Could Wearable Voltage-Triggered Paper Patch Revolutionize Psoriasis Treatment by Enhancing Skin Permeation of Methotrexate?

Could Wearable Voltage-Triggered Paper Patch Revolutionize Psoriasis Treatment by Enhancing Skin Permeation of Methotrexate? | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Could Wearable Voltage-Triggered Paper Patch Revolutionize Psoriasis Treatment by Enhancing Skin Permeation of Methotrexate? Masoud Mehrgardi, Elham Momtaz This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4877240/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This study introduces a wearable iontophoresis transdermal paper patch for delivering methotrexate, a potent anti-inflammatory drug, to treat imiquimod (IMQ)-induced psoriasis-like inflammations. This patch features a paper-based electrode that has been coated with polypyrrole and loaded with the anti-inflammatory medication methotrexate. Incorporating paper into the design enhanced the amount of drug that could be loaded and reduced its unintended release. The most effective release of the drug, was achieved with a voltage of -1.2 V. The specially designed iontophoresis patch ensured that the methotrexate optimally penetrated the mouse skin (19.7 ± 0.5 µg cm − 2 after 2 h). The patch's ability to alleviate psoriasis, which was experimentally induced in BALB/c mice (gender random), was confirmed through successful testing. Histological analysis of the skin and internal organs such as the spleen, lungs, liver, and kidneys showed that methotrexate is highly effective and has minimal adverse effects. Physical sciences/Chemistry/Chemical biology/Drug delivery Biological sciences/Chemical biology Voltage-triggered drug release Transdermal Psoriasis Iontophoresis Paper patch Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Skin diseases are currently the fourth most common non-fatal disease in the world, with a prevalence rate of 25%. 1,2 One of the most widespread skin disorders is chronic inflammatory autoimmune psoriasis, which affects 1–3% of the world's population. 3 This disease is characterized by scaling erythematous plaques, skin shedding, swelling, itching, and painful inflammation. These symptoms significantly impact the patient's quality of life. 4 , 5 Psoriasis can be categorized into three conditions: mild, moderate, and severe, affecting less than 3%, 3–10%, and more than 10% of the body, respectively. Current treatment options are primarily to reduce symptoms, preventing the progression of the disease and improving the patient's quality of life. The methods available for treating psoriasis can be classified into three methods systematic therapy, phototherapy, and dermal therapy. 6 Methotrexate (MTX) is one of the most common anti-psoriasis drugs. This drug has been approved by the United States Food and Drug Administration (U.S. FDA) for the treatment of psoriasis since the 1970s. 7 Despite significant advances in psoriasis therapy, MTX is still one of the first options in the treatment of moderate to severe psoriasis owing to its affordable price and significant therapeutic effects. 8 – 10 For systematic therapy, MTX is administrated in oral and injectable form, which is associated with side effects such as gastrointestinal disturbances, hepatotoxicity, asthma, decreased bone marrow density, vomiting, anemia, and menstrual changes. Therefore, considering all these disadvantages, dermal therapy with methotrexate is judged to be a more desirable method. 11 In transdermal therapy, the drug is placed directly on the damaged skin and reduces systematic side effects. The drug levels are maintained constantly within therapeutic window to reduce drug toxicity and improves patient compliance. 12 – 14 MTX is a polar molecule with a relatively high molecular weight, and the stratum corneum (the outermost layer of the skin) restricts the penetration of the MTX from its hydrophilicity. 15 – 17 Stimulus-responsive drug delivery systems appear to be a promising area of research for the controlled and precise delivery of medications, aiming to minimize drug side effects. Current stimuli, including ultrasound waves, 18 magnetic fields, 19 temperature, 20 light, 21 pH levels, 22 and electric fields, 23 communicate with the responsive system to regulate drug release. This regulation ensures the drug is released in a controlled way, achieving the local drug concentrations necessary for therapeutic impact, and also allows for the timing of drug release to be managed. 24 To design and fabricate a voltage-triggered drug release patch, a power source and drug reservoir facing the surface skin is required. The use of polymers as drug reservoirs plays an essential role in maintaining and controlling targeted drug delivery and providing the desired mechanical strength for transdermal therapy. 25 Conductive polymers are organic materials that respond to electrical stimulation and have electrical and mechanical properties similar to metals and polymers. 26 Polypyrrole (PPy) is one of the most attractive conductive polymers due to its stability, biocompatibility, good electrochemical properties, low actuation voltage, and high electrical conductivity. 27 , 28 The unstable positive charge along the polymer backbone results in relatively high electrical conductivity and the loading of anionic drugs through electrostatic attraction. The controlled release of the drug occurs through the electrochemical reduction of the polymer and volume contraction as well as the expulsion of the anionic drug from the polymer structure. 29 , 30 Although polypyrrole offers many advantages for drug delivery, it has a low drug-loading capacity, especially those with high molecular weight and undesired passive drug leakage. 31 , 32 To address these issues, we explored the application of paper-based electrodes for electrodeposition of PPy, which has the advantage that paper shows a huge drug loading capacity. Paper has a soft and porous structure, excellent biocompatibility for the storage of biomolecules, abundant hydroxyl groups, and a hydrophilic surface for the deposition of water-soluble polymers, so it has attracted the attention of many researchers for the development of wearable drug delivery systems. 33 , 34 We have found that electric stimuli in a voltage-triggered patch not only offers precise control over the timing and location of drug release, but also can enhance the penetration of drug through the skin’s top layer. This mirrors the advantages of iontophoresis techniques, which amplify the therapeutic effect while minimizing unwanted side effects. Iontophoresis, a non-invasive method with minimal skin irritation, is a promising alternative to other physical strategies for transdermal drug delivery. 35 It works by applying a low voltage (less than 10 V) and low electrical current (up to 0.5 mA cm - 2 ) to electrodes on the skin surface, allowing the drug to pass through the stratum corneum. 36 As far as we know, there are no reports of enhancing skin permeation by applying electric stimuli to a transdermal paper-based wearable patch for the release of a high molecular weight drug like MTX. We hypothesize that the integration of paper-based electrodes with battery-powered electrical stimulation provides a robust platform for precise control of dosage timing and location, improving therapeutic efficacy and reducing potential side effects. For this purpose, the drug delivery component includes a paper-based electrode modified with PPy, which serves as a carrier for the anti-psoriasis drug methotrexate. Encapsulating the drug within the electrode significantly increases the drug loading capacity and reduces passive drug leakage. Crucially, the electric signal also promotes the skin permeation of MTX. 2. Results 2.1. Preparation and characterization of patch The morphological structures of Pap, Pap/C/PPy, and Pap/C/PPy/MTX are characterized using the FESEM technique, as shown in Fig. 1 a-c. Figure 1 a clearly displays the cellulose fibers of the paper. Figure 1 b indicates that PPy is effectively deposited on the surface of the electrode. When MTX is present, the morphology of PPy/MTX transforms into spherical nanoparticles. Figure 1 c demonstrates that these nanoparticles are evenly distributed on the surface of the electrode and the remaining pores. FTIR spectra of Pap, Pap/C/PPy, and Pap/C/PPy/MTX are shown in Fig. 1 d. The Pap spectrum displays absorption bands related to cellulose fibers. The peak at 3322 cm - 1 is associated with the O-H stretching vibration, 37 while the peaks at 2895 cm - 1 , 1159 cm - 1 , and 891 cm - 1 are related to the C-H linkage, the asymmetric stretching of the C-O-C bridge, and the CH deformation, respectively. 38 , 39 In the Pap/C/PPy spectrum, the peaks at 820 cm - 1 and 1650 cm - 1 are attributed to C-H stretching and the C = N bonds of PPy, respectively. 