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Preparation of the artificial lipid nanoparticle

CA Cicero P. Albuquerque
FF Fatima R. Freitas
AM Ana Elisa M. Martinelli
JL Josefa H. Lima
RC Rafael F. Coelho
CJ Carlos V. Serrano Jr.
WN Willian C. Nahas
RF Roberto Kalil Filho
RM Raul C. Maranhão
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The radioactively labeled lipid donor nanoparticle was prepared from a lipid mixture described previously by Ginsburg et al. [10] and modified by Maranhão et al. [11]. Two sets of the nanoemulsion were prepared, one labeled with 3H-triglycerides and 14C-cholesterol and the other with 3H-cholesteryl esters and 14C-phospholipids. In a vial, 40 mg cholesteryl oleate, 20 mg egg phosphatidylcholine, 1 mg triolein and 0.5 mg cholesterol, purchased from Sigma Aldrich (St. Louis, MO, USA), were mixed. Trace amounts of glycerol tri [[9, 10](n)-3H] oleate and 4-14C-cholesterol or [1α,2α(n)-3H]-cholesteryl oleate and L-3-phosphatidylcholine,1-stearoyl-2-[1-14C] arachidonyl (Amersham BioSciences, Little Chalfont, Buckinghamshire, UK) were added to the initial solution. The lipids were emulsified by prolonged ultrasonic irradiation in aqueous media for 3 h, and the crude emulsion then ultracentrifugated in a two-step process, with density adjustment by addition of KBr to obtain the nanoparticle. The nanoparticle fraction was dialyzed against a 0.9% NaCl solution.

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