References of "Evrard, Brigitte"
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See detailDevelopment and evaluation of injectable nanosized drug delivery systems for apigenin
Karim, Reatul ULiege; Palazzo, Claudio ULiege; Laloy, Julie et al

in International Journal of Pharmaceutics (2017), 532(2), 757-768

The purpose of this study was to develop different injectable nanosized drug delivery systems (NDDSs) i.e. liposome, lipid nanocapsule (LNC) and polymeric nanocapsule (PNC) encapsulating apigenin (AG) and ... [more ▼]

The purpose of this study was to develop different injectable nanosized drug delivery systems (NDDSs) i.e. liposome, lipid nanocapsule (LNC) and polymeric nanocapsule (PNC) encapsulating apigenin (AG) and compare their characteristics to identify the nanovector(s) that can deliver the largest quantity of AG while being biocompatible. Two liposomes with different surface characteristics (cationic and anionic), a LNC and a PNC were prepared. A novel tocopherol modified poly(ethylene glycol)-b-polyphosphate block-copolymer was used for the first time for the PNC preparation. The NDDSs were compared by their physicochemical characteristics, AG release, storage stability, stability in serum, complement consumption and toxicity against a human macrovascular endothelial cell line (EAhy926). The diameter and surface charge of the NDDSs were comparable with previously reported injectable nanocarriers. The NDDSs showed good encapsulation efficiency and drug loading. Moreover, the NDDSs were stable during storage and in fetal bovine serum for extended periods, showed low complement consumption and were non-toxic to EAhy926 cells up to high concentrations. Therefore, they can be considered as potential injectable nanocarriers of AG. Due to less pronounced burst effect and extended release characteristics, the nanocapsules could be favorable approaches for achieving prolonged pharmacological activity of AG using injectable NDDS. [less ▲]

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See detailSampling only 10 μL of whole blood to study the bioavailability of itraconazole formulations in rats
Kok, Miranda ULiege; Thiry, Justine ULiege; Evrard, Brigitte ULiege et al

Conference (2017, September 12)

Background Volumetric absorptive microsampling (VAMS) offers a unique opportunity to collect small and accurate quantities of biological fluids. This sampling technique is of great interest for volume ... [more ▼]

Background Volumetric absorptive microsampling (VAMS) offers a unique opportunity to collect small and accurate quantities of biological fluids. This sampling technique is of great interest for volume-limited samples and for the collection of multiple samples from the same animal. This serial sample collection may reduce the number of required study animals, thereby fulfilling the three Rs rule (refine, reduce, replace). Here, we demonstrate the applicability of VAMS to study the bioavailability of drug formulations in rats. Methods Four itraconazole-containing formulations were successively administered to rats with a wash-out period of one week. VAMS was used to collect 14 whole blood samples of only 10 μL each within a time frame of 48 hours after administration of the different drug formulations. Particular attention was paid to sample preparation and stability. The extraction of itraconazole and its main metabolite hydroxy-itraconazole was optimized to provide a high recovery and minimal matrix effects. A developed and validated LC–MS/MS method was used for the quantification of the two compounds. Pharmacokinetic profiles for the different formulations were constructed and compared. Results The stability of itraconazole and hydroxy-itraconazole in dried VAMS samples of whole rat blood could not be guaranteed for more than a day when the samples were stored at room temperature. However, samples were stable for at least two weeks when stored at -80°C after sample preparation. Differences in pharmacokinetic profiles were observed for the tested drug formulations. Whole blood concentrations of itraconazole and its main metabolite were significantly higher after administration of three in-house produced formulations compared to concentrations obtained with a commercially available product. Moreover, these in vivo results could be partly related to in vitro dissolution rates of the various formulations. Conclusions VAMS is an attractive approach for bioavailability studies. Due to the low blood volumes per sampling point, the same rats can be used to compare various drug formulations. Therefore, the number of required animals can be drastically reduced. Moreover, this helps to suppress the inter-individual variability and strengthens the validity of results. [less ▲]

