PMID- 25646741 OWN - NLM STAT- MEDLINE DCOM- 20151221 LR - 20150330 IS - 1873-4367 (Electronic) IS - 0927-7765 (Linking) VI - 127 DP - 2015 Mar 1 TI - In vitro characterization of 6-Coumarin loaded solid lipid nanoparticles and their uptake by immunocompetent fish cells. PG - 79-88 LID - S0927-7765(15)00029-6 [pii] LID - 10.1016/j.colsurfb.2015.01.022 [doi] AB - 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 mug/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 mug/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. CI - Copyright (c) 2015 Elsevier B.V. All rights reserved. FAU - Trapani, Adriana AU - Trapani A AD - Department of Pharmacy-Drug Sciences, University of Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy. FAU - Mandracchia, Delia AU - Mandracchia D AD - Department of Pharmacy-Drug Sciences, University of Bari "Aldo Moro", Via Orabona 4, 70125 Bari, Italy. FAU - Di Franco, Cinzia AU - Di Franco C AD - CNR-IFN Bari, Bari via Amendola 173, 70126 Bari, Italy. FAU - Cordero, Hector AU - Cordero H AD - Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Murcia, Spain. FAU - Morcillo, Patricia AU - Morcillo P AD - Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Murcia, Spain. FAU - Comparelli, Roberto AU - Comparelli R AD - 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. FAU - Cuesta, Alberto AU - Cuesta A AD - Department of Cell Biology and Histology, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, 30100 Murcia, Spain. FAU - Esteban, Maria Angeles AU - Esteban MA AD - 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. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150121 PL - Netherlands TA - Colloids Surf B Biointerfaces JT - Colloids and surfaces. B, Biointerfaces JID - 9315133 RN - 0 (Coumarins) RN - 0 (Lipids) RN - 0 (Thiazoles) RN - 0 (coumarin 6) SB - IM MH - Animals MH - Calorimetry, Differential Scanning MH - Cell Line MH - Coumarins/*pharmacology MH - Endocytosis/*drug effects MH - Flow Cytometry MH - Head Kidney/cytology MH - Immunocompetence/*drug effects MH - Lipids/*chemistry MH - Microscopy, Confocal MH - Microspheres MH - Nanoparticles/*chemistry/ultrastructure MH - Sea Bream MH - Spectroscopy, Fourier Transform Infrared MH - Thiazoles/*pharmacology OTO - NOTNLM OT - Confocal microscopy OT - Fish OT - Flow cytometry OT - Microparticles OT - Solid lipid nanoparticles EDAT- 2015/02/04 06:00 MHDA- 2015/12/22 06:00 CRDT- 2015/02/04 06:00 PHST- 2014/11/01 00:00 [received] PHST- 2014/12/22 00:00 [revised] PHST- 2015/01/13 00:00 [accepted] PHST- 2015/02/04 06:00 [entrez] PHST- 2015/02/04 06:00 [pubmed] PHST- 2015/12/22 06:00 [medline] AID - S0927-7765(15)00029-6 [pii] AID - 10.1016/j.colsurfb.2015.01.022 [doi] PST - ppublish SO - Colloids Surf B Biointerfaces. 2015 Mar 1;127:79-88. doi: 10.1016/j.colsurfb.2015.01.022. Epub 2015 Jan 21.