Diffusion Cell Experiments Research Articles

Application of diffusive gradients in thin-films for in-situ monitoring of nitrochlorobenzene compounds in aquatic environments

Nitrochlorobenzene compounds (NCBs) are of key interest in environmental monitoring due to their high toxicity. To better understand the presence and fate of NCBs in aquatic environments, an in-situ sampling technique of diffusive gradients in thin films (DGT) based on hydrophilic-lipophilic-balanced (HLB) resin, combined with gas chromatography, was developed to measure four typical NCBs, e.g. meta-chloronitrobenzene (MNCB), para-chloronitrobenzene (PNCB), ortho-chloronitrobenzene (ONCB), and 2,4-dinitrochlorobenzene (CDNB). The diffusion coefficients of MNCB, PNCB, ONCB, and CDNB in agarose-based gel were firstly determined in diffusion cell experiments and ranged from 7.19 × 10−6 to 7.49 × 10−6 cm/s. The capacities of HLB-DGT for MNCB, PNCB, ONCB, and CDNB were higher than 114.65, 117.52, 117.72, and 37.58 μg/cm2, respectively. The HLB-DGT performance on NCBs determination was demonstrated to be independent of natural fluctuations in pH (3–9), ionic strength (0.001–0.5 M), and dissolved organic matter concentrations (0–20 mg/L) and of deployment time (0–120 h). In the field application, the DGT-based method to measure NCBs not only proved to be accurate and effective, but also performed better than the grab sampling method under the variable conditions. This study demonstrates that the newly developed in-situ method based on DGT can provide an attractive alternative for the routine monitoring of NCBs in aquatic environments.

Permeation-enhancing effects and mechanisms of borneol and menthol on ligustrazine: A multiscale study using in vitro and coarse-grained molecular dynamics simulation methods.

Borneol (BO) and menthol (MEN) are two widely used natural permeation enhancers in the transdermal drug delivery system. In previous studies, their permeation enhancement effects and mechanisms of action on the hydrophobic drug osthole (logP=3.8) and hydrophilic drug 5-fluorouracil (logP=-0.9) have been studied. In this study, ligustrazine (LTZ), whose logP is 1.3, was used as a model drug to provide a comprehensive understanding of the influence of its logP on the permeation-enhancing effects of BO and MEN. Both BO and MEN enhanced the permeation of LTZ through the skin stratum corneum, as determined using the modified Franz diffusion cell experiment. The enhancement mechanisms were illustrated by coarse-grained molecular dynamics simulations as follows: at low concentrations, the enhancing ratio of MEN was higher than that of BO because of the stronger perturbation effects of MEN on the lipid bilayer, making it looser and facilitating LTZ diffusion. However, at high concentrations, in addition to the diffusion mechanism, BO induced the formation of water channels to improve the permeation of LTZ; however, MEN had no significant effects through this mechanism. Their results were different from those found with osthole and 5-fluorouracil and have been discussed in this study.

Evaluation of Amphiphilic Star/Linear-Dendritic Polymer Reverse Micelles for Transdermal Drug Delivery: Directing Carrier Properties by Tailoring Core versus Peripheral Branching.

The reverse micelle self-assembly of lipophile-functionalized poly(ethylene glycol) (PEG) dendrimer hybrids is probed for applications in carrier-mediated transdermal drug delivery. Under investigation are topologically diverse amphiphiles featuring controlled branching motifs at either the polymer core (one-, two-, and four-arm PEG) and the polar/nonpolar interface (peripheral dendritic generations 0-2). Thus, a systematic investigation of the effect of branching location (core vs peripheral) on carrier properties is described. Dye-encapsulation experiments verify these materials are capable of forming well-defined aggregates and solubilizing polar compounds. Further quantification of reverse micelle critical micelle concentration and dye loading capacity for the branched amphiphile library was obtained through spectroscopy characterization. Both core and peripheral branching are shown to significantly influence dynamic encapsulation behavior, with evidence of location-based contributions extending beyond multiplicity of branching alone. Finally, the in vitro transdermal diffusion of the reverse micelle carriers was investigated through Franz diffusion cell experiments using physiologically relevant juvenile porcine dermis. The permeation results, combined with previously reported aggregate size trends, show the complex relationship between polymer branching and transdermal transport, with the lowest core- and highest peripherally-branched amphiphilic analogs exhibiting optimal transdermal permeation characteristics for this set of branched carriers.

