Nanoparticles as a novel delivery system for vitamin C administration in aquaculture.

Abstract

abstract Article history:Received 12 October 2013Received in revised form 16 February 2014Accepted 2 March 2014Available online xxxxKeywords:Chitosan nanoparticlesVitamin CNutritionZFL cellsCytotoxicitySolea senegalensis The potential of chitosan-based nanoencapsulation as a tool for delivering ascorbic acid (AA) to marine andfreshwater organisms was investigated. Polymeric non-loaded and loaded vitamin C nanoparticles (NPs) weremadebyionicgelationandtheparticleswerecharacterized.Invitroperformanceofnanoparticleswasevaluatedin a zebrafish liver cell-line (ZFL) and in vivo studies were carried out in fish (post-metamorphic larvae of Soleasenegalensis) and rotifers (Brachionus plicatilis) to assess the potential use of these NPs to be used as a tool innutritional aquaculture studies. The results showed that NPs are suitable to trap hydrosoluble compounds suchasAAbyformingpositivelychargedcomplexes(30–35mV),inananosizerange(b300nm),withencapsulationefficiency(EE) higher than15%andhighstability (N90%ofloadedAAremainedwithinnanoparticles after2h inseawater). The potential cytotoxicity of the NPs was evaluated in ZFL cells and no decrease in cell viability wasnotedupto2.5 mg/mlofnanoparticleconcentration.TheNPuptakewasanalyzedinZFLcellsbyFACScytometryand confocal laser scanning microscopy (CLSM). Time course and dose–response experiments were performedusing fluorescein isothiocyanate labeled NPs (FITC-NPs). The in vitro endocytosis assays with ZFL cells showeda maximum uptake after 6 h of incubation and a dose-dependent increase of fluorescence intensity directlyproportional to the FITC-NP concentration. The antioxidant properties of vitamin C nanoparticles (AA-NPs)werealsoanalyzedinZFLcellextracts.Lipopolysaccharide(LPS)wasaddedtoZFLcellstoinduceoxidativestress.The total antioxidant capacity of the AA-NP-treated cells showed a statistically significant increase with respectto the control with non-loaded nanoparticles (71.00 ± 9.6 and 25.36 ± 3.96 μM Trolox equivalent; p b 0.05respectively). The NPs' ability to penetrate fish intestinal epithelium was also evaluated. After 2 h, NPs wereable to penetrate through intestinal epithelium in post-metamorphic larvae of S. senegalensis as detected byCLSM.ThepotentialuseofNPs asadditiveto rotiferswasanalyzedusingAA-loaded or non-loadedNPs inrotiferenrichment for 2 h.Rotifers fed with AA-NPs increasedupto2-foldof theirascorbicacid levelsincomparison tocontrolgroups.Asawhole,resultsshowthatthesepolymericNPsmightrepresentaninterestingvehiclefororaladministration of AA and other active compounds in aquaculture.© 2014 Elsevier B.V. All rights reserved.