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See detailIn vitro biphasic dissolution tests and their suitability for establishing in vitro-in vivo correlations: A historical review
Pestieau, Aude ULiege; Evrard, Brigitte ULiege

in European Journal of Pharmaceutical Sciences (2017), 102

For many decades, one of the most critical issues in the pharmaceutical industry has been the poor solubility of some drugs. Indeed, a prerequisite for drug absorption is the presence of dissolved drug at ... [more ▼]

For many decades, one of the most critical issues in the pharmaceutical industry has been the poor solubility of some drugs. Indeed, a prerequisite for drug absorption is the presence of dissolved drug at the absorption site and this can be challenging for compounds with low aqueous solubility such as BCS class II (low solubility, high permeability) and IV (low solubility, low permeability) compounds. If the development of oral delivery formulations of these compounds is frequently challenging to formulation scientists in the pharmaceutical industry, the in vitro evaluation of these new formulations is also a great challenge. One alternative approach to overcome the problems encountered with conventional dissolution methods is the use of biphasic dissolution systems. This review provides an overview of the origin and the evolution over time of the biphasic systems and the growing interest among scientists regarding their suitability for establishing in vitro-in vivo correlations. The evolution of these systems and their applications from the 1960s to the present day, such as in system variants and improvements, analysis of complex formulations, discriminatory power, bio-relevance, precipitation and supersaturation visualization, etc. will be discussed. [less ▲]

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See detailGlobal approach for the validation of an in-line Raman spectroscopic method to determine the API content in real-time during a hot-melt extrusion process
Netchacovitch, Lauranne ULiege; Thiry, Justine ULiege; De Bleye, Charlotte ULiege et al

in Talanta (2017), 171

Since the Food and Drug Administration (FDA) published a guidance based on the Process Analytical Technology (PAT) approach, real-time analyses during manufacturing processes are in real expansion. In ... [more ▼]

Since the Food and Drug Administration (FDA) published a guidance based on the Process Analytical Technology (PAT) approach, real-time analyses during manufacturing processes are in real expansion. In this study, in-line Raman spectroscopic analyses were performed during a Hot-Melt Extrusion (HME) process to determine the Active Pharmaceutical Ingredient (API) content in real-time. The method was validated based on a univariate and a multivariate approach and the analytical performances of the obtained models were compared. Moreover, on one hand, in-line data were correlated with the real API concentration present in the sample quantified by a previously validated off-line confocal Raman microspectroscopic method. On the other hand, in-line data were also treated in function of the concentration based on the weighing of the components in the prepared mixture. The importance of developing quantitative methods based on the use of a reference method was thus highlighted. The method was validated according to the total error approach fixing the acceptance limits at ± 15% and the α risk at ± 5%. This method reaches the requirements of the European Pharmacopeia norms for the uniformity of content of single-dose preparations. The validation proves that future results will be in the acceptance limits with a previously defined probability. Finally, the in-line validated method was compared with the off-line one to demonstrate its ability to be used in routine analyses. [less ▲]

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See detailPromoting vaginal distribution of two active siRNA-complexed in liposomes for cervical cancer treatment
Lechanteur, Anna ULiege; Furst Tania; Evrard, Brigitte ULiege et al

Conference (2017, April 25)

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See detailPromoting vaginal distribution of E7 and MCL-1 siRNA-silencing nanoparticles for cervical cancer treatment
Lechanteur, Anna ULiege; Furst, Tania; Delvenne, Philippe ULiege et al

in Molecular Pharmaceutics (2017)

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See detailBioavailability enhancement of itraconazole-based solid dispersions produced by hot melt extrusion in the framework of the Three Rs rule
Thiry, Justine ULiege; Kok, Miranda ULiege; Collard, Laurence ULiege et al

in European Journal of Pharmaceutical Sciences (2017), 99

Solid dispersion formulations made of itraconazole (ITZ) and Soluplus® (polyethylene glycol, polyvinyl acetate and polyvinylcaprolactame-based graft copolymer abbreviated SOL) were produced using hot melt ... [more ▼]