Ultrasound-Enhanced Ciclopirox Delivery for Treatment of Onychomycosis.

The study aim was to determine ultrasound's efficacy in increasing the permeability of the nail in order to improve treatment outcomes in onychomycosis. Three sets of ultrasound experiments were performed - the luminosity experiment and two sets of diffusion cell experiments. The luminosity experiments assessed dye levels inside the nail after ultrasound application as compared to sham treatments, and the diffusion cell experiments compared changes in nail permeability due to the application of ultrasound. All in vitro experiments used planar ultrasound transducers, frequencies of 400 kHz, 600 kHz, 800 kHz, and 1 MHz, an intensity of 1 W/$cm^{2}$ and a duration of 5 min in a continuous mode. The safety of applying ultrasound to the toe was assessed by performing modeling studies. It was found that application of ultrasound at higher frequencies (800 kHz and 1 MHz) resulted in more (and statistically significant) permeation of the nail, as compared to the control trials.

Diffusion Studies of Glucose and Sucrose in Chitosan Membranes and Beads for Enzymatic Production Processes

AbstractThe diffusion of glucose and sucrose was investigated in membrane and bead experiments. Concentration‐dependent diffusion coefficients of pure glucose and sucrose were determined in precipitated chitosan membranes of varying thickness using diffusion cell experiments. Contrary to fructose, the resulting diffusion coefficients of glucose and sucrose did not reach their free diffusion coefficients at infinite dilution suggesting additional interactions between chitosan and these two sugars. Counter‐diffusion in bead experiments showed a good agreement between predicted and measured data allowing the inclusion of the diffusion data in the simulation of the proposed production process for laminaribiose. In conclusion, the encapsulation in chitosan presented a good trade‐off between increased mass transfer resistance as evaluated by the Thiele modulus and improved thermal stability and antibacterial activity.

Comparison of experimental methods for estimating matrix diffusion coefficients for contaminant transport modeling

Diffusion cell and diffusion wafer experiments were conducted to compare methods for estimating effective matrix diffusion coefficients in rock core samples from Pahute Mesa at the Nevada Nuclear Security Site (NNSS). A diffusion wafer method, in which a solute diffuses out of a rock matrix that is pre-saturated with water containing the solute, is presented as a simpler alternative to the traditional through-diffusion (diffusion cell) method. Both methods yielded estimates of effective matrix diffusion coefficients that were within the range of values previously reported for NNSS volcanic rocks. The difference between the estimates of the two methods ranged from 14 to 30%, and there was no systematic high or low bias of one method relative to the other. From a transport modeling perspective, these differences are relatively minor when one considers that other variables (e.g., fracture apertures, fracture spacings) influence matrix diffusion to a greater degree and tend to have greater uncertainty than effective matrix diffusion coefficients. For the same relative random errors in concentration measurements, the diffusion cell method yields effective matrix diffusion coefficient estimates that have less uncertainty than the wafer method. However, the wafer method is easier and less costly to implement and yields estimates more quickly, thus allowing a greater number of samples to be analyzed for the same cost and time. Given the relatively good agreement between the methods, and the lack of any apparent bias between the methods, the diffusion wafer method appears to offer advantages over the diffusion cell method if better statistical representation of a given set of rock samples is desired.

Ultrasound-Enhanced Drug Delivery for Treatment of Onychomycosis.

