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Biomedical subjects

P Couvreur

Publications and source records attributed to P Couvreur.

At least 37 records · Page 2Linked to original sources

II. Glutamine and glutamate.

Glutamine and glutamate with proline, histidine, arginine and ornithine, comprise 25% of the dietary amino acid intake and constitute the "glutamate family" of amino acids, which are disposed of through conversion to glutamate. Although glutamine has been classified as a nonessential amino acid, in major trauma, major surgery, sepsis, bone marrow transplantation, intense chemotherapy and radiotherapy, when its consumption exceeds its synthesis, it becomes a conditionally essential amino acid. In mammals the physiological levels of glutamine is 650 micromol/l and it is one of the most important substrate for ammoniagenesis in the gut and in the kidney due to its important role in the regulation of acid-base homeostasis. In cells, glutamine is a key link between carbon metabolism of carbohydrates and proteins and plays an important role in the growth of fibroblasts, lymphocytes and enterocytes. It improves nitrogen balance and preserves the concentration of glutamine in skeletal muscle. Deamidation of glutamine via glutaminase produces glutamate a precursor of gamma-amino butyric acid, a neurotransmission inhibitor. L-Glutamic acid is a ubiquitous amino acid present in many foods either in free form or in peptides and proteins. Animal protein may contain from 11 to 22% and plants protein as much as 40% glutamate by weight. The sodium salt of glutamic acid is added to several foods to enhance flavor. L-Glutamate is the most abundant free amino acid in brain and it is the major excitatory neurotransmitter of the vertebrate central nervous system. Most free L-glutamic acid in brain is derived from local synthesis from L-glutamine and Kreb's cycle intermediates. It clearly plays an important role in neuronal differentiation, migration and survival in the developing brain via facilitated Ca++ transport. Glutamate also plays a critical role in synaptic maintenance and plasticity. It contributes to learning and memory through use-dependent changes in synaptic efficacy and plays a role in the formation and function of the cytoskeleton. Glutamine via glutamate is converted to alpha-ketoglutarate, an integral component of the citric acid cycle. It is a component of the antioxidant glutathione and of the polyglutamated folic acid. The cyclization of glutamate produces proline, an amino acid important for synthesis of collagen and connective tissue. Our aim here is to review on some amino acids with high functional priority such as glutamine and to define their effective activity in human health and pathologies.

Animals↗

[Oral tolerance induced by poly (lactide-co-glycolide) containing B lactoglobulin].

Allergies to milk proteins are frequently encountered in the new born population. In order to prevent this allergy by inducing oral tolerance, one of the major allergenic milk protein, B lactoglobulin (BLG) was entrapped into biodegradable Poly (lactide-co-glycolide) microspheres and was then orally given to mice. Microspheres are able to protect proteins against degradation by intestinal proteolytic enzymes and to target the Peyers patches which are one important priming site of the mucosal immune system. Microspheres were prepared by the multiple emulsion solvent evaporation method. The goal of the formulation study was to associate large amounts of proteins to the smallest amount of polymer so that a minimal quantity of microspheres would be administered. It was shown that introducing tween 20 in the formulation was able to increase the encapsulation efficiency and to better control protein release reducing the burst release effect. Moreover, Oral administration of microspheres containing BLG reduced significantly (by 10.000) the amount of protein necessary to decrease both specific anti BLG IgE and DTH response. In conclusion, microspheres appear to be optimal systems to induce oral tolerance.

Drug Carriers↗

PEGylated polycyanoacrylate nanoparticles as vector for drug delivery in prion diseases.

PEGylated polymeric nanoparticles are hereby presented as a potential efficient drug carrier for the delivery of active therapeutic molecules in prion experimental diseases. Based on their blood long-circulating characteristics, these PEGylated particles made by the amphiphilic copolymer poly [methoxy poly(ethylene glycol) cyanoacrylate-co-hexadecyl cyanoacrylate] (PEG-PHDCA), showed comparatively conventional non-PEGylated nanoparticles, a higher uptake by the spleen and the brain which are both the target tissues of PrPres accumulation in scrapie infected animals.

Animals↗

Characterization of oligonucleotide/lipid interactions in submicron cationic emulsions: influence of the cationic lipid structure and the presence of PEG-lipids.

