Progress in Topical and Transdermal Drug Delivery Research-Focus on Nanoformulations.

Skin is the largest organ and a multifunctional interface between the body and its environment. It acts as a barrier against cold, heat, injuries, infections, chemicals, radiations or other exogeneous factors, and it is also known as the mirror of the soul. The skin is involved in body temperature regulation by the storage of fat and water. It is an interesting tissue in regard to the local and transdermal application of active ingredients for prevention or treatment of pathological conditions. Topical and transdermal delivery is an emerging route of drug and cosmetic administration. It is beneficial for avoiding side effects and rapid metabolism. Many pharmaceutical, technological and cosmetic innovations have been described and patented recently in the field. In this review, the main features of skin morphology and physiology are presented and are being followed by the description of classical and novel nanoparticulate dermal and transdermal drug formulations. The biophysical aspects of the penetration of drugs and cosmetics into or across the dermal barrier and their investigation in diffusion chambers, skin-on-a-chip devices, high-throughput measuring systems or with advanced analytical techniques are also shown. The current knowledge about mathematical modeling of skin penetration and the future perspectives are briefly discussed in the end, all also involving nanoparticulated systems.

Influence of design parameters on sustained drug release properties of 3D-printed theophylline tablets.

The application of three-dimensional printing (3DP) in the pharmaceutical industry brings a broad spectrum of benefits to patients by addressing individual needs and improve treatment success. This study investigates the sustained release properties of 3DP tablets containing Theophylline (TPH), which is commonly used to treat respiratory diseases and recently having a comeback due to its potential in the treatment of conditions like Covid-19. Since TPH is a narrow therapeutic window (NTW) drug with serious side effects in the event of overdose, the release properties must be observed particularly closely. We employed a state-of-the-art single screw extrusion 3D printer, which is fed with granules containing the drug. By employing a Taguchi orthogonal array design of experiments (DOE), tablet design parameters and factor related process stability were sought to be evaluated fundamentally. Following this, examinations regarding tailored TPH dosages were undertaken and a relationship between the real printed dose of selected tablet designs and their sustained drug release was established. The release profiles were analyzed using different mathematical model fits and compared in terms of mean dissolution times (MDT). Finally, in-vivo/in-vitro correlation (IVIVC) and physiologically based pharmacokinetic (PBPK) modeling showed that a paradigm patient group could be covered with the dosage forms produced.

Development of a film-forming oleogel with increased substantivity for the treatment of psoriasis.

The aim of this work was the development of a film-forming formulation (FFF) for the topical treatment of psoriasis that shows an increased substantivity compared to conventional semi-solid dosage forms. The developed formulation is an oleogel. It is based on a combination of castor oil and medium chain triglycerides, and the oil-soluble film former MP-30 (Croda GmbH, Nettetal, Germany), a polyamide that upon mixing with a polar oil entraps the oil und thus substantially increases the viscosity of the formulation up to a semisolid state. Betamethasone dipropionate (BDP) and calcipotriole (CA) were used as active pharmaceutical ingredients (APIs). Oleogels of different compositions were evaluated regarding substantivity, rheological properties, ex-vivo penetration into the skin and ex-vivo permeation through the skin. Marketed products were used as controls. It was found that the amount of betamethasone dipropionate penetrating and permeating into and through the skin from the film-forming formulation is at an intermediate value compared to the marketed products. The substantivity of the developed formulation is described by an amount of 57.7 % formulation that remains on the skin surface and is thus significantly higher compared to the marketed products. In the film forming formulation, the proportion of API penetrating the skin remains the same when the skin repetitively brought in contact with a piece of textile during the penetration experiment. In contrast with the in-market formulations tested, this proportion was reduced by up to 97 %. As a result, the developed formulations can lead to an increased patient compliance.

Evaluation of Printability of PVA-Based Tablets from Powder and Assessment of Critical Rheological Parameters.

