Lipid nanocarriers as skin drug delivery systems: Properties, mechanisms of skin interactions and medical applications.

During the past decades, lipid nanocarriers are gaining momentum with their multiple advantages for the management of skin diseases. Lipid nanocarriers enable to target the therapeutic payload to deep skin layers or even to reach the blood circulation making them a promising cutting-edge technology. Lipid nanocarriers refer to a large panel of drug delivery systems. Lipid vesicles are the most conventional, known to be able to carry lipophilic and hydrophilic active agents. A variety of lipid vesicles with high flexibility and deformability could be obtained by adjusting their composition; namely ethosomes, transfersomes and penetration enhancer lipid vesicles which achieve the best results in term of skin permeation. Others are designed with the objective to perform higher encapsulation rate and higher stability, such as solid lipid nanoparticles and nanostructured lipid nanocarriers. In this review, we attempted to give an overview of lipid based nanocarriers developed with the aim to enhance dermal and transdermal drug delivery. A special focus is put on the nanocarrier composition, behavior and interaction mechanisms with the skin. Recent applications of lipid-based nanocarriers for the management of skin diseases and other illnesses are highlighted as well.

Diclofenac Loaded Lipid Nanovesicles Prepared by Double Solvent Displacement for Skin Drug Delivery.

PURPOSE: Herein, we detail a promising strategy of nanovesicle preparation based on control of phospholipid self-assembly: the Double Solvent Displacement. A systematic study was conducted and diclofenac as drug model encapsulated. In vitro skin studies were carried out to identify better formulation for dermal/transdermal delivery. METHODS: This method consists in two solvent displacements. The first one, made in a free water environment, has allowed triggering a phospholipid pre-organization. The second one, based on the diffusion into an aqueous phase has led to liposome formation. RESULTS: Homogeneous liposomes were obtained with a size close to 100 nm and a negative zeta potential around -40 mV. After incorporation of acid diclofenac, we obtained nanoliposomes with a size between 101 ± 45 and 133 ± 66 nm, a zeta potential between 34 ± 2 and 49 ± 3 mV, and the encapsulation efficiency (EE%) was between 58 ± 3 and 87 ± 5%. In vitro permeation studies showed that formulation with higher EE% dispayed the higher transdermal passage (18,4% of the applied dose) especially targeting dermis and beyond. CONCLUSIONS: Our results suggest that our diclofenac loaded lipid vesicles have significant potential as transdermal skin drug delivery system. Here, we produced cost effective lipid nanovesicles in a merely manner according to a process easily transposable to industrial scale. Graphical Abstract ᅟ.

Advances in psoriasis physiopathology and treatments: Up to date of mechanistic insights and perspectives of novel therapies based on innovative skin drug delivery systems (ISDDS).

Psoriasis is a chronic inflammatory disease affecting mainly the skin but which can be complicated by psoriatic arthritis (PsA).This autoimmune skin disorder concerns 2-5% of the world population. To date, the physiopathology of psoriasis is not still completely elucidated but many researches are ongoing which have led for example to the discovery of the Th17/Th22 pathway. The conventional therapeutic approaches (local or systemic route) appeal to various classes of drugs with complex mechanisms of action and non-negligible side effects. Although there is no therapy capable to cure psoriasis, the current goal is to relieve symptoms as longer as possible with a good benefit/risk ratio. That is one of the principal limits of conventional antipsoriatic drugs. New formulations based on nanoencapsulation are a promising opportunity to answer to this limit by offering an optimization of the conventional antipsoriatic drug use (higher activity, lower side effects and frequency of application, etc.). Herein, we tried to put in perspective the mechanistic insights (histological and immunological views) proposed into scientific literature these last years in order to have a better comprehension of psoriasis physiopathology resulting in skin lesions and PsA. The therapeutic armamentarium and the different strategies in the management of psoriasis are discussed in greater details. To finish, the field of encapsulation in nanoparticles is broached in order to put forward recent advances in innovative skin drug delivery systems (ISDDSs) of antipsoriatic active agents for a better efficacy, safety and compliance.

Elaboration of Sterically Stabilized Liposomes for S-Nitrosoglutathione Targeting to Macrophages.

