A novel technique to evaluate nail softening effects of different urea formulations.

Urea has been incorporated into several topical ungual formulations to hydrate and soften the nail plate. In this study, we employed various characterization techniques (visual observation, scanning electron microscopy, measurement of thickness, transonychial water loss, nail electrical resistance, and mechanical study) to investigate the effect of urea concentration on the hydration of bovine hoof membranes - an in vitro model of infected human nails. We obtained inconsistent results in the thickness, transonychial water loss, nail electrical resistance, and scanning electron microscopy studies. In the mechanical study using a modified Texture Analyzer method, we reported an inverse and linear correlation between urea concentrations in the formulations and the force required to puncture the treated membrane (R2 = 0.9582, n ≥ 8). As the urea concentration decreased from 4x to 2x, 1x, and 0x % w/w, the puncture force increased significantly from 0.47 ± 0.07 to 0.77 ± 0.07, 0.91 ± 0.09, and 1.33 ± 0.26 N, respectively (p < 0.05). Thus, urea provided a positive softening effect on the membranes and the puncture force could indicate the urea level in topical formulations. In this study, we provided a novel, efficient, and reliable tool to evaluate the hydration level and physical properties of bovine hoof membranes.

Delivery of Methotrexate and Characterization of Skin Treated by Fabricated PLGA Microneedles and Fractional Ablative Laser.

PURPOSE: This study investigated in vitro transdermal delivery of methotrexate through dermatomed porcine ear and cadaver human skin treated with poly (D,L-lactide-co-glycolide) acid microneedles or fractional ablative laser. METHODS: PLGA microneedles were fabricated and characterized using scanning electron microscopy and mechanical assessment techniques. The integrity of treated skin was evaluated by rheometer, transepidermal water loss, and skin electrical resistance measurements. Successful skin microporation was demonstrated by dye binding, histology, pore uniformity, confocal laser microscopy, and DermaScan studies. In vitro permeation experiment was performed on Franz diffusion cells to determine drug delivery into and across the skin. RESULTS: Both physical treatments resulted in a considerable decrease in skin resistance and an increase in transepidermal water loss value. The laser-created microchannels were significantly larger than those formed by microneedles (p < 0.05). An effective force of 41.04 ± 18.33 N was required to achieve 100% penetration efficiency of the microneedles. For both porcine ear and human skin, laser ablation provided a significantly higher methotrexate permeability into the receptor chamber and skin layers compared to microneedle poration and untreated skin (p < 0.05). CONCLUSIONS: Both fractional ablative laser and polymeric microneedles markedly enhanced in vitro transdermal delivery of methotrexate into and across skin. Graphical Abstract ᅟ.

Polymeric Microneedles Enhance Transdermal Delivery of Therapeutics.

This research presents the efficacy of polymeric microneedles in improving the transdermal permeation of methotrexate across human skin. These microneedles were fabricated from PLGA Expansorb® 50-2A and 50-8A and subjected to comprehensive characterization via scanning electron microscopy, Fourier-transform infrared spectroscopy, and mechanical analysis. We developed and assessed a methotrexate hydrogel for physicochemical and rheological properties. Dye binding, histological examinations, and assessments of skin integrity demonstrated the effective microporation of the skin by PLGA microneedles. We measured the dimensions of microchannels in the skin using scanning electron microscopy, pore uniformity analysis, and confocal microscopy. The skin permeation and disposition of methotrexate were researched in vitro. PLGA 50-8A microneedles appeared significantly longer, sharper, and more mechanically uniform than PLGA 50-2A needles. PLGA 50-8A needles generated substantially more microchannels, as well as deeper, larger, and more uniform channels in the skin than PLGA 50-2A needles. Microneedle insertion substantially reduced skin electrical resistance, accompanied by an elevation in transepidermal water loss values. PLGA 50-8A microneedle treatment provided a significantly higher cumulative delivery, flux, diffusion coefficient, permeability coefficient, and predicted steady-state plasma concentration; however, there was a shorter lag time than for PLGA 50-2A needles, base-treated, and untreated groups (p < 0.05). Conclusively, skin microporation using polymeric microneedles significantly improved the transdermal delivery of methotrexate.

Microneedle-Mediated Transdermal Delivery of Biopharmaceuticals.