Fused deposition modeling (FDM) is a rather new technology in the production of personalized dosage forms. The melting and printing of polymer-active pharmaceutical ingredient (API)-mixtures can be used to produce oral dosage forms with different dosage as well as release behavior. This process is utilized to increase the bioavailability of pharmaceutically relevant active ingredients that are poorly soluble in physiological medium by transforming them into solid amorphous dispersions (ASD). The release from such ASDs is expected to be faster and higher compared to the raw materials and thus enhance bioavailability. Printing directly from powder while forming ASDs from loperamide in Polyvinylalcohol was realized. Different techniques such as a change in infill and the incorporation of sorbitol as a plastisizer to change release patterns as well as a non-destructive way for the determination of API distribution were shown. By measuring the melt viscosities of the mixtures printed, a rheological model for the printer used is proposed.

Freeze-dried nanocrystal dispersion of novel deuterated pyrazoloquinolinone ligand (DK-I-56-1): Process parameters and lyoprotectant selection through the stability study.

Recently, nanocrystal dispersions have been considered as a promising formulation strategy to improve the bioavailability of the deuterated pyrazoloquinolinone ligand DK-I-56-1 (7­methoxy-2-(4­methoxy-d3-phenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one). In the current study, the freeze-drying process (formulation and process parameters) was investigated to improve the storage stability of the previously developed formulation. Different combinations of lyoprotectant (sucrose or trehalose) and bulking agent (mannitol) were varied while formulations were freeze-dried under two conditions (primary drying at -10 or -45 °C). The obtained lyophilizates were characterized in terms of particle size, solid state properties and morphology, while the interactions within the samples were analyzed by Fourier transform infrared spectroscopy. In the preliminary study, three formulations were selected based on the high redispersibility index values (around 95%). The temperature of primary drying had no significant effect on particle size, but stability during storage was impaired for samples dried at -10 °C. Samples dried at lower temperature were more homogeneous and remained stable for three months. It was found that the optimal ratio of sucrose or trehalose to mannitol was 3:2 at a total concentration of 10% to achieve the best stability (particle size < 1.0 µm, polydispersity index < 0.250). The amorphous state of lyoprotectants probably provided a high degree of interaction with nanocrystals, while the crystalline mannitol provided an elegant cake structure. Sucrose was superior to trehalose in maintaining particle size during freeze-drying, while trehalose was more effective in keeping particle size within limits during storage. In conclusion, results demonstrated that the appropriate combination of sucrose/trehalose and mannitol together with the appropriate selection of lyophilization process parameters could yield nanocrystals with satisfactory stability.

Systematic investigation of factors, such as the impact of emulsifiers, which influence the measurement of skin barrier integrity by in-vitro trans-epidermal water loss (TEWL).

Trans-epidermal water loss (TEWL) has been the most widely used method to assess the integrity of the skin barrier and evaluate the irritation potential or the protective properties of topical products for many years. It detects the amount of water that diffuses across the stratum corneum (SC) to the external environment. As one of the most important functions of the skin is to keep water inside the body, an increase in TEWL is used to indicate the skin's impaired barrier function. So far, a variety of commercial instruments are available to measure the TEWL. Their applications mainly focus on the in-vivo TEWL measurements for dermatological examinations or formulation development. Recently, an in-vitro TEWL probe has also been commercially released enabling preliminary tests with excised skin samples. In our study, we first aimed to optimize the experimental procedures for detecting the in-vitro TEWL of porcine skin. Secondly, different kinds of emulsifiers were applied to the skin, including polyethylene glycol-containing emulsifiers (PEG-ylated emulsifiers), sorbitan esters, cholesterol, and lecithin. Sodium lauryl sulfate (SLS) was used as a positive control, and water as a negative control. Based on the findings, we established a protocol for accurately measuring the in-vitro TEWL values, emphasizing that the temperature of the skin sample should be constantly maintained at 32℃. Subsequently, the influences of emulsifiers on the in-vitro TEWL were analyzed. They indicated a significant skin barrier impairment of PEG-20 cetyl ether, PEG-20 stearyl ether, and SLS on in-vitro skin. Furthermore, we interestingly found that there consistently was an alteration of the TEWL values, even after the application of water to the skin. Our findings are of special interest, as the European Medicines Agency (EMA) recommends the use of in-vitro TEWL to determine skin barrier intactness during Franz cell experiments. Thus, this study provides a validated protocol for measuring the in-vitro TEWL and elucidates the impact of emulsifiers on the skin barrier. It also improves the understanding of tolerable variations of in-vitro TEWL and offers recommendations for its use in research.

