Low molecular weight heparin gels, based on nanoparticles, for topical delivery.

A commercial suspension of nanoparticles (Eudragit RS 30D) was used to manufacture a gel for topical application. Gels were prepared by mixing a polycationic polymer (Eudragit(®) RS 30D) and a low molecular weight heparin (LMWH), an antithrombotic agent. Gels formed spontaneously at a ratio of 1:1 as a result of electrostatic interactions between the polyanionic drug and the polycationic polymer. Different types of heparin were used: Bemiparin, Enoxaparin (Lovenox), Nadroparin (Fraxiparin) and Tinzaparin (Innohep). Several LMWH concentrations were tested. Rheological measurements were performed to investigate the gel behavior. Gel formation was confirmed by dynamic rheological measurements as the elastic modulus (G') was higher than the viscous one (G″). The amount of heparin incorporated into the gel matrix was determined. A maximum of incorporation (100%) was reached using a heparin solution of 600 IU/mL. The release kinetics of LMWH from the gel were also studied. Regardless of the LMWH used in the formulation, a biphasic release profile was observed. Accordingly, a burst effect was observed. Afterwards, the release rate became steady. The penetration of the LMWH through the dermal barrier was also investigated.

Rheological and syringeability properties of highly concentrated human polyclonal immunoglobulin solutions.

This study of highly concentrated polyvalent immunoglobulin solutions, IgG, aimed at analyzing the relationships between protein concentration and aggregation on the one hand and viscosity on the other hand. Viscosity variations as a function of IgG concentration showed two well-defined behaviours: a Newtonian behaviour for low-concentrated solutions and a shear-thinning behaviour for highly concentrated ones. The viscosity data fitted very well with the Mooney model, suggesting the absence of intermolecular interactions in the IgG solutions that behaved like a non-interacting suspension of hard particles. The polyclonal nature of IgG seems to prevent intermolecular interaction. The shape factor, determined from Mooney fitting, revealed a non-spherical shape of the polyclonal IgG molecules. The rheological properties were also correlated with the injection force (F) through hypodermic needles by syringeability tests. Here, F was mainly affected by three parameters: the solution viscosity, the injection flow rate, and the needle characteristics. In fact, syringeability tests showed that F increased with IgG concentration and flow rate and decreased with the internal diameter of the needle. A zone for optimal injection conditions was then identified taking into account the different affecting parameters and mainly a maximum force for manual injection, which was fixed at 30N.

Structural and rheological properties of chitosan semi-interpenetrated networks.

The local structure and the viscoelastic properties of semi-interpenetrated biopolymer networks based on cross-linked chitosan and poly(ethylene oxide) (PEO) were investigated by Small Angle Neutron Scattering and rheological measurements. The specific viscosity and the entanglement concentration of chitosan were first determined, respectively, by capillary viscosimetry and steady-state shear rheology experiments performed at different polymer concentrations. Mechanical spectroscopy was then used to study the gelation process of chitosan/PEO semi-interpenetrated networks. By fitting the frequency dependence of the elastic and loss moduli with extended relations of relaxation shear modulus around the sol-gel transition, it was shown that the addition of PEO chains had a significant effect on the viscoelastic properties of aqueous chitosan networks but no effect on the gelation time. The improvement of mechanical properties was in accordance with the correlation length decrease deduced from Small Angle Neutron Scattering experiments.

[Interactions between cyclodextrins and triglycerides: from emulsion stabilisation to the emergence of a new drug delivery system called "beads"]. / Interactions entre les cyclodextrines et les triglycérides : de la stabilisation des émulsions à l'obtention d'un nouveau système galénique appelé << billes >>.

Natural cyclodextrins are cyclic oligosaccharides which can be modified to obtain more water soluble or insoluble derivatives. The main interest of cyclodextrins results from their ability to form an inclusion complex with hydrophobic molecules. Inclusion constitutes a true molecular encapsulation. This property is employed in pharmaceutical industry to facilitate the formulation of poorly water soluble and/or fragile drugs. A more recent application of cyclodextrins consists in their use in the preparation of dispersed systems such as micro- and nanoparticles or even liposomes. When incorporated in dispersed systems, cyclodextrin can enhance drug solubility, drug stability and drug loading. Interestingly, cyclodextrins themselves can also be employed to form or stabilise dispersed systems (material or emulsifying agent). For example, the interactions between cyclodextrins with components of the vegetable oils (more especially with triglycerides) allow to stabilise simple or multiple emulsions but also to form particles called "beads". Very rich in oil, this novel lipid carrier presents an important potential for the encapsulation of highly lipophilic compounds and their delivery by topical and oral routes. These two applications are more particularly developed in the present paper.

