Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive degeneration of spinal motor neurons, for which, to date, there is no cure. A therapeutic approach is the transplantation of mesenchymal stromal cells (MSCs) that are able both to migrate spontaneously to damaged tissues (homing) and to secrete vesicular systems containing trophic and immunomodulant factors. The aim of this work is the development of an innovative integrated pharmaceutical-biological approach to improve the therapeutic effect of potential new drugs, such as curcumin, exploiting homing properties of MSCs. It was created a "carrier-in-carrier" system: curcumin has been conveyed in micellar nanosystems (first carrier) in order to avoid hydrolytic or enzymatic degradation, improve its solubility and protect cells from its possible cytotoxic effects; the nano-encapsulated drug was then vehiculated in MSCs from which were isolated extracellular vesicles (EVs) (second carrier) with potential homing property. This study demonstrated that curcumin loaded in nanosystems is internalized by MSCs in few minutes without cytotoxic effects and that the system loading capability is concentration dependent. The nano-encapsulated drug is effectively trapped inside of MSCs, which, in turn, release it into the surrounding environment through vesicular systems. The application of this innovative biological and technological system allows MSCs to be directly administered as an Advanced Therapy Medicinal Product but also to obtain biological products such as microvesicles, exosomes or conditioned medium, which do not have the same limitations of use of vital cellular products.
La sclerosi laterale amiotrofica (SLA) è una patologia neurodegenerativa, caratterizzata da una progressiva degenerazione dei motoneuroni spinali, per la quale, ad oggi, non esiste una cura. Un approccio terapeutico è rappresentato dal trapianto di cellule stromali mesenchimali (MSCs) che sono in grado sia di migrare spontaneamente ai tessuti danneggiati (homing) sia di secernere sistemi vescicolari contenenti fattori trofici e immunomodulatori. Scopo della tesi è lo sviluppo di un innovativo approccio biologico-farmaceutico integrato al fine di migliorare l’effetto terapeutico di potenziali farmaci, quale la curcumina, sfruttando le proprietà di homing delle MSCs. E’ stato realizzato un sistema “carrier-in-carrier”: la curcumina è stata veicolata in nanosistemi micellari (primo carrier) al fine di evitarne fenomeni di degradazione idrolitica o enzimatica, migliorarne la sua solubilità e proteggere le cellule dai suoi eventuali effetti citotossici; il farmaco nano-incapsulato è stato quindi veicolato nelle MSCs dalle quali sono state isolate vescicole extracellulari (EVs) (secondo carrier) con potenziali proprietà di homing. Questo studio ha dimostrato che la curcumina caricata nei nanosistemi viene internalizzata dalle MSCs in pochi minuti senza effetti citotossici e che la capacità di loading del sistema dipende dalla concentrazione del farmaco. Il farmaco nano-incapsulato viene efficacemente intrappolato all’interno delle MSCs, le quali, a loro volta lo rilasciano nell’ambiente circostante veicolato all’interno di sistemi vescicolari. L’applicazione di questo sistema innovativo biologico-tecnologico permette alle MSCs di poter essere direttamente somministrate in qualità di Prodotto Medicinale per le Terapie Avanzate ma anche di ottenere, prodotti biologici quali microvescicole, esosomi o medium condizionato, che non presentano le stesse limitazioni di utilizzo dei prodotti cellulari vitali.
Cellule mesenchimali stromali e nanoparticelle caricate di curcumina per l’allestimento di un sistema carrier-in-carrier: un approccio innovativo per il trattamento della sclerosi laterale amiotrofica.
MICIELI, LIDIA
2014/2015
Abstract
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive degeneration of spinal motor neurons, for which, to date, there is no cure. A therapeutic approach is the transplantation of mesenchymal stromal cells (MSCs) that are able both to migrate spontaneously to damaged tissues (homing) and to secrete vesicular systems containing trophic and immunomodulant factors. The aim of this work is the development of an innovative integrated pharmaceutical-biological approach to improve the therapeutic effect of potential new drugs, such as curcumin, exploiting homing properties of MSCs. It was created a "carrier-in-carrier" system: curcumin has been conveyed in micellar nanosystems (first carrier) in order to avoid hydrolytic or enzymatic degradation, improve its solubility and protect cells from its possible cytotoxic effects; the nano-encapsulated drug was then vehiculated in MSCs from which were isolated extracellular vesicles (EVs) (second carrier) with potential homing property. This study demonstrated that curcumin loaded in nanosystems is internalized by MSCs in few minutes without cytotoxic effects and that the system loading capability is concentration dependent. The nano-encapsulated drug is effectively trapped inside of MSCs, which, in turn, release it into the surrounding environment through vesicular systems. The application of this innovative biological and technological system allows MSCs to be directly administered as an Advanced Therapy Medicinal Product but also to obtain biological products such as microvesicles, exosomes or conditioned medium, which do not have the same limitations of use of vital cellular products.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/20.500.14239/25907