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Biomedical subjects

D J Crommelin

Publications and source records attributed to D J Crommelin.

At least 19 recordsLinked to original sources
PubMed1995

Physical (in) stability of liposomes upon chemical hydrolysis: the role of lysophospholipids and fatty acids.

As a consequence of chemical hydrolysis of liposomal phospholipids the organization of the lipid assembly can change from a lamellar into a micellar system. Different approaches provided evidence for this conversion: 31P-NMR analysis, turbidity measurements and ultracentrifugation experiments. Two conditions have to be met before this conversion can take place: (1) the liposomes must pass through a gel-to-liquid crystalline phase-transition during a heating or cooling run, and (2) the degree of chemical hydrolysis must exceed a critical hydrolysis percentage (or the phospholipid bilayer must contain critical amounts of lysophospholipid and fatty acid). As monitored by turbidity measurements, this critical level of hydrolysis and the relative change depended on the chain length and on the head group of the liposomal phospholipids. It does not depend on concentration, pH, storage temperature or on size of the liposomes within the experimental range. Addition of cholesterol to bilayers composed of dipalmitoylphosphatidylcholine prevents the lamellar to micellar transformation. Fluorescence anisotropy measurements of the lipophilic probe 1,6-diphenyl-1,3,5-hexatriene in 0.18-microns dipalmitoylphosphatidylcholine/dipalmitoylphosphatidylglycerol (10:1)-liposomes indicated that behavior of the probe below and above the phase-transition temperature was not affected by chemical hydrolysis, or even by formation of micelles. However, the phase-transition temperature range broadened and shifted towards higher temperatures upon hydrolysis.

Fatty Acids↗
PubMed1995

Adhesion molecules: a new target for immunoliposome-mediated drug delivery.

The anti-ICAM-1 monoclonal antibody F10.2 was conjugated to liposomes to target to cells expressing the cell adhesion molecule ICAM-1. We demonstrate that F10.2 immunoliposomes bind to human bronchial epithelial cells (BEAS-2B) and human umbilical vein endothelial cells (HUVEC) in a specific, dose- and time-dependent manner. It appears that the degree of ICAM-1 expression is the limiting factor in the degree of immunoliposome binding to the cells. These results are a first step in the strategy for specific drug delivery to target sites characterised by increased expression of adhesion molecules.

Antibodies, Monoclonal↗
PubMed1995

A mechanistic study of the effects of the 1-alkyl-2-pyrrolidones on bilayer permeability of stratum corneum lipid liposomes: a comparison with hairless mouse skin studies.

The influence of a series of 1-alkyl-2-pyrrolidones (C2-C8) on the transport behavior of lipophilic and polar/ionic permeants across hairless mouse skin was recently investigated by employing a physical model approach that treats the stratum corneum barrier as a diffusional system of parallel lipoidal and pore pathways. In this previous study, the transport enhancement effects (enhancement factor, EHMS) on the lipoidal pathway of the stratum corneum were found to be essentially the same for all steroidal probe permeants investigated at various concentrations of these 1-alkyl-2-pyrrolidones. In the present research, the relationship between solute transport enhancement in the lipoidal pathway of hairless mouse skin and the transport enhancement in the stratum corneum lipid liposome bilayer was studied by comparing the enhancement factor for the lipoidal pathway in the hairless mouse skin, EHMS, with that for the stratum corneum lipid liposome, ESCLL, at equal solution concentrations of the 1-alkyl-2-pyrrolidones. The release rates of D-mannitol, D-glucose, 3-O-methyl-D-glucose, sucrose, and raffinose from stratum corneum lipid liposomes were determined, and the ESCLL values for these permeants were compared with the EHMS values obtained with hairless mouse skin using the steroidal permeants. An important finding in this study was a semiquantitative correlation between the enhancement effects induced by the 1-alkyl-2-pyrrolidones, except 1-ethyl-2-pyrrolidone, with the liposome bilayer using sugar molecules as permeants and those found with the lipoidal pathway in hairless mouse skin using steroid molecules as permeants.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Chemical hydrolysis of phospholipids.