S-nitrosoglutathione (GSNO) is a potential therapeutic for infectious disease treatment because of its pivotal role in macrophage-mediated inflammatory responses and host defense in addition to direct antibacterial activities. In this study, sterically stabilized cationic liposomes (SSCL) and sterically stabilized anionic liposomes (SSAL) were developed as nanocarriers for macrophage targeting. Elaborated liposomes were characterized in terms of size, zeta potential, morphology, encapsulation efficiency, in vitro drug release behavior and cytotoxicity. Their versatility in targeting monocytes/macrophages was determined by confocal laser scanning microscopy and transmission electron microscopy. Flow cytometry revealed that cellular uptake of both SSCL and SSAL was governed by several endocytic clathrin- and caveolae-dependent mechanisms. Quantitative assessments of intracellular nitric oxide demonstrated highly efficient uptake of GSNO-loaded SSCL that was twenty-fold higher than that of GSNO-free molecules. GSNO-loaded SSCL displayed strong bacteriostatic effects on Staphylococcus aureus and Pseudomonas aeruginosa, which can be involved in pulmonary infectious diseases. These results reveal the potential of liposomal GSNO as an anti-infective therapeutic due to its macrophage targeting capacity and direct antibacterial effects.

Essential oils: from extraction to encapsulation.

Essential oils are natural products which have many interesting applications. Extraction of essential oils from plants is performed by classical and innovative methods. Numerous encapsulation processes have been developed and reported in the literature in order to encapsulate biomolecules, active molecules, nanocrystals, oils and also essential oils for various applications such as in vitro diagnosis, therapy, cosmetic, textile, food etc. Essential oils encapsulation led to numerous new formulations with new applications. This insures the protection of the fragile oil and controlled release. The most commonly prepared carriers are polymer particles, liposomes and solid lipid nanoparticles.

Administration strategies for proteins and peptides.

Proteins are a vital constituent of the body as they perform many of its major physiological and biological processes. Recently, proteins and peptides have attracted much attention as potential treatments for various dangerous and traditionally incurable diseases such as cancer, AIDS, dwarfism and autoimmune disorders. Furthermore, proteins could be used for diagnostics. At present, most therapeutic proteins are administered via parenteral routes that have many drawbacks, for example, they are painful, expensive and may cause toxicity. Finding more effective, easier and safer alternative routes for administering proteins and peptides is the key to therapeutic and commercial success. In this context, much research has been focused on non-invasive routes such as nasal, pulmonary, oral, ocular, and rectal for administering proteins and peptides. Unfortunately, the widespread use of proteins and peptides as drugs is still faced by many obstacles such as low bioavailability, short half-life in the blood stream, in vivo instability and numerous other problems. In order to overcome these hurdled and improve protein/peptide drug efficacy, various strategies have been developed such as permeability enhancement, enzyme inhibition, protein structure modification and protection by encapsulation. This review provides a detailed description of all the previous points in order to highlight the importance and potential of proteins and peptides as drugs.

Particles from preformed polymers as carriers for drug delivery.

Biodegradable and biocompatible polymers are widely used for the encapsulation of drug molecules. Various particulate carriers with different sizes and characteristics have been prepared by miscellaneous techniques. In this review, we reported the commonly used preformed polymer based techniques for the preparation of micro and nano-structured materials intended for drug encapsulation. A description of polymer-solvent interaction was provided. The most widely used polymers were reported and described and their related research studies were mentioned. Moreover, principles of each technique and its crucial operating conditions were described and discussed. Recent applications of all the reported techniques in drug delivery were also reviewed.

Gene therapy and DNA delivery systems.

Gene therapy is a promising new technique for treating many serious incurable diseases, such as cancer and genetic disorders. The main problem limiting the application of this strategy in vivo is the difficulty of transporting large, fragile and negatively charged molecules like DNA into the nucleus of the cell without degradation. The key to success of gene therapy is to create safe and efficient gene delivery vehicles. Ideally, the vehicle must be able to remain in the bloodstream for a long time and avoid uptake by the mononuclear phagocyte system, in order to ensure its arrival at the desired targets. Moreover, this carrier must also be able to transport the DNA efficiently into the cell cytoplasm, avoiding lysosomal degradation. Viral vehicles are the most commonly used carriers for delivering DNA and have long been used for their high efficiency. However, these vehicles can trigger dangerous immunological responses. Scientists need to find safer and cheaper alternatives. Consequently, the non-viral carriers are being prepared and developed until techniques for encapsulating DNA can be found. This review highlights gene therapy as a new promising technique used to treat many incurable diseases and the different strategies used to transfer DNA, taking into account that introducing DNA into the cell nucleus without degradation is essential for the success of this therapeutic technique.

