Franz Cell Research Articles

Healing activity of hydrogel containing nanoemulsified β-caryophyllene

β-caryophyllene is a sesquiterpene present in the oil of many plant species, such as Copaifera sp., which has been shown to possesses potent anti-inflammatory action; however, its healing activity remains under study. The objectives of the present study were to produce a nanoemulsion containing β-caryophyllene followed by a hydrogel containing nanoemulsified β-caryophyllene, to evaluate the permeation profile in vitro, and to assess the in vivo healing activity, which is so far unexplored in the literature for pure β-caryophyllene and in pharmaceutical formulation. The nanoemulsion was obtained through high-pressure homogenization and the hydrogel by direct dispersion with hydroxyethylcellulose. Both formulations were characterized according to droplet size, polydispersity index, volume-weighted mean diameters, particle distribution, droplets diameters tracking, zeta potential, viscosity and bioadhesion behavior. β-caryophyllene content was determined by gas chromatography coupled with mass spectrometry (GC/MS). Both formulations presented a nanometric droplet size, negative zeta potential, high β-caryophyllene content, and were stable for 60 days. In agreement with the viscosity results, the hydrogel containing the β-caryophyllene nanoemulsion showed superior bioadhesiveness than the nanoemulsion. The skin permeation study in Franz cells demonstrated that isolated β-caryophyllene was unable to cross the stratum corneum and that its nanoemulsification promoted its permeation. On the other hand, in the simulated deeply wounded skin (dermis), no significant differences were observed between the formulations and isolated β-caryophyllene with respect to the amount of marker retention in the dermis, suggesting saturation of this skin layer. For the study of healing activity, the dorsal wound model was performed with an evaluation of the lesion size, anti-inflammatory markers, and antioxidant activity. The initial closure of the wound was achieved sooner in the group treated with the hydrogel containing the β-caryophyllene nanoemulsion, indicating its anti-inflammatory effect. The histological analysis indicated that on day 12 day of the lesion, the hydrogel presented similar results to those of the positive control group (Dersani® oil), proving effectiveness in cutaneous tissue repair.

Hydrogel-forming microneedle arrays as a therapeutic option for transdermal esketamine delivery

Treatment resistant depression is, by definition, difficult to treat using standard therapeutic interventions. Recently, esketamine has been shown as a viable rescue treatment option in patients in depressive crisis states. However, IV administration is associated with a number of drawbacks and advanced delivery platforms could provide an alternative parenteral route of esketamine dosing in patients. Hydrogel-forming microneedle arrays facilitate transdermal delivery of drugs by penetrating the outer layer of the skins surface, absorbing interstitial skin fluid and swelling. This subsequently facilitates permeation of medicines into the dermal microcirculation. This paper outlines the in vitro formulation development for hydrogel-forming microneedle arrays containing esketamine. Analytical methods for the detection and quantitation of esketamine were developed and validated according to International Conference on Harmonisation standards. Hydrogel-forming microneedle arrays were fully characterised for their mechanical strength and skin insertion properties. Furthermore, a series of esketamine containing polymeric films and lyophilised reservoirs were assessed as drug reservoir candidates. Dissolution testing and content drug recovery was carried out, followed by permeation studies using 350 μm thick neonatal porcine skin in modified Franz cell apparatus. Lead reservoir candidates were selected based on measured physicochemical properties and brought forward for testing in female Sprague-Dawley rats. Plasma samples were analysed using reverse phase high performance liquid chromatography for esketamine. Both polymeric film and lyophilised reservoirs candidate patches achieved esketamine plasma concentrations higher than the target concentration of 0.15–0.3 μg/ml over 24 h. Mean plasma concentrations in rats, 24 h post-application of microneedle patches with drug reservoir F3 and LW3, were 0.260 μg/ml and 0.498 μg/ml, respectively. This developmental study highlights the potential success of hydrogel-forming microneedle arrays as a transdermal drug delivery platform for ESK and supports moving to in vivo tests in a larger animal model.