Hydrolysis kinetics of phospholipids in liposomes composed of dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine/cholesterol (DPPC/CHOL) 10/4 (molar ratio) and egg phosphatidylcholine (EPC) at pH 4.0 and different temperatures could be described by Arrhenius curves without breaks. However, the Arrhenius curves for the hydrolysis of liposomal DPPC and distearoylphosphatidylcholine (DSPC) under the same conditions were biphasic. A break was observed in the curves extending over a broad range before and after the known Tm of each of these phospholipids in liposomes (42 and 56 degrees C, respectively). The activation energy (Ea) for the hydrolysis of liposomal DPPC and DSPC below the Tm was substantially larger than the Ea for liposomal DMPC, DPPC/CHOL 10/4, and EPC and decreased when DPPC was mixed with CHOL in a 10/4 molar ratio. Hardly any influence of the presence of alpha-tocopherol, cryoprotectants (glucose, trehalose, sucrose, and propylene glycol), and the major hydrolysis products lysophospholipids and fatty acids or of the absence of sodium chloride on the hydrolysis kinetics of DPPC at pH 4.0 and 30 degrees C was observed. Changes in fatty acid chains and size did not influence the hydrolysis rate constant (kobs) of liposomal phospholipids at pH 4.0 and 30 degrees C either. The only effects of uncharged compounds on the kobs of liposomal DPPC at pH 4.0 and 30 degrees C were found upon mixing with a high concentration of the detergent Triton X-100 or palmitic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Cryoprotective Agents↗
PubMed1995

Comparison of 24 different adjuvants for inactivated HIV-2 split whole virus as antigen in mice. Induction of titres of binding antibodies and toxicity of the formulations.

The objective of this study was the comparison of the quantity and quality of the humoral immune response against inactivated whole human immunodeficiency virus type 2. Twenty-four different adjuvants were tested. The toxicity of these preparations was determined. Due to the large number of experiments, the animal model used was the NMRI mouse. Sera were assayed for the presence of antibodies by ELISA and Western blot. The toxicity of the adjuvants was determined by observing lethal side-effects occurring within two days after injection of the immunogenic preparations. The results show that polymethylmethacrylate nanoprticles were the best overall adjuvant, inducing the highest titres of antibodies compared with other adjuvants as well as producing a significantly better immune response against a number of individual HIV-2 antigens without any observable toxic side-effects. However, this adjuvant did not induce antibodies against the outer envelope protein gp120, whereas such antibodies were induced by aluminium compounds, Freund's complete and incomplete adjuvants, and fumed silica (Aerosil). This result suggests the necessity of using vaccines with two or more different adjuvants in order to induce the required immune response against physically different antigens.

AIDS Vaccines↗
PubMed1995

Sterically stabilized liposomes labeled with indium-111 to image focal infection.

UNLABELLED: To determine the potential of sterically stabilized liposomes to image infectious and inflammatory foci, the in vivo behavior of 111In-labeled PEGylated (coated with polyethylene glycol) liposomes was studied in a rat model. METHODS: Indium-111-PEGylated lipsomes were administered intravenously to rats infected with S. aureus in the left calf muscle. The distribution of the radiolabel was studied by gamma counting of dissected tissues and gamma camera imaging for 48 hr. As a reference agent, the preparation of 111In-IgG was included in these studies. RESULTS: Clearance of the PEGylated liposomes from the blood compartment was similar to the clearance of 111In-IgG in this model (t1/2 approximately 20 hr). Uptake of the radiolabel in the abscess with the 111In-liposomes was twice as high as the uptake following injection of 111In-IgG (2.7%ID/g versus 1.1%ID/g at 48 hr postinjection). Tissue counting revealed that abscess-to-muscle ratios reached values up to 20 and 34 (24 and 48 p.i., respectively). As early as 1 hr postinjection, the abscess could be visualized scintigraphically. CONCLUSION: The in vivo characteristics of this liposomal formulation in this rat model indicate that sterically stabilized liposomes labeled with gamma emitters might be a valuable addition to the arsenal of radiopharmaceuticals currently used for infection imaging.

