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The comparison of protein-entrapped liposomes and lipoparticles: preparation, characterization, and efficacy of cellular uptake

Authors Chang W, Tai Y, Chiang C, Hu C, Hong P, Yeh M

Published 20 October 2011 Volume 2011:6 Pages 2403—2417


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Video abstract presented by Wei-Kuo Chang and Ming-Kung Yeh

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Wei-Kuo Chang1, Yu-Ju Tai2, Chiao-Hsi Chiang3, Chieh-Shen Hu2, Po-Da Hong2, Ming-Kung Yeh3,4
1Division of Gastroenterology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taiwan, ROC; 2Graduate Institute of Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan, ROC; 3School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, ROC; 4Institute of Preventive Medicine, National Defense Medical Center, Sanhsia, Taipei, Taiwan, ROC

Abstract: Fluorescein isothiocyanate-conjugated bovine serum albumin (FITC-BSA)-loaded polyethylene glycol (PEG)-modified liposomes and lipoparticles with high protein entrapment were developed. The lipid formula of the liposomes contained PEGylated lipids and unsaturated fatty acids for enhancing membrane fluidity and effective delivery into cells. The preparation techniques, lipid content, and PEG-modified lipoparticle ratios were evaluated. The PEG-modified lipoparticles prepared by ethanol injection extrusion (100 nm pore size) achieve a population of blank liposomes with a mean size of 125 ± 2.3 nm and a zeta potential of —12.4 ± 1.5 mV. The average particle size of the PEG-modified lipoparticles was 133.7 ± 8.6 nm with a zeta potential of +13.3 mV. Lipoparticle conformation was determined using transmission electron microscopy and field-emission scanning electron microscopy. The FITC-BSA encapsulation efficiency was dramatically increased from 19.0% for liposomes to 59.7% for lipoparticles. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results confirmed the preparation process, and an 8-hour leaching test did not harm the protein structure. Once prepared, the physical and chemical stability of the PEG-modified lipoparticle formulations was satisfactory over 90 days. In vitro retention tests indicated that the 50% retention time for the protein-containing lipoparticles was 7.9 hours, substantially longer than the liposomes at 3.3 hours. A Caco-2 cell model was used for evaluating the cytotoxicity and cell uptake efficiency of the PEG-modified lipoparticles. At a lipid content below 0.25 mM, neither the liposomes nor the lipoparticles caused significant cellular cytotoxicity (P < 0.01) and FITC-BSA was significantly taken up into cells within 60 minutes (P < 0.01).

Keywords: liposomes, lipoparticles, formulation, protein, stability

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