High amount of lecithin facilitates oral delivery of a poorly soluble pyrazoloquinolinone ligand formulated in lipid nanoparticles: Physicochemical, structural and pharmacokinetic performances.

Preclinical development of deuterated pyrazoloquinolinone ligands, promising drug candidates for various neuropsychiatric disorders, was hindered by unusually low solubility in water and oils. DK-I-60-3 (7-methoxy-d3-2-(4-methoxy-d3-phenyl)-2,5-dihydro-3Hpyrazolo[4,3-c]quinolin-3-one) is one of such pyrazoloquinolinones, and we recently reported about increased oral bioavailability of its nanocrystal formulation (NC). Lipid nanoparticles (LNP) with a high concentration of lecithin, which enhances loading capacity of the lipid matrix, may give rise to further improvement. After preformulation studies by differential scanning calorimetry and polarized light microscopy, LNP were prepared by the hot high pressure homogenization, and characterized in terms of particle size, morphology, and encapsulation efficacy. The layered structure visible on atomic force micrographs was confirmed by nuclear magnetic resonance. Obtained formulations were desirably stable, with small particle size (<100 nm), and high encapsulation efficacy (>99 %). Lecithin was partially fluid and most probably located in the outer shell of the particle, together with DK-I-60-3. While the hydrophobic part of polysorbate 80 was completely immobilized, its hydrophilic part was free in the aqueous phase. In oral neuropharmacokinetic study in rats, an around 1.5-fold increase of area under the curve with LNP compared to NC was noticed both in brain and plasma. In bioavailability study, F value of LNP (34.7 ± 12.4 %) was 1.4-fold higher than of NC (24.5 ± 7.8 %); however, this difference did not reach statistical significance. Therefore, employment of LNP platform in preclinical formulation of DK-I-60-3 imparted an incremental improvement of its physicochemical as well as pharmacokinetic behavior.

Assessment of penetration and permeation of caffeine by confocal Raman spectroscopy in vivo and ex vivo by tape stripping.

OBJECTIVE: Tape stripping is an often-used non-invasive destructive method to investigate the skin penetration of a substance. In recent years, however, the suitability of confocal Raman spectroscopy (CRS) as a non-invasive method of non-destructive examination of the skin has become increasingly apparent. In this study, we compared invasion and depletion penetration and permeation kinetics of a 2% caffeine solution with and without 1,2-pentanediol as a penetration enhancer measured with CRS and tape stripping. METHODS: Porcine skin was used for tape stripping and human skin for CRS. 2% caffeine solution was applied to the skin for different incubation times. Human skin was then examined by CRS while caffeine was extracted from porcine skin and quantified via reverse-phase HPLC. Fluxes were also measured and calculated by sum of the total amounts of caffeine penetrated into the skin. RESULTS: Without penetration enhancers, there is hardly any difference between the penetration profiles of the two measurement methods for invasion, but the curves for depletion are different. Furthermore, the calculated flux values for the invasion are almost identical, but for the depletion the tape stripping values are about twice as high as the CRS values. CONCLUSION: The relevance of conducting invasion and depletion studies became clear and was able to show the still existing problems in the comparability of CRS and tape stripping.

OBJECTIF: Le décapage au ruban adhésif est une méthode destructive non invasive fréquemment utilisée pour examiner la pénétration cutanée d'une substance. Cependant, ces dernières années, la pertinence de la spectroscopie raman confocale, une méthode non invasive pour examiner la peau de manière non destructive, a été de plus en plus mise en évidence. Dans cette étude, nous avons comparé les cinétiques d'invasion et de déplétion de la pénétration et de la perméation d'une solution de caféine à 2 % avec et sans 1,2-pentanediol en tant qu'amplificateur de pénétration, mesurées par spectroscopie raman confocale et par décapage au ruban adhésif. MÉTHODES: La peau de porc a été utilisée pour le décapage au ruban adhésif et la peau humaine pour la spectroscopie raman confocale. Une solution de caféine à 2 % a été appliquée sur la peau pendant différents temps d'incubation. La peau humaine a ensuite été examinée par spectroscopie raman confocale tandis que la caféine a été extraite de la peau porcine et quantifiée par HPLC en phase inverse. Les flux ont également été mesurés et calculés par la somme des quantités totales de caféine ayant pénétré dans la peau. RÉSULTATS: Sans amplificateurs de pénétration, il n'y a presque aucune différence entre les profils de pénétration des deux méthodes de mesure pour l'invasion, mais les profils pour la déplétion sont différents. De plus, les valeurs de flux calculées pour l'invasion sont presque identiques, mais pour la déplétion, les valeurs de ruban adhésif sont environ deux fois plus élevées que les valeurs CRS. CONCLUSION: La pertinence de conduire des études d'invasion et de déplétion est devenue claire et a pu montrer les problèmes toujours existants dans la comparabilité du CRS et du décapage au ruban adhésif.