The aim of our study was to determine the effectiveness of using ultrasound (US) to increase the permeability of the nail, with the goal of improving outcomes in the treatment of onychomycosis. Porcine nails were used because of their similarity to human nails. A hydrophilic blue dye was used as a drug-mimicking compound. Two sets of experiments were performed: luminosity experiments to assess the dye levels inside the nail after US and sham treatments and diffusion cell experiments for determination of changes in nail permeability due to US application. In both sets of experiments, planar US transducers were used to sonicate the nails at frequencies of 400, 600, and 800 kHz and 1 MHz, an intensity of 1 W/cm2 , and a duration of 5 min in a continuous mode. Modeling studies were also performed to assess the safety of US application to the human toe for later clinical studies. In the luminosity experiments, application of US at frequencies of 600 and 800 kHz led to statistically significant results (P < .05), with an increase in dye delivery into the nail of up to 95% compared to control values. The diffusion cell results found statistical significance (P < .05) at all applied frequencies, with up to a 70% increase in the nail permeability compared to the control. Safety modeling studies found a maximal temperature increase of 4.4 °C in the bone. Our proposed US method may offer an alternative for improved treatment of onychomycosis. The current maximal temperature increase was found to be at the safety limit, and so pulsing and other alternatives will be investigated to minimize this temperature increase.

Sorption-desorption equilibrium and diffusion of tetracycline in poultry litter and municipal biosolids soil amendments

Tetracycline (TET) is commonly used to treat bacterial diseases in humans and chickens (Gallus gallus domesticus), is largely excreted, and is found at elevated concentrations in treated sewage sludge (biosolids) and poultry litter (excrement plus bedding materials). Routine application of these nutrient-and carbon-enriched materials to soils improves fertility and other characteristics, but the presence of antibiotics (and other pharmaceuticals) in amendments raises questions about potential adverse effects on biota and development of antibiotic resistance in the environment. Hazard risks are largely dictated by sorption-desorption and diffusion behavior in amendments, so these processes were evaluated from sorption-desorption equilibrium isotherm and diffusion cell experiments with four types amendments (biosolids, poultry manure, wood chip litter, and rice hull litter) at three temperatures (8 °C, 20 °C and 32 °C). Linear sorption-desorption equilibrium distribution constants (Kd) in native amendments ranged between 124–2418 L kg−1. TET sorption was significantly increased after treatment with alum, and there was a strong exponential relationship between Kd and the concentration of bound Al3+ in amendments (R2 = 0.94), which indicated that amendments contained functional groups capable of chelating Al3+ and forming metal bridges with TET. Effective diffusion coefficients of TET in amendments ranged between 0.1 and 5.2 × 10−6 cm2 s−1, which were positively related to temperature and inversely related to Kd by a multiple regression model (R2 = 0.86). Treatment of organic amendments with alum greatly increased Kd, would decrease Ds, and so would greatly reduce hazard risks of applying these organic amendments with this antibiotic to soils.

Dendrimer pre-treatment enhances the skin permeation of chlorhexidine digluconate: Characterisation by in vitro percutaneous absorption studies and Time-of-Flight Secondary Ion Mass Spectrometry

Skin penetration and localisation of chlorhexidine digluconate (CHG) within the skin have been investigated in order to better understand and optimise the delivery using a nano polymeric delivery system of this topically-applied antimicrobial drug. Franz-type diffusion cell studies using in vitro porcine skin and tape stripping procedures were coupled with Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) to visualise the skin during various treatments with CHG and polyamidoamine dendrimers (PAMAM). Pre-treatment of the skin with PAMAM dendrimers significantly increased the amount and depth of permeation of CHG into the skin in vitro. The effect observed was not concentration dependant in the range 0.5–10mM PAMAM. This could be important in terms of the efficiency of treatment of bacterial infection in the skin. It appears that the mechanism of enhancement is due to the PAMAM dendrimer disrupting skin barrier lipid conformation or by occluding the skin surface. Franz-type diffusion cell experiments are complimented by the detailed visualisation offered by the semi-quantitative ToF-SIMS method which provides excellent benefits in terms of sensitivity and fragment ion specificity. This allows a more accurate depth profile of chlorhexidine permeation within the skin to be obtained and potentially affords the opportunity to map the co-localisation of permeants with skin structures, thus providing a greater ability to characterise skin absorption and to understand the mechanism of permeation, providing opportunities for new and more effective therapies.