Paraquat detoxication with multiple emulsions.

In this study, we show that detoxifying W/O/W multiple emulsions, prepared with an appropriate extractant/trapping couple, represent a promising technology for quick and safe poisoning treatments, with application to the highly toxic herbicide Paraquat, responsible of poisonings from low-dose exposure leading to several deaths every year. In vitro tests led to the choice of an appropriate extractant/trapping couple system with significant detoxication performance. In vivo tests showed (i) that rats receiving high doses of Paraquat, then a detoxifying emulsion, presented an increase from 50% to 100% of the MST (median survival time) and (ii) that no mortality was observed during 30 days with rats dosed with emulsions initially loaded with Paraquat at a concentration much higher than the lethal dose, proving the stability and the inocuity of the detoxifying multiple emulsion in the gastrointestinal tract.

Formulation and characterisation of beads prepared from natural cyclodextrins and vegetable, mineral or synthetic oils.

A continuous external shaking for 2.5 days of a mixture composed of alpha-cyclodextrin (6%), soybean oil (19.6%) and water (74.4%) resulted in a calibrated lipid carrier namely bead with a high fabrication yield. The purpose of this work was to explore the possibility to substitute alpha-cyclodextrin by other natural cyclodextrins, i.e. beta- and gamma-cyclodextrin and then soybean oil by mineral (Primol) 352 and Marcol 82) or synthetic (Silicon 200) fluid 10, 50 or 100cSt) oils. Beads can be successfully prepared using Marcol 82 with alpha-cyclodextrin and Silicon 50 or 100cSt with gamma-cyclodextrin. The area inside oil/cyclodextrin/water ternary diagram corresponding to bead occurrence was superior for the Marcol 82/alpha-cyclodextrin couple compared to that observed with soybean oil/alpha-cyclodextrin couple. Only a few ratios of Silicon 50 and 100cSt/gamma-cyclodextrin/water led to beads. The combinations which did not induce bead occurrence gave either emulsions, two non-miscible liquids or a solid mixture. Whatever the materials used, beads exhibited similarities: presence of a crystalline organisation and viscoelastic properties. Manufacturing process of paraffin- and silicon-based beads need further optimisation to increase fabrication yield and later on, to take advantages from the high stability of both oils for the formulation of drugs with beads.

In vitro and in vivo characteristics of a thermogelling and bioadhesive delivery system intended for rectal administration of quinine in children.

The aim of this work was to improve the rectal bioavailability of quinine hydrochloride by designing thermosensitive and mucoadhesive gels intended for rectal delivery. The rheological and mucoadhesive properties of poloxamer 407 solutions have been modulated by addition of hydroxypropylmethycellulose (HPMC) and propanediol-1,2. In vitro release and rectal absorption of quinine have been highlighted by a dialysis dissolution testing method and by the determination of bioavailability of the different formulations in rabbits. Increasing the proportions of HPMC and poloxamer in the formulations resulted in a prolonged release of quinine. Indeed, compared to the DT 50% of a rectal solution and a simple HPMC gel (27 and 65 min, respectively) the DT 50% of thermosensitive ternary systems was increased and ranged between 80 and 138 min, depending on the system composition. The release rate depended strongly on the elasticity of the gels after thermogelation. The absolute rectal bioavailability of quinine determined in rabbits was significantly improved with these thermosensitive and adhesive systems. It increased from 62% for the rectal solution to 98% for a ternary system 16/0.5/30 (poloxamer (16%)/HPMC (0.5%)/propanediol-1,2 (30%)). As a result of combined bioadhesion and prolonged release of quinine in vivo, higher average values of MRT and t(max) (9.1+/-0.2h and 30 min, respectively) were obtained compared to the rectal solution (6.9+/-0.9h and 15 min, respectively). Moreover, these formulations presented a very good rectal tolerance. Modulation by HPMC of the viscoelastic and mucoadhesive properties of poloxamer 407 thermogelling solutions allowed a prolonged release of quinine hydrochloride and an improvement of bioavailability in rabbit.

Self-assembling cyclodextrin based hydrogels for the sustained delivery of hydrophobic drugs.