Phase homogeneity in ternary amorphous solid dispersions and its impact on solubility, dissolution and supersaturation - Influence of processing and hydroxypropyl cellulose grade.

As performance of ternary amorphous solid dispersions (ASDs) depends on the solid-state characteristics and polymer mixing, a comprehensive understanding of synergistic interactions between the polymers in regard of dissolution enhancement of poorly soluble drugs and subsequent supersaturation stabilization is necessary. By choosing hot-melt extrusion (HME) and vacuum compression molding (VCM) as preparation techniques, we manipulated the phase behavior of ternary efavirenz (EFV) ASDs, comprising of either hydroxypropyl cellulose (HPC)-SSL or HPC-UL in combination with Eudragit® L 100-55 (EL 100-55) (50:50 polymer ratio), leading to single-phased (HME) and heterogeneous ASDs (VCM). Due to higher kinetic solid-state solubility of EFV in HPC polymers compared to EL 100-55, we visualized higher drug distribution into HPC-rich phases of the phase-separated ternary VCM ASDs via confocal Raman microscopy. Additionally, we observed differences in the extent of phase-separation in dependence on the selected HPC grade. As HPC-UL exhibited decisive lower melt viscosity than HPC-SSL, formation of partially miscible phases between HPC-UL and EL 100-55 was facilitated. Consequently, as homogeneously mixed polymer phases were required for optimal extent of solubility improvement, the manufacturing-dependent differences in dissolution performances were smaller using HPC-UL, instead of HPC-SSL, i.e. using HPC-UL was less demanding on shear stress provided by the process.

Novel Approach to Pharmaceutical 3D-Printing Omitting the Need for Filament-Investigation of Materials, Process, and Product Characteristics.

The utilized 3D printhead employs an innovative hot-melt extrusion (HME) design approach being fed by drug-loaded polymer granules and making filament strands obsolete. Oscillatory rheology is a key tool for understanding the behavior of a polymer melt in extrusion processes. In this study, small amplitude shear oscillatory (SAOS) rheology was applied to investigate formulations of model antihypertensive drug Metoprolol Succinate (MSN) in two carrier polymers for pharmaceutical three-dimensional printing (3DP). For a standardized printing process, the feeding polymers viscosity results were correlated to their printability and a better understanding of the 3DP extrudability of a pharmaceutical formulation was developed. It was found that the printing temperature is of fundamental importance, although it is limited by process parameters and the decomposition of the active pharmaceutical ingredients (API). Material characterization including differential scanning calorimetry (DSC) and thermogravimetric analyses (TGA) of the formulations were performed to evaluate component miscibility and ensure thermal durability. To assure the development of a printing process eligible for approval, all print runs were investigated for uniformity of mass and uniformity of dosage in accordance with the European Pharmacopoeia (Ph. Eur.).

ex vivo-in vivo comparison of drug penetration analysis by confocal Raman microspectroscopy and tape stripping.

When it comes to skin penetration analysis of a topically applied formulation, the number of suitable methods is limited, and they often lack in spatial resolution. In vivo studies are pivotal, especially in the approval of a new product, but high costs and ethical difficulties are limiting factors. For that reason, good ex vivo models for testing skin penetration are crucial. In this study, caffeine was used as a hydrophilic model drug, applied as a 2% (w/w) hydrogel, to compare different techniques for skin penetration analysis. Confocal Raman microspectroscopy (CRM) and tape stripping with subsequent HPLC analysis were used to quantify caffeine. Experiments were performed ex vivo and in vivo. Furthermore, the effect of 5% (w/w) 1,2-pentanediol on caffeine skin penetration was tested, to compare those methods regarding their effectiveness in detecting differences between both formulations.