Numerical simulation of Franz diffusion experiment: Application to drug loaded soft contact lenses

Drug loaded soft contact lenses (SCLs) have been considered as an alternative to drug administration via eye-drops to improve bioavailability and patient comfort. Understanding the drug delivery characteristics is key to the design of therapeutic drug delivery systems. In this study, Franz diffusion cell experiments were carried out in order to analyse such characteristics of hydrogel used in SCLs. Two types of experiments were carried out. In one case, the donor compartment was loaded with an initial concentration of diclofenac while in the other case the lens material was pre-loaded with diclofenac. Experiments were carried out with varied initial concentration and temperature. In order to simulate the experimental data and estimate relevant physical parameters of the SCL material, we introduce a general mathematical multi-layer model that takes into account diffusion within each layer as well as the effects of both partition and barriers between layers. The model includes a novel treatment of the discontinuity in drug concentration between layers in an accurate manner. We succeeded in simulating all these experiments. However, the donor-loaded experiments proved more consistent than the pre-loaded case as the same set of estimated parameters could consistently explain varied starting conditions in the simulation of donor-loaded experiments.

Tetramethylpyrazine-Loaded Hydrogels: Preparation, Penetration Through a Subcutaneous-Mucous-Membrane Model, and a Molecular Dynamics Simulation.

Tetramethylpyrazine (TMP) was extracted from Ligusticum chuanxiong hort. The compound is known to have a variety of medicinal functions; in particular, it is used for the treatment of cerebral ischemic diseases. TMP-loaded hydrogels offer an excellent preparation with the capacity to bypass the blood-brain barrier, allowing treatment of the brain through intranasal administration. We prepared TMP-loaded hydrogels using carbomer 940 and evaluated the release of TMP from the hydrogel. We determined the release rate using Franz-type diffusion cell experiments with a subcutaneous-mucous-membrane model and also by a molecular dynamics (MD) simulation. In general, the former method was more complicated than the latter was. The dynamic behavior of TMP release from the hydrogel was revealed by analysis of the mean square displacement of the trajectory in the MD simulation. The coefficient of TMP diffusion from the hydrogel was calculated at different temperatures (277, 298, and 310K) by using MD software. The results showed that the coefficient of diffusion increased with an increase in temperature. This trend was observed both experimentally and in the MD simulation. Therefore, the MD simulation was a complementary method to verify the experimental data.

The Correlative Approach of 3D Imaging Techniques, Simulation and Diffusion Experiments to Explore Transport Properties in Porous Otm Support Material

Solid state electrochemical energy devices such as batteries, fuel cells and oxygen transport membranes (OTMs) have the potential to revolutionize distributed low-carbon power supply. However, in order to accelerate their commercialization across a range of applications, a much improved understanding of the underlying material microstructure is required. In particular, the rate limiting step of an OTM membrane applied for syngas production is found in concentration losses caused by mass transport limitations. In this field, microstructural parameters, such as tortuosity, porosity and average pore diameter, play a vital role in characterising, comparing and evaluating porous materials. However, unlike the latter two parameters, tortuosity cannot be measured directly which is why different methods have been developed for this purpose. These differ considerably from each other in terms of calculation approach and data preparation techniques. Here, we utilise X-ray nano computed tomography (CT) techniques to reveal the complex microstructure and resolve geometric features affecting mass transport of tubular porous support layers of OTMs to examine the relationship between the pore structure and mass transport (Figure 1). This is achieved via image based tortuosity calculation methods: a fast marching computational technique and a Laplace solver using MATLAB is employed to examine the geometrical tortuosity, and StarCCM+ and COMSOL software packages are used to model the heat flux and effective transport parameters of the porous phase, respectively. In turn, the results are validated using diffusion cell experiments to evaluate the bulk transport properties of the same samples for different binary gas mixtures at different temperatures. Tortuosity is then extracted by applying equimolar and equimass diffusion models [1]. We conclude, that image based calculation methodologies arrive at lower tortuosity values in comparison to diffusion cell experiments (Figure 2). Even the widely used Bruggeman [2] and Maxwell relations prove to be unfit to estimate tortuosity values in the analysed microstructure. The observed difference may stem from Knudsen diffusion effects which are not accounted for in the entirety of tomography based modelling techniques. Secondly, tortuosity values extracted via diffusion cell experiments are a function of gas mixture, temperature and microstructural characteristics rather than a constant value. Consequently, care must be taken in applying purely continuum interpretations of diffusion processes in these complex porous media.