We have recently described how oligonucleotide (ON) stability and release from O/W cationic emulsions are governed by the lipid composition. The aim of the present paper was to investigate the properties of the ON/lipid complexes through fluorescence resonance energy transfer (FRET), size, surface tension measurements and cryomicroscopy. Starting from a typical emulsion containing stearylamine as a cationic lipid, the influence of the lipid structure (monocationic molecules bearing mono or diacyl chains, or polycations) as well as of the presence of PEGylated lipids, were studied. The presence of a positive charge on the droplet surface clearly contributed to enhance the ON interaction with lipid monolayers and to bring the ON molecules closer to the interface. Hydrophobic interactions through the acyl chains were shown to further enhance the anchorage of the ON/lipid complexes. In contrast, the incorporation of PEGylated lipids acted as a barrier against the establishment of electrostatic bindings, the polyethyleneglycol chains acting themselves as interaction sites for the ON leading to hydrophilic complexes. Similar features were observed for the polycationic lipid, and cryomicroscopy revealed the existence of bridges of various intensities between the droplets of the emulsion containing either PEG or the polycation, probably because of the configuration of the ON at the interface.

Algorithms↗

Nanoparticulate systems for the delivery of antisense oligonucleotides.

Antisense oligonucleotides are molecules that are able to inhibit gene expression being therefore potentially active for the treatment of viral infections or cancer. However, because of their poor stability in biological medium and their weak intracellular penetration, colloidal drugs carriers such as nanoparticles were developed for the delivery of oligonucleotides (ODN). ODN associated to nanoparticles were shown to be protected against degradation and to penetrate more easily into different types of cells. As a consequence, nanoparticles were shown to improve the efficiency of ODNs for the inhibition of the proliferation of cells expressing the point mutated Ha-ras gene. In vivo, polyalkylcyanoacrylate (PACA) nanoparticles were able to efficiently distribute the ODNs to the liver whereas the alginate nanosponges could concentrate the ODNs in the lungs. Finally, ODN loaded to PACA nanoparticles were able to improve in mice, the treatment of RAS cells expressing the point mutated Ha-ras gene.

Animals↗

Polyisobutylcyanoacrylate nanocapsules containing an aqueous core for the delivery of oligonucleotides.

Antisense oligonucleotides (ODNs) with base sequences complementary to a specific RNA can, after binding to intracellular mRNA, selectively modulate the expression of a gene. However, these molecules are poorly stable in biological fluids and are characterized by a low intracellular penetration. In view of using ODNs as active molecules, the development of nanocapsules containing ODNs in their aqueous core was considered. Nanocapsules were prepared by interfacial polymerization of isobutylcyanoacrylate (IBCA) in a W/O emulsion. After ultracentrifugation and re-suspension in water, the nanocapsules displayed a size of 350+/-100 nm. Oligonucleotide loading did not significantly influence the zeta potential, suggesting that they were located within the core of the nanocapsules. Fluorescence quenching assays confirmed this localization. When encapsulated in the nanocapsules and incubated in the presence of serum, the ODNs were efficiently protected from degradation by nucleases, whereas ODNs adsorbed onto nanospheres were less efficiently protected. This paper describes, for the first time, a nanotechnology able to encapsulate ODNs, rather than adsorbing them at the surface of a solid support. Such a formulation has great potential for oligonucleotide delivery.

Cyanoacrylates↗

Tamoxifen encapsulation within polyethylene glycol-coated nanospheres. A new antiestrogen formulation.

When dealing with solid tumors in vivo, pegylated long-circulating carrier systems show, after intravenous administration, an attractive extravasation profile with an enhanced localization in the tumoral interstitium. These systems could be of help for the delivery of cancer fighting drugs, such as Tamoxifen, a well known antiestrogen used in breast cancer therapy that possesses an extended biodistribution in vivo. This work aimed at encapsulating Tamoxifen in long-circulating poly(MePEGcyanoacrylate-co-hexadecylcyanoacrylate) 1:4 nanospheres. Tamoxifen-loaded poly(MePEGcyanoacrylate-co-hexadecylcyanoacrylate) nanospheres were successfully synthesized and characterized in terms of hydrophilicity/hydrophobicity by a model made up from near infrared spectra using principal component analysis. Zeta potential, drug loading, encapsulation efficiency, as well as biological effect, in vitro release and nanospheres integrity were also investigated. Even though near infrared spectroscopy could not detect Tamoxifen, it revealed that Pluronic F68 was associated with the pegylated nanospheres. HPLC measurements demonstrated that Tamoxifen was encapsulated in the pegylated nanospheres following a partition equilibrium between the polymeric and the aqueous phases. The Tamoxifen encapsulated in the nanospheres still showed a transcription inhibitory activity in ex vivo experiments. However, zeta potential and in vitro release suggested that Tamoxifen was essentially localized at the nanoparticles surface, resulting in an important and immediate drug release.