40 , 41 The band at 1518 cm - 1 is attributed to the C = C/C-C stretching vibration of the PPy rings. 42 The shift of this band in Pap/C/PPy to a lower frequency indicates the presence of a longer conjugation length than in pure PPy. The 3400 cm - 1 peak, which corresponds to the NH stretching vibration of PPy, 43 , 44 disappears because of the strong interaction between the NH group of PPy and the OH group of MTX. This interaction leads to the formation of hydrogen bonds, which results in the disappearance of the NH stretching vibration peak. In the spectrum of Pap/C/PPy/MTX, the peaks at 1023 cm - 1 , 1143 cm - 1 , and 1671 cm - 1 correspond to the CH 3 bending vibration, the C = O and NH stretching vibration, respectively. 45 The peak at 3400 cm - 1 , attributed to the OH stretching vibration, has a low intensity. This is caused by the strong interaction between the OH group of MTX and the NH group of PPy and the placement of MTX in the pores and the PPy skeleton. The strong interaction between MTX and the film reduces MTX passive leakage effectively. 2.2. Investigation of passive and active drug release The effect of various factors such as monomer concentration (Py), supporting electrolyte concentration (NaCl), and MTX concentration on drug release was investigated (Supplementary Fig. 1). Different Py concentrations can lead to the different film thicknesses, and the optimum value of 0.1 M was selected by evaluating various concentrations (0.025, 0.05, 0.1, and 0.15 M). The amount of released drug decreases at concentrations higher than 0.1 M, which can be attributed to the formation of polymers in the solution instead of electrode surface. 46 By investigating the effect of sodium chloride concentrations (0.025, 0.05, 0.1, and 0.15 M) on drug release, the concentration of 0.1 M was selected as the optimum concentration. The decrease of MTX release at higher concentrations than 0.1 M can be attributed to the competition between MTX and chloride upon entry into the polymer skeleton. 46 The effect of MTX concentration on the amount of released MTX was also studied at the concentrations (0.0025, 0.005, 0.0075, 0.01 M). According to the results, by increasing the MTX concentration of 0.005 M, the release decrease which can be attributed to the weak involvement of the drug in the polymer skeleton due to the filling of the pores. MTX, an anionic drug, can be loaded into PPy due to the electrostatic interaction with its cationic backbone. The application of a negative potential leads to the reduction of PPy, which in turn triggers the release of the negatively charged MTX. One of the main challenges associated with PPy/drug systems is the passive leakage of the drug, which occurs due to passive ion exchange at the film surface with the solution. 47 Arbabian and co-workers addressed this issue by using a protective silicone oil-PDMS gel on a nanoparticle-coated film to minimize the passive leakage of the drug. 47 In the current study, we employed a paper substrate to overcome the problem of passive leakage. We explored the ability of paper to reduce leakage and its potential impact on the quantity of loading and released MTX. Figure 2 a displays the quantities of released MTX, calculated using the calibration curve, in the absence of potential and at different negative potentials (-0.5, -1, -1.2, and − 1.5 V). The results demonstrate a significant decrease in the amount of passive leakage, which can be attributed to the strong interaction between PPy and MTX by trapping in the substrate’s pores. As shown in Fig. 2 a, the release increases by increasing the potential up to -1.2 V, after which it decreases. This decrease in release can be attributed to gas production due to water splitting. 47 Therefore, we selected − 1.2 V for subsequent experiments. The absorbance measurement was conducted both prior to and after loading MTX onto the prepared electrode. An estimated drug loading capacity of 814 ± 28 µg g⁻¹ indicates the amount of MTX that the electrode can retain per gram of its weight. 2.3. Effect of time and pH on drug release The impact of time on the release of MTX was examined at various time intervals. As shown in Fig. 2 b, approximately one third of the total MTX is released within the first 5 minutes, with the release rate slowing thereafter. This suggests that a higher amounts of released MTX on the electrode surface lead to an decrease in the reduction rate of PPy, resulting in a quicker release of MTX during the initial stages. 47 PPy, as a pH-responsive polymer, 48 influences the drug release from itself, which depends on the nature and composition of the surrounding solution. The in-vitro release of MTX was studied at four different pH values (4.4, 5.4, 6.4, and 7.4). As depicted in Fig. 2 c, the release of MTX is higher at lower pH values. This can be attributed to the increased conductivity of PPy at lower pH values and the competition between the incorporation of cations and the expulsion of anions into and out of the polymer during the reduction process. 49 , 50 2.4. Evaluation of electrically controlled drug delivery Two advantages of the electrically responsive system are the temporal and spatial resolution of drug release. 51 Pulsed release is utilized to determine the ideal timing and dosage of the drug to achieve the desired therapeutic effects. For this, pulses of 1.5 minutes with a voltage of -1.2 V were applied, and the amounts of the released drug were calculated. The device was turned off for 1.5 minutes between each pulse, and the passive drug leakage was analyzed (Fig. 3 a). The results indicate that during a 1.5-minute stimulation at -1.2 V, MTX is released by the expansion of the polymer skeleton. In subsequent stimulations, the quantity of released MTX decreases due to the reduction in the amount of loaded drug on the surface. 52 A satisfactory amount of drug was released during four consecutive stimulations, confirming that the current electrodes can function as a voltage-triggered drug delivery system. Further assessment of released MTX was conducted using a potentiostat and a battery as well, to compare the performance of the drug release system at -1.2 V for 5 minutes (Supplementary Fig. 2). The results of active releases were consistent in both modes, indicating that it is preferable to use a battery as an electrical driver for the designed electrodes. Due to the small size of the battery, the designed iontophoresis patch is portable and can be used to treat diseases without specialized personnel. 2.5. Transdermal delivery of the designed iontophoresis patch One of the most intriguing aspects of this study is the utilization of the identical electrical signal that triggers the release of MTX, for a concurrent iontophoresis effect. This effect can notably amplify skin permeation. To evaluate skin permeation, the penetration of MTX through the skin of a hairless rat was studied using a diffusion Franz cell. The rate of drug penetration through the skin via this patch (ITP) was compared to that of an MTX solution (Top) and a MTX solution applied to voltage-stimulated skin (Stimuli-Top). The patterns of skin permeation after two hours are illustrated in Fig. 3 b. This permeation profile helps in determining the total MTX amount that has penetrated. The maximum quantity of MTX permeated for the ITP group was 19.7 ± 0.5 µg cm - 2 . In contrast, the quantities for the Top and Stimuli-Top groups were 8.5 ± 0.7 and 15.1 ± 0.5 µg cm - 2 , respectively. These findings underscore the present patch as a more effective method for enhancing drug permeation through the skin and makes it a highly efficient method for transdermal drug delivery. 