Solid dispersion formulations made of itraconazole (ITZ) and Soluplus® (polyethylene glycol, polyvinyl acetate and polyvinylcaprolactame-based graft copolymer abbreviated SOL) were produced using hot melt extrusion. Since ITZ possesses a water solubility of less than 1 ng/mL, the aim of this work was to enhance the aqueous solubility of ITZ, and thereby improve its bioavailability. The three formulations consisted of a simple SOL/ITZ amorphous solid dispersion (ASD), an optimized SOL/ITZ/AcDiSol® (super-disintegrant) ASD and an equimolar inclusion complex of ITZ in hydroxypropyl-β-cyclodextrin (substitution degree = 0.63, CD) with SOL. The three formulations were compared in vitro and in vivo to the marketed product Sporanox®. The in vitro enhancement of dissolution rate was evaluated using a biphasic dissolution test. In vitro dissolution results showed that all three formulations had a higher percentage of ITZ released than Sporanox® with the following ranking: SOL/ITZ/CD > SOL/ITZ/AcDiSol® > SOL/ITZ > Sporanox®. The bioavailability of these four formulations was evaluated in rats. The bioanalytical method was optimized so that only 10 μL of blood was withdrawn from the rats using specific volumetric absorptive microsampling devices. This enabled to keep the same rats during the whole study, which was in accordance with the Three Rs rules (reduction, refinement and replacement). Furthermore, this technique allowed the suppression of inter-individual variability. Higher Cmax and AUC were obtained after the administration of all three formulations compared to the levels after the use of Sporanox® as follows: SOL/ITZ/AcDiSol® > SOL/ITZ/CD > SOL/ITZ > Sporanox®. The inversion in the ranking between SOL/ITZ/CD and SOL/ITZ/AcDiSol® made impossible the establishment of an in vitro–vivo correlation. Indeed, very different release rates were obtained in vitro and in vivo for the two optimized formulations. These results suggest that ITZ would be protected inside the core of the SOL micelles even during the absorption step at the intestine, while some agents present in the intestinal fluids could displace ITZ from the hydrophobic cavity of CD by competition. [less ▲]

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See detailEvaluation of different in vitro dissolution tests based on level A in vitro–in vivo correlations for fenofibrate self-emulsifying lipid-based formulations
Pestieau, Aude ULiege; Lebrun, Sonia; Cahay, Bernard et al

in European Journal of Pharmaceutics & Biopharmaceutics (2017), 112

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See detailHot-Melt Extrusion as a Continuous Manufacturing Process to Form Ternary Cyclodextrin Inclusion Complexes
Thiry, Justine ULiege; Krier, Fabrice; Ratwatte, Shenelka et al

in European Journal of Pharmaceutical Sciences (2017), 96

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See detailSampling only ten microliters of whole blood for the quantification of poorly soluble drugs: Itraconazole as case study
Thiry, Justine ULiege; Evrard, Brigitte ULiege; Nys, Gwenaël ULiege et al

in Journal of Chromatography. A (2017), 1479

Nowadays in animal studies, it is important to comply with the so-called Three Rs rule by replacing or reducing the number of tested animals. Volumetric absorptive microsampling (VAMS) can be used to ... [more ▼]

Nowadays in animal studies, it is important to comply with the so-called Three Rs rule by replacing or reducing the number of tested animals. Volumetric absorptive microsampling (VAMS) can be used to collect small quantities (10 or 20 µL) of whole blood, thereby limiting the amount of animals needed. In this study, a quantitative method was developed and subsequently validated for the poorly soluble drug itraconazole (ITZ) using VAMS and ultra-high performance liquid chromatography (UHPLC) coupled to tandem mass spectrometry (MS). A proof of concept study showed that the optimized method is applicable to test the bioavailability of drug formulations containing ITZ. Using VAMS, smaller blood volumes can be taken per sampling point (10-20 µL instead of the conventional 0.2-0.5 mL) avoiding the sacrifice of animals. Moreover, the same rats can be used to compare different drug formulations which strengthens the validity of the results. In long-term bioavailability studies, it is necessary to guarantee the stability of the tested drugs supported on VAMS devices. In this study, we show that ITZ was only stable for 24 hours after collection with VAMS, but for at least two weeks by the storage of extracted samples at -80°C. [less ▲]

