Romano, Eugenia (2021) Engineered Extracellular Vesicles: Processing and testing of cell-derived Exos. [Tesi di dottorato]
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| Tipologia del documento: | Tesi di dottorato |
|---|---|
| Lingua: | English |
| Titolo: | Engineered Extracellular Vesicles: Processing and testing of cell-derived Exos |
| Autori: | Autore Email Romano, Eugenia eugenia.romano1@gmail.com |
| Data: | 15 Luglio 2021 |
| Numero di pagine: | 132 |
| Istituzione: | Università degli Studi di Napoli Federico II |
| Dipartimento: | Ingegneria Chimica, dei Materiali e della Produzione Industriale |
| Dottorato: | Ingegneria dei prodotti e dei processi industriali |
| Ciclo di dottorato: | 33 |
| Coordinatore del Corso di dottorato: | nome email D'Anna, Andrea anddanna@unina.it |
| Tutor: | nome email Torino, Enza [non definito] Netti, Paolo Antonio [non definito] |
| Data: | 15 Luglio 2021 |
| Numero di pagine: | 132 |
| Parole chiave: | Exosomes, Extracellular Vesicles, drug delivery, personalized medicine |
| Settori scientifico-disciplinari del MIUR: | Area 09 - Ingegneria industriale e dell'informazione > ING-IND/34 - Bioingegneria industriale |
| Depositato il: | 20 Lug 2021 12:48 |
| Ultima modifica: | 07 Giu 2023 11:24 |
| URI: | http://www.fedoa.unina.it/id/eprint/13590 |
Abstract
In recent decades, endogenous nanocarrier-exosomes have received considerable scientific interest as drug delivery systems. The unique proteo-lipid architecture allows the crossing of various natural barriers and protects Exosomes cargo from degradation in the bloodstream. However, the presence of this bilayer membrane as well as their endogenous content make production and loading of exogenous molecules challenging. In the present work, we will investigate how to promote the manipulation of cellular. And vesicles curvature by a high pressure microfluidic system as ground-breaking method for both Exosomes production and encapsulation. First of all, we exploited this approach to isolate the Exosomes derived by Glioblastoma U87-MG tumoral cell line. An increased yield and purity of Exosomes have been obtained. Furthermore, we proposed a complete protein-profiling comparing traditional isolation method in cell medium and isolation by Dynamic High-Pressure Homogenization. To validate our approach for Exosomes encapsulation, we tested in vitro the prodrug Irinotecan (IRI) in U87-MG Exosomes. As a result, we obtained a high EE, up to 45%, comparable to the principal industrial methodologies used for polymer nanoparticles, shortening the processing time for the encapsulation to several days to 1 hr, improving the drug uptake, and entirely avoiding the use of permeabilization enhancers. Also, this new approach has been tested on Doxorubicin and validated on a different cell lines and 3D cells model. Finally, we performed in vitro preliminary analysis to further understand Exosomes fate and nanobiointeraction with biological environment.
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