Drug Delivery Systems↗

Design of triptorelin loaded nanospheres for transdermal iontophoretic administration.

Triptorelin is a decapeptide analog of luteinizing hormone releasing hormone, currently used for the treatment of sex-hormones dependents diseases. The aim of this work was to prepare triptorelin-loaded nanospheres useful for transdermal iontophoretic administration. Nanospheres were prepared with the double emulsion/solvent evaporation technique. The effect of three parameters on the encapsulation efficiency has been determined: the role of the pH of the internal and external aqueous phases, the nature of the organic solvent and the effect of three different poly(lactide-co-glycolide) (PLGA) co-polymers. Particle size, zeta potential and release kinetics were also determined. The encapsulation efficiency varied from 4 to 83% reaching the maximum value when both the internal and the external water phases were brought to pH 7 (isoelectric point of the peptide), methylene chloride was used as solvent of the copolymers and PLGA rich in free carboxylic groups was employed. The release profiles obtained with this co-polymer were characterized by the absence of burst effect. This behavior as well as the high encapsulation efficiency was explained by an ionic interaction occurring between the peptide and the co-polymer. This supports the already expressed theory that the release of peptides and proteins from PLGA nanospheres is also governed by the affinity of the encapsulated molecule versus the polymer. The obtained nanoparticles, regarding their size, amount encapsulated and zeta potential, were shown to be suitable for transdermal iontophoretic administration.

Administration, Cutaneous↗

Factors influencing the oligonucleotides release from O-W submicron cationic emulsions.

We recently described a positively charged O-W emulsion as a delivery system for oligonucleotides (ON) [Teixeira et al., Pharm. Res. 16 (1999) 30-36]. The present paper investigates the role of the main formulation parameters that may have an influence on the release-rate of a model ON in a protein-containing medium, i.e. the nature of the oily core, the presence of pegylated lipids, the lipid phase transition temperature, and the cationic lipid structure. The use of cationic lipids bearing diacyl chains (and especially polycations) appeared as the only efficient strategy to reduce the ON release rate. In order to have a better insight on the nature of the interactions between the ON and the interfacial lipids, adsorption isotherms at the air-water interface, fluorescence resonance energy transfer and zeta-potential measurements have been performed. Electrostatic interactions were found to play a crucial role. In contrast, the incorporation of PEG-phospholipids acted as a barrier and maintained the ON molecules distant from the interface, leading to a more rapid release. Finally, ON integrity was assessed by a competitive hybridization assay. The results suggest the existence of a transient ion-pair (ON-cationic lipids) protecting ON against nuclease degradation even after its release from the emulsions.

Cells, Cultured↗

Novel microparticulate system made of poly(methylidene malonate 2.1.2).

Formulation of PMM 2.1.2 microparticles entrapping ovalbumin as a model protein was achieved by using a double emulsion solvent evaporation method. Parameters such as the nature of the solvent, polymer concentration and polymer molecular weight were investigated. Preparation process led to the formation of spherical and smooth particles with a mean diameter of 5 microm, and an encapsulation efficiency and protein loading level of up to 16 and 2.9% w/w, respectively. After an initial burst of approximately 10%, the protein was released at a rate of less than 1% per day. This slow release kinetics of encapsulated ovalbumin in phosphate buffer indicates that most of the protein was encapsulated within the polymer matrix. Degradation of PMM 2.1.2 microparticles in the presence of esterases indicated that side chain hydrolysis of the polymer was the rate-determining step in bioerosion; cleavage of the ester side chain, which was further hydrolyzed to glycolic acid and ethanol, led to an acrylic acid and subsequent solubilization of the polymer. However, slow polymer backbone solubilization after degradation was observed.

Biocompatible Materials↗

Gliding resistance after repair of partially lacerated human flexor digitorum profundus tendon in vitro.

OBJECTIVE: This study reports the gliding resistance between repaired, partially lacerated tendon and pulley in human cadaver digits, using several commonly employed repair techniques. BACKGROUND: Suture techniques with multi-strands and locking loops have been recommended to reduce the risk of rupture of the repair tendon with early active motion. Such sutures may increase the gliding resistance, and the gliding resistance after tendon repair is also an important factor influencing the rehabilitation. METHOD: 105 specimens of second, third, or fourth fingers from 36 adult human hands were tested for the gliding resistance between flexor digitorum profundus tendon and A2 pulley in the normal condition. After an 80% laceration, each tendon was repaired with one of the following suture techniques: (1) Kessler; (2) modified Kessler; (3) Savage; (4) Lee; (5) Tsuge; and (6) Becker. All suture techniques were reinforced with a circumferential epitenon simple running suture. After tendon repair, the gliding resistance was remeasured. RESULTS: The gliding resistance of the Becker repair was significantly greater than each of the other four repairs (P<0.05). The resistance of the modified Kessler repair was significantly less than that of the Kessler, Savage, or Tsuge repairs. CONCLUSIONS: We conclude that the type of tendon repair can significantly affect the gliding resistance between the tendon and pulley system after tendon repair. RELEVANCE: The design of the tendon repair, through its effect on friction, may have an adverse effect on the clinical results of tendon mobilization.