A proposed eye ex vivo permeation approach to evaluate pesticides: Case dimethoate

This study aims to propose different ex vivo methodologies for pesticide evaluation when in contact with ocular mucosa. The first ex vivo study was performed using vertical Franz cells with fresh and refrigerated excised bovine corneas. The second evaluated the permeation through the cornea by using fresh, refrigerated and damaged whole bovine eyes. Both experiments were evaluated by applying 50 μl (of 5 mg/ml solution) of an emulsifiable pesticide formulation (Dimetoato 500 EC®). In the first study, dimethoate profiles and permeation fluxes showed no statistical differences (p < .05) between fresh and refrigerated excised corneas, probably due to the excision procedure generating damage to the cornea by altering stromal structure, irrespective of the refrigeration procedure. In relation to the results obtained for developed ex vivo permeation method in whole eyes, there was a statistical difference (p < .05) between experiments performed with fresh eyes and those performed with refrigerated/damaged eyes. Damage generated by both the refrigeration process and exposure to irritant products led to a five-fold reduction in the amount of corneal-permeated pesticide as well as a two-fold increase in retention. Thus, the ex vivo permeation approach with whole eyes seems to be a simple and reproducible method to evaluate pesticides. It is evident that further evaluations need to be performed using other pesticides with distinct physicochemical characteristics.

Nanoscale investigation of human skin and study of skin penetration of Janus nanoparticles.

Janus nanoparticles (JNP) are innovative nanocarriers with an interesting pharmaceutical and cosmetic potential. They are characterized by the presence of a lipid compartment associated with an aqueous compartment delimited by a phospholipid bilayer containing phospholipids and non-ionic surfactants. The hydrodynamic diameter of JNP varies between 150 and 300nm. The purpose of this study was to answer the following questions: after cutaneous application, are JNP penetrating? If so, how deep? And in which state, intact or degraded? It was essential to understand these phenomena in order to control the rate and kinetics of diffusion of active ingredients, which can be encapsulated in this vehicle for pharmaceutical or cosmetic purposes. An innovative technique called AFM-IR, was used to elucidate the behavior of JNP after cutaneous application. This instrument, coupling atomic force microscopy and IR spectroscopy, allowing to perform chemical analysis at the nanometer scale thanks to local absorption measurements. The identification of organic molecules at the nanoscale is possible without any labelling. Before cutaneous application of JNP, the nano-structure of untreated human skin was investigated with AFM-IR. Then, in vitro human skin penetration of JNP was studied using Franz cells, and AFM-IR allowed us to perform ultra-local information investigations.

Determination of the Permeation and Penetration of Flurbiprofen into Cadaveric Human Pharynx Tissue.

ObjectiveFlurbiprofen 8.75 mg spray and lozenge have a rapid onset of action for sore throat relief, suggesting local action, although tissue penetration and the mechanism of local relief have not been determined. This investigation aimed to quantify the permeation and penetration of flurbiprofen, applied as local pharmaceutical forms, into full-thickness cadaveric human mucosal pharynx tissue, representing the clinical scenario as far as possible.MethodsA validated high-performance liquid chromatography method quantified the permeation and penetration of flurbiprofen (spray and lozenge formulations) into human cadaveric pharynx tissue using a micro Franz cell model mimicking physiological and anatomical conditions. Full-thickness mucosal pharynx tissue, consisting of oral epithelium, basement membrane, and lamina propria, was utilized to imitate the in vivo setting. Flurbiprofen was analyzed on the surface of the pharynx tissue, within the pharynx tissue and in receiver fluid, over 60 mins.ResultsFlurbiprofen was detected in receiver fluid from 10 mins following spray application and was quantifiable from 20 mins. Flurbiprofen from lozenge was detected from 10 mins and was above the limit of quantitation in receiver fluid from 40 mins. Flurbiprofen recovered from the surface of the pharynx tissue was 24.45% and 8.48% of applied dose for spray and lozenge, respectively. Flurbiprofen recovered within pharynx tissue was 46.50% and 54.65% of applied dose for spray and lozenge, respectively. For flurbiprofen lozenge, recovery within pharynx tissue was 6-fold higher relative to recovery from the pharynx tissue surface.ConclusionFlurbiprofen from spray and lozenge formulations penetrated human cadaveric pharynx tissue, indicating that flurbiprofen can reach all layers of the pharynx mucosal tissue, including the underlying lamina propria, which contains blood vessels and nerve fibers that contribute to pain during sore throat. This suggests that flurbiprofen may have a local mechanism of action for sore throat, although this has yet to be determined.

Assessment of the nail penetration of antifungal agents, with different physico-chemical properties.