2.6. Therapeutic efficiency of the designed iontophoresis patch To assess the effectiveness of transdermal therapy, we employed the battery-powered patch to treat psoriatic mouse, as shown schematically in Fig. 4 . Throughout the treatment duration, we closely observed psoriasis symptoms (thickening of the skin layer, scaling, erythema, and weight loss) (Supplementary Fig. 3). Encouragingly, after just 5 days of using the patch, the psoriasis symptoms vanished. On the fifth day, samples of skin, spleen, kidney, liver, and lung tissues were gathered from the mice. Macroscopic and microscopic studies of the skin were performed to check the signs of psoriasis in microscopic images (increased thickness of epidermis (irregular acanthosis), scaling (hyperkeratosis), the presence of Monro’s abscess (subcorneal pustule) in the stratum corneum, and intermittent loss of the granular layer) 53 (Fig. 5 a). In the microscopic images of psoriatic skin, we observed psoriasis symptoms. In the Stimuli group, psoriasis effects are still observable (irregular acanthosis), but there was minimal scaling of mouse skin during the treatment. In samples from mice that received oral treatment, psoriasis-related changes persist (including irregular acanthosis, subcorneal pustules, hyperkeratosis, and granular layer loss). Topical treatment shows slight improvement, with noticeable hyperkeratosis and inflammation. However, in ITP group, symptoms are significantly alleviated, and only mild hyperkeratosis remains. The findings are also supported by the images captured of both ill and treated mice (Fig. 5 b). The epidermal thickness was assessed using imageJ software 54 (Fig. 5 c). The findings revealed that psoriasis induction led to an increase in epidermal thickness, whereas treatment with MTX resulted in a decrease. The thickness values for the Oral, Top, and ITP groups were obtained about 123, 108, and 20 µm, respectively. Based on the results obtained, the epidermal thickness returned to normal in the patch group, while the epidermis was thinner in the topical group compared to the Oral group. Consequently, the transdermal form is superior to the topical and oral forms. Microscopic images of the spleens in psoriatic mice (Control group) showed an increase in red pulps size, elevated white pulps counts, and their close proximity to each other, confirming spleen enlargement. 55 Additionally, extramedullary hematopoiesis was initiated within the spleens. 56 The Stimuli group continued to exhibit signs of psoriasis. The Oral group displayed a more pronounced degree of extramedullary hematopoiesis compared to the Control psoriasis group. This could be ascribed to the influence of MTX. Conversely, the Top group showed a reduction in both extramedullary hematopoiesis and the enlargement of red pulps. Meanwhile, the ITP group demonstrated a slight separation of white pulps, a lesser expansion of red pulps, and low hemorrhage. Upon examining the liver's histological images, it was noted that psoriasis induced lobular inflammation within the liver's tissues. 57 The Oral group experienced an intensification of inflammation from the effect of MTX, 58 while the Top and ITP groups showed the accumulation of fat in the liver cells. The histological analysis of lung tissues indicated that psoriasis led to scant necrosis, an increase in inflammatory cells, and bleeding within the lung parenchyma. The findings align with the outcomes reported by the researchers, which stem from the correlation between psoriasis and respiratory diseases. 59 , 60 The group receiving oral treatment showed symptoms of bronchitis, further bleeding, and air space ruptures due to destructive effect of MTX. 61 The Top group retained signs of psoriasis, including tissue congestion and bleeding. However, the group treated with the iontophoresis patch displayed normal lung tissue, suggesting successful treatment in the mice. The histological examination of the kidneys indicates that psoriasis triggers inflammatory cells to infiltrate the tubules and interstitium, revealing preliminary symptoms of nephrotoxicity. 57 The Oral group exhibited initial kidney injury and extravasation of lymphoplasma cells into the kidney parenchyma. This could be ascribed to the influence of MTX. 62 The Top group displayed minor kidney damage and nephrotoxicity, albeit less severe than that observed in the psoriasis group. In contrast, the ITP group demonstrated a resurgence of the kidneys' typical operational capacity (Fig. 6 ). 3. Discussion Iontophoresis patches are safe and non-invasive devices for delivering therapeutic agents through the skin, minimizing skin irritation compared to other transdermal strategies like microneedling or electroporation. One of the main advantages of iontophoresis is its ability to significantly promote the release of various therapeutic agents, including high molecular weight drugs, and provide better control over their release through electrical voltage. This method ensures a high local concentration of the drug while keeping its systemic concentration minimal, thereby reducing side effects. To achieve effective therapeutic concentrations, materials with high loading capacity should be used, and portable iontophoresis patches are preferable as they are easier to handle and can be used without specialist assistance. In this study, a portable, battery-powered, wearable iontophoresis transdermal paper patch was developed for treating psoriasis. The flexible paper patch features an electrode coated with polypyrrole, which contains the anti-inflammatory drug methotrexate. This patch offers several advantages for better-controlled and effective treatment of psoriasis: it uses polypyrrole to achieve high treatment efficiency by adjusting the electrical variable, releases an appropriate amount of drug during consecutive electrical stimulations, benefits from the flexibility and biodegradability of paper, increases drug amount while reducing passive leakage, and is easy to apply due to its portable battery. The therapeutic efficacy of the designed patch was tested in-vivo on mice with psoriasis, showing that after five days of treatment, the symptoms of psoriasis, such as redness, dryness, folds, and thickening of the skin, disappeared, and the skin returned to normal. The side effects on various organs were minimal compared to other treatment methods, indicating that this method has fewer unwanted complications. The designed paper patch can assist patients with personal care in their treatment regimen. 4. Conclusion This study has successfully demonstrated the potential of a battery-powered iontophoresis paper patch for the effective delivery of MTX for the treatment of psoriasis. The patch, which uses an electrical signal as a stimulus for drug release, showed a significant enhancement in skin permeation. The study also highlighted the patch’s ability to control the timing and dosage of the drug, leading to a more efficient and targeted treatment approach. Furthermore, the patch demonstrated a significant reduction in passive drug leakage, a common issue with traditional drug delivery systems. Histological examinations of various tissues from treated psoriatic mice showed promising results, with significant improvements observed in the skin, spleen, liver, lungs, and kidneys. The ITP group displayed normal lung tissue and a resurgence of the kidneys’ typical operational capacity, suggesting successful treatment of the psoriatic mice. Overall, the findings of this study underscore the potential of simultaneous voltage triggered drug release and iontophoresis as a more effective method for enhancing released drug permeation through the skin, paving the way for further research and development in this field. The designed iontophoresis patch, due to its compact size and battery-powered operation, is portable and can be used to treat diseases without the need for specialized personnel, making it a promising candidate for future transdermal drug delivery systems. 4. Methods 4.1. Materials Pyrrole (98%), lactic acid (85%), sodium chloride (NaCl), potassium chloride (KCl), disodium hydrogen phosphate (Na 2 HPO 4 ), sodium dihydrogen phosphate (NaH 2 PO 4 ), urea, and graphite powder were purchased from Sigma-Aldrich. MTX injectable solution (100 mg mL − 1 ), Ringer's injection serum, imiquimod cream (5%), and Formalin (10%) were purchased from Nanoalvand Co., Iran, Daru-Pakhsh Co., Iran, and Kimia Kala Razi Pharmaceutical Co., Iran, Alzahra hospital, Isfahan, Iran, respectively. Conductive silver ink was purchased from Electroninks Incorporation. All chemicals were selected to be of analytical grade or better. Throughout the process, deionized water was utilized using a Millipore Q water purification system. 4.2. Design and fabrication of the drug delivery patch The MTX-loaded drug delivery patch is constructed using paper-based electrodes. These electrodes, which include working, counter, and reference electrodes, are made by applying carbon and silver inks onto a paper substrate, forming a paper-based screen-printed electrode (P-SPE). The first layer of PPy is deposited on the working electrode by applying a solution (200 µL, 0.2 M pyrrole, 0.2 M sodium chloride) under a steady potential of 0.9 V for 250 seconds. Following this, a second layer of the drug-loaded PPy film is deposited by using a mixture (200 µL, 0.1 M pyrrole, 0.1 M sodium chloride, and 0.005 M MTX). When a constant voltage of 0.75 V is applied for 1200 seconds, the monomer (pyrrole) undergoes oxidation to form polypyrrole, and MTX is loaded into the film. Subsequently, the electrodes are rinsed to eliminate any MTX that might have been physically absorbed onto the electrode surface. Finally, the electrode is preserved at a temperature of 4°C for storage. 