Aged↗

Long-circulating PEGylated polycyanoacrylate nanoparticles as new drug carrier for brain delivery.

PURPOSE: The aim of this study was to evaluate the ability of long-circulating PEGylated cyanoacrylate nanoparticles to diffuse into the brain tissue. METHODS: Biodistribution profiles and brain concentrations of [14C]-radiolabeled PEG-PHDCA, polysorbate 80 or poloxamine 908-coated PHDCA nanoparticles, and uncoated PHDCA nanoparticles were determined by radioactivity counting after intravenous administration in mice and rats. In addition, the integrity of the blood-brain barrier (BBB) after nanoparticles administration was evaluated by in vivo quantification of the diffusion of [14C]-sucrose into the brain. The location of fluorescent nanoparticles in the brain was also investigated by epi-fluorescent microscopy. RESULTS: Based on their long-circulating characteristics, PEGylated PHDCA nanoparticles penetrated into the brain to a larger extent than all the other tested formulations. Particles were localized in the ependymal cells of the choroid plexuses, in the epithelial cells of pia mater and ventricles, and to a lower extent in the capillary endothelial cells of BBB. These phenomena occurred without any modification of BBB permeability whereas polysorbate 80-coated nanoparticles owed, in part, their efficacy to BBB permeabilization induced by the surfactant. Poloxamine 908-coated nanoparticles failed to increase brain concentration probably because of their inability to interact with cells. CONCLUSIONS: This study proposes PEGylated poly (cyanoacrylate) nanoparticles as a new brain delivery system and highlights two requirements to design adequate delivery systems for such a purpose: a) long-circulating properties of the carrier, and b) appropriate surface characteristics to allow interactions with BBB endothelial cells.

Acrylic Resins↗

Study of the breakup under shear of a new thermally reversible water-in-oil-in-water (W/O/W) multiple emulsion.

PURPOSE: Thickening of the external aqueous phase of W/O/W multiple emulsions is essential to increase the release under shear. However, it leads to globules bursting during fabrication. To reduce this problem, we have tested a novel thermally reversible hydrogel, EMP hydrogel. This way, the corresponding multiple emulsion (EMPME) would gel only at skin temperature, which may increase the active ingredient delivery when topically applied. METHODS: Samples were sheared at different shear rates and temperatures (20, 30, and 35 degrees C) with a controlled rheometer. A granulometric analysis was then performed with a laser diffraction granulometer, to assess the break up as a function of the shear rate at the three temperatures. Conductometric measurements (CDM 230 conductometer) provided the corresponding release curves. RESULTS: As we expected, EMPME exhibited a thermally reversible behavior. Compared to a reference emulsion thickened by carbopol, this new thermo-sensitive multiple emulsion displayed higher break up and fraction released at 35 degrees C. CONCLUSION: The first thermally reversible multiple emulsion has been developed in the present work. This one presents interesting advantages: (1) an easy fabrication process with a higher entrapment yield and (2) a higher fraction released at 35 degrees C compared with the reference emulsion.

Acrylic Resins↗

The effect of suture technique on adhesion formation after flexor tendon repair for partial lacerations in a canine model.

BACKGROUND: Adhesion formation is a serious problem after flexor tendon repair. Many repair techniques have been developed to increase the suture strength after tendon repair surgery. The purpose of this study was to assess adhesion formation with different suture techniques in an in vivo canine model. METHODS: Sixty flexor digitorum profundus tendons were partially lacerated (80%) and repaired with either a modified Kessler (MK) or Becker (MGH) suture technique and supplemented with a simple running suture. The dogs were sacrificed at 1 week, 3 weeks, or 6 weeks after surgery and the repaired tendons were evaluated for adhesion breaking strength. RESULTS: At 1 week there was no significant difference between the two repair groups (p > 0.05). At 3 and 6 weeks, the adhesion breaking strength in the MK suture group was significantly less than that of the MGH suture group (p < 0.05). CONCLUSION: High friction suture techniques may cause more adhesion formation than the lower friction suture techniques under passive postoperative therapy.