Topically applied lipid-containing emulsions based on PEGylated emulsifiers: Formulation, characterization, and evaluation of their impact on skin properties ex vivo and in vivo.

PEGylated emulsifiers have been largely used in topical formulations for skin research. They have been a continuous study focus in our group as well. According to our previous studies, severe interruptions of the skin barrier were observed with certain types of emulsifiers. To restore the skin barrier function and counteract the effects of emulsifiers, we considered topically delivering lipids into the lipid matrix of the SC. Herein, PEG-20 cetyl ether (C20) -based oil-in-water (O/W) emulsions were developed owing to the stronger interactions of C20 with skin. The lipids containing ceramides (Cers), palmitic acids (PA), and cholesterol with different ratios and combinations were merged into the base emulsion. PEG-40 stearyl ether (S40)-based emulsion was used as a reference as S40 showed negligible impact on SC lipids. The evaluations were conducted ex vivo with confocal Raman spectroscopy (CRS) regarding the SC lipid, SC thickness, and skin penetration properties. In parallel, the in vivo irritation studies were also implemented including the transepidermal-water-loss (TEWL), skin hydration, and erythema index. The results indicated less SC lipid extraction of topically delivered lipids on ex vivo porcine skin with the addition and ratio of incorporated Cers influencing the extent of formulations counteracting the skin interruption by C20. The ex vivo penetration study showed a similar trend in drug penetration depths. In regards to the in vivo studies, TEWL was demonstrated to be suitable for differentiating the impact on skin barrier properties. The in vivo observations were generally correlated with the ex vivo results. The exact findings in this research can lead us to a better selection of applied lipid components and compositions. Future research will elucidate which type of Cer was predominantly extracted by C20, advancing future formulation development.

The Impact of the Oil Phase Selection on Physicochemical Properties, Long-Term Stability, In Vitro Performance and Injectability of Curcumin-Loaded PEGylated Nanoemulsions.

A nanotechnology-based approach to drug delivery presents one of the biggest trends in biomedical science that can provide increased active concentration, bioavailability, and safety compared to conventional drug-delivery systems. Nanoemulsions stand out amongst other nanocarriers for being biodegradable, biocompatible, and relatively easy to manufacture. For improved drug-delivery properties, longer circulation for the nanoemulsion droplets should be provided, to allow the active to reach the target site. One of the strategies used for this purpose is PEGylation. The aim of this research was assessing the impact of the oil phase selection, soybean or fish oil mixtures with medium chain triglycerides, on the physicochemical characteristics and injectability of curcumin-loaded PEGylated nanoemulsions. Electron paramagnetic resonance spectroscopy demonstrated the structural impact of the oil phase on the stabilizing layer of nanoemulsions, with a more pronounced stabilizing effect of curcumin observed in the fish oil nanoemulsion compared to the soybean oil one. The design of the experiment study, employed to simultaneously assess the impact of the oil phase, different PEGylated phospholipids and their concentrations, as well as the presence of curcumin, showed that not only the investigated factors alone, but also their interactions, had a significant influence on the critical quality attributes of the PEGylated nanoemulsions. Detailed physicochemical characterization of the NEs found all formulations were appropriate for parenteral administration and remained stable during two years of storage, with the preserved antioxidant activity demonstrated by DPPH and FRAP assays. In vitro release studies showed a more pronounced release of curcumin from the fish oil NEs compared to that from the soybean oil ones. The innovative in vitro injectability assessment, designed to mimic intravenous application, proved that all formulations tested in selected experimental setting could be employed in prospective in vivo studies. Overall, the current study shows the importance of oil phase selection when formulating PEGylated nanoemulsions.

Novel aspects of Raman spectroscopy in skin research.