Drug carriers in osteoporosis: preparation, drug encapsulation and applications.

Carriers are largely used to enhance therapy efficiency via the encapsulation of active molecules. The encapsulation enhances the stability of drug molecules, improves the targeting properties and prolongs pharmacological activity via continuous local release of active molecules. The aim of this review is to report the carrier systems used in osteoporosis therapy. This state of the art research has mainly focused on describing all types of carriers used in this area, their elaboration and properties, the drug characteristics used in such specific application, and drug release and efficiency. In this field, various processes have been used in order to obtain well-defined capsules, spheres and more complex carriers. In this exhaustive review, each process is described, illustrated and discussed.

Individual inorganic nanoparticles: preparation, functionalization and in vitro biomedical diagnostic applications.

Inorganic nanoparticles have become the focus of modern materials science due to their potential technological importance, particularly in bionanotechnology, which stems from their unique physical properties including size-dependent optical, magnetic, electronic, and catalytic properties. The present article provides an overview on the currently used individual inorganic nanoparticles for in vitro biomedical domains. These inorganic nanoparticles include iron oxides, gold, silver, silica, quantum dots (QDs) and second harmonic generation (SHG) particles. For each of these interesting nanoparticles, the main issues starting from preparation up to bio-related applications are presented.

Argan oil nanoemulsions as new hydrophobic drug-loaded delivery system for transdermal application.

This research work deals with the development of argan oil-based nanoemulsions as vehicle of hydrophobic drugs such as diclofenac used as model. Nanoemulsions of oil in water were prepared using the ultrasonication method in order to obtain submicron size colloidal dispersion. The size, zeta potential and encapsulation efficiency of the dispersions obtained were investigated. In addition, the ability of sorbitan ester derivatives to form nanovesicles (niosomes), which in turn were used for encapsulating drug in oily solutions forming stable nanoemulsions, was particularly examined. Thus, additional stabilizing agents were not required in the recipe and formulations using only sorbitan monolaurate, argan oil and water lead to attractive results. Their submicronic size (<250 nm), high negative zeta potential (between -40 and -50 mV) and drug-encapsulation efficiency (higher than 85%) allow predicting both a good physical stability and a good performance as drug carriers.

Influence of process and formulation parameters on the formation of submicron particles by solvent displacement and emulsification-diffusion methods critical comparison.

Solvent displacement and emulsification-diffusion are the methods used most often for preparing biodegradable submicron particles. The major difference between them is the procedure, which results from the total or partial water miscibility of the organic solvents used. This review is devoted to a critical and a comparative analysis based on the mechanistic aspects of particle formation and reported data on the influence of operating conditions, polymers, stabilizing agents and solvents on the size and zeta-potential of particles. In addition, a systematic study was carried out experimentally in order to obtain experimental data not previously reported and compare the data pertaining to the different methods. Thus the discussion of the behaviors reported in the light of the results obtained from the literature takes into account a wide range of theoretical and practical information. This leads to discussion on the formation mechanism of the particles and provides criteria for selecting the adequate method and raw materials for satisfying specific objectives in submicron particle design.

Stimuli-responsive magnetic particles for biomedical applications.

In recent years, magnetic nanoparticles have been studied due to their potential applications as magnetic carriers in biomedical area. These materials have been increasingly exploited as efficient delivery vectors, leading to opportunities of use as magnetic resonance imaging (MRI) agents, mediators of hyperthermia cancer treatment and in targeted therapies. Much attention has been also focused on "smart" polymers, which are able to respond to environmental changes, such as changes in the temperature and pH. In this context, this article reviews the state-of-the art in stimuli-responsive magnetic systems for biomedical applications. The paper describes different types of stimuli-sensitive systems, mainly temperature- and pH sensitive polymers, the combination of this characteristic with magnetic properties and, finally, it gives an account of their preparation methods. The article also discusses the main in vivo biomedical applications of such materials. A survey of the recent literature on various stimuli-responsive magnetic gels in biomedical applications is also included.