4.3. Patch Characterization The surface structure of the paper (Pap), and the prepared films (Pap/C/PPy, Pap/C/PPy/MTX) were examined using field emission scanning electron microscopy (FESEM) (MIRA3 TESCAN, Czech Republic) at an accelerating voltage of 25 KV. The presence of PPy and MTX was confirmed using attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) with a Jasco FTIR-6300 spectrometer in the wavenumber range of 650–4000 cm - 1 . 4.4. In-Vitro Release Studies MTX release in response to an electrical stimulus was investigated in a laboratory environment. This involved placing Ringer’s serum (200 µL) on the P-SPE to cover all three electrodes. The released MTX concentration was calculated using absorbance measurements at 303 nm and the MTX calibration curve. The study also explored the effect of loading parameters, such as Py monomer, dopant (NaCl), and MTX concentrations, on the amount of released MTX in Ringer’s serum (200 µL) at an applied potential of -1 V. Furthermore, the release of the drug under optimal conditions was assessed by applying different potentials (-0.5, -1, -1.2, and − 1.5 V) for a duration of 5 minutes. The passive leakage of MTX was measured under the same conditions, but without the application of an electrical stimulus. The release profiles of MTX at various time intervals were examined by adding Ringer’s serum (200 µL) to the P-SPE to cover all three electrodes, followed by the application of an appropriate potential (-1.2 V) for a specific duration. The release of MTX was examined under various pH levels of artificial sweat at the optimal potential of -1.2 V for a duration of 5 minutes. To create artificial sweat, we dissolved specific weight/volume ratios of materials (0.5% NaCl, 0.1% KCl, 0.1% lactic acid, and 0.1% urea) in deionized water. 63 We then adjusted the pH by adding ammonium hydroxide (0.001 M) using a laboratory pH meter (827 pH lab, Metrohm, Switzerland). In our study, we tested the impact of four different pH values (4.4, 5.4, 6.4, and 7.4) on drug release. The choice of sweat over other fluids was made to optimize the patch for transdermal drug delivery. Following the release, the amount of released MTX was measured for 200 µL of the prepared solution placed on the P-SPE. The absorbance of the resulting solutions was then recorded. Pulsed release was studied by adding sweat (300 µL, pH = 5.4) to the P-SPE. Four consecutive electrical stimulations were applied at -1.2 V in 1.5-minute intervals. The potentiostat was turned off for 1.5 minutes between each stimulation. After each stimulation, sample of the stimulated solution (200 µL) was taken and its absorbance was recorded. Then fresh sweat solution (200 µL) was added to the remaining solution on the P-SPE, and the potentiostat was turned off for 1.5 minutes. The absorbances of the solutions were recorded at 1.5-minute intervals. Lastly, the performance of the prepared P-SPE was evaluated under different electrical actuator conditions (potentiostat and battery) by applying a voltage of -1.2 V. For tests using a battery as an electrical driving source, a 9 V battery equipped with a voltage-reducing module was used to apply a voltage of -1.2 V. The release was carried out under the same conditions (200 µL of sweat at pH 5.4 for 5 min), and the absorbance of the resulting solution was measured at 303 nm. 4.5. In-vitro skin Permeation Test A cell resembling the Franz diffusion cell (with a penetration area of 1.54 cm² and a receptor volume of 10 mL) was created and utilized for skin permeation experiments using normal hairless rat skin. Male Wistar rats (weighing 180–230 g) were anesthetized with a mixed isopentane gas (1–3%). The dorsal skin of the rat was removed, and then the subcutaneous fat was separated. The skin was subsequently divided into appropriate segments. To keep it hydrated, the rat skin was placed on a Franz cell and left for 30 minutes. The temperature within the cell containing PBS (pH = 7.4) was maintained at 37°C, with normal rat skin secured on top. Three groups of skin permeability tests were conducted (The experiment was repeated three times in each group). In the first group (ITP), an iontophoresis patch containing MTX was applied to the skin (donor cell) and the drug was released for 5 minutes. In the second group (Stimuli-Top), the designed patch without MTX was initially placed on the skin, followed by the application of voltage (-1.2 V) using artificial sweat (pH = 5.4) for 5 minutes. Subsequently, MTX solution (200 µL, 160 µg mL - 1 ) dissolved in artificial sweat were applied to the skin to assess its permeability. In the third group (Top), MTX solution (200 µL, 200 µg mL - 1 ) was applied topically to the skin, and the drug’s penetration rate was assessed. In each group, we collected samples (200 µL) at various time intervals (0.25, 0.5, 0.75, 1, and 2 hours) from receptor cells and measured their absorption. Same volume of buffer solution (200 µL) was periodically added to the receptor during sampling intervals to maintain volume. Detection was performed using UV absorbance at 303 nm and a graph illustrating the cumulative drug permeation per unit area over time was created. 4.6. Animal Tests (In-Vivo Experiment) Eighteen BALB/c mice, aged 5–6 weeks and of random gender, were obtained from the animal house at the School of Pharmacy, Isfahan University of Medical Sciences. The mice were kept in groups of three per cage, with libitum access to food and water, in a room maintained at 22 ± 2°C and 50% humidity. A 12-hour light-dark cycle was followed. Ethical guidelines from the Ethical Committee of University of Isfahan (IR.UI.REC.1402.132) were strictly adhered to during the animal studies. All procedures were performed under anesthesia, with efforts made to minimize pain. The mice’s dorsal hair was shaved (area: 2×2.5 cm), and psoriasis was induced by applying a 5% IMQ cream at a dose of 65 mg cm - ² for 6 consecutive days. Throughout the treatment, the cream application was continued to sustain the psoriasis.). Six groups of mice randomly were established, each containing three mice: Normal group (healthy mice), Control group (Consisting of psoriatic mice without any treatment), Stimuli group (Psoriatic mice subjected to electrical stimulation (-1.2 V) using a specially designed drug-free patch for 5 minutes daily), Oral group (Psoriatic mice that received MTX tablets via gavage. The oral MTX tablet dosage was 5.143 mg kg - 1 week - 1 (equivalent to a clinical dose of 25 mg for a 60 kg human). 64 The MTX solution (300 µL, 0.5 mg mL - 1 ) was administered once during the treatment period using gavage tubes), Top group (Psoriatic mice treated with topical MTX solution (150 µg mL - 1 ) once a day), and ITP group (Psoriatic mice treated with a battery-powered iontophoresis patch using electrical stimulation (-1.2 V) for five minutes once a day). Lesion images were captured daily prior to treatment. On the fifth day, the mice were euthanized, and samples of treated skin, spleen, and kidney, liver, and lung tissues were collected for immunohistochemical analysis. These tissues were preserved in formalin solution (10%) at 4°C separately. for the purpose of analysis and comparison, tissues were stained using hematoxylin and eosin colors (H&E). subsequently, microscopic images were captured using a microscope equipped with a digital camera. 4.7. Statistics and reproducibility The data presented in this study underwent statistical analysis using Tukey , s multiple-comparison post hoc test and One-way analysis of variance (ANOVA) in GraphPad Prism software (version 8). The data in the figures were marked by * for **P < 0.01, ***P < 0.001, ****p < 0.0001, ns for no significances. The results were expressed as the mean ± standard deviation (SD). No animals were excluded from the analysis. No data were excluded from the analyses. Declarations The Ethical Committee of the University of Isfahan approved the study. All animal studies conducted in this research strictly adhered to the ethical guidelines set forth by the Ethical Committee of the University of Isfahan (IR.UI.REC.1402.132). The welfare of the animals was a primary concern, and all procedures were designed to minimize suffering and ensure humane treatment throughout the study. Reporting summary Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article. Data availability The datasets generated during the current study are available from the corresponding author (Masoud A. Mehrgardi) on reasonable request. Acknowledgments This work was funded by Isfahan University Research Council and Stanford University.The authors express their deep sincere thanks to Azar Baradaran for her constructive comments and guidance throughout this project. Author contributions E.M.Conceptualization, Investigation, Validation, Methodology, Visualization, Formal analysis, Data Curation, Writing - Original Draft; M A.M. Conceptualization, Validation, Formal analysis, Resources, Writing - Review & Editing, Supervision, Project administration, Funding acquisition. Competing interests The authors declare no competing financial interest. Additional Information Supplementary information . The supplementary information contains supplementary figures available at http://www.nature.com. Corresponding author . correspondence to Masoud A. Mehrgardi. References Qu F, Geng R, Liu Y, Zhu J (2022) Advanced nanocarrier-and microneedle-based transdermal drug delivery strategies for skin diseases treatment. Theranostics 12:3372 Jee MH, Mraz V, Geisler C, Bonefeld C (2020) M. $ γ$$δ $ T cells and inflammatory skin diseases. Immunol Rev 298:61–73 Parnami N, Garg T, Rath G, Goyal AK (2014) Development and characterization of nanocarriers for topical treatment of psoriasis by using combination therapy. Artif cells Nanomed Biotechnol 42:406–412 Dubey V et al (2007) Dermal and transdermal delivery of an anti-psoriatic agent via ethanolic liposomes. J Control release 123:148–154 Sarkar R, Chugh S, Bansal S (2016) General measures and quality of life issues in psoriasis. 