The magnetic iron oxide nanoparticles (NPs), such as magnetite (Fe3O4), thanks to their chemical-physical and superparamagnetic properties are extensively used in many industrial (audio speaker, sensory of position, water purification from heavy metals) and medical (agents of contrast in MRI, drug or gene delivery and hyperthermia) applications. Although there are experimental evidences that Fe3O4NPs can reach the CNS, the effects of these nanoparticles on human cerebral cells are still to be clarified. The aim of this study was to assess in vitro the polyvinylpyrrolidone coated magnetite nanoparticles (PVP-Fe3O4NPs) toxicological profile on two human cerebral cell lines (astrocytoma: D384 and neuroblastoma: SH-SY5Y) after short and prolonged exposure to increasing doses of PVP-Fe3O4NPs. For short-term exposure (4, 24 and 48 hours) (1-100 µg/ml PVP-Fe3O4NPs) were evaluated the following endpoints: mitochondrial activity (MTT assay), membrane integrity (double staining calcein-AM/Iodide Propidium) and uptake (Perls’ staining). For long-term exposure (up to 10 days) (0.05-10 µg/ml PVP-Fe3O4NPs), the growth and the cellular proliferation were appraised by clonogenic assay. The results suggest that astrocytes (D384) are more susceptible (greater toxicity) to PVP-Fe3O4NPs’ treatment than neurons (SH-SY5Y) both after short and prolonged exposure. Considering the crucial role of astrocytes in the metal balance regulation, prolonged exposure to such NPs could then alter iron homeostasis, induce toxicity and impair the normal function of the CNS.
Le nanoparticelle (NP) magnetiche di ossido di ferro, come la magnetite (Fe3O4), grazie alle loro proprietà chimico-fisiche e superparamagnetiche, sono ampiamente utilizzate in numerose applicazioni industriali (audio speaker, sensori di posizione, purificazione dell’acqua da metalli pesanti) e mediche (agenti di contrasto nella MRI, drug o gene delivery e ipertermia). Nonostante vi siano evidenze sperimentali che le Fe3O4NP possano raggiungere il SNC, gli effetti di tali nanoparticelle a livello cerebrale non sono ancora noti. Lo scopo principale di questo studio è stato quello di caratterizzare il profilo tossicologico in vitro di nanoparticelle di magnetite rivestite con polivinilpirrolidone (PVP-Fe3O4NP) su due linee cellulari cerebrali umane (astrocitoma: D384 e neuroblastoma: SH-SY5Y) dopo breve e prolungata esposizione a dosi crescenti di PVP-Fe3O4NP. Per l’esposizione a breve termine (4, 24 e 48 ore) (1-100 µg/ml PVP-Fe3O4NP) sono stati valutati i seguenti endpoint: funzionalità mitocondriale (MTT test), integrità di membrana (doppia colorazione calceina AM/Ioduro di Propidio) e uptake (colorazione di Perls’). Per l’esposizione prolungata (fino a 10 giorni) (0.05-10 µg/ml) (PVP-Fe3O4NP) è stata invece valutata la crescita e la proliferazione cellulare mediante test clonogenico. I risultati ottenuti indicano che gli astrociti (D384) sono più suscettibili (maggiore tossicità) al trattamento con PVP-Fe3O4NP rispetto ai neuroni (SH-SY5Y) sia dopo breve che lunga esposizione. Considerando il ruolo fondamentale degli astrociti nella regolazione dell’equilibrio dei metalli, l’esposizione prolungata a tali NP potrebbe quindi alterare l’omeostasi del ferro, indurre tossicità e compromettere il normale funzionamento del SNC.
STUDIO COMPARATIVO DELLA TOSSICITÀ DI NANOPARTICELLE DI MAGNETITE (Fe3O4) IN DUE LINEE CELLULARI CEREBRALI UMANE (Astrocitoma: D384 e Neuroblastoma: SH-SY5Y)
GIUDICI, MARTINA
2015/2016
Abstract
The magnetic iron oxide nanoparticles (NPs), such as magnetite (Fe3O4), thanks to their chemical-physical and superparamagnetic properties are extensively used in many industrial (audio speaker, sensory of position, water purification from heavy metals) and medical (agents of contrast in MRI, drug or gene delivery and hyperthermia) applications. Although there are experimental evidences that Fe3O4NPs can reach the CNS, the effects of these nanoparticles on human cerebral cells are still to be clarified. The aim of this study was to assess in vitro the polyvinylpyrrolidone coated magnetite nanoparticles (PVP-Fe3O4NPs) toxicological profile on two human cerebral cell lines (astrocytoma: D384 and neuroblastoma: SH-SY5Y) after short and prolonged exposure to increasing doses of PVP-Fe3O4NPs. For short-term exposure (4, 24 and 48 hours) (1-100 µg/ml PVP-Fe3O4NPs) were evaluated the following endpoints: mitochondrial activity (MTT assay), membrane integrity (double staining calcein-AM/Iodide Propidium) and uptake (Perls’ staining). For long-term exposure (up to 10 days) (0.05-10 µg/ml PVP-Fe3O4NPs), the growth and the cellular proliferation were appraised by clonogenic assay. The results suggest that astrocytes (D384) are more susceptible (greater toxicity) to PVP-Fe3O4NPs’ treatment than neurons (SH-SY5Y) both after short and prolonged exposure. Considering the crucial role of astrocytes in the metal balance regulation, prolonged exposure to such NPs could then alter iron homeostasis, induce toxicity and impair the normal function of the CNS.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/20.500.14239/21638