Parkinson's disease (PD) is a progressive neurological disorder that affects movements and is characterized by symptoms such as tremors, rigidity, and slowness of movement and it is caused by the degeneration of dopaminergic neurons in the brain. The idea behind this study is to use the nasal route as access to the Central Nervous System to deliver the neurotransmitter dopamine (DA) in order to bypass the blood-brain barrier. In this perspective, the first barrier that a drug has to cross is the nasal epithelium; therefore, in this study, we decided to use the RPMI2650 human cell line, which is often chosen as a model of the nasal epithelial barrier and for this reason has been shown to be suitable for use in drug permeation studies. On this cell line, we conducted in vitro viability tests using drug delivery systems composed of solid lipid nanoparticles (SLNs) loaded with DA and grape seed extract (GSE)1 with antioxidant function, synthesized with the aim of reducing oxidative stress and finding a remedy for neurotransmitter deficiency in dopaminergic neurons of PD patients. Cell viability assays were conducted using two different methods, Resazurin and MTT. Resazurin is a blue dye commonly used in cell biology experiments to monitor the growth or viability of cells. MTT, or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, is another commonly used dye in biological assays; it is a yellow dye that is reduced by living cells to form a purple formazan product and is often used as a measure of cell viability, metabolic activity, or proliferation. Some different formulations of SLNs containing DA and GSE have been synthesized and used: DA-SLNs co-encapsulating (CO) GSE, DA-SLNs adsorbing (ADS) GSE, DA-SLNs CO and ADS GSE in gel Poloxamer/Carbopol and AlgOx/Hydroxypropyl Methylcellulose, DA-SLNs CO and ADS GSE in cryoprotectant sucrose and methyl beta cyclodextrin, DA-FITC-SLNs CO and ADS GSE, DA-SLNs without GSE. After treatment for 6, 12, 24 hours with various nanoformulations, no significant decrease in viable cells number was observed as compared to control condition.
In vitro model to study different nanoformulations in human nasal epithelial cell line (RPMI2650)
Antonello Caponio;Rosanna Mallamaci;Rosa Angela Cardone;Maria Luana Poeta;Stefano Castellani;Lorenzo Guerra;Adriana Trapani.
2023-01-01
Abstract
Parkinson's disease (PD) is a progressive neurological disorder that affects movements and is characterized by symptoms such as tremors, rigidity, and slowness of movement and it is caused by the degeneration of dopaminergic neurons in the brain. The idea behind this study is to use the nasal route as access to the Central Nervous System to deliver the neurotransmitter dopamine (DA) in order to bypass the blood-brain barrier. In this perspective, the first barrier that a drug has to cross is the nasal epithelium; therefore, in this study, we decided to use the RPMI2650 human cell line, which is often chosen as a model of the nasal epithelial barrier and for this reason has been shown to be suitable for use in drug permeation studies. On this cell line, we conducted in vitro viability tests using drug delivery systems composed of solid lipid nanoparticles (SLNs) loaded with DA and grape seed extract (GSE)1 with antioxidant function, synthesized with the aim of reducing oxidative stress and finding a remedy for neurotransmitter deficiency in dopaminergic neurons of PD patients. Cell viability assays were conducted using two different methods, Resazurin and MTT. Resazurin is a blue dye commonly used in cell biology experiments to monitor the growth or viability of cells. MTT, or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, is another commonly used dye in biological assays; it is a yellow dye that is reduced by living cells to form a purple formazan product and is often used as a measure of cell viability, metabolic activity, or proliferation. Some different formulations of SLNs containing DA and GSE have been synthesized and used: DA-SLNs co-encapsulating (CO) GSE, DA-SLNs adsorbing (ADS) GSE, DA-SLNs CO and ADS GSE in gel Poloxamer/Carbopol and AlgOx/Hydroxypropyl Methylcellulose, DA-SLNs CO and ADS GSE in cryoprotectant sucrose and methyl beta cyclodextrin, DA-FITC-SLNs CO and ADS GSE, DA-SLNs without GSE. After treatment for 6, 12, 24 hours with various nanoformulations, no significant decrease in viable cells number was observed as compared to control condition.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