The analytical technology of Raman spectroscopy has an almost 100-year history. During this period, many modifications and developments happened in the method like discovery of laser, improvements in optical elements and sensitivity of spectrometer and also more advanced light detection systems. Many types of the innovative techniques appeared (e.g. Transmittance Raman spectroscopy, Coherent Raman Scattering microscopy, Surface-Enhanced Raman scattering and Confocal Raman spectroscopy/microscopy). This review article gives a short description about these different Raman techniques and their possible applications. Then, a short statistical part is coming about the appearance of Raman spectroscopy in the scientific literature from the beginnings to these days. The third part of the paper shows the main application options of the technique (especially confocal Raman spectroscopy) in skin research, including skin composition analysis, drug penetration monitoring and analysis, diagnostic utilizations in dermatology and cosmeto-scientific applications. At the end, the possible role of artificial intelligence in Raman data analysis and the regulatory aspect of these techniques in dermatology are briefly summarized. For the future of Raman Spectroscopy, increasing clinical relevance and in vivo applications can be predicted with spreading of non-destructive methods and appearance with the most advanced instruments with rapid analysis time.

Ex vivo-In vivo correlation of retinol stratum corneum penetration studies by confocal Raman microspectroscopy and tape stripping.

Skin penetration studies of topically applied drugs are a challenging topic in the development of semisolid formulations. The most commonly used methods can be differentiated by their character into ex vivo/in vivo, invasive/non-invasive and offline/in-line measurements. In this study, we compare ex vivo tape stripping, an invasive technique, which is often used, to confocal Raman microspectroscopy (CRM), ex and in vivo, to establish a correlation between those methods. Retinol was used as a model drug, applied in an oil-in-water emulsion, to compare the skin penetration profiles obtained by the different methods.

Les études de pénétration cutanée des médicaments appliqués par voie topique constituent un véritable défi dans le développement de formulations semi-solides. Les méthodes les plus couramment utilisées peuvent être différenciées selon que leurs mesures ont lieu ex vivo/in vivo, leur caractère invasif/non invasif, ou encore selon qu'elles ont lieu en ligne/hors ligne. Dans cette étude, nous comparons le tape stripping ex vivo, une technique invasive et souvent utilisée, à la microspectroscopie confocale Raman (confocal Raman microspectroscopy, CRM), ex et in vivo, pour établir une corrélation entre ces méthodes. Le rétinol a été utilisé comme médicament modèle et appliqué dans une émulsion huile dans l'eau afin de comparer les profils de pénétration cutanée obtenus à l'aide des différentes méthodes.

Confocal Raman spectroscopy at different laser wavelengths in analyzing stratum corneum and skin penetration properties of mixed PEGylated emulsifier systems.

Emulsifier mixtures are widely used in cosmetics and pharmaceutics and thus, brought extensive studies for their performances on skin applications. PEG-20cetyl ether (C20) is recently proposed to induce skin irritation and is of interest to study its skin interactions when mixed with other emulsifiers. PEG-2oleyl ether (O2) and PEG-20stearyl ether (S20) are selected and in specific, 50 mM of C20, O2, S20 as well as Mix1 (50 mM C20 mixed with 50 mM O2) and Mix2 (50 mM C20 mixed with 50 mM S20) solutions were applied on skin samples. Confocal Raman spectroscopy (CRS) analyses of stratum corneum (SC) thickness and SC lipid content were performed after 4 h skin treatments. In parallel, skin penetration properties were also evaluated via CRS by applying procaine solutions with/without emulsifiers on skin samples for 24 h. In terms of the CRS measurements, two excitation wavelengths of 532 nm and 785 nm are both utilized in this study and we secondly aimed to compare their results and suitability in SC and skin analyses. Based on the experimental observations, comparable results are obtained by using both excitation wavelengths of 532 nm and 785 nm demonstrating their suitability in analyzing SC and skin samples. Thereinto, 785 nm laser wavelength shows the advantage of deeper skin penetration and allows the measurements of fluorescent skin samples; 532 nm laser wavelength enables simple measurement performance without substrate and coverslip interference. With regards to the results of emulsifier mixtures, the addition of S20 and O2 reduced the skin interactions and penetration enhancing ability of C20, giving us the hint to build milder systems with emulsifier mixtures. Besides, the CRS results of stronger skin interruption were also correlated with the higher critical micelle concentration (CMC) values of emulsifiers and their mixtures, which may provide evidence in explaining the interactions between emulsifiers and skin.

Critical parameters for accurate monitoring of caffeine penetration in porcine skin using confocal Raman spectroscopy.