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See detailDevelopment of an analytical method for crystalline content determination in amorphous solid dispersions produced by Hot-Melt Extrusion using transmission Raman spectroscopy: A feasibility study.
Netchacovitch, Lauranne ULiege; Dumont, Elodie ULiege; Cailletaud, Johan ULiege et al

in International Journal of Pharmaceutics (2017), 530(1-2), 249-255

The development of a quantitative method determining the crystalline percentage in an amorphous solid dispersion is of great interest in the pharmaceutical field. Indeed, the crystalline Active ... [more ▼]

The development of a quantitative method determining the crystalline percentage in an amorphous solid dispersion is of great interest in the pharmaceutical field. Indeed, the crystalline Active Pharmaceutical Ingredient transformation into its amorphous state is increasingly used as it enhances the solubility and bioavailability of Biopharmaceutical Classification System class II drugs. One way to produce amorphous solid dispersions is the Hot-Melt Extrusion (HME) process. This study reported the development and the comparison of the analytical performances of two techniques, based on backscattering and transmission Raman spectroscopy, determining the crystalline remaining content in amorphous solid dispersions produced by HME. Principal Component Analysis (PCA) and Partial Least Squares (PLS) regression were performed on preprocessed data and tended towards the same conclusions: for the backscattering Raman results, the use of the DuoScan™ mode improved the PCA and PLS results, due to a larger analyzed sampling volume. For the transmission Raman results, the determination of low crystalline percentages was possible and the best regression model was obtained using this technique. Indeed, the latter acquired spectra through the whole sample volume, in contrast with the previous surface analyses performed using the backscattering mode. This study consequently highlighted the importance of the analyzed sampling volume. [less ▲]

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See detailChanges in biophysical membrane properties induced by the Budesonide/ Hydroxy-β-cyclodextrin complex
dos Santos, Andreia; Bayiha, Jules; Dufour, Gilles et al

in BBA Biomembranes (2017)

Budesonide (BUD), a poorly soluble anti-inflammatory drug, is used to treat patients suffering from asthma and COPD (Chronic Obstructive Pulmonary Disease). Hydroxypropyl-β-cyclodextrin (HPβCD), a ... [more ▼]

Budesonide (BUD), a poorly soluble anti-inflammatory drug, is used to treat patients suffering from asthma and COPD (Chronic Obstructive Pulmonary Disease). Hydroxypropyl-β-cyclodextrin (HPβCD), a biocompatible cyclodextrin known to interact with cholesterol, is used as a drug-solubilizing agent in pharmaceutical formulations. Budesonide administered as an inclusion complex within HPβCD (BUD:HPβCD) required a quarter of the nominal dose of the suspension formulation and significantly reduced neutrophil induced inflammation in a COPD mouse model exceeding the effect of each molecule administered individually. This suggests the role of lipid domains enriched in cholesterol for inflammatory signaling activation. In this context, we investigated the effect of BUD:HPβCD on the biophysical properties of membrane lipids. On cellular models (A549, lung epithelial cells), BUD:HPβCD extracted cholesterol similarly to HPβCD. On large unilamellar vesicles (LUVs), by using the fluorescent probes diphenylhexatriene (DPH) and calcein, we demonstrated an increase in membrane fluidity and permeability induced by BUD:HPβCD in vesicles containing cholesterol. On giant unilamellar vesicles (GUVs) and lipid monolayers, BUD:HPβCD induced the disruption of cholesterol-enriched raft-like liquid ordered domains as well as changes in lipid packing and lipid desorption from the cholesterol monolayers, respectively. Except for membrane fluidity, all these effects were enhanced when HPβCD was complexed with budesonide as compared with HPβCD. Since cholesterol-enriched domains have been linked to membrane signaling including pathways involved in inflammation processes, we hypothesized the effects of BUD:HPβCD could be partly mediated by changes in the biophysical properties of cholesterol-enriched domains. [less ▲]

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See detailEnhancement of the internalization of lipid nanocapsules in human glioblastoma cells: Effect of surface concentration of NFL peptide
Karim, Reatul ULiege; Lepeltier, Elise; Palazzo, Claudio ULiege et al

Poster (2017)