Analysis of Variance↗

In vitro and in vivo evaluation of poly(methylidene malonate 2.1.2) microparticles behavior for oral administration.

The goal of this paper was to investigate the fate of novel poly(methylidene malonate 2.1.2) microparticles with different surface properties, i.e. prepared with or without polyvinylalcohol (PVA), after oral administration, using in vitro cell culture and an in vivo mice model. Incubation of particles with Caco-2 cells induced no cytotoxicity except for the microparticles prepared without PVA at high concentrations. At subtoxic concentrations, microparticles were highly associated to cells, independently of particles concentrations, particles surface properties (with or without PVA) or incubation time. Confocal microscopy analysis revealed that adsorption was the main phenomenon leading to the association of particles to cells. However, association was greater at 37 degrees C than at 4 degrees C, suggesting that an active process, such as endocytosis, could also occur. In vivo, radiolabeled particles were mainly found in luminal content and also adsorbed onto the epithelium. After 24 hours, more than 15% of PVA-free microparticles were still present in the gastrointestinal tract, compared to 5% for particles prepared with PVA. However, histological evaluation revealed low uptake of particles by Peyer's patches. As a conclusion, this study provided a good correlation between in vitro and in vivo evaluation. These particles could be useful for oral sustained release and delivery of drugs to intestinal and colon epithelium.

Administration, Oral↗

[Drug vectorization or how to modulate tissular and cellular distribution of biologically active compounds].

Drug vectorization has undergone considerable development over the last few years. This review focuses on the intravenous route of administration. Colloid formulations allow a modulation of drug tissue distribution. Using liposomes and nanoparticles with unmodified surfaces, drugs can be targeted to macrophages of the reticulum endothelium system. When the liposomes or nanoparticles are covered with hydrophilic or flexible polymers, the vascular phase can be favored in order, for example, to facilitate selective extravasation at a tumor site. Therapeutic applications of these systems are presented. The development of "intelligent" vectors capable of modulating intracellular distribution of an active compounds is an equally interesting approach, for example pH-sensitive liposomes or nanoparticles decorated with folic acid capable of targeting intracellular cytoplasm.

Animals↗

EWS fli-1 antisense nanocapsules inhibits ewing sarcoma-related tumor in mice.

EWS Fli-1, a fusion gene resulting from a t(11;22) translocation is found in 90% of both Ewing's sarcoma and primitive neuroectodermal tumor (PNET). In the present study, we show that recently developed polyisobutylcyanoacrylate nanocapsules with an aqueous core were able to encapsulate efficiently high amounts of phosphorothioate oligonucleotides (ODN) directed against EWS Fli-1 chimeric RNA. Release of these ODN in serum medium was shown to be biphasic which was explained by the presence of two types of nanocapsules able to release ODN with different kinetics. In addition, nanocapsules were found to provide protection of these oligonucleotides from the degradation in serum. These ODN nanocapsules permitted to obtain inhibition of Ewing sarcoma-related tumor in mice after intratumoral injection of a cumulative dose as low as 14.4 nanomoles. This new type of non viral vector shows great potential for in vivo administration of oligonucleotides.

3T3 Cells↗

pH-sensitive liposomes as a carrier for oligonucleotides: a physico-chemical study of the interaction between DOPE and a 15-mer oligonucleotide in excess water.

The cytoplasmic delivery of drugs encapsulated into pH-sensitive liposomes is under the control of a lamellar-to-hexagonal transition. In a previous study, under anhydrous conditions, oligonucleotides (ODN) encapsulated in pH-sensitive liposomes composed of dioleoylphosphatidylethanolamine (DOPE)/oleic acid (OA)/cholesterol (CHOL) were shown to modify the phase behaviour of DOPE. In the present study, the lipid/ODN interactions were evaluated in fully hydrated samples by surface tension measurements, differential scanning calorimetry, X-ray diffraction and turbidimetry. Concerning the lipids, it was shown that OA provoked a disorganisation of DOPE lamellar phases and led to the complete disappearance of hexagonal transition along with heating. The addition of CHOL further decreased the lipid packing in the bilayers. Concerning ODN, these molecules provoked an increase in the surface pressure of a DOPE/OA/CHOL monolayer, indicating the existence of molecular interactions with the lipids. At a supramolecular level, ODN induced a more ordered organisation of DOPE molecules in the lamellar and hexagonal phases, and completely abolished the disorganisational effect of OA and CHOL.

Calorimetry, Differential Scanning↗