Ethyl cellulose nanocarriers and nanocrystals differentially deliver dexamethasone into intact, tape-stripped or sodium lauryl sulfate-exposed ex vivo human skin - assessment by intradermal microdialysis and extraction from the different skin layers

Understanding penetration not only in intact, but also in lesional skin with impaired skin barrier function is important, in order to explore the surplus value of nanoparticle-based drug delivery for anti-inflammatory dermatotherapy.Herein, short-term ex vivo cultures of (i) intact human skin, (ii) skin pretreated with tape-strippings and (iii) skin pre-exposed to sodium lauryl sulfate (SLS) were used to assess the penetration of dexamethasone (Dex). Intradermal microdialysis was utilized for up to 24h after drug application as commercial cream, nanocrystals or ethyl cellulose nanocarriers applied at the therapeutic concentration of 0.05%, respectively. In addition, Dex was assessed in culture media and extracts from stratum corneum, epidermis and dermis after 24h, and the results were compared to those in heat-separated split skin from studies in Franz diffusion cells.Providing fast drug release, nanocrystals significantly accelerated the penetration of Dex. In contrast to the application of cream and ethyl cellulose nanocarriers, Dex was already detectable in eluates after 6h when applying nanocrystals on intact skin. Disruption of the skin barrier further accelerated and enhanced the penetration. Encapsulation in ethyl cellulose nanocarriers delayed Dex penetration. Interestingly, for all formulations highly increased concentrations in the dialysate were observed in tape-stripped skin, whereas the extent of enhancement was less in SLS-exposed skin. The results were confirmed in tissue extracts and were in line with the predictions made by in vitro release studies and ex vivo Franz diffusion cell experiments. The use of 45kDa probes further enabled the collection of inflammatory cytokines. However, the estimation of glucocorticoid efficacy by Interleukin (IL)-6 and IL-8 analysis was limited due to the trauma induced by the probe insertion.Ex vivo intradermal microdialysis combined with culture media analysis provides an effective, skin-sparing method for preclinical assessment of novel drug delivery systems at therapeutic doses in models of diseased skin.

Chemical penetration enhancers in stratum corneum — Relation between molecular effects and barrier function

Skin is attractive for drug therapy because it offers an easily accessible route without first-pass metabolism. Transdermal drug delivery is also associated with high patient compliance and through the site of application, the drug delivery can be locally directed. However, to succeed with transdermal drug delivery it is often required to overcome the low permeability of the upper layer of the skin, the stratum corneum (SC). One common strategy is to employ so-called penetration enhancers that supposedly act to increase the drug passage across SC. Still, there is a lack of understanding of the molecular effects of so-called penetration enhancers on the skin barrier membrane, the SC. In this study, we provide a molecular characterization of how different classes of compounds, suggested as penetration enhancers, influence lipid and protein components in SC. The compounds investigated include monoterpenes, fatty acids, osmolytes, surfactant, and Azone. We employ natural abundance 13C polarization transfer solid-state nuclear magnetic resonance (NMR) on intact porcine SC. With this method it is possible to detect small changes in the mobility of the minor fluid lipid and protein SC components, and simultaneously obtain information on the major fraction of solid SC components. The balance between fluid and solid components in the SC is essential to determine macroscopic material properties of the SC, including barrier and mechanical properties. We study SC at different hydration levels corresponding to SC in ambient air and under occlusion. The NMR studies are complemented with diffusion cell experiments that provide quantitative data on skin permeability when treated with different compounds. By correlating the effects on SC molecular components and SC barrier function, we aim at deepened understanding of diffusional transport in SC, and how this can be controlled, which can be utilized for optimal design of transdermal drug delivery formulations.

