Calorimetry and cryo-transmission electron microscopic studies of PEG2000-grafted liposomes.

The phase behavior of poly(ethylene glycol) grafted liposomes (PEG-liposomes) was investigated by differential scanning calorimetry (DSC), dynamic light scattering (DLS) and cryo-transmission electron microscopy (cryo-TEM). PEG-liposomes were prepared from mixtures of dipalmitoyl phosphatidylcholine (DPPC) and distearoyl phosphatidylethanolamine with a covalently attached PEG molecular weight of 2000 (DSPE-PEG2000). From the results of DLS measurements, the coexistence of PEG-liposomes and small molecular assemblies were confirmed at mole fractions of DSPE-PEG2000 above about 0.1. Moreover, it was confirmed that small molecular assemblies were disk micelles by cryo-TEM. However, the phase transition enthalpies of PEG-liposomes were hardly changed according to the DSC measurement, though the mole fraction of the PEG lipid increased. From these results, it was suggested that the phase transition enthalpies hardly changed despite mixed micelles being formed because the bilayer structure of the disk micelle maintains high cooperativity between the DPPC molecules.

Carboxyfluorescein leakage from poly(ethylene glycol)-grafted liposomes induced by the interaction with serum.

The effects of fetal bovine serum (FBS) on carboxyfluorescein (CF) leakage from poly(ethylene glycol)-grafted liposomes (PEG-liposomes) were investigated. PEG-liposomes were prepared from dipalmitoylphosphatidylcholine (DPPC) and distearoyl-N-monomethoxy poly(ethylene glycol)-succinyl-phosphatidylethanolamines (DSPE-PEG) having PEG molecular weights of 1000, 2000, 3000 and 5000. The presence of FBS dramatically increased CF leakage from liposomes near the gel-liquid crystalline phase transition temperature, but had little effect at lower and higher temperatures. The CF leakage from PEG-liposomes whose molecular weight in PEG units was above 2000 was suppressed compared with that of liposomes without PEG. And, there was hardly any difference in the effect of the PEG molecular weight of the PEG-lipids on CF leakage from PEG-liposomes with FBS when PEG-lipids with a molecular weight in PEG units above 2000 were used. On the other hand, the leakage of CF from liposomes containing 0.145 mol fractions of DSPE-PEG1000 was larger than that of liposomes without PEG. Furthermore, the effects of FBS on the cooperative units of lipid molecules during the gel-liquid crystalline phase transition of liposomes were examined. However, the cooperative units of liposomes with FBS had little change compared with that of liposomes without FBS.

Preparation and clinical application of indomethacin gel for medical treatment of stomatitis.

The preparation and clinical applications of indomethacin (IM) gel were investigated in the treatment of stomatitis resulting from chemotherapy and radiotherapy for cancer. IM gel was prepared by adding various water-soluble polymers [hydroxypropyl cellulose (HPC), etc.] to IM aqueous solution. The release rate of IM from IM gels was found to decrease with increasing polymer concentration and viscosity and to follow a first-order reaction rate equation. The release rate of IM from the IM gel with HPC was decreased gradually with increasing polymer concentration and to be easily controllable compared with gels with other polymers. The time before pain relief occurred after application of the IM gel was slightly shorter and the duration of pain relief was longer compared with the IM aqueous solution. It was confirmed that IM gel is useful in the treatment of stomatitis.

Effects of poly(ethylene glycol) (PEG) concentration on the permeability of PEG-grafted liposomes.

Poly(ethylene glycol)-grafted liposomes (PEG-liposomes) were prepared from dipalmitoylphosphatidylcholine (DPPC) with various amounts of distearoyl-N-monomethoxy poly(ethylene glycol)-succinyl-phosphatidylethanolamines (DSPE-PEG) with PEG molecular weights of 1000, 2000, 3000 and 5000. The effects of DSPE-PEG concentration on the permeability of PEG-liposomes were investigated using carboxyfluorescein (CF). In the gel state, the CF leakage from PEG-liposomes was decreased with increasing mole fractions of DSPE-PEG for all PEG molecular weights. In the liquid-crystalline state, the CF leakage from PEG-liposomes containing DSPE-PEG1000 gradually increased with increasing mole fractions of DSPE-PEG, while that of PEG-liposomes whose molecular weight in PEG units was above 2000 rapidly decreased by the addition of DSPE-PEG. Furthermore, no effect of PEG molecular weight on CF leakage was observed. The relationship between the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) (or 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH)) and the mole fraction of DSPE-PEG for PEG-liposomes was also investigated. No significant changes in fluorescence polarization of DPH for liposomal bilayer membranes was observed in the gel and liquid-crystalline states due to the addition of DSPE-PEG, while that of TMA-DPH was decreased compared with that of liposomes without DSPE-PEG in both states.

