pH-sensitive paramagnetic liposomes as MRI contrast agents: in vitro feasibility studies.

A novel type of pH-sensitive paramagnetic contrast agent is introduced; a low molecular weight gadolinium (Gd) chelate (GdDTPA-BMA) encapsulated within pH-sensitive liposomes. The in vitro relaxometric properties of the liposomal Gd chelate were shown to be a function of the pH in the liposomal dispersion and the membrane composition. Only a minor pH-dependency of the T1 relaxivity (r1) was observed for liposomal GdDTPA-BMA composed of the unsaturated lipids dioleoyl phosphatidyl ethanolamine (DOPE) and oleic acid (OA). On the other hand, the r1 of GdDTPA-BMA encapsulated within saturated dipalmitoyl phosphatidyl ethanolamine/palmitic acid (DPPE/PA) liposomes demonstrated a strong pH-dependency. At physiological pH and above, the r1 of this system was significantly lowered compared to that of non-liposomal Gd chelate, which was explained by an exchange limited relaxation process. Lowering the pH below physiological value, however, gave a sharp and 6-7 fold increase in r1, due to liposome destabilisation and subsequent leakage of entrapped GdDTPA-BMA. The pH-sensitivity of the DPPE/PA liposome system was confirmed in an in vitro magnetic resonance imaging (MRI) phantom study.

Chelating agents.

The antibacterial activity of metal ions, metal chelates, and molecules with chelating ability for polyvalent cations have been evaluated. The chelator N, N'-ethylenebis[2-(2-hydroxyphenyl)-glycine] (EHPG) exerted moderate-to-good activity against isolates of pathogenic bacteria and fungi. Other chelating agents such as ethylenediamine-tetraacetic acid (EDTA) and diethylene-triamine-pentaacetic acid (DTPA) revealed weak-to-moderate activity. Metal chelation of ligands reduced the activity of EDTA and DTPA.

Tensile properties of a bone cement containing non-ionic contrast media.

The addition of contrast media such as BaSO4 or ZrO2 to bone cement has adverse effects in joint replacements, including third body wear and particle-induced bone resorption. Ground PMMA containing particles of the non-ionic water-soluble iodine-based X-ray contrast media, iohexol (IHX) and iodixanol (IDX), has, in bone tissue culture, shown less bone resorption than commercial cements. These water-soluble non-ceramic contrast media may change the mechanical properties of acrylic bone cement. The static mechanical properties of bone cement containing either IHX or IDX have been investigated. There was no significant difference in ultimate stress between Palacos R (with 15.0 wt % of ZrO2) and plain cement with 8.0 wt % of IHX or IDX with mass median diameter (MMD) of 15.0 or 16.0 microm, while strain to failure was higher for the latter (p < 0.02). The larger particles (15.0 or 16.0 microm) gave significantly higher (p < 0.001) ultimate tensile strengths and strains to failure than smaller sizes (2.4 or 3.6 microm). Decreasing the amount of IHX from 10.0 wt % to 6.0 wt % gave a higher ultimate tensile strength (p < 0.001) and strain to failure (p < 0.02). Scanning electron microscopy (SEM) showed the smaller contrast media particles attached to the surface of the polymer beads, which may prevent areas of the acrylate bead surface from participating in the polymerization. In conclusion, the mechanical properties of bone cement were influenced by the size and amount of contrast medium particles. By choosing the appropriate amount and size of particles of water-soluble non-ionic contrast media the mechanical properties of the new radio-opaque bone cement can be optimized, thus reaching and surpassing given regulatory standards.

Organobismuth compounds: activity against Helicobacter pylori.

BACKGROUND AND METHODS: The antibacterial activity of originally devised and synthesized organobismuth compounds was tested against fresh clinical isolates of Helicobacter pylori and compared with clinically well-established inorganic bismuth salts currently used in triple antibacterial treatment to eradicate H. pylori. The test conditions in vitro were standard for determination of minimum inhibitory concentrations (MICs). RESULTS: Organic compounds with covalently bound bismuth showed stronger and more consistent antibacterial activity than inorganic bismuth salts. Whereas the most active among the standard therapeutic inorganic compounds showed MICs against the test organisms of 4-8 mg/l (bismuth salicylate) and 0.5-64 mg/l, the most active neosynthesized organic substance, tris(2.6-dimethylphenyl)-bismuthine, consistently showed an MIC of 4 mg/l against all bacterial strains. CONCLUSIONS: The new line of organobismuth compounds might offer a therapeutic potential against the bacteria causing peptic ulcer disease.