In this research, we addressed a challenge while measuring the penetration performance of caffeine (CAF) using confocal Raman spectroscopy (CRS). Normally in the process of CRS analysis, skin sample was moved from an incubation setup to a specified CRS-measuring sample holder. Accurate data collection may be questioned due to the variation of the environment the skin placed in. Therefore, two critical parameters including the CRS measuring temperature and proper skin hydration were focused; accordingly, four different conditions were designed. First, the skin was incubated in a real-time device with the skin placing onto PBS-filled chamber where the temperature was adjusted to 32℃. This device can be fixed under the CRS microscope, enabling simultaneous skin incubation and dynamic CRS measurements (condition i, reference). The other conditions referred to skins incubated in Franz diffusion cells for simulating the common experimental procedures. In order to control variables of CRS measuring condition, skins were transferred from cells to real-time device and open device. In real-time device, proper skin hydration was maintained and the skin temperature was adjusted to 32℃ (condition ii) and room temperature (condition iii). In open device, the skin was in a less hydrated state by moving onto a PBS-soaked filter paper and wrapped with aluminum foil at room temperature (condition iv). The skin penetration performances measured in these conditions were compared with reference. Caffeine solution and gel formulation were separately applied to the skin. The results showed in both cases that the decrease of skin temperature and hydration in condition iii and iv would apparently induce the decrease of detected caffeine signal, resulting in the inaccurate data collection. To this point, it indicates the reduction of solubilized caffeine in skin layer. We suggest the forming of caffeine crystallization at varied skin conditions to be the factor. Achieved video image, CRS spectrum collection and surface scan demonstrated the caffeine crystallization process on superficial skin layer. Polarized microscopic images exemplified the crystalline drug on tape stripped skin layers. It can be a potential support of caffeine crystallization inside skin. In short, we suggest the consideration of these parameters during CRS measurements for accurate monitoring of topical drug delivery. Meanwhile, the use of real-time device for dynamic skin incubation and data collection provides advantages in saving time and efforts in this study.

Film-Forming Systems for Dermal Drug Delivery.

Film-forming formulations represent a novel form of sustained release dermatic products. They are applied to the skin as a liquid or semi-solid preparation. By evaporation of the volatile solvent on the skin, the polymer contained in the formulation forms a solid film. Various film-forming formulations were tested for their water and abrasion resistance and compared with conventional semi-solid formulations. Penetration and permeation studies of the formulations indicate a potential utility as transdermal therapeutic systems. They can be used as an alternative to patch systems to administer a variety of drugs in a topical way and may provide sustained release characteristics.

Profiling skin penetration using PEGylated emulsifiers as penetration enhancers via confocal Raman spectroscopy and fluorescence spectroscopy.

Non-ionic emulsifiers have been continuous research focus in skin analysis. With the aim of finding their role as penetration enhancers in dermal drug delivery systems, PEGylated emulsifiers of polyethylene glycol (PEG) ethers were targeted to be investigated ex-vivo. The effectiveness of them in the enhancement of skin penetration was examined by conventional tape stripping method and confocal Raman spectroscopy (CRS). Fluorescein sodium salt (Fluo-Na) and procaine HCl were respectively used as model drugs. The drug delivery performances were compared in the aspects of penetration amount and depth. Based on the results from both analyses, all investigated emulsifiers have the ability to enhance the amount of drug penetration. PEG-20 ethers showed higher ability than PEG-2 oleyl ether (O2) in promoting drug distribution by depth, especially PEG-20 cetyl ether (C20) showed a distinct effect. According to this study, their penetration enhancing performances seem to be linked to their interruption of intercellular lipids, which can be considered as the underlying mechanism for governing the ability of PEGylated emulsifiers as penetration enhancers. Further instrumental comparison highlighted the benefits of using CRS as an alternative in skin penetration analysis.

Formulation of topical acidic products and acidification of the skin - Contribution of glycolic acid.