Abscess↗
PubMed1995

Thrombolytic treatment with tissue-type plasminogen activator (t-PA) containing liposomes in rabbits: a comparison with free t-PA.

In this study, we aimed at improving the therapeutic index of tissue-type Plasminogen Activator (t-PA) as thrombolytic agent in the treatment of myocardial infarction. Liposome-encapsulated t-PA was tested in a rabbit jugular vein thrombosis model: administration of free t-PA (t-PA) as a bolus injection in the ear vein was compared to a similar administration of liposomal t-PA (t-PA-lip), liposomal t-PA in plasminogen-coated liposomes (Plg-t-PA-lip), a mixture of free t-PA and empty liposomes (t-PA+ empty lip) and a saline-blank (blank) in terms of thrombolytic activity and side effects. Liposomal t-PA (t-PA-lip/Plg-t-PA-lip) showed a significantly better thrombolysis efficiency than equimolar doses of free t-PA (t-PA/ t-PA+ empty lip): about 0.24 mg/kg of liposomal t-PA practically equalled the lysis-activity of a dose of free t-PA of 1.0 mg/kg (t-PA1mg/kg). On the other hand, liposome encapsulation did not affect the systemic activation of alpha 2-antiplasmin and plasminogen by t-PA. We conclude that for this model an improvement in thrombolytic efficacy of t-PA is achieved by liposome encapsulation of t-PA. As t-PA-lip and Plg-t-PA-lip -treatment induced similar results, targeting of liposomal t-PA by coupled glu-Plg remains a topic to be optimized in future studies.

Animals↗
PubMed1994

Effect of immunomodulators on specific tumor immunity induced by liposome-encapsulated tumor-associated antigens.

Reconstituted membranes consist of liposomal structures formed by removal of detergent from solubilized membrane constituents. The membrane-like configuration of reconstituted membranes makes them attractive as vehicles for presentation of tumor-associated antigens and induction of immune responses. In this study the potential of immunomodulators was assessed to enhance the specific immune response induced by immunization with reconstituted membranes prepared from SL2 lymphosarcoma cells. Reconstituted membranes containing muramyl tripeptide phosphatidylethanolamine (MTP-PE) provided better protection against a challenge with SL2 cells than did reconstituted membranes containing alternative immunomodulators. Local administration of IL-2 at the immunization sites further augmented the protection induced by reconstituted membranes with MTP-PE, but was ineffective when administered with plain reconstituted membranes. Immunity elicited by the triple modality of reconstituted SL2 membranes with MTP-PE and IL-2 was specific for SL2 cells. Systemic immunity was obtained against a challenge with a 100-fold higher number of SL2 cells than was reached after immunization with reconstituted membranes alone (10(5) vs. 10(3) SL2 cells). Macrophages isolated from the peritoneal cavity of immunized mice 5 to 7 days after tumor challenge expressed high in vitro cytotoxicity. However, in contrast to the observed specificity of the systemic immunity, macrophages killed both SL2 cells and non-related P815 cells. Neither major cytotoxic lymphocyte activity nor substantial cytotoxic antibody titers were detectable. These results clearly indicate that the approach using reconstituted membranes combined with particular immunomodulators warrants further exploration for the development of safe, well-characterized cancer vaccines.

Adjuvants, Immunologic↗
PubMed1994

A novel technique for the one-step preparation of liposomes and nonionic surfactant vesicles without the use of organic solvents. Liposome formation in a continuous gas stream: the 'bubble' method.

A novel technique for the preparation of liposomes without the use of organic solvents, detergents, or high shear forces is described. Introduction of inert gas bubbles into a coarse dispersion of nonhydrated (phospho)lipids (hydrogenated soybean phosphatidylcholine and/or dicetylphosphate as charge-inducing agent) or nonionic surfactant (C18EO3)/cholesterol mixtures produces stable liposome/niosome dispersions with a mean particle size in the range between 0.2 and 0.5 microns. The influence of addition of cholesterol and the effect of the ionic strength of the aqueous medium on the preparation process of liposomes was investigated. An hypothesis for the mechanism of liposome formation under the chosen conditions is presented and discussed.