This study aims to investigate the rheological properties of self-assembling gels containing cyclodextrins with potential application as injectable matrix for the sustained delivery of poorly soluble drugs. The ability of these gels to entrap two hydrophobic molecules, benzophenone (BZ) and tamoxifen (TM), and to allow their in vitro sustained release was evaluated. In view of their future pharmaceutical use, gels were sterilized by high hydrostatic pressures (HHP) and tested for their biocompatibility. The gels formed instantaneously at room temperature, by mixing the aqueous solutions of two polymers: a beta-cyclodextrin polymer (pbetaCD) and a hydrophobically modified dextran by grafting alkyl side chains (MD). MD-pbetaCD gels presented a viscoelastic behavior under low shear, characterized by constant values of the loss modulus G'' and the storage modulus G'. The most stable gels were obtained for a total polymer concentration C(p) of 6.6% and 7.5% (w/w), and a polymer ratio MD/pbetaCD of 50/50 and 33/67 (w/w). BZ and TM were successfully incorporated into MD-pbetaCD gels with loading efficiencies as high as 90%. In vitro, TM and BZ were released gradually from the gel matrix with less than 25% and 75% release, respectively, after 6 days incubation. HHP treatment did not modify the rheological characteristics of MD-pbetaCD gels. Moreover, the low toxicity of these gels after intramuscular administration in rabbits makes them promising injectable devices for local delivery of drugs.

Assessment of oil polarity: comparison of evaluation methods.

In multiple emulsion systems, oily or aqueous transfers may occur between the dispersed droplets through the continuous phase. These transfers are controlled by both the surfactant system (micellar transport), and the partial solubility of one phase in another (molecular transport). The latter could be anticipated from the knowledge of oil polarity, if this information could easily be obtained. In this work, the relative polarity of eight oils used for various purposes has been evaluated from the comparison of their dielectric requirement for solubilization, their interfacial tension and chromatographic analysis. The results showed the complementarities of HPLC analysis and interfacial tension measurements and their superiority over the solubilization method for classifying oils as a function of their polarity.

alpha-Cyclodextrin/oil beads: an innovative self-assembling system.

The aim of this work was to characterise a new type of particulate system, named beads, prepared by a straightforward technique starting from a mixture of alpha-cyclodextrin aqueous solution and soybean oil without the use of any organic solvent or surface-active agent. Mechanisms involved in bead formation were also investigated. Optimal ratio between alpha-cyclodextrin (6%, w/w), soybean oil (19.6%, w/w) and water (74.4%, w/w) led to homogeneous bead size (1.6 mm) with a fabrication yield superior to 80% after a continuous external shaking during 2.5 days. After freeze-drying, oil and alpha-cyclodextrin contents were estimated at 80% (w/w) and 20% (w/w), respectively. X-ray diffraction studies revealed that beads presented a crystalline organisation and microscopic techniques showed that their inner structure was constituted by a matrix containing oily compartments. Beads offer interesting prospects for the microencapsulation of lipophilic and poorly stable molecules. Due to their semi-solid consistency and their ability to be freeze-dried, these beads have great potentialities for pharmaceutical (oral and topical routes) and cosmetic applications.

Modelization of the release from a tetradecane/water/hexadecane multiple emulsion: evidence of significant micellar diffusion.

The release of tetradecane from a multiple emulsion of the type tetradecane/water/hexadecane was studied experimentally using the differential scanning calorimetry technique. The kinetics of the tetradecane release was measured for three formulations containing different concentrations of hydrophilic surfactant (2%, 4%, and 7%). A new mass transfer model derived from the shrinking core model was developed. The values of the model parameters deduced from the least-squares fittings led to the determination of the tetradecane diffusivity. Thus, the preponderant mechanism of mass transfer was proved to be micellar diffusion and not molecular diffusion. This conclusion was confirmed by considering the effect of the change in the hydrophilic surfactant concentration.

Modulation of the rheological and mucoadhesive properties of thermosensitive poloxamer-based hydrogels intended for the rectal administration of quinine.

The aim of this work was to formulate and characterize thermosensitive gels based on poloxamer 407, a thermosensitive polymer, and hydroxypropylmethylcellulose (HPMC), a bioadhesive polymer, intended for the rectal delivery of quinine in children. In order to avoid the macroscopic phase separation between the two polymers it was necessary to add propanediol-1,2. After the formulation of stable ternary systems, their rheological properties were studied as a function of temperature. It was thus possible to obtained the gelation temperature of the systems as well as their elastic modulus, G', at 37 degrees C. It appeared that HPMC in the presence of propanediol-1,2 had a synergistic effect on the gelation of poloxamer 407. Tests on the rabbit rectal mucous membrane, founded on a technique of traction of the adhesive/adhered joint, made it possible to characterize the bioadhesive properties of the gels by measuring the work of adhesion, W, and the maximum detachment force, F(max). Over small concentration ranges of poloxamer and HPMC, very important variations of the viscoelastic and mucoadhesives properties were observed. It was shown that the viscoelastic and the bioadhesive properties were tightly correlated. Indeed, an empirical equivalence relation was established between the shear frequency and the rate of deformation in traction and allowed to describe all our samples by a master curve. This master curve would make it possible to predict the values of W at various rates of traction by the simple non-destructive measurement of the elastic modulus, G', at an equivalent shear frequency.