Le glioblastome multiforme (GBM) est un des cancers les plus fatal, avec une médiane de survie de 14 mois après traitement. Il est donc nécessaire de développer de nouvelles thérapies plus efficaces. La ... [more ▼]

Le glioblastome multiforme (GBM) est un des cancers les plus fatal, avec une médiane de survie de 14 mois après traitement. Il est donc nécessaire de développer de nouvelles thérapies plus efficaces. La fonctionnalisation en surface de nanocapsules lipidiques (LNC) avec le peptide NFL-TBS.40-63 (NFL) a déjà montré une amélioration de leur internalisation dans des cellules de glioblastome murin. Le but de cette étude a été d’évaluer l’impact de la concentration en NFL présente en surface des LNC sur l’internalisation de ces dernières dans des cellules humaines de GBM U87MG. De plus, le mécanisme d’internalisation LNC-NFL a été étudié. Une sonde fluorescente (DiA) a été encapsulé dans : des LNC (F1), des LNC avec 0.86 % et 2.58 % (w/w) de NFL adsorbé à la surface (F2 et F3 respectivement). Des analyses par cytométrie en flux (FACS) ont révélé une internalisation cellulaire de F3 plus importante de 46.4 et 6.8 fois après 30 min, de 21.6 et 6.1 fois après 1 heure, de 31.5 et 1.6 fois après 6 heures et de 7.3 et 1.1 fois après 24 heures, comparés à F1 et F2 respectivement. L’internalisation de F3 dans les cellules U87MG s’est révélée être énergie-dépendant, avec comme mécanisme principal la macropinocytose. Les cinétiques de désorption du peptides (obtenues par dialyse de F3 dans du Tris-Buffer pH 7.4 à 37°C suivi par une HPLC analytique) ont montré que 66 % de NFL restaient à la surface des LNC après 6h de dialyse, montrant une désorption lente. De plus, les trois formulations ont montré une faible activation du complément. Du fait d’une internalisation significativement plus prononcée et rapide, F3 semble être prometteur pour améliorer l’efficacité des thérapies antiGBM. De plus, la lente désorption du NFL de la surface des LNC et la faible consommation du complément font de F3 une thérapie ciblé prometteuse contre le GBM. [less ▲]

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See detailPEGylated and Functionalized Aliphatic Polycarbonate Polyplex Nanoparticles for Intravenous Administration of HDAC5 siRNA in Cancer Therapy
Frère, Antoine ULiege; Baroni, Alexandra; Hendrick, Elodie ULiege et al

in ACS Applied Materials and Interfaces (2017), 25(9(3)), 2181-2195

Guanidine and morpholine functionalized aliphatic polycarbonate polymers are able to efficiently deliver histone deacetylase 5 (HDAC5) siRNA into the cytoplasm of cancer cells in vitro leading to a ... [more ▼]

Guanidine and morpholine functionalized aliphatic polycarbonate polymers are able to efficiently deliver histone deacetylase 5 (HDAC5) siRNA into the cytoplasm of cancer cells in vitro leading to a decrease of cell proliferation were previously developed. To allow these biodegradable and biocompatible polyplex nanoparticles to overcome the extracellular barriers and be effective in vivo after an intravenous injection, polyethylene glycol chains (PEG750 or PEG2000) were grafted on the polymer structure. These nanoparticles, showed an average size of about 150 nm and a slightly positive zeta potential with complete siRNA complexation. Behavior of PEGylated and non-PEGylated polyplexes were investigated in the presence of serum, in terms of siRNA complexation (Fluorescence Correlation Spectroscopy), size (Dynamic Light Scattering and Single-Particle Tracking), interaction with proteins (Isothermal Titration Calorimetry) and cellular uptake. Surprisingly, both PEGylated and non-PEGylated formulations presented relatively good behavior in the presence of fetal bovine serum (FBS). Hemocompatibility tests showed no effect of these polyplexes on hemolysis and coagulation. In vivo biodistribution in mice was performed and showed a better siRNA accumulation at the tumor site for PEGylated polyplexes. However, cellular uptake in protein-rich conditions showed that PEGylated polyplex lost their ability to interact with biological membranes and enter into cells, showing the importance to perform in vitro investigations in physiological conditions closed to in vivo situation. In vitro, the efficiency of PEGylated nanoparticles decreases compared to non-PEGylated particles, leading to the loss of the antiproliferative effect on cancer cells. [less ▲]