Cholesterol↗
PubMed1994

Dissolution kinetics of griseofulvin in mixed micellar solutions.

The dissolution kinetics of the poorly water soluble, lipophilic model drug substance griseofulvin were studied in solutions containing simple and mixed micelles of sodium cholate (BS) and phosphatidylcholine (PC). Variables under investigation were the concentration of BS and the ratio PC/BS. A rotating disk apparatus was used to monitor the dissolution process. With this method, (mixed) micellar diffusion coefficients could be calculated and some detailed information about the physicochemical behavior and composition of mixed PC/BS micelles could be derived.

Bile Acids and Salts↗
PubMed1994

Immunoliposomes in vivo.

Attachment of antibodies to the surface of liposomes was performed to confer specificity for a certain cell or organ expressing the targeted antigenic determinant. These so-called immunoliposomes are expected to be applied as targeted drug carriers. In this article, the literature concerning in vivo studies of the targeting of immunoliposomes to various sites in the body is reviewed. The anatomical, physiological, and pathological constraints and current progress are described. Moreover, perspectives on the therapeutic feasibility of this drug targeting system are discussed.

Amphotericin B↗
PubMed1993

A new application for liposomes in cancer therapy. Immunoliposomes bearing enzymes (immuno-enzymosomes) for site-specific activation of prodrugs.

We have tested a new type of immunoliposomes which may effectively mediate the targeting of enzymes to be used for site-specific prodrug activation (immuno-enzymosomes). The enzyme beta-glucuronidase, capable of activating the prodrug epirubicin-glucuronide (epi-glu), was coupled to the external surface of immunoliposomes directed towards ovarian cancer cells. A significant increase in cytotoxicity of the prodrug epi-glu was shown when the in vitro cultured cancer cells were pretreated with these immuno-enzymosomes.

Antibodies, Monoclonal↗
PubMed1993

The role of protein charge in protein-lipid interactions. pH-dependent changes of the electrophoretic mobility of liposomes through adsorption of water-soluble, globular proteins.

The role of electrostatics in the adsorption process of proteins to preformed negatively-charged (phosphatidylcholine/phosphatidylglycerol) and neutral (phosphatidylcholine) liposomes was studied. The interaction was monitored at low ionic strength for a set of model proteins as a function of pH. The adsorption behavior of trypsin inhibitor (pI = 4.6), myoglobin (pI = 7.4), ribonuclease (pI = 9.6), and lysozyme (pI = 10.7) with preformed liposomes was investigated, along with changes in the electrophoretic mobility of liposomes through the adsorption of charged proteins. Mean protein charge was determined by acid/base titration. Significant adsorption of the proteins to negatively-charged liposomes was only found at pH values where the number of positive charge moieties exceeds the number of negative charge moieties on the protein by at least three charge units. Negligible adsorption to liposomes composed of zwitterionic lipids was observed in the pH range tested (4-9). The absolute value of the electrophoretic mobilities of negatively-charged, empty liposomes decreased after adsorption of positively-charged proteins. With increasing protein to phospholipid ratio, the drop in the electrophoretic mobility leveled off and reached a plateau; protein adsorption profiles showed a similar shape. Analysis of the data demonstrated that neutralization of the liposome charge due to the adsorption of the positively-charged proteins is the controlling factor in their adsorption. The plateau level reached depended on the type of protein and the pH of the incubation medium. This pH dependency could be ascribed to the mean positive charge of the protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Adsorption↗
PubMed1993

The effect of surface charge on the hydrolysis kinetics of partially hydrogenated egg phosphatidylcholine and egg phosphatidylglycerol in aqueous liposome dispersions.