Onychomycosis, or fungal nail infection, is a common fungal infection largely caused by dermatophyte fungi, such as Trichophyton rubrum or Trichophyton mentagrophytes, which affects a significant number of people. Treatment is either through oral antifungal medicines, which are efficacious but have significant safety concerns, or with topical antifungal treatments that require long treatment regimens and have only limited efficacy. Thus, an efficacious topical therapy remains an unmet medical need. Among the barriers to topical delivery through the nail are the physico-chemical properties of the antifungal drugs. Here, we explore the ability of a range of antifungal compounds with different hydrophilicities to penetrate the nail. Human nail discs were clamped within static diffusion (Franz) cells and dosed with equimolar concentrations of antifungal drugs. Using LC-MS/MS we quantified the amount of drug that passed through the nail disc and that which remained associated with the nail. Our data identified increased drug flux through the nail for the more hydrophilic compounds (caffeine as a hydrophilic control and fluconazole, with LogP -0.07 and 0.5, respectively), while less hydrophilic efinaconazole, amorolfine and terbinafine (LogP 2.7, 5.6 and 5.9 respectively) had much lower flux through the nail. On the other hand, hydrophilicity alone did not account for the amount of drug associated with/bound to the nail itself. While there are other factors that are likely to combine to dictate nail penetration, this work supports earlier studies that implicate compound hydrophilicity as a critical factor for nail penetration.

Voriconazole-loaded self-nanoemulsifying drug delivery system (SNEDDS) to improve transcorneal permeability

The aim of this study was to develop self- nanoemulsifying drug delivery system (SNEDDS) to improve the transcorneal permeability of voriconazole. A ‘mixture design around a reference mixture’ approach was applied. This latter included four components, namely, isopropyl myristate, PEG 400, Tween® 80 and Span® 80 as oil, co-solvent, surfactant and co-surfactant, respectively. Droplet size was selected as response. The effect of mixture components on droplet size was analyzed by means of response trace method. Optimal formulation was subjected to stability studies and characterized for droplet size, polydispersity index (PDI), pH, osmolarity, viscosity and percentage of transmittance. Ex-vivo transcorneal permeation of the optimal and the marketed formulations was carried out on excised bovine cornea using Franz cell diffusion apparatus. Optimal voriconazole loaded-SNEDDS showed moderate emulsification efficiency and was characterized by a droplet size of 21.447 ± 0.081 nm, a PDI of 0.156 ± 0.004, a pH of 7.205 ± 0.006, an osmolarity of 310 mosmol/Kg and a viscosity of 8.818 ± 0.076 cP. Moreover, it presented an excellent stability and exhibited a significant improvement (p < 0.05) in apparent permeability coefficient (1.982 ± 0.187 × 10−6 cm/s) when compared to commercialized formulation (1.165 ± 0.106 × 10−6 cm/s). These results suggest that SNEDDS is a promising carrier for voriconazole ocular delivery.

Enzymatically derived oil-based L-ascorbyl esters: Synthesis, antioxidant properties and controlled release from cosmetic formulations

Lipase-catalyzed transesterification of vitamin C by natural triglycerides is promising approach for cost-efficient synthesis of liposoluble food and cosmetic antioxidants. Nevertheless, application of these alternative acyl donors is insufficiently explored, despite of their low price, wide abundance and possibility for obtaining versatile products. Within current study, fatty acid ascorbyl esters were synthesized from vitamin C and natural triglycerides (lard, sunflower, coconut and linseed oil) in a process catalyzed by immobilized lipase Novozym® 435 and their controlled release was examined. High conversions were achieved at optimized conditions, even at high substrate concentrations, thus high concentrations of acsorbyl-ester were synthesized using these natural lipids. All ester mixtures exhibited very high capacity for scavenging of DPPH radicals, among which linseed oil derived esters, with ascorbyl linolenate as prevailing compound, were the most efficient (IC50 0.663 μM), while coconut oil derived ester mixture (IC50 0.739 μM), composed predominantly of molecules with medium side chains, was second best. Coconut oil derived esters were successfully incorporated in typical cosmetic formulations for controlled release of bioactive compounds - O/W emulsion and gel-emulsion. Franz cell diffusion study demonstrated that esters release from mixture and two carrier systems was formulation dependent and revealed decrease of effective diffusivities with ester side acyl chain length increase. According to calculated effective diffusivity coefficients, faster trans-membrane delivery of the same molecule was achieved from gel-emulsion comparing to O/W emulsion. Applied approach enabled highly efficient cost-saving production of fatty acid ascorbyl esters, which could find direct application in various lipophilic products.