Investigation of lanthanide-based starch particles as a model system for liver contrast agents.

Gadolinium and dysprosium diethylenetriamine pentaacetic acid-labeled starch microparticles (Gd-DTPA-SP and Dy-DTPA-SP) were investigated as model liver contrast agents. The liver contrast efficacy of particles with low and high metal contents was compared in two imaging models: in vivo rat liver and ex vivo perfused rat liver. The biodistribution of intravenously injected particles was also assessed by ex vivo relaxometry and inductively coupled plasma atomic emission spectrophotometry of tissues. All particles reduced the liver signal intensity on T2-weighted spin-echo and gradient-recalled echo images as a result of susceptibility effects. Because of their higher magnetic susceptibility, the Dy-DTPA-SP were more effective negative contrast enhancers than the Gd-DTPA-SP. On T1-weighted spin-echo images, only the Gd-DTPA-SP with low metal content significantly increased the liver signal intensity. In addition, these low-loading Gd-DTPA-SP markedly reduced the blood T1. The two latter observations were not consistent with the anticipated blood circulation time of microparticles, but were a result of the lower stability of these particles in blood compared with Gd-DTPA-SP, which has a high metal content. Regardless of stability or imaging conditions, the paramagnetic starch particles investigated showed potential as negative liver contrast enhancers. However, the observed accumulation of particles in the lungs represented a biological limitation for their use as contrast agents.

Paramagnetic liposomes as MRI contrast agents: influence of liposomal physicochemical properties on the in vitro relaxivity.

The in vitro contrast efficacy of liposome encapsulated gadolinium-[10-(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane-1, 4,7-triacetic acid] (GdHPDO3A) has been assessed by relaxometry. The internal concentrations were 150 and 250 mM Gd. Two types of liposome compositions were investigated: a phospholipid blend consisting of both hydrogenated phosphatidylcholine (HPC) and phosphatidylserine (HPS) with a gel-to-liquid crystalline phase transition temperature (Tm) of 50 degrees C, and a mixture of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) with a Tm of 41 degrees C. The investigated liposome size range was 70-400 nm. The T1 and T2 relaxivities (r1 and r2) of liposome encapsulated GdHPDO3A were significantly reduced at 37 degrees C and 0.47 T, compared to those of non-liposomal metal chelate, due to an exchange limitation of the dipolar relaxation process. The highest relaxivity values were obtained for the DPPC/DPPG liposomes, and were attributed to a higher liposome water permeability and to a more efficient water exchange across the membrane. A reduction in liposome size increased the r1, confirming the exchange limited dipolar relaxation. The increased r1 with increasing temperature demonstrated the prerequisite of rapid water exchange between the interior and exterior of the liposome for efficient dipolar relaxation enhancement. Susceptibility effects were present in the liposome systems as the r2/r1 ratio increased with increasing liposome size and internal Gd concentration. In summary, the current work has shown the influence of key physicochemical properties, such as liposome size, membrane composition and permeability, on the in vitro relaxivity of liposome encapsulated GdHPDO3A.

Paramagnetic liposomes as magnetic resonance imaging contrast agents. Assessment of contrast efficacy in various liver models.

RATIONALE AND OBJECTIVES: Liposomal gadolinium (Gd)-HP-DO3A has been evaluated as a contrast agent for liver magnetic resonance imaging. The influence of various liposomal physicochemical properties on the liver uptake and contrast efficacy was investigated in various ex vivo and in vivo liver models. METHODS: Liposomes of different size and membrane properties were prepared. The liposome size ranged from 74 to 304 nm. Two types of phospholipid compositions were studied; a mixture of hydrogenated phosphatidylcholine (HPC) and hydrogenated phosphatidylserine (HPS) with a phase transition temperature (Tm) of 51 degrees C and, a blend composed of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) displaying a Tm of 41 degrees C. Ex vivo tissue relaxometry and in vivo liver imaging were used to study the influence of liposome composition on the liver uptake and contrast efficacy of intravenously injected liposomes. The influence of liposome size and composition on the kinetics of liver uptake and imaging effect was assessed ex vivo in the perfused rat liver. RESULTS: The HPC/HPS preparations showed generally a higher and faster liver uptake than the DPPC/DPPG preparations due to a higher stability in blood/perfusate (high Tm) and to the HPS component. The liposome size modulated the extent and kinetics of liver uptake; the larger the size, the faster and more extensive was the liver uptake. Both types of liposome preparations were shown to be efficient liver susceptibility agents both ex vivo and in vivo due to their uptake by the Kupffer cells of liver. The lack of full correlation between the extent of liver uptake and degree of contrast enhancement might be attributed to different regimes of susceptibility-based relaxation. CONCLUSIONS: The present study has demonstrated the influence of key liposomal physicochemical properties on the liver uptake and contrast efficacy of liposome-encapsulated Gd chelates, exemplified by Gd-HP-DO3A.