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Commun Biol 3:95 Kolosnjaj-Tabi J, Gibot L, Fourquaux I, Golzio M, Rols M-P (2019) Electric field-responsive nanoparticles and electric fields: physical, chemical, biological mechanisms and therapeutic prospects. Adv Drug Deliv Rev 138:56–67 Bansal M et al (2020) Conducting polymer hydrogels for electrically responsive drug delivery. J Control Release 328:192–209 Borandeh S, van Bochove B, Teotia A, Seppälä J (2021) Polymeric drug delivery systems by additive manufacturing. Adv Drug Deliv Rev 173:349–373 Guimard NK, Gomez N, Schmidt CE (2007) Conducting polymers in biomedical engineering. Prog Polym Sci 32:876–921 Puiggal\’\i-Jou A, Valle D, L. J., Alemán C (2019) Drug delivery systems based on intrinsically conducting polymers. J Control Release 309:244–264 Tat’yana VV, Efimov ON (1997) Polypyrrole: a conducting polymer; its synthesis, properties and applications. Russ Chem Rev 66:443 Yu C et al (2014) All-solid-state flexible supercapacitors based on highly dispersed polypyrrole nanowire and reduced graphene oxide composites. ACS Appl Mater \& interfaces 6:17937–17943 Fielding LA, Hillier JK, Burchell MJ, Armes SP (2015) Space science applications for conducting polymer particles: synthetic mimics for cosmic dust and micrometeorites. Chem Commun 51:16886–16899 Jin J, Huang Z, Yin G, Yang A, Tang S (2015) Fabrication of polypyrrole/proteins composite film and their electro-controlled release for axons outgrowth. Electrochim Acta 185:172–177 Ryan EM, Breslin CB (2019) The incorporation of drug molecules with poor water solubility into polypyrrole as dopants: Indomethacin and sulindac. Electrochim Acta 296:848–855 Liu J et al (2020) Wireless, battery-free and wearable device for electrically controlled drug delivery: sodium salicylate released from bilayer polypyrrole by near-field communication on smartphone. Biomed Microdevices 22:1–10 Xu Y et al (2020) Pencil–paper on-skin electronics. Proc. Natl. Acad. Sci. 117, 18292–18301 Garland MJ, Caffarel–Salvador E, Migalska K, Woolfson AD, Donnelly RF (2012) Dissolving polymeric microneedle arrays for electrically assisted transdermal drug delivery. J Control release 159:52–59 Banga AK, Bose S, Ghosh TK (1999) Iontophoresis and electroporation: comparisons and contrasts. Int J Pharm 179:1–19 Sain M, Panthapulakkal S (2006) Bioprocess preparation of wheat straw fibers and their characterization. Ind Crops Prod 23:1–8 Chen W et al (2011) Isolation and characterization of cellulose nanofibers from four plant cellulose fibers using a chemical-ultrasonic process. Cellulose 18:433–442 He Q, Huang Z, Liu Y, Chen W, Xu T (2007) Template-directed one-step synthesis of flowerlike porous carbonated hydroxyapatite spheres. Mater Lett 61:141–143 Zhu C, Zhai J, Wen D, Dong S (2012) Graphene oxide/polypyrrole nanocomposites: one-step electrochemical doping, coating and synergistic effect for energy storage. J Mater Chem 22:6300–6306 Ghanbari K, Bonyadi S (2018) An electrochemical sensor based on reduced graphene oxide decorated with polypyrrole nanofibers and zinc oxide–copper oxide p–n junction heterostructures for the simultaneous voltammetric determination of ascorbic acid, dopamine, paracetamol, and trypto. New J Chem 42:8512–8523 Gemeiner P et al (2015) Polypyrrole-coated multi-walled carbon nanotubes for the simple preparation of counter electrodes in dye-sensitized solar cells. Synth Met 210:323–331 Cai Z et al (2017) Electrochemical synthesis of graphene/polypyrrole nanotube composites for multifunctional applications. Synth Met 227:100–105 Liu S et al (2010) Green electrochemical synthesis of Pt/graphene sheet nanocomposite film and its electrocatalytic property. J Power Sources 195:4628–4633 Alizadeh N, Shamaeli E (2014) Electrochemically controlled release of anticancer drug methotrexate using nanostructured polypyrrole modified with cetylpyridinium: Release kinetics investigation. Electrochim Acta 130:488–496 Jalal NR, Madrakian T, Afkhami A, Ghoorchian A (2021) Graphene oxide nanoribbons/polypyrrole nanocomposite film: Controlled release of leucovorin by electrical stimulation. Electrochim Acta 370:137806 Wang ML et al (2022) On-demand electrochemically controlled compound release from an ultrasonically powered implant. RSC Adv 12:23337–23345 Samanta D, Meiser JL, Zare R (2015) N. Polypyrrole nanoparticles for tunable, pH-sensitive and sustained drug release. Nanoscale 7:9497–9504 Xie Q, Kuwabata S, Yoneyama H (1997) EQCM studies on polypyrrole in aqueous solutions. J Electroanal Chem 420:219–225 Gandhi MR, Murray P, Spinks GM, Wallace GG (1995) Mechanism of electromechanical actuation in polypyrrole. Synth Met 73:247–256 Vinchhi P, Rawal SU, Patel MM (2021) External stimuli-responsive drug delivery systems. in Drug Delivery Devices and Therapeutic Systems 267–288Elsevier Lee H, Hong W, Jeon S, Choi Y, Cho Y (2015) Electroactive polypyrrole nanowire arrays: synergistic effect of cancer treatment by on-demand drug release and photothermal therapy. Langmuir 31:4264–4269 Zanvit P et al (2015) Antibiotics in neonatal life increase murine susceptibility to experimental psoriasis. Nat Commun 6:8424 Panonnummal R, Sabitha M (2018) Anti-psoriatic and toxicity evaluation of methotrexate loaded chitin nanogel in imiquimod induced mice model. Int J Biol Macromol 110:245–258 Liang L et al (2021) Improved imiquimod-induced psoriasis like dermatitis using microneedles in mice. Eur J Pharm Biopharm 164:20–27 Liu Z et al (2024) Biguanide chitosan microneedles with cell-free DNA scavenging ability for psoriasis therapy. Bioact Mater 33:497–505 Su Y-J et al (2022) A study on MDA5 signaling in splenic B cells from an imiquimod-induced lupus mouse model with proteomics. Cells 11:3350 Conway R, Carey JJ (2017) Risk of liver disease in methotrexate treated patients. World J Hepatol 9:1092 Fang H-Y, Liao W-C, Lin C-L, Chen C-H, Kao C-H (2015) Association between psoriasis and asthma: a population-based retrospective cohort analysis. Br J Dermatol 172:1066–1071 Kao L-T, Lee C-Z, Liu S-P, Tsai M-C, Lin H-C (2014) Psoriasis and the risk of pneumonia: a population-based study. PLoS ONE 9:e116077 Jakubovic BD, Donovan A, Webster PM, Shear NH (2013) others. Methotrexate-induced pulmonary toxicity. Can Respir J 20:153–155 Kitamura M et al (2018) Methotrexate-induced acute kidney injury in patients with hematological malignancies: three case reports with literature review. Ren Replace Ther 4:1–8 Liu G et al (2015) Real-time sweat analysis via alternating current conductivity of artificial and human sweat. Appl Phys Lett 106:133702 Rashid SA et al (2021) Olive oil based methotrexate loaded topical nanoemulsion gel for the treatment of imiquimod induced psoriasis-like skin inflammation in an animal model. Biology (Basel) 10:1121 Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryInformation.