The topic of my thesis is the investigation of Chitosan-TPP nanoparticles as model compounds for protein delivery. The aim of the project is the preparation and characterization of chitosan-TPP nanoparticles loaded with five selected model proteins. A set of proteins (Nrf2,Lysozyme,BSA,OVA, IgG) characterized by different molecular weight and isoelectric point, is studied in order to analyse the impact of these parameters on the encapsulation efficiency. Chitosan-Tripolyphosphate unloaded nanoparticles were prepared using ionic gelation method and characterized in terms of colloidal properties and morphology. After loading with proteins, the encapsulation efficiency was found to be affected by different parameters such as the initial amount of proteins loaded in NPs, the size and isoelectric point of proteins. Finally stability and cytotoxicity of the develop carrier system were also tested. MTT assay on human cells from alveolar epithelium (A549 cell line) was used to evaluate any toxic effect.
L’argomento della mia tesi consiste nella caratterizzazione di nanoparticelle di chitosano-TPP come carriers per i farmaci proteici. Nanoparticelle di chitosano e tripolifosfato sono state preparate con il metodo di gelificazione ionotropica. L’obbiettivo principale è l’incapsulazione di un set di 5 proteine caratterizzate da diverso peso molecolare e punto isoelettrico utilizzando nanoparticelle di chitosano come vettori. Le 5 proteine selezionate sono : il fattore di trascrizione nucleare eritroide-2 (Nrf2), Lisozima,Albumina di siero bovino (BSA), Ovalbumina (OVA), e Immunoglobulina G (IgG). Il goal consiste nel verificare e analizzare l’impatto di queste caratteristiche delle proteine sull’efficienza di incapsulazione. Tre fattori influiscono principalmente sull’efficienza di incapsulazione: il peso molecolare,il punto isoelettrico e la quantità di proteina inizialmente caricata. Infine,la citotossicità delle nanoparticelle chitosano-TPP sia vuote che caricate con OVA è stata determinata con il saggio MTT,eseguito su cellule umane dell’epitelio alveolare (linea cellulare A549).
Valutazione di Nanoparticelle di Chitosano-TPP come carriers di farmaci proteici. Investigation of Chitosan-TPP Nanoparticles as carriers for protein delivery.
MAGNANO, GRETA CAMILLA
2014/2015
Abstract
The topic of my thesis is the investigation of Chitosan-TPP nanoparticles as model compounds for protein delivery. The aim of the project is the preparation and characterization of chitosan-TPP nanoparticles loaded with five selected model proteins. A set of proteins (Nrf2,Lysozyme,BSA,OVA, IgG) characterized by different molecular weight and isoelectric point, is studied in order to analyse the impact of these parameters on the encapsulation efficiency. Chitosan-Tripolyphosphate unloaded nanoparticles were prepared using ionic gelation method and characterized in terms of colloidal properties and morphology. After loading with proteins, the encapsulation efficiency was found to be affected by different parameters such as the initial amount of proteins loaded in NPs, the size and isoelectric point of proteins. Finally stability and cytotoxicity of the develop carrier system were also tested. MTT assay on human cells from alveolar epithelium (A549 cell line) was used to evaluate any toxic effect.È consentito all'utente scaricare e condividere i documenti disponibili a testo pieno in UNITESI UNIPV nel rispetto della licenza Creative Commons del tipo CC BY NC ND.
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https://hdl.handle.net/20.500.14239/24189