The hydrolysis kinetics of partially hydrogenated egg phosphatidylcholine (PHEPC) and egg phosphatidylglycerol (EPG) were investigated as a function of surface pH at 60 degrees C. The bulk pH of the liposome dispersions was varied from pH 3.0 to 8.0. Charge was introduced to the liposomal bilayers by addition of EPG. Different electrostatic profiles were obtained at the bilayer-water interface by varying the surface charge density and the ionic strength of the buffer solutions. With the Gouy-Chapman equation the surface potential and with the Boltzmann equation the resulting shift in the surface pH compared to the bulk pH was estimated. The hydrolysis rate of both phospholipids increased with surface charge density in acidic medium; in alkaline medium the opposite was true. The hydrolysis rate constants, however, plotted as a function of surface pH agreed reasonably well with the results found with neutral liposomes.

Hydrogen-Ion Concentration↗
PubMed1993

Hydrolysis of saturated soybean phosphatidylcholine in aqueous liposome dispersions.

Hydrolysis kinetics of saturated soybean phosphatidylcholine (PC) were investigated as a function of pH, temperature, buffer concentration, buffer species, and ionic strength in aqueous liposome dispersions. A high-performance liquid chromatography method was used to separate PC and the hydrolysis products. Hydrolysis of PC followed pseudo first-order kinetics. Hydrolysis of fatty acid esters at sn-1 and sn-2 positions initially resulted in the formation of 2-acyl lyso-phosphatidylcholine and 1-acyl lyso-phosphatidylcholine; because of acyl migration, the most stable isomer of lyso-phosphatidylcholine, 1-acyl lyso-phosphatidylcholine, was formed predominantly. General acid-base catalysis was observed for acetate and Tris ions. The pH profiles at 40 and 70 degrees C for a buffer concentration of 0.05 M showed a minimum hydrolysis rate at about pH 6.5. The relationship between the observed rate constant and temperature could be described adequately by the Arrhenius equation below and above the phase transition temperature of saturated soybean PC (52 degrees C).

Buffers↗
PubMed1993

Critical factors for liposome-incorporated tumour-associated antigens to induce protective tumour immunity to SL2 lymphoma cells in mice.

Physical and immunogenic properties of reconstituted membranes designed for the presentation of tumour-associated antigens (TAA) to the immune system are described. Proteins and lipids of crude membranes of SL2 murine lymphosarcoma cells were partially solubilized with octylglucoside. Reconstituted membranes, consisting mainly of unilamellar vesicles with a diameter of 0.03-0.15 microns, were formed by detergent removal and were purified by floatation in a discontinuous sucrose gradient to remove non-lipid-bound protein. Subcutaneous immunization of syngeneic mice with reconstituted membranes or with purified reconstituted membranes induced protection against an intraperitoneal challenge with 10(3) viable SL2 cells. Reconstituted membranes were more immunogenic than crude membranes in immunoprotection experiments when compared on the basis of protein dose. Detergent removal was required to obtain an immunogenic presentation form of SL2 membrane antigens and to avoid toxicity associated with the detergent. Reconstitution of SL2 membranes in the presence of exogenous phospholipid slightly increased the fraction of protein that associated with the reconstituted membranes. However, the immunogenicity of the solubilized membrane TAA was not significantly affected by the presence of exogenous phospholipid. The reconstitution procedure described may be useful in identifying membrane factors required for the induction of immune responses against TAA. The versatility of the system may be employed to develop safe alternatives for whole-cell vaccines.

Animals↗
PubMed1993

Chemical stability of liposomes: implications for their physical stability.

In the first part of this article, chemical and physical stability of aqueous liposome dispersions have been addressed. Chemical stability of phospholipids has been considered in two parts: oxidation and hydrolysis. Major attention has been paid to hydrolysis kinetics of phospholipids as a function of pH, temperature, buffer concentration and ionic strength. Furthermore, the effect of chain length, head group, state of aggregation, addition of cholesterol and presence of charge on the hydrolysis kinetics of phospholipids has been dealt with. In the second part physical stability of chemically degraded liposome dispersions has been evaluated. In the final part quality control assays for liposome dispersions is presented and a HPLC method with a refractive index detector for the analysis of phospholipids from aqueous liposome dispersions is described.

Drug Stability↗