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4877240","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":340578048,"identity":"adc8cf94-b6f6-412e-b8c7-e5f0f4267dde","order_by":0,"name":"Masoud Mehrgardi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA2klEQVRIiWNgGAWjYJACZgiV2PgASPLwkaKl2QCkhY0ELQlsEiCKoBbdBu7Ex4V7auXN25PbKr/m2MmwMTA/fHQDjxazA7ybjWc8O24458zDttuy25KBDmMzNs7Br2WbNM+BY4wzJBLbbktuYwZq4WGTJqBl+2+gFnuQlmLJbfVEaQGafKAmEaSF8eO2w0RoOcy7GeiwA8kzeB42SzNuO87DxkzIL8d7N37mOVBnO4M9/eHHn9uq7fnZmx8+xqcFGimHIWwehAhBUAcmGX8Qp3oUjIJRMApGGAAAxudHTegM5t0AAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0002-1544-7257","institution":"Stanford University","correspondingAuthor":true,"prefix":"","firstName":"Masoud","middleName":"","lastName":"Mehrgardi","suffix":""},{"id":340578049,"identity":"f82a2bf5-3536-405f-a4d7-de0a87f23cfa","order_by":1,"name":"Elham Momtaz","email":"","orcid":"","institution":"University of Isfahan","correspondingAuthor":false,"prefix":"","firstName":"Elham","middleName":"","lastName":"Momtaz","suffix":""}],"badges":[],"createdAt":"2024-08-08 00:05:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4877240/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4877240/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":62874218,"identity":"b01a8ad0-1a58-45af-b84a-467da7c151b4","added_by":"auto","created_at":"2024-08-20 13:32:45","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":687449,"visible":true,"origin":"","legend":"\u003cp\u003eFESEM image of (\u003cstrong\u003ea\u003c/strong\u003e) Pap, (\u003cstrong\u003eb\u003c/strong\u003e) Pap/C/PPy, and (\u003cstrong\u003ec\u003c/strong\u003e) Pap/C/PPy/MTX electrodes. \u003cstrong\u003ed\u003c/strong\u003e FTIR spectra of different electrodes (Pap, Pap/C/PPy, and Pap/C/PPy/MTX).\u003c/p\u003e","description":"","filename":"image1.png","url":"https://assets-eu.researchsquare.com/files/rs-4877240/v1/981e6ddf0075249bf39e4f18.png"},{"id":62874811,"identity":"3de551dd-8183-40ca-a465-35b907591426","added_by":"auto","created_at":"2024-08-20 13:40:45","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":31025,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e The passive leakage (Control) and released MTX amounts (mean ± SD) at different applied potentials (-0.5, -1, -1.2, and -1.5 V). \u003cstrong\u003eb\u003c/strong\u003eRelease (mean ± SD) profile of MTX in Ringer’s serum at optimal potential (-1.2 V). \u003cstrong\u003ec\u003c/strong\u003e The amount of Released MTX (mean ± SD) at various pH levels of artificial sweat (4.4, 5.4, 6.4, and 7.4). n=3 samples per group for all measurements.\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-4877240/v1/b4478d35c83ca296051b4197.png"},{"id":62874215,"identity":"5d26489c-cfd1-41e1-99aa-773b9fa021c1","added_by":"auto","created_at":"2024-08-20 13:32:45","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":31569,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e Cumulative release of MTX (mean ± SD) during pulsed release (-1.2 V for 1.5 min) after four cycles of on/off electrical stimulation (n=3, independent experiments). \u003cstrong\u003eb\u003c/strong\u003e In-vitro skin permeation (mean ± SD) profile of MTX for different groups (Top, Stimuli-Top, and ITP), with n=3 samples per group.\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-4877240/v1/dbe2fd59b88d3f60d259f2ed.png"},{"id":62874220,"identity":"4fdd04eb-73d6-4ea8-a3c5-4abbbe53f940","added_by":"auto","created_at":"2024-08-20 13:32:45","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":245674,"visible":true,"origin":"","legend":"\u003cp\u003eSchematic illustration of a wearable iontophoresis patch.\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-4877240/v1/d3f78ce4c9956ba92e1f03de.png"},{"id":62874221,"identity":"1d1c45a2-da1a-4f0a-a2eb-5f3ce07a0a0e","added_by":"auto","created_at":"2024-08-20 13:32:45","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":868494,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ea\u003c/strong\u003e Hematoxylin and eosin (H\u0026amp;E) staining of psoriasis mouse skin under different treatments, scale bar: 100 µm. \u003cstrong\u003eb\u003c/strong\u003e Corresponding photos of normal, psoriatic (Control) skins, and treatment process (Oral, Top, Stimuli, and ITP). \u003cstrong\u003ec\u003c/strong\u003e Epidermis thickness (mean ± SD) of psoriatic mice under different treatments. (n=3 per group). **P\u0026lt;0.01, ***P\u0026lt;0.001, ****p\u0026lt;0.0001, ns for no significances, P-value was obtained through one-way analysis of variance (ANOVA), followed by Tukey\u003csup\u003e,\u003c/sup\u003es multiple-comparison post hoc tests.\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-4877240/v1/f3f94ffd77f75bb4d8329a9d.png"},{"id":62874222,"identity":"4bed487a-3ff9-4ed7-97b3-d1cd69a1e2e9","added_by":"auto","created_at":"2024-08-20 13:32:45","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":560821,"visible":true,"origin":"","legend":"\u003cp\u003eH\u0026amp;E staining images of spleen, kidney, liver, lung under different treatments.\u003c/p\u003e","description":"","filename":"image6.png","url":"https://assets-eu.researchsquare.com/files/rs-4877240/v1/726ed8a428e3b493b711635b.png"},{"id":62875903,"identity":"23ed8945-0d7e-44e2-a611-c2573018778b","added_by":"auto","created_at":"2024-08-20 13:56:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3587772,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4877240/v1/57c3b3fe-c8c9-4cb1-a408-671dcfe9c6ed.pdf"},{"id":62874219,"identity":"358a1a55-3891-4713-ae05-900d072b15ab","added_by":"auto","created_at":"2024-08-20 13:32:45","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":116040,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"SupplementaryInformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-4877240/v1/7df51c8c8c25bb23389f4dff.docx"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Could Wearable Voltage-Triggered Paper Patch Revolutionize Psoriasis Treatment by Enhancing Skin Permeation of Methotrexate?","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eSkin diseases are currently the fourth most common non-fatal disease in the world, with a prevalence rate of 25%.\u003csup\u003e1,2\u003c/sup\u003e One of the most widespread skin disorders is chronic inflammatory autoimmune psoriasis, which affects 1\u0026ndash;3% of the world's population.\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e This disease is characterized by scaling erythematous plaques, skin shedding, swelling, itching, and painful inflammation. These symptoms significantly impact the patient's quality of life.\u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e,\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e Psoriasis can be categorized into three conditions: mild, moderate, and severe, affecting less than 3%, 3\u0026ndash;10%, and more than 10% of the body, respectively. Current treatment options are primarily to reduce symptoms, preventing the progression of the disease and improving the patient's quality of life. The methods available for treating psoriasis can be classified into three methods systematic therapy, phototherapy, and dermal therapy.\u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eMethotrexate (MTX) is one of the most common anti-psoriasis drugs. This drug has been approved by the United States Food and Drug Administration (U.S. FDA) for the treatment of psoriasis since the 1970s.\u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e Despite significant advances in psoriasis therapy, MTX is still one of the first options in the treatment of moderate to severe psoriasis owing to its affordable price and significant therapeutic effects.\u003csup\u003e\u003cspan additionalcitationids=\"CR9\" citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e For systematic therapy, MTX is administrated in oral and injectable form, which is associated with side effects such as gastrointestinal disturbances, hepatotoxicity, asthma, decreased bone marrow density, vomiting, anemia, and menstrual changes. Therefore, considering all these disadvantages, dermal therapy with methotrexate is judged to be a more desirable method.\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e In transdermal therapy, the drug is placed directly on the damaged skin and reduces systematic side effects. The drug levels are maintained constantly within therapeutic window to reduce drug toxicity and improves patient compliance.\u003csup\u003e\u003cspan additionalcitationids=\"CR13\" citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e MTX is a polar molecule with a relatively high molecular weight, and the stratum corneum (the outermost layer of the skin) restricts the penetration of the MTX from its hydrophilicity.\u003csup\u003e\u003cspan additionalcitationids=\"CR16\" citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eStimulus-responsive drug delivery systems appear to be a promising area of research for the controlled and precise delivery of medications, aiming to minimize drug side effects. Current stimuli, including ultrasound waves,\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e magnetic fields,\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e temperature,\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e light,\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e pH levels,\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e and electric fields,\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e communicate with the responsive system to regulate drug release. This regulation ensures the drug is released in a controlled way, achieving the local drug concentrations necessary for therapeutic impact, and also allows for the timing of drug release to be managed.