Transdermal delivery provides numerous benefits over conventional routes of administration. However, this strategy is generally limited to a few molecules with specific physicochemical properties (low molecular weight, high potency, and moderate lipophilicity) due to the barrier function of the stratum corneum layer. Researchers have developed several physical enhancement techniques to expand the applications of the transdermal field; among these, microneedle technology has recently emerged as a promising platform to deliver therapeutic agents of any size into and across the skin. Typically, hydrophilic biomolecules cannot penetrate the skin by passive diffusion. Microneedle insertion disrupts skin integrity and compromises its protective function, thus creating pathways (microchannels) for enhanced permeation of macromolecules. Microneedles not only improve stability but also enhance skin delivery of various biomolecules. Academic institutions and industrial companies have invested substantial resources in the development of microneedle systems for biopharmaceutical delivery. This review article summarizes the most recent research to provide a comprehensive discussion about microneedle-mediated delivery of macromolecules, covering various topics from the introduction of the skin, transdermal delivery, microneedles, and biopharmaceuticals (current status, conventional administration, and stability issues), to different microneedle types, clinical trials, safety and acceptability of microneedles, manufacturing and regulatory issues, and the future of microneedle technology.

Comparing combined laser iridoplasty and surgical iridectomy with trabeculectomy in treatment of refractory acute primary angle closure without significant cataract: a randomized controlled trial.

OBJECTIVES: To compare the safety and efficacy of combined laser iridoplasty followed by surgical iridectomy (LI-SI) versus trabeculectomy in the management of medically unresponsive acute primary angle closure (APAC) with minimal cataract. PATIENTS AND METHODS: This was a randomized controlled trial conducted among patients with medically unresponsive APAC without significant cataract. Study participants were randomized into: LI-SI or unaugmented trabeculectomy. Primary outcome of the study was the rate of post-operative surgical complications in the first 3 months after surgery. Secondary outcome assessed at 1 year was whether treatment was completely successful (IOP < 21 mmHg without IOP lowering drops), or partially successful (IOP < 21 mmHg with IOP lowering drops). Failure was defined as IOP ≥ 21 mmHg with IOP lowering drops. RESULTS: The study included 67 eyes of 67 patients (59 females/8 males = 7.4/1) who were randomized into 2 groups: LI-SI (Group 1, 37 eyes), and trabeculectomy (Group 2, 30 eyes). There was no statistical difference between the two groups at baseline. Overall, there were more post-operative complications in Group 1 versus Group 2 (45.9% versus 33.3% - p = 0.23), although all responded well to medical treatment and resolved without sequelae. Complete success was found in 97.1% (34/35 eyes) in Group 1 and 92.6% in group 2 (p = 0.19, Fisher's exact test). CONCLUSIONS: There was a higher rate of post-operative complications after LI-SI compared to trabeculectomy performed for medically unresponsive APAC with minimal cataract. Both procedures had similar surgical outcomes at 1 year.

Characterization of microneedles and microchannels for enhanced transdermal drug delivery.

Microneedle (MN)-based technologies are currently one of the most innovative approaches that are being extensively investigated for transdermal delivery of low molecular weight drugs, biotherapeutic agents and vaccines. Extensive research reports, describing the fabrication and applications of different types of MNs, can be readily found in the literature. Effective characterization tools to evaluate the quality and performance of the MNs as well as for determination of the dimensional and kinetic properties of the microchannels created in the skin, are an essential and critical part of MN-based research. This review paper provides a comprehensive account of all such tools and techniques.

Electrically and Ultrasonically Enhanced Transdermal Delivery of Methotrexate.

In this study, we used sonophoresis and iontophoresis to enhance the in vitro delivery of methotrexate through human cadaver skin. Iontophoresis was applied for 60 min at a 0.4 mA/sq·cm current density, while low-frequency sonophoresis was applied at a 20 kHz frequency (2 min application, and 6.9 W/sq·cm intensity). The treated skin was characterized by dye binding, transepidermal water loss, skin electrical resistance, and skin temperature measurement. Both sonophoresis and iontophoresis resulted in a significant reduction in skin electrical resistance as well as a marked increase in transepidermal water loss value (p < 0.05). Furthermore, the ultrasonic waves resulted in a significant increase in skin temperature (p < 0.05). In permeation studies, the use of iontophoresis led to a significantly higher drug permeability than the untreated group (n = 4, p < 0.05). The skin became markedly more permeable to methotrexate after the treatment by sonophoresis than by iontophoresis (p < 0.01). A synergistic effect for the combined application of sonophoresis and iontophoresis was also observed. Drug distribution in the skin layers revealed a significantly higher level of methotrexate in the sonicated skin than that in iontophoresis and untreated groups. Iontophoresis and low-frequency sonophoresis were found to enhance the transdermal and intradermal delivery of methotrexate in vitro.

