Association of liposomes with the isolated perfused rabbit heart

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Abstract
Liposomes were prepared from a mixture of either phosphatidylcholine (PC) and cholesterol (Ch) (7∶2) (neutral), PC, dicetylphosphate and Ch (4∶1∶3) (negative), or PC, stearylamine and Ch (4∶1∶3) (positive). In addition, as the lipid phaseliposomal marker,3H- or14C-Ch was added. Alternatively,14C-labelled mannitol, glucose or inulin was used as the aqueous phase-marker in some experiments. The liposomes were purified by Sephadex gel chromatography and by using microfilter. After 4 h of sonication, 95% of the total liposomes were found to be smaller than 1.2 μm. The entrapment volume was calculated to be 0.9–1.2 μl/μmole of lipid. The ratio of lipid radioactivity and aqueous phase-radioactivity, which were found in a non-filtrable portion of the perfusate, remained constant during a heart-perfusion period of 30 min, indicating that the liposomes were stable during the experimental period. The wash-out experiment indicated that the liposomes were distributed in a space of the perfused heart nearly as large as the mannitol space. The liposomes were rapidly taken up by the heart during perfusion in a Langendorff manner. The positive liposomes were taken up by the perfused nonischemic heart at a greater rate than were the negative liposomes. The results with perfused ischemic hearts were equivocal. The liposomal label was found to be distributed unequally in the subcellular fractions, namely, relatively greater amounts (per mg protein) of liposome-bound radioactivity of Ch or mannitol were found in the microsomal fraction than in the mitochondrial or cytosolic fractions of the perfused heart. This distribution pattern was not influenced by the electrical charge of liposomes or by the oxygenation state of the heart perfusion. The accumulation of the liposomal label in the microsomal fraction found in the heart perfusion experiment could not be observed in the experiment in which tissue slices were incubated in the presence of liposomes, or in the experiment in which free mannitol was administered in the heart perfusion.

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