title: In vitro characterization of 6-Coumarin loaded solid lipid nanoparticles and their uptake by immunocompetent fish cells.
abstract: The primary aim of the present work was to evaluate the in vitro uptake of 6-Coumarin (6COUM) loaded solid lipid nanoparticles (SLN) by two gilthead seabream (Sparus aurata L.) cell types: an established cell line (SAF-1 cells) and the primary cultures of head-kidney (HK)-the main haemopoietic organ in fish, equivalent to mammalian bone marrow-leucocytes. For this purpose, after the physicochemical characterization of SLN, the uptake by those immunocompetent fish cells was evaluated using flow cytometry and confocal microscopy. Concomitantly, the uptake of 6-COUM loaded SLN was compared with that achieved with 6-COUM loaded pectin microparticles (MPs), which were selected as a competitor of the delivery carriers. After SLN and MP physicochemical characterization, the results demonstrated that SAF-1 cells were able to internalize high percentages of 6-COUM SLNs when incubated for 4, 8 and 24h, with the highest SLN concentration tested (10 μg/ml). The ability of HK leucocytes to internalize SLN was also found to vary depending on both incubation time and SLN concentration. The highest values of HK leucocytes internalizing SLN particles (around 16%) were detected at the maximum SLN concentration (20 μg/ml) at incubation times of 4 or 8h. Conversely, HK leucocytes were unable to internalize MPs at any tested concentration and incubation time. A possible mechanism explaining the uptake into cells is proposed. The present work constitutes the first approximation to consider SLN as nanocarriers for delivering biologically active substances to fish.
authors: Department of Pharmacy-Drug Sciences, University of Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy.
authors: Adriana
authors: A
authors: Trapani
authors: Department of Pharmacy-Drug Sciences, University of Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy.
authors: Delia
authors: D
authors: Mandracchia
authors: CNR-IFN Bari, Bari via Amendola 173, 70126 Bari, Italy.
authors: Cinzia
authors: C
authors: Di Franco
authors: Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Murcia, Spain.
authors: Héctor
authors: H
authors: Cordero
authors: Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Murcia, Spain.
authors: Patricia
authors: P
authors: Morcillo
authors: CNR-IPCF, Istituto per i Processi Chimici e Fisici, UOS Bari, c/o Dipartimento di Chimica, University of Bari "Aldo Moro", Via Orabona 4, 70126 Bari, Italy.
authors: Roberto
authors: R
authors: Comparelli
authors: Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Murcia, Spain.
authors: Alberto
authors: A
authors: Cuesta
authors: Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Murcia, Spain. Electronic address: aesteban@um.es.
authors: Maria Angeles
authors: MA
authors: Esteban
mesh_terms: D000818:Animals; D002152:Calorimetry, Differential Scanning; D002460:Cell Line; D003374:Coumarins / Q000494:pharmacology*; D004705:Endocytosis / Q000187:drug effects*; D005434:Flow Cytometry; D060226:Head Kidney / Q000166:cytology; D007121:Immunocompetence / Q000187:drug effects*; D008055:Lipids / Q000737:chemistry*; D018613:Microscopy, Confocal; D008863:Microspheres; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D021541:Sea Bream; D017550:Spectroscopy, Fourier Transform Infrared; D013844:Thiazoles / Q000494:pharmacology*
chemical_list: D003374:Coumarins; D008055:Lipids; D013844:Thiazoles; C517282:coumarin 6
keywords: Confocal microscopy; Fish; Flow cytometry; Microparticles; Solid lipid nanoparticles