OBJECTIVE: The acidic skin pH is one of the regulating factors of skin barrier homeostasis. Topical products as extrinsic factors which influence skin pH could be used for acidification of the skin and consequent beneficial effect. To formulate stabile and safe topical emulsion product with low pH is on-going challenge and areas interesting to explore are related to the effect of acidic products on the skin pH together with development of protocols for these studies. Aim of our work was to investigate formulations of acidic topical products with glycolic acid (GA) stabilized with long chain alkyl polyglucoside emulsifier, in regard to the specific colloidal structure of the vehicle, together with effect of products with different concentration of acidic active on skin pH. METHODS: Investigated formulations were basic vehicle and two creams with glycolic acid (concentration 2 and 10 wt%). Microstructure was investigated by polarization microscopy, Raman spectral imaging, thermal analysis and rheological measurements. Effects on the skin were assessed by measurement of biophysical skin parameters in vivo studies (5-hour, 24-hour and 7-days). In vitro screening of antimicrobial activity was performed against bacteria Staphylococcus epidermidis. RESULTS: Polarization micrographs and Raman images have shown that GA does not disturb the specific colloidal structure. Together with rheological and thermal analysis obtained results have shown that GA in higher concentrations contributes to vehicles' lamellar structure. In 5-hour study the mean values of skin pH ranged from 3.98-4.25 and 3.89-4.10 after application of products with smaller and higher GA concentration. GA samples lowered skin surface pH to 5 and less in 24-hour and 7-day study, with stronger effect of sample with more GA. Sample with 10% of GA had significant inhibitory effect on growth of S. epidermidis in 1:1 concentration. CONCLUSIONS: Investigated APG emulsifier could be used as a stabilizer for acidic topical products with GA which are characterized by satisfactory safety profile. Topical products induce acidification of the skin after short- and long-term application without barrier impairment or sign of irritation. Acidification of the skin depends on presence of ingredients which are proton donors and their concentrations.

OBJECTIF: Le pH acide de la peau est l'un des facteurs de régulation de l'homéostasie de la barrière cutanée. Les produits topiques pourraient être utilisés en tant que facteurs extrinsèques d'influence du pH cutané pour permettre l'acidification de la peau et obtenir l'effet bénéfique qui en résulte. Formuler des émulsions topiques stables et sûres à faible pH représente un défi constant et les domaines d'étude dignes d'intérêt portent sur l'effet des produits acides sur le pH cutané et sur l'élaboration de protocoles pour ces études. L'objectif de notre travail était d'étudier des formulations de produits topiques acides à base d'acide glycolique (AG) stabilisé à l'aide d'un émulsionnant à base d'alkylpolyglucoside (APG) à longue chaîne, par rapport à la structure colloïdale spécifique de l'excipient, ainsi que l'effet des produits à différentes concentrations d'acide actif sur le pH cutané. MÉTHODES: Les formulations étudiées étaient un excipient de base et deux crèmes à base d'acide glycolique (concentration égale à 2 % et 10 % de la fraction massique). La microstructure a été étudiée par microscopie à polarisation, par spectroscopie Raman, par analyse thermique et par mesures rhéologiques. Les effets cutanés ont été évalués par la mesure des paramètres cutanés biophysiques dans des études in vivo (5 heures, 24 heures et 7 jours). Un dépistage in vitro de l'activité antimicrobienne a été effectué sur la bactérie Staphylococcus epidermidis. RÉSULTATS: Les micrographies après polarisation et les images obtenues par spectroscopie Raman ont montré que l'AG ne perturbe pas la structure colloïdale spécifique. Avec les analyses rhéologique et thermique, les résultats obtenus ont montré que l'AG à des concentrations plus élevées joue un rôle dans la structure lamellaire des excipients. Dans l'étude de 5 heures, les valeurs moyennes du pH cutané allaient de 3,98 à 4,25 et de 3,89 à 4,10 après l'application des produits présentant une concentration d'AG plus faible et plus élevée. Grâce aux échantillons d'AG, le pH de la surface cutanée a diminué, passant ainsi à une valeur de 5 et à des valeurs inférieures dans les études de 24 heures et de 7 jours, et l'échantillon contenant davantage d'AG a eu un effet plus important. L'échantillon contenant 10 % d'AG a eu un effet inhibiteur significatif sur la croissance de la bactérie S. epidermidis à une concentration de 1:1. CONCLUSION: L'émulsionnant à base d'APG étudié pourrait être utilisé comme stabilisateur pour les produits topiques acides à base d'AG caractérisés par un profil d'innocuité satisfaisant. Les produits topiques induisent une acidification de la peau après une application à court et à long terme sans altération de la barrière cutanée ou signe d'irritation. L'acidification de la peau dépend de la présence de donneurs de proton parmi les composants et de leurs concentrations.