New lipid formulation of amphotericin B: spectral and microscopic analysis.

UV-visible and dichroic spectrum analysis and electron microscopy have been used to characterize a new amphotericin B (AmB) lipid formulation prepared by a solvent displacement process. The composition was dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylglycerol (DMPG) in molar ratio DMPC/DMPG/AmB 7:3:5, a similar composition to that of Abelcet. Although the latter has a "ribbon-like" structure, our process gave a thin disc-like structure. Analysis of circular dichroism (CD) and UV-visible spectra of formulations containing different percentages of AmB revealed that a minimum of AmB self-association was observed with 7:3:5 molar ratio. Varying the lipid ratio (DMPC/DMPG) while maintaining the fixed ratio of AmB yielded similar results when DMPC was in excess (DMPC/DMPG from 10:0 to 6:4). However, when the ratio was between 5:5 to 3:7, AmB self-aggregation increased. For compositions rich in DMPG (2:8 and 0:10), inversion of the CD spectrum was observed. The influence of the lipid composition on the morphology of the complex was also evident in electron microscopy. DMPC/DMPG/AmB (10:0:5) gave large unfracturable lamellae. The presence of DMPG shortened the lamellae, which often appeared as disc-like structures. AmB content, the presence of DMPG and the preparation process all contribute to generating these original structures with particular CD spectra.

Laser-induced disruption of systemically administered liposomes for targeted drug delivery.

Liposomal formulations of drugs have been shown to enhance drug efficacy by prolonging circulation time, increasing local concentration and reducing off-target effects. Controlled release from these formulations would increase their utility, and hyperthermia has been explored as a stimulus for targeted delivery of encapsulated drugs. Use of lasers as a thermal source could provide improved control over the release of the drug from the liposomes with minimal collateral tissue damage. Appropriate methods for assessing local release after systemic delivery would aid in testing and development of better formulations. We use in vivo bioluminescence imaging to investigate the spatiotemporal distribution of luciferin, used as a model small molecule, and demonstrate laser-induced release from liposomes in animal models after systemic delivery. These liposomes were tested for luciferin release between 37 and 45 degrees C in PBS and serum using bioluminescence measurements. In vivo studies were performed on transgenic reporter mice that express luciferase constitutively throughout the body, thus providing a noninvasive readout for controlled release following systemic delivery. An Nd:YLF laser was used (527 nm) to heat tissues and induce rupture of the intravenously delivered liposomes in target tissues. These data demonstrate laser-mediated control of small molecule delivery using thermally sensitive liposomal formulations.

MRI of intravenously injected bone marrow cells homing to the site of injured arteries.

The aim of this study was to test the feasibility of using MRI to detect magnetically labeled, intravenously injected bone marrow (BM) cells homing to injured arteries. In the first phase, BM cells from LacZ-transgenic or green fluorescent protein (GFP)-transgenic mice were transplanted into eight recipient mice. The left femoral arteries of recipient mice were injured using a cuff-constriction or endothelium-damage approach, and the right femoral arteries were uninjured to serve as controls. The location and distribution of migrated LacZ-BM or GFP-BM cells were confirmed with histology. In the second phase, BM-derived cells from LacZ-transgenic mice were labeled with superparamagnetic iron oxide (Feridex) and then transplanted into eight recipient mice with cuff-induced injuries in the left femoral arteries. Migrated Feridex/LacZ-BM cells were monitored in vivo using a 4.7 T MR scanner. Subsequently, high-resolution ex vivo MRI was performed on 9.4 T and 11.7 T. LacZ-positive or GFP-positive cells in the thickened adventitia of the injured arteries were evident on histology. Both in vivo and ex vivo MRI showed larger regions of hypointensity with Feridex-labeled cells at the sites of the injured arteries compared with control arteries (P < 0.01). This study provides initial evidence that may support the potential use of MRI to detect homing of intravenously injected BM cells to injured arteries.

Toxicity and antileishmanial activity of a new stable lipid suspension of amphotericin B.

The aim of the present study was to evaluate the toxicity and the activity of a new lipid complex formulation of amphotericin B (AMB) (LC-AMB; dimyristoyl phosphatidylcholine, dimyristoyl phosphatidylglycerol, and AMB) that can be produced by a simple process. Like other lipid formulations, this new complex reduced both the hemolytic activity of AMB (the concentration causing 50% hemolysis of human erythrocytes, >100 microg/ml) and its toxicity toward murine peritoneal macrophages (50% inhibitory concentration, >100 microg/ml at 24 h). The in vivo toxicity of the new formulation (50% lethal dose, >200 mg/kg of body weight for CD1 mice) was similar to those of other commercial lipid formulations of AMB. The complex was the most effective formulation against the DD8 strain of Leishmania donovani. It was unable to reverse the resistance of an AMB-resistant L. donovani strain. In vivo LC-AMB was less efficient than AmBisome against L. donovani.