Applications and delivery mechanisms of hyaluronic acid used for topical/transdermal delivery - A review.

The purpose of this review is to introduce the functionalities of hyaluronic acid (HA) and its potential application as an effective carrier for topical/transdermal delivery. Specifically, several delivery mechanisms of HA were summarized here in order to explain its potential permeation-enhancing roles for the skin, which includes receptor-based delivery pathway, skin hydration, hydrophobic interaction with stratum corneum, bioadhesive properties, and viscoelastic properties. To achieve the optimum delivery efficacy for bioactive compounds at different target layers of the skin, HA with various molecular weights and chemical modifications were applied to design different delivery systems, including hydrogel, nanoemulsion, microemulsion, prodrug, microneedle, liposome/hyalurosome. Delivery efficacy has been evaluated using in vitro Franz Cell Diffusion method and/or in vivo animal models. Throughout this review, it was confirmed that HA could be an effective carrier for both topical and transdermal deliveries due to its unique viscoelasticity, biocompatibility, biodegradability, non-immunogenicity, and biomedical benefits for the skin.

Influence of the ethanol/dichloromethane ratio on the preparation of microsponges composed of ethylcellulose and Eudragit or HPMCphthalate for hydrophilic drug delivery

Microsponges (MS) have shown great potential for the incorporation of relatively higher amounts of drugs. This is due to the presence of both surface pores and interconnected channels that provide a greater contact area for the absorption of active agents. However, the use of MS for carrying hydrophobic drugs is limited because the preparation methodology can totally remove hydrophilic drugs. Therefore, this work aimed to investigate possible changes in the proportion of organic solvents used to produce MS containing water-soluble drugs. A modified quasi-emulsion solvent diffusion technique was used to formulate MS with ethylcellulose and Eudragit RS 100 (MS-EE) or ethylcellulose and HPMCphthalate (MS-EH) using different combinations of ethanol/dichloromethane ratios. The effects of the polymeric composition and organic solvent ratio were evaluated on the morphology, size, swelling, yield, drug content, entrapment efficiency and in vitro release of methylene blue (MB), a hydrophilic drug model. Both polymer combinations resulted in spherical and porous particles. MS-EE was larger (8–13 μm) and displayed higher product yield (62–77%), but MS-EH showed a higher drug entrapment (11–41%), which was higher with an increase in the amount of ethanol amount used. Only MS-EH displayed in vitro MB release during 24 h (70%). ATR-FTIR and FT-Raman analyses did not demonstrate chemical interactions between MB and the polymers. Differential scanning calorimetry, thermogravimetry, and x-ray diffraction confirmed the molecular dispersion of MB in the MS polymer matrix. Ex vivo permeation studies by Franz cells and photoacoustic spectroscopy showed that MB could diffuse out from the polymer matrix and permeate both pig skin and mucosa. The particles were shown to be good carriers for MB and they provided controlled drug release and permeation through skin and mucous tissue; however, the MS particles themselves are not able to permeate these membranes. The use of ethanol in the solvent system constitutes a good strategy to obtain MS of ethylcellulose and HPMCphthalate for delivery of MB and suggests that MS is worthy of investigation as carrier for delivery other hydrophilic drugs.

Study of Melatonin as Preventive Agent of Gastrointestinal Damage Induced by Sodium Diclofenac.

Safety profile of nonsteroidal anti-inflammatory drugs (NSAIDs) has been widely studied and both therapeutic and side effects at the gastric and cardiovascular level have been generally associated with the inhibitory effect of isoform 1 (COX-1) and 2 (COX-2) cyclooxygenase enzymes. Now there are evidences of the involvement of multiple cellular pathways in the NSAIDs-mediated-gastrointestinal (GI) damage related to enterocyte redox state. In a previous review we summarized the key role of melatonin (MLT), as an antioxidant, in the inhibition of inflammation pathways mediated by oxidative stress in several diseases, which makes us wonder if MLT could minimize GI NSAIDs side effects. So, the aim of this work is to study the effect of MLT as preventive agent of GI injury caused by NSAIDs. With this objective sodium diclofenac (SD) was administered alone and together with MLT in two experimental models, ex vivo studies in pig intestine, using Franz cells, and in vivo studies in mice where stomach and intestine were studied. The histological evaluation of pig intestine samples showed that SD induced the villi alteration, which was prevented by MLT. In vivo experiments showed that SD altered the mice stomach mucosa and induced tissue damage that was prevented by MLT. The evaluation by quantitative reverse transcription PCR (RT-qPCR) of two biochemical markers, COX-2 and iNOS, showed an increase of both molecules in less injured tissues, suggesting that MLT promotes tissue healing by improving redox state and by increasing iNOS/NO that under non-oxidative condition is responsible for the maintenance of GI-epithelium integrity, increasing blood flow and promoting angiogenesis and that in presence of MLT, COX-2 may be responsible for wound healing in enterocyte. Therefore, we found that MLT may be a preventive agent of GI damages induced by NSAIDs.

