Interactions of liposome bilayers composed of 1,2-diacyl-3-succinylglycerol with protons and divalent cations.
Bilayer liposomes were prepared by using pure DOSG (1,2-dioleoyl-3-succinylglycerol) or DPSG (1,2-dipalmitoyl-3-succinylglycerol) at pH 7.4 or above. These liposomes undergo destabilization upon incubation with acid. When calcein was used as an entrapped aqueous marker, half maximal content leakage was observed between pH 5.8-6.3. Differential scanning calorimetry showed that at pH 7.4, the chain-melting temperature (Tm) of DPSG was 60.4 degrees C, and increased with decreasing pH (Tm = 57.0 degrees C and 62.7 degrees C at pH 8.9 and 6.7, respectively). Below pH 6.7, extensive phase separation occurred as the major chain melting peak split into three peaks. These three peaks coalesced into one peak below pH 5. Freeze fracture electron micrographs of DOSG liposomes at pH 4 showed the formation of non-bilayer as well as hexagonal phase structures. The effects of divalent cations, such as Ca2+ and Mg2+, on the destabilization of DASG bilayers have also been studied. Differential scanning calorimetry studies of bilayers composed of DPSG showed that both Ca2+ and Mg2+ could increase the Tm of DPSG with increasing concentrations. However, under identical conditions Mg2+ was more effective than Ca2+ in increasing the Tm of DPSG. X-ray diffraction indicated that both Ca2+ and Mg2+ could induce DPSG bilayers to undergo a complete lamellar to hexagonal phase transition. There was a size-dependency on the plasma stability of DOSG liposomes. DOSG liposomes that were smaller in size were more stable in plasma than the larger ones. After incubation with plasma, DOSG liposomes became less acid-sensitive. DOSG immunoliposomes entrapping diphtheria toxin A chain were used as a model for cytoplasmic delivery of the novel pH-sensitive liposomes. The delivery activity was comparable to that of the conventional pH-sensitive liposomes containing unsaturated phosphatidylethanolamine. Our data indicate that the mechanism of liposome destabilization involves extensive bilayer phase separation as well as the formation of non-bilayer structures.
Department of Biochemistry, University of Tennessee, Knoxville 37996-0840.
A M
AM
Tari
N
N
Fuller
L T
LT
Boni
D
D
Collins
P
P
Rand
L
L
Huang
D002152:Calorimetry, Differential Scanning; D002413:Cations, Divalent / Q000737:chemistry*; D005452:Fluoresceins; D005614:Freeze Fracturing; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry; D008854:Microscopy, Electron; D011522:Protons; D014280:Triglycerides / Q000737:chemistry*; D014961:X-Ray Diffraction
D002413:Cations, Divalent; D005452:Fluoresceins; D008051:Lipid Bilayers; D008081:Liposomes; D011522:Protons; D014280:Triglycerides; C088171:1,2-dipalmitoyl-3-succinylglycerol; C088169:1,2-dioleoyl-3-succinylglycerol; C007740:fluorexon