\u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e To design and fabricate a voltage-triggered drug release patch, a power source and drug reservoir facing the surface skin is required. The use of polymers as drug reservoirs plays an essential role in maintaining and controlling targeted drug delivery and providing the desired mechanical strength for transdermal therapy.\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eConductive polymers are organic materials that respond to electrical stimulation and have electrical and mechanical properties similar to metals and polymers.\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e Polypyrrole (PPy) is one of the most attractive conductive polymers due to its stability, biocompatibility, good electrochemical properties, low actuation voltage, and high electrical conductivity.\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e,\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e The unstable positive charge along the polymer backbone results in relatively high electrical conductivity and the loading of anionic drugs through electrostatic attraction. The controlled release of the drug occurs through the electrochemical reduction of the polymer and volume contraction as well as the expulsion of the anionic drug from the polymer structure.\u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e,\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e Although polypyrrole offers many advantages for drug delivery, it has a low drug-loading capacity, especially those with high molecular weight and undesired passive drug leakage.\u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e,\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e To address these issues, we explored the application of paper-based electrodes for electrodeposition of PPy, which has the advantage that paper shows a huge drug loading capacity. Paper has a soft and porous structure, excellent biocompatibility for the storage of biomolecules, abundant hydroxyl groups, and a hydrophilic surface for the deposition of water-soluble polymers, so it has attracted the attention of many researchers for the development of wearable drug delivery systems.\u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e,\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eWe have found that electric stimuli in a voltage-triggered patch not only offers precise control over the timing and location of drug release, but also can enhance the penetration of drug through the skin\u0026rsquo;s top layer. This mirrors the advantages of iontophoresis techniques, which amplify the therapeutic effect while minimizing unwanted side effects. Iontophoresis, a non-invasive method with minimal skin irritation, is a promising alternative to other physical strategies for transdermal drug delivery.\u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e It works by applying a low voltage (less than 10 V) and low electrical current (up to 0.5 mA cm\u003csup\u003e-\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e) to electrodes on the skin surface, allowing the drug to pass through the stratum corneum.\u003csup\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e As far as we know, there are no reports of enhancing skin permeation by applying electric stimuli to a transdermal paper-based wearable patch for the release of a high molecular weight drug like MTX. We hypothesize that the integration of paper-based electrodes with battery-powered electrical stimulation provides a robust platform for precise control of dosage timing and location, improving therapeutic efficacy and reducing potential side effects. For this purpose, the drug delivery component includes a paper-based electrode modified with PPy, which serves as a carrier for the anti-psoriasis drug methotrexate. Encapsulating the drug within the electrode significantly increases the drug loading capacity and reduces passive drug leakage. Crucially, the electric signal also promotes the skin permeation of MTX.\u003c/p\u003e"},{"header":"2. Results","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003e2.1. Preparation and characterization of patch\u003c/h2\u003e\n \u003cp\u003eThe morphological structures of Pap, Pap/C/PPy, and Pap/C/PPy/MTX are characterized using the FESEM technique, as shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ea-c. Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ea clearly displays the cellulose fibers of the paper. Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eb indicates that PPy is effectively deposited on the surface of the electrode. When MTX is present, the morphology of PPy/MTX transforms into spherical nanoparticles. Figure \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ec demonstrates that these nanoparticles are evenly distributed on the surface of the electrode and the remaining pores.\u003c/p\u003e\n \u003cp\u003eFTIR spectra of Pap, Pap/C/PPy, and Pap/C/PPy/MTX are shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003ed. The Pap spectrum displays absorption bands related to cellulose fibers. The peak at 3322 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e is associated with the O-H stretching vibration,\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e while the peaks at 2895 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, 1159 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, and 891 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e are related to the C-H linkage, the asymmetric stretching of the C-O-C bridge, and the CH deformation, respectively.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e38\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e In the Pap/C/PPy spectrum, the peaks at 820 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e and 1650 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e are attributed to C-H stretching and the C\u0026thinsp;=\u0026thinsp;N bonds of PPy, respectively.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e40\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e41\u003c/span\u003e\u003c/sup\u003e The band at 1518 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e is attributed to the C\u0026thinsp;=\u0026thinsp;C/C-C stretching vibration of the PPy rings.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e The shift of this band in Pap/C/PPy to a lower frequency indicates the presence of a longer conjugation length than in pure PPy. The 3400 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e peak, which corresponds to the NH stretching vibration of PPy,\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e43\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e44\u003c/span\u003e\u003c/sup\u003e disappears because of the strong interaction between the NH group of PPy and the OH group of MTX. This interaction leads to the formation of hydrogen bonds, which results in the disappearance of the NH stretching vibration peak. In the spectrum of Pap/C/PPy/MTX, the peaks at 1023 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, 1143 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, and 1671 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e correspond to the CH\u003csub\u003e3\u003c/sub\u003e bending vibration, the C\u0026thinsp;=\u0026thinsp;O and NH stretching vibration, respectively.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e The peak at 3400 cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, attributed to the OH stretching vibration, has a low intensity. This is caused by the strong interaction between the OH group of MTX and the NH group of PPy and the placement of MTX in the pores and the PPy skeleton. The strong interaction between MTX and the film reduces MTX passive leakage effectively.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003e2.2. Investigation of passive and active drug release\u003c/h2\u003e\n \u003cp\u003eThe effect of various factors such as monomer concentration (Py), supporting electrolyte concentration (NaCl), and MTX concentration on drug release was investigated (Supplementary Fig.\u0026nbsp;1). Different Py concentrations can lead to the different film thicknesses, and the optimum value of 0.1 M was selected by evaluating various concentrations (0.025, 0.05, 0.1, and 0.15 M). The amount of released drug decreases at concentrations higher than 0.1 M, which can be attributed to the formation of polymers in the solution instead of electrode surface.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e By investigating the effect of sodium chloride concentrations (0.025, 0.05, 0.1, and 0.15 M) on drug release, the concentration of 0.1 M was selected as the optimum concentration. The decrease of MTX release at higher concentrations than 0.1 M can be attributed to the competition between MTX and chloride upon entry into the polymer skeleton.