The application of nanomaterial science in the formulation a novel antibiotic: Assessment of the antifungal properties of mucoadhesive clotrimazole loaded nanofiber versus vaginal films.

Candidiasis is the origin of several chronic diseases and causes a wide range of symptoms from mucosal to systemic and deadly infections. Vaginal patches are one of the best drug delivery systems for the treatment of fungal infections in the vaginal environment, so a mucoadhesive film containing drugs such as clotrimazole and metronidazole is commercially available for patients. In the present study, a physicochemical comparison is made between clotrimazole loaded film and nanofiber fabricated with the new hybrid mucoadhesive formulation of dextran and alginate. Toxicity testing was performed using the MTT assay. Bioadhesion and antifungal effects were investigated for fibers and films. The release behavior of clotrimazole from two systems was evaluated by Franz cell in each case. The most important difference between nanofibrous and film mats were obtained in antifungal, mucoadhesive, Young's modulus and morphology. The nanofiber has a higher antifungal effect and two-fold adhesive to the mouse tissue, than film. The inherent flexibility of nanofiber obviated the need for a plasticizer, which may have cytotoxic side effects. The Clotrimazole loaded nanofibrous of Alginate/Dextran mats were successfully electrospun. They exhibited more bioadhesive with higher and faster antifungal properties versus similar formulation film. Further in vivo investigation is required for their application in vaginal candidiasis.

Pesticide dermal absorption: Case study x in vitro study

In order to evaluate dermal absorption during typical working conditions in family farming, the amount of dimethoate on clothing and in the stratum corneum (SC) was measured in three rural workers. This was achieved by using cotton patches on the worker’s clothes and SC quantification by the tape stripping approach. To mimic the above study, an in vitro approach was performed using Franz cells by applying dimethoate (0.4 and 1.8 μg) direct to pig skin or, on a section of cotton before contact with pig skin. The in vivo case results demonstrated the high levels and variability of agrochemicals to which these farmers are subjected, with the total potential dermal absorption between 0.87–2.85 mg/person/h and the estimated SC penetration factor (PF) between 0–54.0 and 0–28.9 % for the back of the neck and the arms respectively. This probably demonstrates the impact of correct protective clothing. For the in vitro study, the amount of pesticide retained in the SC was 52.63 ± 10.73and 135.15 ± 31.8 ng/cm2 after applying 0.4 and 1.8 μg of pesticide directly on SC, and demonstrated close agreement with the in vivo approach. Further studies performed with this and other pesticides with different characteristics will contribute to the understanding of their transport through the skin.

Development and Percutaneous Permeation Study of Escinosomes, Escin-Based Nanovesicles Loaded with Berberine Chloride.

Escin is a natural saponin, clinically used for the anti-edematous and anti-inflammatory effects. The aim of the study was to explore the possibility of converting escin into vesicle bilayer-forming component. The hyaluronidase inhibition activity of escin was evaluated after its formulation in escinosomes. Berberine chloride, a natural quaternary isoquinoline alkaloid isolated from several medicinal plants that is traditionally used for various skin conditions was loaded in the vesicles. The developed nanovesicles were characterized in terms of diameter, polydispersity, ζ-potential, deformability, recovery, encapsulation efficiency, stability, and release kinetics. Nanovesicle permeation properties through artificial membranes and rabbit ear skin were investigated using skin-PAMPATM and Franz cells were also evaluated. Escinosomes, made of phosphatidylcholine and escin, were loaded with berberine chloride. These nanovesicles displayed the best characteristics for skin application, particularly optimal polydispersity (0.17) and deformability, high negative ζ-potential value, great encapsulation efficiency (about 67%), high stability, and the best release properties of berberine chloride (about 75% after 24 h). In conclusion, escinosomes seem to be new vesicular carriers, capable to maintain escin properties such as hyaluronidase inhibition activity, and able to load other active molecules such as berberine chloride, in order to enhance or expand the activity of the loaded drug.