Polymer-based nanocapsules for drug delivery.

A review of the state of knowledge on nanocapsules prepared from preformed polymers as active substances carriers is presented. This entails a general review of the different preparation methods: nanoprecipitation, emulsion-diffusion, double emulsification, emulsion-coacervation, polymer-coating and layer-by-layer, from the point of view of the methodological and mechanistic aspects involved, encapsulation of the active substance and the raw materials used. Similarly, a comparative analysis is given of the size, zeta-potential, dispersion pH, shell thickness, encapsulation efficiency, active substance release, stability and in vivo and in vitro pharmacological performances, using as basis the data reported in the different research works published. Consequently, the information obtained allows establishing criteria for selecting a method for preparation of nanocapsules according to its advantages, limitations and behaviours as a drug carrier.

Force measurements between emulsion droplets-ssDNA conjugates: a new tool for medical diagnostics.

We describe here a new system involving direct force measurements between biomolecules that could be used in biomedical diagnostics. The method consists in the use of magnetic emulsion droplets bearing immobilized single stranded DNA fragments (ssDNA, Deoxyribo Nucleic Acid). The immobilized ssDNA fragments are able to recognize complementary DNA molecules via specific hydrogen binding (hybridization process). The ssDNA used in this study are 32 bases oligonucleotides functionalized at their 5' extremity with biotin and then immobilized onto the magnetic nanodroplets via interactions with streptavidin previously chemically grafted onto the nanomagnetic support. The aim of this work is to evaluate the possible detection of captured nucleic acid targets via single force measurements as an alternative to classical ELOSA (Enzyme Linked Oligo Sorbent Assay). The obtained results are discussed mainly in terms of electrostatic interactions.

Adsorption of oligonucleotides on PMMA/PNIPAM core-shell latexes: polarity of the PNIPAM shell probed by fluorescence.

The adsorption of a rhodamine X labeled oligonucleotide composed of 25-mers of thymine (dT(25)-ROX) onto the thermosensitive shell of PMMA/PNIPAM core-shell latex particles was studied at 22 and 40 degrees C, below and above the T(VPT) (volume phase transition temperature) of the PNIPAM shell, respectively. The experimental binding isotherms were well fitted with the cooperative Hill model. The Hill coefficient is lower than 1 at both temperatures showing that the adsorption is anticooperative. The polarity of the shell was probed by both the lifetimes and solvatochromic shifts of the zwitterionic form of rhodamine X. For temperatures below the shell T(VPT) has a polarity similar to that of water, while for temperatures above the transition the polarity is equivalent to that of a water/dioxane mixture with 30% (v/v) water.

Activity, conformation and dynamics of cutinase adsorbed on poly(methyl methacrylate) latex particles.

The adsorption of a recombinant cutinase from Fusarium solani pisi onto the surface of 100 nm diameter poly(methyl methacrylate) (PMMA) latex particles was evaluated. Adsorption of cutinase is a fast process since more than 70% of protein molecules are adsorbed onto PMMA at time zero of experiment, irrespective of the tested conditions. A Langmuir-type model fitted both protein and enzyme activity isotherms at 25 degrees C. Gamma(max) increased from 1.1 to 1.7 mg m(-2) and U(max) increased from 365 to 982 U m(-2) as the pH was raised from 4.5 to 9.2, respectively. A decrease (up to 50%) in specific activity retention was observed at acidic pH values (pH 4.5 and 5.2) while almost no inactivation (eta(act) congruent with 87-94%) was detected upon adsorption at pH 7.0 and 9.2. Concomitantly, far-UV circular dichroism (CD) spectra evidenced a reduction in the alpha-helical content of adsorbed protein at acidic pH values while at neutral and alkaline pH the secondary structure of adsorbed cutinase was similar to that of native protein. Fluorescence anisotropy decays showed the release of some constraints to the local motion of the Trp69 upon protein adsorption at pH 8.0, probably due to the disruption of the tryptophan-alanine hydrogen bond when the tryptophan interacts with the PMMA surface. Structural data associated with activity measurements at pH 7.0 and 9.2 showed that cutinase adsorbs onto PMMA particles in an end-on orientation with active site exposed to solvent and full integrity of cutinase secondary structure. Hydrophobic interactions are likely the major contribution to the adsorption mechanism at neutral and alkaline pH values, and a higher amount of protein is adsorbed to PMMA particles with increasing temperature at pH 9.2. The maximum adsorption increased from 88 to 140 mg cutinase per g PMMA with temperature raising from 25 to 50 degrees C, at pH 9.2.