Low molecular weight lanthanide contrast agents: in vitro studies of mechanisms of action.

The MR contrast properties of a series of structurally dissimilar low molecular weight (LMW) gadolinium (Gd) and dysprosium (Dy) chelates have been investigated under controlled experimental conditions in various in vitro test systems. Relaxation analysis (water, pH = 5.8, 37 degrees C, .47 T) demonstrated the high dipolar relaxation efficacy of the tested Gd chelates. The T1 and T2 relaxivities of both metal chelate series decreased with decreasing hydration number, confirming the strong correlation between metal chelate structure and dipolar relaxivity. Susceptibility-induced T2 relaxation, commonly known as the susceptibility effect, is modulated primarily by the magnetic susceptibility and compartmentalization of the contrast agent. The influence of these parameters on the susceptibility effect of Dy diethylenetriamine penta-acetic acid bis-methylamide (DTPA-BMA) and GdDTPA-BMA was investigated in two-compartment in vitro models. In red blood cell suspensions (45% hematocrit, 37 degrees C, .47 T, 2 and 3 mM metal ion concentration), the T2 relaxation efficacy of DyDTPA-BMA was markedly improved due to susceptibility effects that were shown to depend on compartmentalization. As the relaxation ability of GdDTPA-BMA was modulated by the dipolar interactions, compartmentalization was not a prerequisite for its T2 relaxation efficacy. In a coaxial glass system with no intercompartmental water exchange, which eliminated the dipolar relaxation mechanism, DyDTPA-BMA was shown to be the most efficient susceptibility agent because of its higher magnetic susceptibility. The reported one- and two-compartment model studies have demonstrated the different mechanism of action of LMW Gd- and Dy-based contrast agents. Gd chelates are predominantly dipolar relaxation enhancers, whereas Dy chelates are efficient susceptibility agents only in compartmentalized systems.

Low-molecular weight lanthanide contrast agents: evaluation of susceptibility and dipolar effects in red blood cell suspensions.

Red blood cell (RBC) suspensions, containing low-molecular weight (LMW) dysprosium (Dy) and gadolinium (Gd) chelates, were selected as a two-compartment system for the evaluation of the magnetic dipolar and susceptibility contributions to the transverse (T2) relaxation of solvent water protons. The influence of RBC geometry and degree of metal chelate compartmentalization on T2 was investigated by variation of the osmolality and hematocrit (HC), respectively. The T2-relaxation ability of Dy-chelates was markedly improved in RBC suspensions, in comparison to aqueous solutions, due to the presence of susceptibility effects that more than compensated for the low dipolar relaxation efficacy. Despite a smaller susceptibility effect, the Gd-chelates were still the most efficacious in shortening T2 due to their comparatively larger dipolar relaxation contribution. The results obtained with the Dy-chelates allowed the evaluation of the relative contributions of susceptibility and dipolar mediated relaxation for the Gd-chelates. The RBC geometry and degree of compartmentalization influenced strongly the T2 relaxation efficacy of Dy-chelates, as opposed to the Gd-chelates. Hemolysis eliminated the susceptibility effect, essentially removing the T2 relaxation ability of Dy-chelates. The T2 relaxation efficacy of Gd-chelates was improved by hemolysis due to enhancement of the dipolar relaxation. As a conclusion, RBC suspensions have clearly been shown to be a suitable ex vivo model with which to distinguish the different contrast mechanisms of LMW Dy- and Gd-based MRI contrast agents.

Ether, carbonate and urethane deoxynucleoside derivatives as prodrugs.