Effect of ablative laser on in vitro transungual delivery.

Topical therapy of nail psoriasis using methotrexate has not been realized due to the high molecular weight and low permeability of the compound. In this study, we used a 2940nm fractional ablative laser to disrupt the nail barrier to enhance the in vitro transungual delivery of methotrexate. Bovine hoof membrane-an in vitro model of the human nail-was treated by the laser at different energy levels and pore densities. A successful microporation was characterized by mechanical properties, scanning electron microscopy, Fourier transform infrared spectrophotometer, dye binding, histology, pore uniformity, confocal laser microscopy, nail integrity measurement, and permeation studies. No significant difference in the pore dimension was found in different treatment groups (p>0.05). Increases in pore depth corresponded with increases in the laser energy. Laser ablation was found to affect the mechanical properties of the hoof membrane. In in vitro permeation studies, laser ablation resulted in a significant increase in the drug cumulative delivery, flux, and permeability coefficient as compared to the untreated group (n=3, p<0.05). A change in the laser energy and pore density was found to alter the drug permeability. Thus, transungual methotrexate delivery was enhanced by the fractional laser ablation.

Qualitative and quantitative analysis of lateral diffusion of drugs in human skin.

This study aimed to qualitatively and quantitatively analyze lateral diffusion of drugs in dermatomed human skin. Lateral diffusion of calcein and methylene blue dyes in skin was investigated using confocal laser microscopy, calcein imaging, and histology studies. In in vitro permeation studies, two linear microdialysis probes were inserted into the dermis of untreated, poly lacto-glycolic acid microneedle-treated, and ablative laser-treated skin such that one was in the center of the diffusion area and the other was parallel, at 8 mm from the central probe. Skin was mounted on Franz cells, sandwiched between donor containing diclofenac sodium solution and receptor containing phosphate buffered saline, pH 7.4. Qualitative techniques revealed faster lateral diffusion of the dyes in microneedle-treated skin than laser-treated skin. Rate of drug diffusion in the central probe in the microneedle-treated skin (11.8 ±â€¯2.5 µg/h) was significantly higher than untreated and laser-treated skin (p  <  0.05). Rate of lateral diffusion in untreated group (0.7 ±â€¯0.1 µg/h) was significantly lower than microneedle and laser-treated skin (p  <  0.05). Overall, in vitro microdialysis was demonstrated as a novel and valuable tool that can be employed for quantitative investigation of rate of vertical and lateral diffusion of drugs in intact and microporated skin.

Poly (vinyl alcohol) microneedles: Fabrication, characterization, and application for transdermal drug delivery of doxorubicin.

Poly (vinyl alcohol) microneedles were fabricated, characterized, and applied to enhance in vitro transdermal delivery of doxorubicin. The microneedles were fabricated using the micromolding technique with the drug load in different locations within the needle array. The polymer solution was assessed for rheological properties, drug dissolution, and chemical structurestudies. Microneedles (unloaded) and drug-loaded microneedles were characterized by optical microscopy, fluorescent microscopy, scanning electron microscopy, and drug release kinetics. Successful microporation of dermatomed human cadaver skin was demonstrated by dye binding, pore uniformity, histology, confocal laser microscopy, and skin integrity studies. The microneedles-mediated transdermal delivery of doxorubicin was investigated using vertical Franz diffusion cells. The fabricated microneedles were sharp, strong, and uniform. In vitro permeation studies showed that the microneedle-treated skin (4351.55 ±â€¯560.87 ng/sq.cm) provided a significantly greater drug permeability than the untreated group (0.00 ±â€¯0.00 ng/sq.cm, n = 4, p < 0.01). The drug location within the needle array was found to affect the drug release profile as well as its permeation into and across human skin. Skin microporation achieved by poly (vinyl alcohol) microneedles was found to enhance transdermal delivery of doxorubicin in vitro.

Fabrication, characterization and application of sugar microneedles for transdermal drug delivery.

AIM: This study aimed to fabricate, characterize and use maltose microneedles for transdermal delivery of doxorubicin. MATERIALS & METHODS: Microneedles were fabricated by micromolding technique and evaluated for dimensions, mechanical properties and in situ dissolution. Microporation of human cadaver skin was confirmed by dye binding, histology, pore uniformity, confocal laser microscopy and skin integrity measurement. In vitro permeation studies were performed on vertical Franz diffusion cells. RESULTS: Maltose microneedles were sharp, mechanically uniform and rapidly dissolvable. Microneedle insertion resulted in a marked decrease in lag time and a significant increase in the permeation across and into human skin (p < 0.05). The skin delivery profile was used to predict the steady-state plasma concentration. CONCLUSION: Maltose microneedles are a promising physical technique to increase skin delivery.