Phase transfer and characterization of poly(epsilon-caprolactone) and poly(L-lactide) microspheres.

A method suitable for transfer of poly(epsilon-caprolactone) and poly(L-lactide) microspheres (synthesized by pseudoanionic dispersion polymerization of epsilon-caprolactone and L-lactide in heptane-1,4-dioxane mixed solvent) from heptane to water was developed. This method consists of treating the microspheres with KOH-ethanol in the presence of surfactants (nonionic Triton X-405, anionic sodium dodecyl sulfate (SDS), and zwitterionic ammonium sulfobetaine-2 (ASB)). Partial hydrolysis of polyesters results in the formation of hydroxyl and carboxyl groups in the surface layer of microspheres and enhances their stability in water-based media. Minimal concentrations of surfactants, needed to obtain stable suspensions of particles, were equal to 3 x 10(-2) and 6 x 10(-2), and 3 x 10(-2) mol l(-1) for Triton X-405. SDS, and ASB, respectively. In the case of poly(epsilon-caprolactone) microspheres, suspensions in water were stable for all three surfactants for pH values ranging from 3 to 11. Suspensions of poly(L-lactide) were stable in the same range of pH values only for ASB. Surface charge density determined by electrophoretic mobility varied for poly(epsilon-caprolactone) microspheres from 2.6 x 10(-7) to 8.9 x 10(-7) mol m(-2), for particles stabilized with Triton X-405 and ASB. respectively. In the case of poly(L-lactide) microspheres, surface charge density varied from 3.9 x 10(-7) (stabilizer: Triton X-405) to 7.4 x 10(-7) mol m(-2) (stabilizer: ASB). Carboxyl groups located in the surface layer of poly(L-lactide) microspheres were used for covalent immobilization of 6-aminoquinoline, a fluorophore with an amino group. Maximum surface concentration of immobilized 6-aminoquinoline was equal to 1.9 x 10(-6) mol m(-2). Poly(epsilon-caprolactone) microspheres transferred into water were loaded with ethyl salicylate. Loading up to 38% (w/w) was obtained.

Electrostatically driven immobilization of peptides onto (Maleic anhydride-alt-methyl vinyl ether) copolymers in aqueous media.

The covalent immobilization of a model peptide onto the MAMVE copolymer, via the formation of amide bonds, occurred in moderate yields in aqueous conditions. The improvement of the grafting reaction was achieved by adding at the amino terminus of the model peptide a sequence (tag) of three positively charged amino acids, lysine or arginine, and by taking profit of electrostatic attractive interactions between the negatively charged copolymer and the tagged peptides. The arginine tag was more efficient than the lysine tag for enhancing the immobilization reaction, proving that the effect was due to an electrostic driving force. On the basis of these results, a tentative mechanism is discussed, and Scatchard plots pointed out two regimes of binding. With the first, at low polymer load (up to 50% of saturation for a lysine tag and 60-70% for an arginine tag), the binding occurred with a positive cooperative effect, the already bound peptide participating to the binding of others. A second one for higher coverages, for which the binding occurred with a negative cooperativity, and saturation was reached in the presence of a large excess of peptide.

Covalent grafting of polythymidylic acid onto amine-containing polystyrene latex particles.

The covalent grafting of a 35-base poly(thymidylic) acid (dT35) onto aminated polystyrene latex has been studied. The oligonucleotide was activated by 1,4-phenylene diisothiocyanate (PDC) prior to grafting reaction. The influence of various experimental conditions related either to the medium (pH, ionic strength) or to the latex (nature and surface charge density, presence of adsorbed nonionic surfactant) was studied. Discrimination between covalent grafting and adsorption was achieved by desorbing the non-covalently grafted dT35 induced by consecutive washings with a suitable buffer solution. The amount of covalently grafted oligonucleotide directly correlated to the concentration and accessible functional groups at the particle surface.