Cardinal Role of Intraliposome Doxorubicin-Sulfate Nanorod Crystal in Doxil Properties and Performance

The uniqueness of Doxil can be attributed, to a large extent, to its intraliposomal doxorubicin-sulfate nanorod crystal. We re-examine these nanocrystal features and their mechanism of the formation by studying pegylated liposomal doxorubicins (PLDs) of the same lipid composition, size distribution, and extraliposome medium that were prepared at different ammonium sulfate (AS) concentrations. This study includes a comparison of the thermotropic behavior, morphology, and in vitro ammonia-induced doxorubicin release (relevant to Doxil’s in vivo performance) of these PLDs. In this study, we confirm that a transmembrane ammonium gradient is critical for doxorubicin remote loading, and we demonstrate that the intraliposomal concentration of sulfate counteranions and ammonium ions determine to a large extent the physical state and stability of the PLDs’ remote loaded doxorubicin. “Fully-developed” intraliposome doxorubicin-sulfate nanorod crystals (as defined by cryogenic transmission electron microscopy imaging) develop only when the ammonium sulfate (AS) concentration used for PLD preparation is ≥150 mM. Less than 10% of PLDs prepared with 100 mM AS show fully developed nanorod crystals. Intraliposomal AS concentration ≥200 mM is required to support the stable nanocrystallization in PLDs. The presence of nanocrystals and their melting enthalpy and phase transition co-operativity strongly affect the ammonia-induced doxorubicin release of PLDs. A quick, biphasic release occurs for PLDs that lack the nanorod crystals or have crystals of poor crystallinity, whereas PLDs prepared with ≥200 mM AS show a monophasic, zero-order slow release. This study also demonstrates that after remote loading, residual intraliposomal ammonium concentration and the transmembrane pH gradient related to it also play an important role in doxorubicin-sulfate intraliposomal crystallization and ammonia-induced doxorubicin release

Nematic Ordering of SWNT in Meso-Structured Thin Liquid Films of Polystyrenesulfonate

The formation of nematic-like islands of single-walled carbon nanotubes (SWNT) in polystyrenesulfonate (PSS) dispersions confined into nanometrically thin films is reported. The SWNT are observed to assemble into orientationally ordered phases, where the intertube distance, as measured via transmission electron microscopy at cryogenic temperatures, matches the polyelectrolyte’s bulk correlation length deduced from X-ray scattering. The micrometers-long islands of orientationally ordered carbon nanotubes are observed in both SWNT and double-walled carbon nanotubes (DWNT) but not in specimens prepared from similar dispersions of multiwalled carbon nanotubes (MWNT). These observations, together with relaxation and rheological experiments, suggest that the orientational ordering may result from coupling between confinement of the polymer-wrapped SWNT and DWNT and the microstructure of the solvated polyelectrolyte