Scognamiglio, Laura Sara (2020) A novel bioengineered cystic fibrosis model. [Tesi di dottorato]


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Item Type: Tesi di dottorato
Resource language: English
Title: A novel bioengineered cystic fibrosis model
Scognamiglio, Laura
Date: 12 October 2020
Institution: Università degli Studi di Napoli Federico II
Department: Ingegneria Chimica, dei Materiali e della Produzione Industriale
Dottorato: Ingegneria dei prodotti e dei processi industriali
Ciclo di dottorato: 32
Coordinatore del Corso di dottorato:
Netti, Paolo AntonioUNSPECIFIED
Date: 12 October 2020
Settori scientifico-disciplinari del MIUR: Area 09 - Ingegneria industriale e dell'informazione > ING-IND/06 - Fluidodinamica
Area 09 - Ingegneria industriale e dell'informazione > ING-IND/22 - Scienza e tecnologia dei materiali
Area 09 - Ingegneria industriale e dell'informazione > ING-IND/31 - Elettrotecnica
Area 09 - Ingegneria industriale e dell'informazione > ING-IND/34 - Bioingegneria industriale
Date Deposited: 14 Oct 2020 12:36
Last Modified: 28 Oct 2021 12:48

Collection description

Cystic fibrosis (CF) is one of the most common genetic diseases in the world; it is an autosomal recessive disease affecting various organs, in particular bronchi and alveoli in the lung. The pathology is caused by a mutation in the CFTR gene, which encodes a protein termed CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) that functions as a channel for chlorine. The most common mutation present in 70% of worldwide CF cases is the ΔF508, a deletion (Δ meaning suppression) of three nucleotides, which results in a loss of phenylalanine in the 508 position of the protein. CFTR is a chloride channel located at the apical membrane of epithelial cells of different organs where it plays an important role in transepithelial electrolyte and fluid transport 1. This CFTR gene is mainly expressed in the respiratory epithelium, where the CFTR protein is found at the apical level of the epithelial cells. In the presence of the ΔF508 mutation, the CFTR protein cannot be glycosylated and folded or properly folded. Therefore, the protein gets stuck in the cell membrane and is not be able to reach the apical side of the respiratory epithelium. This causes a decompensation of chlorine, liquids and consequently the formation of a thick and viscous mucus that promotes bacterial colonization of the airways 2. As a consequence of this infected and compromised environment, inflammation, fibrosis and remodeling of the extracellular matrix occur at the baso-lateral side of the epithelium; thus inducing the formation of a fibrotic connective tissue as a secondary response to the pathology 3 4. Furthermore, some studies have also revealed an alteration of the submucous glands present in the CF respiratory connective tissue 5. Most of the studies about CF are focused on epithelial cells because they express CFTR and are directly compromised by the channel dysfunction. For this reason, the large majority of the in vitro models of CF are represented by epithelial cells. Currently, the most used models to study CF are animal models (murine) and epithelial cells models on filter. However, experiments with animals have always the drawback of ethical issues and, particularly in the case of murine models, of the scarce representativity of the human lung pathology, which is the most clinically relevant. As for the models of epithelia on porous membranes, although they are useful for many applications, they do not completely recapitulate the conditions of the tissue in vivo. Indeed, the absence of an extracellular matrix means that the epithelium / connective tissue crosstalk is missing, which influences the differentiation and function of the epithelium in vitro and the response / adaptation to physiological stimuli or harmful events in vivo. In this perspective, in the IIT@CRIB lab we established a novel tissue engineering approach to build-up organ in vitro. By following such strategy, we produced different organ models (i.e. skin 6,7, cervix 8). Our results clearly demonstrated the fundamental role of the connective tissue in guiding in vitro epithelium morphogenesis. Using a similar method, as first aim of this PhD project, we developed a full thickness CF airway model, a 3D human bronchial tissue never made before, consisting of bronchial epithelium on an endogenous pulmonary extracellular matrix, thus produced exclusively by lung fibroblasts. This innovative model is extraordinarily useful for studying the effects of therapeutic strategies focused not only on epithelial response but also on stromal response in a dynamic environment representative of the native human condition. Moreover, the second aim of this PhD project was to design and manufacture a microfluidic chip in which to insert the full tickness airway (normal and CF) or the epithelial model on porous membrane. This microfluidic device represents a new platform for tissue culture, which best summarizes the structure of the human lung and its environment under dynamic conditions. Furthermore, in addition to offering a new cell culture method, this chip allows a more realistic drug administration, by the systemic and air route, with the use of an engineered aerosol system directly on the chip. Finally, the microfluidic platform allows live monitoring of the tissue conditions by means of electrical measurements and the insertion of gold electrodes.


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