Fibrosis is one of the main causes of death in the United State, can affect different organs and the initialization and progression of the disease may be due to several factors. Among others, idiopathic pulmonary fibrosis (IPF) and gut fibrosis (GF) are prevalent in the population. IPF is a chronic fibrotic disease of the lung that leads to the progressive impaire of lung function and to respiratory failure, while intestinal fibrosis is an inevitable complication in patients with inflammatory Bowel disease (IBD), which manifests itself in ulcerative colitis and Crohn’s disease. For both diseases, fibroblasts, together with different epithelial and endothelial cells, have been reported to acquire a myofibroblast phenotype and to play an important role in the subsequent formation of ECM. Since the exact mechanisms of the pathology onset are not entirely clear, pre-clinical in vitro models are needed. Among them, OOC emerged as very promising. In this work, a market analysis was performed to identify the most suitable model between IPF and GF to be introduced in the BiomimX’s R&D pipeline from a business perspective, by considering the market space, the competition and the versatility of the technological advances that such model can introduce. Results of the analyses evidenced that IPF 3D human model was more suitable, so a preliminary biological campaign was performed to establish cell culture protocols for the independent culture of epithelial (i.e. A549) and somatic compartment (i.e. normal human lung fibroblasts, NHLF). Cell viability, cell functionality (i.e. barrier formation) and the effect of a mechanical stress (i.e. 10% uniaxial strain or 30% confined compression) on fibrotic traits onset (i.e. fibroblast to myofibroblast transition and ECM deposition) were evaluated. Specifically, results indicated that epithelial cells showed a better cell viability in dynamic (i.e. 10% uniaxial strain) condition by using gelatin coating. The cells viability for NHLF was good in all conditions tested and the fibrotics traits mainly appeared by using fibroblast growth medium (FGM) in combination with a 10% uniaxial strain. The results allowed to achieve a new design of the platform to create the complete model (epithelial and stromal compartment) and its technical validation. The platform will be used to screen molecules with antifibrotic potentials.
La fibrosi è una delle principali cause di morte negli Stati Uniti, può colpire diversi organi, e la causa e la progressione della malattia può essere dovuta a diversi fattori. Tra gli altri, la fibrosi polmonare idiopatica (IPF) e la fibrosi intestinale sono prevalenti nella popolazione. IPF è una malattia fibrotica cronica del polmone che porta al progressivo declino della funzionalità polmonare all'insufficienza respiratoria, mentre la fibrosi intestinale è una complicanza inevitabile in pazienti con malattia infiammatoria intestinale (IBD) che si manifesta nella colite ulcerosa e nella malattia di Crohn. Per entrambe le malattie i fibroblasti, insieme alle cellule epiteliali ed endoteliali, acquistano un fenotipo miofibroblastico e svolgono un ruolo importante nella successiva formazione di ECM. Il meccanismo esatto non è del tutto chiaro, per questo motivo sono necessari modelli pre-clinici in vitro e tra questi gli OOC sono molto promettenti. In questo lavoro, è stata eseguita un'analisi di mercato per identificare il modello più adatto tra IPF e IF da introdurre nella pipeline R&D di BiomimX da una prospettiva di business, considerando lo spazio di mercato, la concorrenza e la versatilità dei progressi tecnologici che tale modello può introdurre. I risultati delle analisi hanno evidenziato che il modello umano 3D IPF era il più adatto, quindi è stata eseguita una campagna biologica preliminare per stabilire i protocolli di coltura cellulare per lo sviluppo indipendente del compartimento epiteliale (cioè A549) e somatico (cioè fibroblasti nomrali polmonari umani, NHLF). Sono state valutate la vitalità e la funzionalità cellulare (ad es. la formazione di una barriera) e l'effetto di una sollecitazione meccanica (ad es. tensione uniassiale al 10% o compressione limitata al 30%) sull'insorgenza di tratti fibrotici (ad es. la transizione fibroblasta in miofibroblasto e la deposizione dell’ECM). Specificatamente, i risultati indicano che le cellule epiteliali hanno una vitalità cellulare migliore in dinamica (i.e. 10% tensione uniassiale) condizione utilizzando rivestimento gelatina. La vitalità delle cellule per NHLF era buona in tutte le condizioni testate e i tratti fibrotici si sono formati principalmente utilizzando il mezzo di coltura fibroblasto (FGM) in combinazione con uno strain uniassiale al 10%. I risultati hanno permesso di arrivare ad un nuovo design della piattaforma per creare il modello completo (epiteliale e stromale) e la sua validazione tecnica. La piattaforma sarà utilizzata per fare screen con molecole con potenziali antifibrotici.
In vitro 3D human microfluidic model of idiopathic pulmonary fibrosis : from market analysis to a microfluidic device prototype
SITTA, SARA
2020/2021
Abstract
Fibrosis is one of the main causes of death in the United State, can affect different organs and the initialization and progression of the disease may be due to several factors. Among others, idiopathic pulmonary fibrosis (IPF) and gut fibrosis (GF) are prevalent in the population. IPF is a chronic fibrotic disease of the lung that leads to the progressive impaire of lung function and to respiratory failure, while intestinal fibrosis is an inevitable complication in patients with inflammatory Bowel disease (IBD), which manifests itself in ulcerative colitis and Crohn’s disease. For both diseases, fibroblasts, together with different epithelial and endothelial cells, have been reported to acquire a myofibroblast phenotype and to play an important role in the subsequent formation of ECM. Since the exact mechanisms of the pathology onset are not entirely clear, pre-clinical in vitro models are needed. Among them, OOC emerged as very promising. In this work, a market analysis was performed to identify the most suitable model between IPF and GF to be introduced in the BiomimX’s R&D pipeline from a business perspective, by considering the market space, the competition and the versatility of the technological advances that such model can introduce. Results of the analyses evidenced that IPF 3D human model was more suitable, so a preliminary biological campaign was performed to establish cell culture protocols for the independent culture of epithelial (i.e. A549) and somatic compartment (i.e. normal human lung fibroblasts, NHLF). Cell viability, cell functionality (i.e. barrier formation) and the effect of a mechanical stress (i.e. 10% uniaxial strain or 30% confined compression) on fibrotic traits onset (i.e. fibroblast to myofibroblast transition and ECM deposition) were evaluated. Specifically, results indicated that epithelial cells showed a better cell viability in dynamic (i.e. 10% uniaxial strain) condition by using gelatin coating. The cells viability for NHLF was good in all conditions tested and the fibrotics traits mainly appeared by using fibroblast growth medium (FGM) in combination with a 10% uniaxial strain. The results allowed to achieve a new design of the platform to create the complete model (epithelial and stromal compartment) and its technical validation. The platform will be used to screen molecules with antifibrotic potentials.| File | Dimensione | Formato | |
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https://hdl.handle.net/10589/186544