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Biophysical studies and intracellular destabilization of pH-sensitive liposomes

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Lipids

Abstract

We examined changes in membrane properties upon acidification of dioleoylphosphatidylethanolamine/cholesterylhemisuccinate liposomes and evaluated their potential to deliver entrapped tracers in cultured macrophages. Membrane permeability was determined by the release of entrapped calcein or hydroxypyrene-1,3,6-trisulfonic acid (HPTS)-p-xylene-bis-pyridinium bromide (DPX); membrane fusion, by measuring the change in size of the liposomes and the dequenching of octadecylrhodamine-B fluorescence; and change in lipid organization, by31P nuclear magnetic resonance spectroscopy. Measurement of cell-associated fluorescence and confocal microscopy examination were made on cells incubated with liposomes loaded with HPTS or HPTS-DPX. The biophysical studies showed (i) a lipid reorganization from bilayer to hexagonal phase progressing from pH 8.0 to 5.0, (ii) a membrane permeabilization for pH<6.5, (iii) an increase in the mean diameter of liposomes for pH<6.0, and (iv) a mixing of liposome membranes for pH<5.7. The cellular studies showed (i) an uptake of the liposomes that were brought from pH 7.5–7.0 to 6.5–6.0 and (ii) a release of ∼15% of the endocytosed marker associated with its partial release from the vesicles (diffuse localization). We conclude that the permeabilization and fusion of pH-sensitive liposomes occur as a consequence of a progressive lipid reorganization upon acidification. These changes may develop intracellular after phagocytosis and allow for the release of the liposome content in endosomes associated with a redistribution in the cytosol.

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Abbreviations

CHEMS:

cholesterylhemisuccinate

DOPC:

dioleoylphosphatidylcholine

DOPE:

dioleoylphosphatidylethanolamine

DPX:

p-xylene

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Authors and Affiliations

  1. Unité de Pharmacologie Cellulaire et Moléculaire, 70 avenue E. Mounier 73, 1200, Brussels, Belgium

    Fran¢oise Van Bambeke, Anne Kerkhofs, Paul M. Tulkens & Marie-Paule Mingeot-Leclercq

  2. Unité de Chimie Physique Moléculaire et de Cristallographie, Louvain-La-Neuve

    André Schanck

  3. Laboratoire de Biologie Cellulaire, Unité de Biologie Animale, Louvain-La-Neuve

    Claude Remacle

  4. Unité de Chimie Pharmaceutique et de Radiopharmacie, Université Catholique de Louvain, Brussels, Belgium

    Etienne Sonveaux

Authors
  1. Fran¢oise Van Bambeke
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  2. Anne Kerkhofs
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  3. André Schanck
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  4. Claude Remacle
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  7. Marie-Paule Mingeot-Leclercq
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Correspondence to Fran¢oise Van Bambeke.

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Van Bambeke, F., Kerkhofs, A., Schanck, A. et al. Biophysical studies and intracellular destabilization of pH-sensitive liposomes. Lipids 35, 213–223 (2000). https://doi.org/10.1007/BF02664772

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  • DOI: https://doi.org/10.1007/BF02664772

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