Fractional Laser-Mediated siRNA Delivery for Mitigating Psoriasis-like Lesions via IL-6 Silencing

The poor permeability of topically applied macromolecules such as small interfering RNA (siRNA) has inhibited the translation to clinical application. In this study, the fractional CO2 laser-assisted approach was developed to describe siRNA permeation enhancement mediated by the created microchannels for silencing the gene to treat psoriasiform lesions. In vitro permeation using Franz cell and in vivo interleukin (IL)-6 silencing using psoriasis-like plaque in mice were evaluated to verify the impact of the laser irradiation. Low-fluence laser exposure enabled a significant increase in skin transport of siRNA, peptide, and 5-fluorouracil (5-FU). The laser treatment resulted in the enhancement of siRNA flux by 33- and 14-fold as compared to the control in nude mouse and pig skin, respectively. The laser exposure also promoted siRNA penetration across psoriatic and photoaging skins with the deficient barrier, although the enhancement level was minor compared to that of intact skin. The 3D images of confocal microscopy revealed a diffusion of macromolecules into the laser-created microchannels; the radial and vertical distribution to the surrounding and deep tissues followed this. A single laser treatment and the following topical siRNA administration were able to reduce IL-6 expression by 64% in the psoriatic skin model. Laser assistance led to the marked improvement in the plaque and the reduction of specific cytokine expression, keratinocyte proliferation, and neutrophil infiltration. Our data support the use of the fractional laser for delivery of functional nucleic acid into the skin and the target cells.

Development and application of an osthole microemulsion hydrogel for external drug evaluation

The aim of the study is to provide a new type of drug delivery, combining the advantages of a microemulsion with the transdermal administration of osthole. Appropriate oil phases, surfactants, and cosurfactants were screened by their solubility, transmittance, etc. The appropriate microemulsion ratio formula was selected by measuring the area of the formed microemulsion region from the pseudoternary phase diagram among the different proportions. After measuring the pH, zeta potential, particle size, etc., a 2% osthole drug-containing microemulsion was prepared, and transdermal experiments were performed using a Franz cell. To select a microemulsion formula with high stability and a high skin transmittance rate, 65% distilled water (w/w) was embedded into the microemulsion (7% IPM (w/w), 14% Tween 80 (w/w) and 14% Labrasol (w/w)), and the microemulsion gel was made with different proportions of carbomer. The structure, water content and molecular stability of the microemulsion gels were measured by TEM and DSC. Transdermal experiments were then performed using Franz cells, and these microemulsion gels were applied to determine the subcutaneous residual amounts and optimal skin penetration microemulsion formulations. Thereby, the best microemulsion gel formulation was selected.

Design and Development of Liquid Drug Reservoirs for Microneedle Delivery of Poorly Soluble Drug Molecules.

The poor aqueous solubility of existing and emerging drugs is a major issue faced by the pharmaceutical industry. Water-miscible organic solvents, termed co-solvents, can be used to enhance the solubility of poorly soluble substances. Typically, drugs with poor aqueous solubility and Log P > 3 are not amenable to delivery across the skin. This study investigated the use of co-solvents as reservoirs to be used in combination with hydrogel-forming microneedles to enhance the transdermal delivery of hydrophobic compounds, namely Nile red, olanzapine and atorvastatin. A custom-made Franz cell apparatus was fabricated to test the suitability of a liquid drug reservoir in combination with polymeric microneedles. A co-solvency approach to reservoir formulation proved effective, with 83.30% ± 9.38% of Nile red dye, dissolved in 1 mL poly(ethylene glycol) (PEG 400), permeating neonatal porcine skin over 24 h. PEG 400 and propylene glycol were found to be suitable reservoir media for olanzapine and atorvastatin, with approximately 50% of each drug delivered after 24 h. This work provides crucial proof-of-concept evidence that the manipulation of microneedle reservoir properties is an effective method to facilitate microneedle-mediated delivery of hydrophobic compounds.

Formulation of survival acceptor medium able to maintain the viability of skin explants over in vitro dermal experiments.

