiente per questa applicazione, senza mostrare effetti citotossici sulle cellule coltivate. Il design finale ha raggiunto con successo un’efficace compartimentazione durante la procedura di semina cellulare e ha mantenuto prestazioni stabili in condizioni operative. Complessivamente, questo lavoro di tesi stabilisce una solida base per le applicazioni di modellamento della NMJ e per attività correlate.
A novel DLP-fabricated compartmentalized platform for modeling neuromuscular junction in vitro
Leonardo, Michael
2024/2025
Abstract
The neuromuscular junction (NMJ) serves as a synapse between the nervous and muscle systems, which is essential for proper muscle contraction and motor control. The NMJ dysfunction leads to severe neuromuscular diseases, and thus, an advanced platform is needed to model the NMJ for understanding disease mechanisms and developing effective treatments. The emergence of 3D printing, digital light processing (DLP) in particular, enables cost-effectiveness, the ability to produce complex designs and architecture, and patient-specificity. The thesis work presents the design of a compartmentalized in vitro platform for NMJ modeling. The platform was fabricated using the DLP manufacturing method. The device design consisted of two parts, i.e., the top part as the culture chamber and the bottom part as support. The culture chamber consists of two compartments, each for a different cell type, connected by microchannels in between, allowing physical separation and connection at the same time. The DLP method was carried out using the LC OPUS printer and the commercially available resins. Prior to device fabrication, preliminary validation tests were performed on the printer to assess resolution capabilities and on the resins to evaluate biocompatibility according to ISO 10993 standard. Subsequently, the functionality of the fabricated was evaluated through the development and optimization of culture protocol for the neuronal stem cells. Results demonstrate that the selected commercially available resins provide enough printing resolution for this application while exhibiting no cytotoxic effect on cultured cells. The final design successfully achieved effective compartmentalization during the cell seeding procedure and maintained stable performance under operating conditions. Overall, this Thesis work study establishes a robust foundation for NMJ modelling applications and related endeavors.| File | Dimensione | Formato | |
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https://hdl.handle.net/10589/243383