A modified method using static head-space gas chromatography for determining the stability constants of 1-alkanol/alpha-cyclodextrin complexation.

A modification of the conventional static head-space gas chromatography method (SHSGC method) to determine stability constants for 1-alkanol/alpha-CD inclusion complexes was investigated. The 1 : 1 stability constants determined by this modified SHSGC method are in reasonable agreement with the corresponding values reported previously. The modified SHSGC method precludes the necessity of the calibration curve by the use of Henry's law constant of guest. Consequently, the modified SHSGC method is more advantageous than the conventional SHSGC method because the experimental time required for determination of the stability constant is markedly reduced.

Effects of poly(ethylene glycol) (PEG) chain length of PEG-lipid on the permeability of liposomal bilayer membranes.

The effects of poly(ethylene glycol) (PEG) chain length of PEG-lipid on the membrane characteristics of liposomes were investigated by differential scanning calorimetry (DSC), freeze-fracture electron microscopy (FFEM), fluorescence polarization measurement and permeability measurement using carboxyfluorescein (CF). PEG-liposomes were prepared from mixtures of dipalmitoyl phosphatidylcholine (DPPC) and distearoyl phosphatidylethanolamines with covalently attached PEG molecular weights of 1000, 2000, 3000 and 5000 (DSPE-PEG). DSC and FFEM results showed that the addition of DSPE-PEG to DPPC in the preparation of liposomes caused the lateral phase separation both in the gel and liquid-crystalline states. The fluidity in the hydrocarbon region of liposomal bilayer membranes was not significantly changed by the addition of DSPE-PEG, while that in the interfacial region was markedly increased. From these results, it was anticipated that the CF leakage from PEG-liposomes is accelerated compared with DPPC liposomes. However, CF leakage from liposomes containing DSPE-PEG with a 0.060 mol fraction was depressed compared with regular liposomes, and the leakage decreased with increasing PEG chain length. Furthermore, the CF leakage from liposomes containing DSPE-PEG with a 0.145 mol fraction was slightly increased compared with that of liposomes containing DSPE-PEG with a 0.060 mol fraction. It is suggested that the solute permeability from the PEG-liposomes was affected by not only properties of the liposomal bilayer membranes such as phase transition temperature, phase separation and membrane fluidity, but also the PEG chain of the liposomal surface.

Complexation of butylbenzenes with 2-hydroxypropyl-cyclodextrins in aqueous solution.

The stability constants for the inclusion of butylbenzenes (BBs) with 2-hydroxypropyl-cyclodextrins (2HP-CDs) in aqueous solution have been determined using the static head-space method. The 1:1 stability constants were obtained by this method. For the complexation of BBs by 2HP-alpha-CD, the order of stability constants was n-BB > isoBB > tert-BB. In contrast, the order for complexation of BBs by 2HP-beta-CD was the opposite: tert-BB > isoBB > n-BB. The order of the stability constants by 2HP-gamma-CD was similar to that of 2HP-beta-CD, but the values of the stability constants were not significantly different in the BB types. In addition, the binding effects are discussed in relation to the different alkyl groups of BBs.

Solubilization of (+)-limonene by anionic/cationic mixed surfactant systems.

The combined effects of sodium n-octyl sulfate (SOS) and cetyltrimethylammonium bromide (CTAB) on the solubilization of (+)-limonene in aqueous solution were studied using a headspace gas chromatography technique. The findings showed the mixing of SOS and CTAB resulted in positive synergistic effects on the solubility of (+)-limonene. The positive synergistic effects are explained from the perspective of the phase behavior of this mixed surfactant system.