Rheological properties of three different vitamin D ointments and their clinical perception by patients with mild to moderate psoriasis.

BACKGROUND: Ointments, classically used for the treatment of dermatological diseases, are monophasic viscous semisolid formulations. According to the proportion of their compounds, they have physicochemical and organoleptic properties and when applied on skin show a specific behaviour allowing to be spread more or less easily. OBJECTIVE: To measure in vitro rheological characteristics of three vitamin D derivative ointments prescribed for the treatment of psoriasis, and to compare their viscosity and clinical acceptability when applied on the diseased skin. METHODS: Rheological characteristics of tacalcitol 4 microg/g, calcipotriol 50 microg/g and calcitriol 3 microg/g ointments were assessed by measuring the oscillatory viscoelastic parameters and the permanent flow analysis. Clinical acceptability was studied in 20 psoriatic male or female subjects, aged 18 years or older. A survey evaluated the acceptability of calcitriol vs. tacalcitol and calcipotriol. Questions included information about fluidity, spreading capacity and stickiness after application. RESULTS: We demonstrated that viscoelastic parameters were four times higher for ointment tacalcitol than for calcipotriol and calcitriol, corresponding to a higher consistency of ointment tacalcitol compared to calcipotriol and calcitriol showing both similar results; better fluidity was demonstrated by calcitriol than by tacalcitol and calcipotriol. Comparable results were obtained for the quality to be spread. The sensation of stickiness, significantly different between tacalcitol and calcitriol, was not different between calcipotriol and calcitriol. CONCLUSION: The above results confirm the relationship between rheological in vitro and sensorial in vivo results: variations between different formulations may have an important influence on non-adherence and treatment failure.

Study of mass transfer in oil-water-oil multiple emulsions by differential scanning calorimetry.

A multiple emulsion of the type O1/W/O2 is studied experimentally by means of differential scanning calorimetry (DSC). The aim of this work is to characterize and measure the time-dependent changes within the emulsion. In particular, interest is focused to quantify the concentration changes in the internal and external phases of the O1/W/O2 multiple emulsion. In order to accomplish the objective, the measurement and analysis carried out by DSC are based on the crystallization behavior of the emulsion. A volume of a few mm3 is periodically removed from the O1/W/O2 multiple emulsion. The sample is submitted to steady cooling and the crystallization thermogram is recorded. The experimental data provided by the crystallization thermogram makes it possible to quantify the crystallized mass for both phases, the internal and the external. In addition, the composition in each phase can also be deduced from the thermogram. To deduce the composition, a diagram of crystallization temperatures is elaborated, employing several mixtures of known composition. In addition to the main objective previously mentioned, the influence of formulation parameters such as surfactant concentration in the aqueous phase and the mass ratio of the internal and external phases are also analyzed. The experimental results made it possible to conclude that a mass transfer took place from the internal phase toward the external phase; this transfer is caused by the composition difference on both sides of the aqueous membrane. In this work we analyzed the mass transfer in the multiple emulsion carried out by a composition gradient through the aqueous membrane. The most likely mechanism of mass transfer through the aqueous membrane is a solution-diffusion of tetradecane enhanced by the micelles of the surfactant Tween 20. The model of mass transfer confirms that the osmotic pressure difference controls the kinetics of tetradecane transfer. It is also confirmed that an increment of surfactant concentration in the aqueous phase allows a faster kinetics of the tetradecane transfer.

A very promising new glucolipidic surfactant: Lipowheat.

Lipowheat is an entirely biodegradable 100% natural active ingredient, extracted from non-transgenic wheat. Thanks to its very interesting properties, it can integrate the composition of most cosmetic and pharmaceutical products. The aim of this work was first to realize a large range of stable simple or multiple emulsions, in order to determine and evaluate the ability of a new glucolipidic surfactant Lipowheat to form and stabilize emulsions. The rheological properties of these emulsions were tested during a 30-day storage period at three different storage conditions (cold, room temperature and at 40 degrees C). In addition to dynamic and static rheological tests, droplet size distribution of the cream was also determined. Furthermore, a stable simple emulsion was selected to realize percutaneous absorption and evaluate the properties of Lipowheat.

Release of antiseptics from the aqueous compartments of a w/o/w multiple emulsion.

