Enhanced oral bioavailability of poorly aqueous soluble drugs encapsulated in a number of lipid-based formulations, including emulsions, micellar systems, self-emulsifying drug delivery systems, liposomes and solid lipid nanoparticles (SLN) via lymphatic delivery has been documented (1). In the present work, SLN were designed for the oral delivery of heparin in order to take advantage from the lymphatic intestinal transport pathway. In order to improve the incorporation of a high hydrophilic compound in a lipid matrix, heparin was “insolubilized” by the coupling with chitosan. In this aim we have developed chitosan/heparin Polyelectrolyte complexes (PEC). Such as systems are able to complex stably heparin (up to pH < 6.8) (2) and after pelletization by centrifugation were embedded in SLN obtaining a hybrid system lipid/chitosan nanoparticles (PEC-SLN). Since no in-vitro lymphoid tissue is currently available, CaCo-2 cell monolayer could be considered an alternative in vitro model to be used as a screening tool before animal studies are undertaken (1). Aim of the work In this work naked PEC, hybrid PEC-SLN as well as heparin-loaded SLN (Hep-SLN) were characterized as regard as the size, zeta potential, morphological characteristics, drug loading and in vitro drug release. Moreover FITC labeled PEC along with Red Nile labeled PEC-SLN and empty SLN were evaluated on CaCo-2 cell line in order to study their cytotoxicity by MTT test and their cell internalization ability by cytometric and confocal analysis. Experimentals Size and zeta potential were measured by Zetasizer Nano ZS (Malvern), morphological characteristics by SEM-FEI (SEM, Nova NanoSEM 450, Fei) and by AFM (Park Autoprobe Atomic Force Microscope (Park Instruments) Intenalization extent was visualized by confocal laser scanning microscopy (CLSM) (Leica DM IRE2) Results and discussion Results demonstrated that PEC size is highly influenced by pH, being 322 ±30 nm at the optimal pH 6.5. PEC-SLN and Hep-SLN displayed a size between 150 and 400 nm and a zeta potential from -20 to -36 mV. The drug loading was higher for PEC-SLN respect to Hep-SLN (50.4 ± 6 and 32.4 ± 4 UI/100 mg, respectively) while the heparin release rate was faster for Hep-SLN than for PEC-SLN, achieving a percentage of heparin released of 100% and 20%, respectively, in 6 h in simulated intestinal fluid. These data indicate that heparin was not well encapsulated in the Hep-SLN while after conjugation with chitosan stable heparin encapsulation was achieved in SLN. Finally naked PEC labeled with FITC along with PEC-SLN labeled with Red Nile was evaluated in vitro on CaCo-2 cells. MTT test showed that all the samples were poor cytotoxic even for long incubation time (overnight). Internalization data showed that PEC complexes (A) achieved a poor internalization level in the cells while PEC-SLN (B) have proved to be able to entry CaCo-2 cells in a time-dependent manner. Conclusions PEC-SLN being able to enter in the CaCo-2 cells unlike the naked PEC, probably due to their lipidic nature, can be considered promising carrier to be further studied as promoter for the lymphatic intestinal transport of heparin
In vitro evaluation on CaCo-2 cells of hybrid lipid/chitosan nanoparticles for the oral delivery of heparin / Sacchetti, Francesca; Aracri, Raffaella; Maretti, Eleonora; Iannuccelli, Valentina; Montanari, Monica; Leo, Eliana Grazia. - (2014). ((Intervento presentato al convegno CRS italian chapter tenutosi a Firenze nel November 6th – 8th 2014.
|Data di pubblicazione:||2014|
|Autore/i:||Sacchetti, Francesca; Aracri, Raffaella; Maretti, Eleonora; Iannuccelli, Valentina; Montanari, Monica; Leo, Eliana Grazia|
|Titolo:||In vitro evaluation on CaCo-2 cells of hybrid lipid/chitosan nanoparticles for the oral delivery of heparin.|
|Nome del convegno:||CRS italian chapter|
|Luogo del convegno:||Firenze|
|Data del convegno:||November 6th – 8th 2014|
|Citazione:||In vitro evaluation on CaCo-2 cells of hybrid lipid/chitosan nanoparticles for the oral delivery of heparin / Sacchetti, Francesca; Aracri, Raffaella; Maretti, Eleonora; Iannuccelli, Valentina; Montanari, Monica; Leo, Eliana Grazia. - (2014). ((Intervento presentato al convegno CRS italian chapter tenutosi a Firenze nel November 6th – 8th 2014.|
|Tipologia||Abstract in Atti di Convegno|
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