3'-Deoxythymidine and its 3'-azido derivative, 2',3'-dideoxycytidine, 2',3'-dideoxyinosine and 2',3'-dideoxyadenosine have been acylated to form carbonates and urethanes in chemoselective reactions. The nucleosides have been N- and/or O-alkylated by alpha-chloroethyl or chloromethyl alkyl carbonates to form alpha-alkyloxycarbonyloxyethyl or alkyloxycarbonyloxymethyl derivatives. The products are lipophilic in order to facilitate transport through biological membranes and are designed to be cleaved by esterases with liberation of the bioactive nucleoside. Initial esterase cleavage of the alkylated derivatives produces hemiacetals or -aminals which subsequently dissociate to the active nucleoside.

Lanthanide-based susceptibility contrast agents: assessment of the magnetic properties.

The T2* contrast efficacy of paramagnetic contrast agents is dependent on their magnetic properties. Vibrating sample magnetometry (VSM) and the Live Chan NMR method have been used to evaluate the influence of ligand structure on the bulk magnetic susceptibility (BMS) of low-molecular weight (LMW) lanthanide chelates. VSM was also used for the BMS assessment of LMW lanthanide chelates covalently attached to cross-linked starch particles. The ligand structure had no influence on the BMS of the gadolinium (Gd) and dysprosium (Dy) chelates. The mean BMS value of the Dy-chelates was 1.8 fold higher than that of the Gd-chelates. The holmium (Ho) DTPA-BMA chelate had a similar BMS to that of Dy-DTPA-BMA while the lowest BMS was found for europium (Eu(III)) DTPA-BMA. The covalent attachment of Gd-DTPA and Dy-DTPA to a cross-linked starch particle had no impact on their intrinsic magnetic properties. The BMS data were in good accordance with those obtained for non-particulate bound LMW Dy- and Gd-chelates. The magnetic susceptibility of the Gd-DTPA labeled particles was described by the Curie law, indicative of no magnetic interactions between Gd-DTPA molecules. The magnetic susceptibility of the Dy-DTPA labeled particles followed the Curie-Weiss law with a Curie-Weiss temperature of about-2 K, indicating magnetic interactions. The magnetic susceptibility of Dy-DTPA will, however, not be affected by such magnetic interactions at physiological temperatures.

A comparison between IEEC, a new biodegradable particulate contrast medium, and iohexol in a tumor model of computed tomography imaging of the liver.

RATIONALE AND OBJECTIVES: Higher contrast between normal and pathologic tissues in the liver may enable detection of smaller lesions in computed tomography (CT). This can be obtained using a liver-specific contrast medium. The authors evaluate a new agent, IEEC (1'-Ethyloxycarbonyloxy)-ethyl-5-acetylamino-3-(N-methyl-acetylami no)-2,4,6- triiodo-benzenecarboxylate), in an animal model, as a potential contrast agent for CT scanning of the liver. The IEEC particulate contrast medium used is based on a prodrug ester design of metrizoic acid and accumulates rapidly in the liver. The particles are quickly degraded into well-known metabolites and excreted from the body. METHODS: Two groups of rabbits were inoculated with VX2-carcinoma directly into the liver by laparotomy. Computed tomography imaging studies were carried out 9 and 11 days after the inoculation. The investigation was designed as a crossover study. The first group was imaged both as controls (without contrast medium) and with the particulate contrast medium on the 9th day and with iohexol on the 11th day. The second group was imaged with iohexol on the 9th day and as controls, and with the particulate contrast medium on the 11th day. The contrast medium was administered in a dose of 100 mgI/kg. Iohexol was administered in a dose of 570 mgI/kg according to a standard clinical scheme in use at a radiology department for dynamic CT. Changes in normal liver/lesion contrast and the conspicuity of tumors were assessed. On completion of imaging studies on day 11, all animals were killed. The liver was removed and evaluated for the presence of tumors. RESULTS: At macroscopic inspection, all rabbits were found to have tumors ranging from 2 to 14 mm in diameter. The size and location of the tumors corresponded well with the CT images. In the images where the particulate contrast medium was used, the attenuation in the normal liver parenchyma and the contrast between normal liver and lesion was significantly higher compared with the images where iohexol was used or the controls. For all tumor sizes, the lesion detection capability with the particulate contrast medium was significantly higher compared with iohexol (P < .005) and controls (P < .05). CONCLUSIONS: VX2-carcinoma in rabbit liver is a useful model for studying the efficacy of contrast media in CT imaging. The particulate contrast medium IEEC improved visualization of liver tumors.

Water-soluble polysaccharides as carriers of paramagnetic contrast agents for magnetic resonance imaging: synthesis and relaxation properties.