A w/o/w multiple emulsion drug carrier system has been developed for local vaginal therapy. To improve its efficacy and to extend the antimicrobial spectrum activity of benzalkonium chloride (CBZ), which is introduced in the external aqueous phase, chlorhexidine digluconate (CHD) was added to the internal aqueous phase of the multiple emulsions. The minimal bactericidal concentrations (MBC) for the association of CHD and CBZ in emulsion were determined towards Escherichia coli and Staphylococcus aureus. The main release mechanism considered for the CHD encapsulated in the inner phase was a swelling-breakdown phenomenon which followed dilution of the emulsion under hypo-osmotic conditions. In order to demonstrate this release, the bactericidal effect of multiple emulsions undiluted and diluted 1-5 and 1-10 in hypo-osmotic conditions at two CHD concentrations was evaluated. To validate and quantify this release, rheological and release kinetics studies were used. The bactericidal activity of combination CBZ-CHD in the emulsion was synergistic on the two bacterial strains and the release of encapsulated CHD in the internal phase was obtained following its dilution in hypo-osmotic conditions. Vaginal administration could be carried out following dilution at 1-5 in sterile water for multiple emulsions containing the lower concentration of CHD.

Physicochemical characterization and in vitro release of salicylic acid from O/W emulsions prepared with Montanov 68: effect of formulation parameters.

Montanovs are surfactants consisting of a combination of alkylpolyglucosides and long chain saturated alcohols. They are used to formulated oil in water (O/W) emulsions where they generate liquid crystals. Emulsions containing 5% Montanov 68 with 40% Lanol 1688 were prepared and salicylic acid (SA) was incorporated at different stages of the O/W emulsion preparation. This study highlights the effects of formulation parameters on the microscopic characteristics, particle size and rheologic properties of Montanov 68 O/W emulsions. Diffusion studies with these emulsions showed the influence of SA incorporation at different steps on the release kinetics. Montanov enabled the release of SA to be controlled when it was solubilized in the internal phase. The presence of a physical barrier formed by the Montanov at the interface between the oil and water appeared to modulate the SA passage to the external phase.

In vitro microbicidal activity of W/O/W multiple emulsion for vaginal administration.

The microbicidal activity of a W/O/W multiple emulsion destined for vaginal application, containing lactic acid in the internal aqueous phase, octadecylamine (ODA) in the oily phase and benzalkonium chloride (CBZ) in the external aqueous phase was evaluated against three microbial strains: Escherichia coli, Staphylococcus aureus and Candida albicans. The results were different depending on the procedure used. Interpretable results were obtained if only a gentle agitation was used just after the introduction of the microbial suspension to the product. This suggested that vigorous agitation lead to a variable fraction of CBZ or ODA entrapped in the micelles of ethylene and propylene oxide copolymer (COE).

Water state characterization, swelling behavior, thermal and mechanical properties of chitosan based networks.

Two kinds of chitosan-based hydrogels, a crosslinked chitosan reference gel and a chitosan-poly(ethylene oxide) semi-interpenetrating network (semi-IPN), with potential pH-sensitive swelling and drug delivery properties are characterized. Swelling studies were performed on the two kinds of hydrogels by differential scanning calorimetry (DSC) at pH 1.2 and by the gravimetric method at pH 1.2 and pH 7.2. Both methods lead to similar results. If pH-dependent swelling properties were observed with both hydrogels, they were however improved for the semi-IPN. The amount of bound water in the xerogels could be determined from DSC measurements and a thermogravimetric analysis. The results obtained by both techniques were in good agreement and indicated that the semi-IPN contained more bound water than the reference gel probably due to the presence of the hydrophilic poly(ethylene oxide) chains. Young modulus of the swollen hydrogels was determined by indentation analysis. The semi-IPN displayed improved mechanical properties compared to the reference gel.

pH compartmented w/o/w multiple emulsion: a diffusion study.

In order to develop w/o/w emulsions characterized by two separate aqueous phases of different pH, a preliminary study was carried out to obtain a better insight into the possible diffusion processes taking place between an inner acidic aqueous phase and an external phase of higher pH (pH approximately 6). In fact, such systems could be of great interest for pharmaceutical use. For this purpose, a model emulsion was formulated. The study of pH and conductivity showed that acidic species transport take place between the two aqueous compartments. The three main release mechanisms that might be responsible for this passage across the oil phase were investigated: breakdown of oil globules, facilitated transport by surfactant micelles across the oil phase or by Fickian diffusion. It appears that this last mechanism was involved. In order to control this diffusion process, an alkaline species, octadecylamine was introduced in the oil phase. This compound could form an ion pair with the lactate ion at the interface of the external aqueous phase and the oil phase, thus, limiting the acidification of the external aqueous phase.