In vitro assessments of skin absorption of xenobiotics are essential for toxicological evaluations and bioavailability studies of cosmetic and pharmaceutical ingredients. Since skin metabolism can greatly contribute to xenobiotic absorption, experiments need to be performed with skin explants kept viable in suitable survival media. Existing protocols for non-viable skin are modified to consider those conditions. The objective was to design a survival medium used as an acceptor fluid in Franz cells for testing cutaneous penetration of hydrophilic or lipophilic molecules. Their metabolism inside skin may be investigated under the same conditions. The determining factors involved in survival mechanisms in vitro are discussed. The consequences of short-term skin preservation at 4°C were also evaluated. The metabolic activity of fresh skin samples mounted in Franz cells was studied by measurement of lactate release over 24h in order to assess the impacts of pH, buffering, osmolality, ionic strength, initial glucose supply and the addition of ethanol or non-ionic surfactant in the acceptor part of Franz cells. Survival media must maintain physiological pH (>5.5) be isotonic with skin cells (300mOsmkg-1 ) and contain at least 0.5gL-1 glucose. Several compositions able to preserve skin metabolism are reported. Storage of skin explants overnight at 4°C impairs skin metabolic activity. The present work provides guidelines for designing survival media according to constraints related to the scientific requirements of the experiments.

Preparation and characterization of indomethacin loaded films by piezoelectric inkjet printing: a personalized medication approach

The purpose of this study was to investigate the application of piezoelectric inkjet technology in the preparation of custom-made indomethacin (IMC) films. Indomethacin solutions with and without PVP were printed onto polymeric sheets using a commercial inkjet printer. Drug loading was varied by selecting a machine parameter different dots per inches (DPIs). The printed patches were evaluated for particulate morphologies, drug loading, in vitro release and ex vivo skin permeation and anti-inflammatory effects using hind paw inflammation model. Calculated drug loaded in 2 × 2 cm2 patches of IMC of 96, 300, and 600 DPIs were in the range of 40, 60, and 65 μg, respectively. Patches loaded with IMC alcoholic solution showed crystalline structures observed by scanning electron microscopy and the addition of PVP in solution turned it to amorphous form. The drug release profile showed 60–70% of total drug released in 3 h. Permeation studies showed 40–50% of total drug loaded permeated through rat skin using Franz cells. Patches with higher printing density 600 DPI showed anti-inflammatory effect in hind paw inflammation model studies. This study has shown the potential of personalized medicine in which a calculated amount of drug can be delivered to patients by piezoelectric technology.

Microstructure and biopharmaceutical performances of curcumin-loaded low-energy nanoemulsions containing eucalyptol and pinene: Terpenes’ role overcome penetration enhancement effect?

The objective of this work was to develop low-energy nanoemulsions for enhanced dermal delivery of curcumin, using monoterpene compounds eucalyptol (EUC) and pinene (PIN) as chemical penetration enhancers.Spontaneous emulsification was the preparation method. All formulations contained 10% of the oil phase (medium-chain triglycerides (MCT), or their mixture with EUC or PIN). Formulations were stabilized by the combination of polysorbate 80 and soybean lecithin (surfactant-to-oil-ratio=1). Concentration of curcumin was set to 3 mg/ml.Average droplet diameter of all tested formulations ranged from 102 nm to 132 nm, but the ones containing monoterpenes had significantly smaller size compared to the MCT formulation. Such finding was profoundly studied through electron paramagnetic resonance spectroscopy, which proved that the presence of monoterpenes modified the nanoemulsions’ interfacial environment, resulting in droplet size reduction. The release study of curcumin (using Franz cells) demonstrated that the cumulative amount released after 6 h of the experiment was 10.1 ± 0.2% for the MCT nanoemulsions, 13.9 ± 0.1% and 14.0 ± 0.2% for PIN and EUC formulations, respectively. In vivo tape stripping revealed their performances in delivering curcumin into the skin, indicating the following order: EUC>MCT>PIN. The formulation with EUC was clearly the most successful, giving the highest cumulative amount of curcumin that penetrated per surface unit: 34.24±5.68 µg/cm2. The MCT formulation followed (30.62±2.61 µg/cm2) and, finally, the one with PIN (21.61±0.11 µg/cm2). These results corelated with curcumin's solubility in the chosen oils: 4.18±0.02 mg/ml for EUC, 1.67±0.04 mg/ml for MCT and 0.21±0.01 mg/ml for PIN. Probably, higher solubility in the oil phase of the nanoemulsion promoted curcumin's solubility in the superficial skin layers, providing enhanced penetration.