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See detailContinuous Production of Itraconazole-based Solid Dispersions by Hot Melt Extrusion: Preformulation, Optimization and Design Space Determination.
Thiry, Justine ULiege; Lebrun, Pierre; Vinassa, Chloé et al

in International Journal of Pharmaceutics (2016), 515(1-2), 114-124

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See detailNEW INJECTABLE LIPOSOME AND DRUG-IN-CYCLODEXTRIN-IN LIPOSOME SYSTEMS ENCAPSULATING ESTETROL FOR THE TREATMENT OF ISCHEMIA DISEASES IN PREMATURE BABIES
Palazzo, Claudio ULiege; Laloy, Julie; Delvigne, Anne-Sophie et al

Poster (2016, December)

Purpose: In 2010, almost 15 million of babies in the world are prematurely borned, 11.1 % of the total amount of alive children. Despite the better neonatology techniques, the number of preterm babies ... [more ▼]

Purpose: In 2010, almost 15 million of babies in the world are prematurely borned, 11.1 % of the total amount of alive children. Despite the better neonatology techniques, the number of preterm babies with motor, vision, hearing or mental deficiencies is still constant along the last twenty years. Moreover, no efficacy treatment is available to the present day. The estetrol (E4) has an important role in the brain development and protection. The aim of this study is to develop new injectable liposome and drug-in-cyclodextrin-in-liposome (DCL) formulations, encapsulating E4 in order to enhance its crossing through the blood-brain barrier (BBB). Methods: Cyclodextrins (CD) were used to increase E4 aqueous solubility. Liposome and DCL (E4-CD complex) formulations were prepared by thin-film hydration technique. The formulations were physicochemical characterized. LDH and MTS tests on endothelial, neuronal and BBB model cells were performed in vitro on the liposome formulation. Hemocompatibility of the formulations was evaluated on red blood cells, platelet aggregation and coagulation. BBB passage tests were performed using human BBB cell line (hCMEC/D3). Results: E4-CD complexes proportionally increased the solubility of the hormone. Liposomes and DCL encapsulating E4 were prepared. All the formulations had average particle size below 150 nm, polydispersity index below 0.10 and ζ potential around + 30 mV. The encapsulation efficacy for liposomes was between 3% and 10% while those of DCL are between 15% and 35%. Moreover, the formulations are capable to release 80 % (liposome) and 90 % (DCL) of encapsulated E4 after 3 h at 37°C. The effect of liposome and DCL formulations on cell viability and integrity was evaluated. The results showed no toxic effects on all the tested cell lines. Hemocompatibility tests showed no hemolysis, platelet aggregation or effects on coagulation, confirming the possibility of the formulations to be intravenously administrated. BBB passage tests highlighted the capability of the formulations to pass the BBB and reach the brain. Conclusions: New non-toxic, hemocompatible liposome and DCL formulations encapsulating E4 were prepared. The formulations are promising drug delivery system to target estrogens to the brain, due to their physiochemical characteristics. [less ▲]

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See detailDevelopment and comparison of liposomes and nanocapsules as injectable nanocarriers for poorly aqueous soluble drugs
Karim, Reatul ULiege; Palazzo, Claudio ULiege; Laloy, Julie et al

Poster (2016, December)

About 90% of drugs in development phase have poor aqueous solubility. Liposomes and nanocapsules are promising approaches that enable parenteral administration of these drugs with possibilities of site ... [more ▼]

About 90% of drugs in development phase have poor aqueous solubility. Liposomes and nanocapsules are promising approaches that enable parenteral administration of these drugs with possibilities of site specific delivery. The objective of the study was to develop different liposomes and lipid nanocapsules entrapping a hydrophobic model molecule (apigenin (AG)), and to characterize and compare them as potential injectable nanocarriers (NCs) for drugs with low aqueous solubility. [less ▲]

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