Methods to simulate rubbing of topical formulation for in vitro skin permeation studies.

Rubbing a topical formulation on skin is generally assumed to enhance drug penetration. The aim of this study was to demonstrate different techniques such as using glass rod, rheometer, and gloved finger for rubbing a 2% salicylic acid gel on skin and investigate their effect on in vitro permeation of salicylic acid through dermatomed porcine ear skin. The studies included evaluation of the gel's rheological properties, gel distribution on skin surface, in vitro permeability, drug distribution in skin, skin extraction recovery, and mass balance. Rubbing with a gloved finger resulted in a uniform gel layer with a thickness of 49.61±15.33µm on the skin surface. No significant difference between the different test groups was observed in terms of the cumulative amount of drug that permeated in 24h (p>0.05). Drug levels in stratum corneum, epidermis, and dermis were also analyzed. Rubbing with gloved finger delivered significantly higher amount of drug into the skin layers as compared to other test groups (p<0.05). Amount of drug extracted from skin was reliably correlated to the actual drug levels in skin (R2=0.99). Considering drug amounts in different compartments, mass balance ranged from 75.86±2.90% to 80.44±2.99%.

Dihydroergotamine mesylate-loaded dissolving microneedle patch made of polyvinylpyrrolidone for management of acute migraine therapy.

Migraine is a widespread neurological disease with negative effects on quality of life and productivity. Moderate to severe acute migraine attacks can be treated with dihydroergotamine mesylate (DHE), an ergot derivative that is especially effective in non-responders to triptan derivatives. To overcome limitations of current DHE formulations in subcutaneous injection and nasal spray such as pain, adverse side effects and poor bioavailability, a new approach is needed for DHE delivery enabling painless self-administration, quick onset of action, and high bioavailability. In this study, we developed a dissolving microneedle patch (MNP) made of polyvinylpyrrolidone, due to its high aqueous solubility and solubility enhancement properties, using a MNP design previously shown to be painless and simple to administer. DHE-loaded MNPs were shown to have a content uniformity of 108±9% with sufficient mechanical strength for insertion to pig skin ex vivo and dissolution within 2min. In vivo pharmacokinetic studies were carried out on hairless rats, and DHE plasma levels were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The area under curve (AUC) value after DHE delivery by MNP (1259±917ng/mL min) was not significantly different (p>0.05) as compared to subcutaneous injection, with a relative bioavailability of 97%. Also, appreciable plasma levels of DHE were seen within 5min for both delivery methods and tmax value of MNPs (38±23min) showed no significant difference (p>0.05) compared to subcutaneous injection (24±13min). These results suggest that DHE-loaded MNPs have promise as an alternative DHE delivery method that can be painlessly self-administered with rapid onset and high bioavailability.

Enhanced skin delivery of vismodegib by microneedle treatment.

The present study investigated the effects of microneedle treatment (maltose microneedles, Admin Pen™ 1200, and Admin Pen™ 1500) on in vitro transdermal delivery of vismodegib with different needle lengths, skin equilibration times, and microneedle insertion durations. The influence of microneedle treatment on the dimensions of microchannels (dye binding, calcein imaging, histology, and confocal microscopy studies), transepidermal water loss, and skin permeability of vismodegib was also evaluated. Skin viscoelasticity was assessed using a rheometer, and microneedle geometry was characterized by scanning electron microscopy. Permeation studies of vismodegib through dermatomed porcine ear skin were conducted using vertical Franz diffusion cells. Skin irritation potential of vismodegib formulation was assessed using an in vitro reconstructed human epidermis model. Results of the in vitro permeation studies revealed significant enhancement in permeation of vismodegib through microneedle-treated skin. As the needle length increased from 500 to 1100 and 1400 µm, drug delivery increased from 14.50 ± 2.35 to 32.38 ± 3.33 and 74.40 ± 15.86 µg/cm(2), respectively. Positive correlation between drug permeability and microneedle treatment duration was observed. The equilibration time was also found to affect the delivery of vismodegib. Thus, changes in microneedle length, equilibration time, and duration of treatment altered transdermal delivery of vismodegib.