Water-soluble, carbohydrate-based, paramagnetic metal chelate derivatives have been investigated as potential organ-selective contrast media for magnetic resonance imaging (m.r.i.). The in vitro proton spin-lattice relaxation properties of compounds with different paramagnetic metals, chelating agents, and carbohydrate matrixes have been studied. Typically, these complexes were 60-260% more efficient proton-relaxation agents than the corresponding low-molecular-weight metal chelates at 10 MHz, but less efficient than the corresponding protein derivatives. As expected, carbohydrates that contained manganese or gadolinium were more effective relaxation agents than iron, copper, erbium, or nickel derivatives.

Cross-linked, degradable starch microspheres as carriers of paramagnetic contrast agents for magnetic resonance imaging: synthesis, degradation, and relaxation properties.

Biodegradable particles were produced by the cross-linking of starch with epichlorohydrin. Diethylenetriaminepenta-acetic acid (DTPA) was covalently linked to the particles by using DTPA bisanhydride. The small, gadolinium-labelled particles were 40-260% more efficient in vitro proton relaxation agents than the corresponding unbound chelate gadolinium-DTPA. The relaxation properties were dependent on the metal chelate, the particle size, the metal content, and the degree of substitution (d.s.). For the small gadolinium-DTPA particles, an increased d.s. decreased the rate of degradation by alpha-amylase.

Structure activity relationship of magnetic particles as MR contrast agents.

Structure activity relationship (SAR) of superparamagnetic MR contrast agents is discussed based on physicochemical properties and relaxivity data of 16 different particles. All the magnetic particles reduce both relaxation times, T1 and T2. The effect on T2 is stronger than the effect on T1. The relaxation efficacy varies over a wide range. Minor modifications in the preparation of the magnetic particles result in products with different susceptibility properties. The T2 relaxivity is dependent upon the magnetic susceptibility as well as particle size. Small particles reduce the relaxation times to a larger extent than the larger particles. No significant difference in relaxivity is observed between compact and porous particles. Magnetic particles coated with nonmagnet polymer are effective relaxation agents, while nonmagnetic monodisperse particles show no effect on the relaxivity.

MRI contrast media for the liver. Efficacy in conditions of acute biliary obstruction.

The authors investigated in a rat model the efficacy of magnetic resonance imaging (MRI) contrast media for evaluating the liver in conditions of acute biliary obstruction. Two liver-specific MRI contrast media, Cr-DEHIDA and Mn-DPDP, and the nonspecific agent Gd-DTPA were studied in normal rats and in rats whose bile ducts had been ligated before administration of the contrast medium. Images were made using a 2.4 T animal MRI system, and intensity enhancement of liver after contrast medium injection was calculated. Metal analyses of serum and liver tissue and T1 and T2 measurements on liver samples in vitro were performed. The differences in image intensity enhancement of liver between normal rats and rats with ligated bile ducts were not significant for any of the three contrast media. Imaging with Mn-DPDP resulted in the highest intensity enhancement of the liver compared with Cr-DEHIDA and Gd-DTPA. Contrast media concentrations in liver tissue were not significantly different between normal rats and rats with ligated bile ducts; however, Cr-DEHIDA concentrations in serum were higher after bile duct ligation. In vitro measurements of liver tissue indicated unique relaxation properties for Mn-DPDP. This investigation indicates that the contrast media studied may be useful in situations where suspected liver pathology is complicated by acute biliary obstruction.

Magnetic starch microspheres, biodistribution and biotransformation. A new organ-specific contrast agent for magnetic resonance imaging.

The biodistribution and elimination of magnetic starch microspheres (MSM) were studied qualitatively and quantitatively by radioiron tracer studies and relaxation time measurements. One hour after injection of MSM (1 mg/kg of Fe), 85% +/- 5% of the dose was accumulated in the liver and 6.5% +/- 1.3% [corrected] in the spleen. The hepatic clearance led to 50% reduction in the T2 relaxation time of liver tissue. This T2 effect was halved after 24 hours and T2 reversed to baseline value within 5 days after injection. The radioiron was gradually cleared from the liver with a t1/2 of 4 to 5 days. Six weeks after injection of MSM, 72% +/- 7% of the radioiron dose was detected in the circulation in a nonsuperparamagnetic form associated with the erythrocytes. The results indicate a redistribution of iron from the liver and spleen via the erythroid bone marrow to the erythrocytes after injection of MSM.