The application of 3D imaging techniques, simulation and diffusion experiments to explore transport properties in porous oxygen transport membrane support materials

Tortuosity plays a vital role in diffusive mass transport through porous layers of electrochemical devices. However, tortuosity cannot be measured directly which is why different methods have been developed to determine the tortuosity of porous samples. These methods differ considerably from each other in terms of calculation approach and applied data sets.Here, we utilise advanced tomography techniques, simulation and diffusion cell experiments to calculate tortuosity of a porous support layer of an oxygen transport membrane (OTM). The results are then compared and the suitability of these different approaches is evaluated.We conclude, that image and simulation based methodologies arrive at lower tortuosity values in comparison to diffusion cell experiments; the difference, depending on the calculation method, can be close to a factor of two. The difference may stem from Knudsen diffusion effects which currently are not accounted for in the image based modelling techniques. Moreover, tortuosity values extracted via diffusion cell experiments are a function of gas mixture and temperature. As a result, one has to distinguish between geometric tortuosity and diffusive tortuosity used for quantifying the resistance of a structure to a flux.

Double emulsions based on silicone-fluorocarbon-water and their skin penetration

Double emulsions have significant potential in pharmacy and cosmetics due to the feasibility of combining incompatible substances in one product and the protection of sensitive compounds by incorporating them into their innermost phase. However, a major drawback of double emulsions is their thermodynamic instability and their strong tendency to coalesce. In the present study, the physicochemical stability, the skin permeation and the skin penetration potential of modified semi-solid double emulsions was investigated. The double emulsions were prepared of the cosmetically applied perfluoropolyethers Fomblin® HC/04 or Fomblin® HC-OH, silicone, carbomer and water. Measurement of the droplet size and examination of the microscopic images confirmed their physicochemical stability over the observation period of eight weeks. Franz-type diffusion cell experiments revealed no increase in curcumin permeation due to the employed perfluoropolyethers compared to the respective control formulations. The formulations used as control were O/W macroemulsions with or without a Polysorbate 80/Sorbitane monooleate 80 surfactant combination. Likewise, tape stripping studies showed no penetration enhancing effect of the employed perfluoropolyethers which is desirable as both perfluoropolyethers are commonly applied components in human personal-care products.

Simultaneous determination of active component and vehicle penetration from F-DPPC liposomes into porcine skin layers

Liposomes have been used as innovative delivery vehicles on skin for a number of years due to their positive influence on skin penetration. However, until now it is not entirely clear how and by which mechanism enhancement is achieved. In the present study, the skin permeation of a model substance incorporated into liposomes and a control formulation was compared to study the influence of the vehicle in Franz-type diffusion cell experiments. Furthermore, the penetration depths of both components were studied by simultaneous determination of the active substance and the vehicle component during tape stripping studies and horizontal sectioning. For these purposes we prepared liposomes with 1-palmitoyl-2-(16-fluoropalmitoyl)-sn-glycero-3-phosphocholine (F-DPPC), the monofluorinated analogue of dipalmitoylphosphaditylcholine (DPPC) loaded with sodium fluorescein (SoFl). A sodium-fluorescein solution was used as control formulation. While the semi-solid F-DPPC liposomes and the SoFl-solution performed equally well with similar permeation profiles during skin diffusion experiments, superior penetrated amounts of SoFl into the stratum corneum (SC) from F-DPPC liposomes compared to the SoFl-solution were observed possibly due to a “push” exerted by the vehicle F-DPPC. We also showed that SoFl penetrated through SC into the viable epidermis.

