Design and biophysical characterization of 5-fluorouracil-loaded nanosystem for topical skin cancer therapy: in vitro, ex vivo and in vivo studies

NATALIA CALIENNI; JIMENA PRIETO; SILVIA ALONSO; JORGE MONTANARI

San Miguel de Tucumán

Congreso; III Latin American Federation of Biophysical Societies (LAFeBS) - IX IberoAmerican Congress of Biophysics - XLV Reunion Anual SAB 2016; 2016

Ultradeformable liposomes (UL) are a drug delivery nanosystem with an elastic modulus lower than conventional liposomes. UL are capable to penetrate the stratum corneum (SC) of the skin and release their content into the viable epidermis, where neoplastic events occur in skin cancer. 5-Fluorouracil (5FU) is an antineoplastic drug, administered parenterally, with severe side effects. Therefore, the incorporation of 5FU in UL improves specific-site delivery and aims to reduce side effects.UL were prepared by resuspension with a 5FU solution of a thin lipid film (soy phosphatidylcholine and sodium cholate as border activator), formed by vacuum rotary evaporation. Vesicles were sonicated to reduce in size and lamellarity. Non-encapsulated 5FU was removed by molecular exclusion chromatography. Size was determined by DLS and NTA and the stability by ζ potential. The interaction drug-lipid was assessed by DSC, using Laurdan and MC540 probes. Drug-lipid ratio, encapsulation efficiency and release in the time were determined. Size and lamellarity were also corroborated by AFM and TEM. In order to study the elastic properties, force distance measurements by AFM and a deformability test in an automated extruder were performed. Free radical scavenging activity was evaluated using DPPH free radical.In vitro cytotoxicity studies were carried out in HaCaT and SK-Mel-28 human cell-lines by MTT, Crystal Violet and Neutral Red. Cellular uptake at 4º and 37ºC was assessed by fluorescent microscopy. Ex vivo penetration studies were performed on a Saarbrücken Penetration Model device with human skin explants. Intact skin and transversal sections were studied by confocal microscopy and penetration profile was quantified in the SC after tape stripping.Finally, in vivo nanotoxicological and teratogenic studies were carried out in the animal model zebrafish (Danio rerio). ReferencesCevc (1992). Biochim Biophys Acta.Montanari (2007). Int J Pharm.Montanari (2010). J Control Release.