Development and in vitro evaluation of lipid nanoparticle-based dressings for topical treatment of chronic wounds

This research addresses the development and in vitro evaluation of lipid nanoparticle (NP)-based dressings to optimize the delivery of human recombinant epidermal growth factor (rhEGF) for the topical treatment of chronic wounds. The systems investigated were rhEGF-loaded solid lipid nanoparticles (rhEGF-SLN) and rhEGF-loaded nanostructured lipid carriers (rhEGF-NLC) formulated in wound dressings comprising either semi-solid hydrogels or fibrin-based solid scaffolds. Following detailed characterisation of the NP, in vitro diffusion cell experiments (coupled with dermatopharmacokinetic measurements), together with confocal microscopic imaging, conducted on both intact skin samples, and those from which the barrier (the stratum corneum) had been removed, revealed that (a) the particles remained essentially superficially located for at least up to 48h post-application, (b) rhEGF released on the surface of intact skin was unable to penetrate to the deeper, viable layers, and (c) sustained release of growth factor from the NP “drug reservoirs” into barrier-compromised skin was observed. There were no significant differences between the in vitro performance of rhEGF-SLN and rhEGF-NLC, irrespective of the formulation employed. It is concluded that, because of their potentially longer-term stability, the fibrin-based scaffolds may be the most suitable approach to formulate rhEGF-loaded lipid nanoparticles.

Skin Concentrations of Topically Applied Substances in Reconstructed Human Epidermis (RHE) Compared with Human Skin Using in vivo Confocal Raman Microscopy.

Detailed knowledge about the skin concentration of topically applied substances is important to understand their local pharmacological activity. In particular since in vitro models of reconstructed human epidermis are increasingly used as models for diseased skin. In general, diffusion cell experiments are performed to determine the diffusion flux of test substances through either skin models or excised skin both from humans and animals. Local concentrations of the test substances within the skin are then calculated applying diffusion laws and suitable boundary conditions. In this study we used a direct approach to reveal the local concentrations of test substances within skin using confocal Raman microscopy. This non-invasive method can also be applied in vivo and therefore we directly compared in vivo concentrations with those obtained from commercially available reconstructed human epidermis (RHE). Hydrophilic and lipophilic test substances with log Pow from -0.07 to 5.91 were topically applied on human skin in vivo and RHE from SkinEthic was used as the commercial skin model. Local concentration profiles in the stratum corneum (SC) showed substantial differences between the RHE model and the in vivo situation. Differences between RHE models and human skin in vivo were also observed in their molecular composition, in particular in terms of their water profile, lipid content and the presence of natural moisturizing factor (NMF). Confocal Raman is shown to be a powerful non-invasive method for qualitative and quantitative comparative studies between RHE models and human skin in vivo. This method can also be applied to validate RHE models for future use in clinical studies.

Cell penetrating peptide conjugated liposomes as transdermal delivery system of Polygonumaviculare L. extract

In this study, Polygonum aviculare L. extract, which has superior antioxidative and cellular membrane protective activity, was loaded onto cell penetrating peptide (CPP) conjugated liposomes to enhance transdermal delivery. The physical characteristics of typical liposomes and CPP-conjugated liposomes containing P. aviculare extract were evaluated. The particle sizes of both liposomes were approximately 150nm. Whereas the zeta potential of typical liposomes was −45mV, that of CPP-conjugated liposomes was +42mV. The loading efficiency of P. aviculare extract in both liposomes was calculated to be about 83%. Fluorescent-labeled liposomes were prepared to evaluate cellular uptake and skin permeation efficiency. Using flow cytometry, we found that CPP-conjugated liposomes improved cellular uptake of the fluorescent dye as compared with the typical liposomes. In addition, the skin permeation of CPP-conjugated liposomes was proved higher than that of typical liposomes by confocal laser scanning microscopy studies and Franz diffusion cell experiments. The improved cellular uptake and skin permeation of the CPP-conjugated liposomes were due to the cationic arginine-rich peptide. In vivo studies also determined that the CPP-conjugated liposomes were more effective in depigmentation and anti-wrinkle studies than typical liposomes. These results indicate that the CPP-conjugated liposomes could be effective for transdermal drug delivery of antioxidant and anti-aging therapeutics.