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e The effect of MTX concentration on the amount of released MTX was also studied at the concentrations (0.0025, 0.005, 0.0075, 0.01 M). According to the results, by increasing the MTX concentration of 0.005 M, the release decrease which can be attributed to the weak involvement of the drug in the polymer skeleton due to the filling of the pores.\u003c/p\u003e\n \u003cp\u003eMTX, an anionic drug, can be loaded into PPy due to the electrostatic interaction with its cationic backbone. The application of a negative potential leads to the reduction of PPy, which in turn triggers the release of the negatively charged MTX. One of the main challenges associated with PPy/drug systems is the passive leakage of the drug, which occurs due to passive ion exchange at the film surface with the solution.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e Arbabian and co-workers addressed this issue by using a protective silicone oil-PDMS gel on a nanoparticle-coated film to minimize the passive leakage of the drug.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e In the current study, we employed a paper substrate to overcome the problem of passive leakage. We explored the ability of paper to reduce leakage and its potential impact on the quantity of loading and released MTX. Figure \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ea displays the quantities of released MTX, calculated using the calibration curve, in the absence of potential and at different negative potentials (-0.5, -1, -1.2, and \u0026minus;\u0026thinsp;1.5 V). The results demonstrate a significant decrease in the amount of passive leakage, which can be attributed to the strong interaction between PPy and MTX by trapping in the substrate\u0026rsquo;s pores. As shown in Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ea, the release increases by increasing the potential up to -1.2 V, after which it decreases. This decrease in release can be attributed to gas production due to water splitting.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e Therefore, we selected \u0026minus;\u0026thinsp;1.2 V for subsequent experiments. The absorbance measurement was conducted both prior to and after loading MTX onto the prepared electrode. An estimated drug loading capacity of 814\u0026thinsp;\u0026plusmn;\u0026thinsp;28 \u0026micro;g g⁻\u0026sup1; indicates the amount of MTX that the electrode can retain per gram of its weight.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n \u003ch2\u003e2.3. Effect of time and pH on drug release\u003c/h2\u003e\n \u003cp\u003eThe impact of time on the release of MTX was examined at various time intervals. As shown in Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eb, approximately one third of the total MTX is released within the first 5 minutes, with the release rate slowing thereafter. This suggests that a higher amounts of released MTX on the electrode surface lead to an decrease in the reduction rate of PPy, resulting in a quicker release of MTX during the initial stages.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003cp\u003ePPy, as a pH-responsive polymer,\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/sup\u003e influences the drug release from itself, which depends on the nature and composition of the surrounding solution. The in-vitro release of MTX was studied at four different pH values (4.4, 5.4, 6.4, and 7.4). As depicted in Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003ec, the release of MTX is higher at lower pH values. This can be attributed to the increased conductivity of PPy at lower pH values and the competition between the incorporation of cations and the expulsion of anions into and out of the polymer during the reduction process.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e49\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003e2.4. Evaluation of electrically controlled drug delivery\u003c/h2\u003e\n \u003cp\u003eTwo advantages of the electrically responsive system are the temporal and spatial resolution of drug release.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e51\u003c/span\u003e\u003c/sup\u003e Pulsed release is utilized to determine the ideal timing and dosage of the drug to achieve the desired therapeutic effects. For this, pulses of 1.5 minutes with a voltage of -1.2 V were applied, and the amounts of the released drug were calculated. The device was turned off for 1.5 minutes between each pulse, and the passive drug leakage was analyzed (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003ea).\u003c/p\u003e\n \u003cp\u003eThe results indicate that during a 1.5-minute stimulation at -1.2 V, MTX is released by the expansion of the polymer skeleton. In subsequent stimulations, the quantity of released MTX decreases due to the reduction in the amount of loaded drug on the surface.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e52\u003c/span\u003e\u003c/sup\u003e A satisfactory amount of drug was released during four consecutive stimulations, confirming that the current electrodes can function as a voltage-triggered drug delivery system.\u003c/p\u003e\n \u003cp\u003eFurther assessment of released MTX was conducted using a potentiostat and a battery as well, to compare the performance of the drug release system at -1.2 V for 5 minutes (Supplementary Fig.\u0026nbsp;2). The results of active releases were consistent in both modes, indicating that it is preferable to use a battery as an electrical driver for the designed electrodes. Due to the small size of the battery, the designed iontophoresis patch is portable and can be used to treat diseases without specialized personnel.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003e2.5. Transdermal delivery of the designed iontophoresis patch\u003c/h2\u003e\n \u003cp\u003eOne of the most intriguing aspects of this study is the utilization of the identical electrical signal that triggers the release of MTX, for a concurrent iontophoresis effect. This effect can notably amplify skin permeation. To evaluate skin permeation, the penetration of MTX through the skin of a hairless rat was studied using a diffusion Franz cell. The rate of drug penetration through the skin via this patch (ITP) was compared to that of an MTX solution (Top) and a MTX solution applied to voltage-stimulated skin (Stimuli-Top). The patterns of skin permeation after two hours are illustrated in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eb. This permeation profile helps in determining the total MTX amount that has penetrated. The maximum quantity of MTX permeated for the ITP group was 19.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 \u0026micro;g cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. In contrast, the quantities for the Top and Stimuli-Top groups were 8.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7 and 15.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 \u0026micro;g cm\u003csup\u003e-\u003cspan class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e, respectively. These findings underscore the present patch as a more effective method for enhancing drug permeation through the skin and makes it a highly efficient method for transdermal drug delivery.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003e2.6. Therapeutic efficiency of the designed iontophoresis patch\u003c/h2\u003e\n \u003cp\u003eTo assess the effectiveness of transdermal therapy, we employed the battery-powered patch to treat psoriatic mouse, as shown schematically in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e. Throughout the treatment duration, we closely observed psoriasis symptoms (thickening of the skin layer, scaling, erythema, and weight loss) (Supplementary Fig.\u0026nbsp;3). Encouragingly, after just 5 days of using the patch, the psoriasis symptoms vanished. On the fifth day, samples of skin, spleen, kidney, liver, and lung tissues were gathered from the mice. Macroscopic and microscopic studies of the skin were performed to check the signs of psoriasis in microscopic images (increased thickness of epidermis (irregular acanthosis), scaling (hyperkeratosis), the presence of Monro\u0026rsquo;s abscess (subcorneal pustule) in the stratum corneum, and intermittent loss of the granular layer)\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003ea). In the microscopic images of psoriatic skin, we observed psoriasis symptoms. In the Stimuli group, psoriasis effects are still observable (irregular acanthosis), but there was minimal scaling of mouse skin during the treatment. In samples from mice that received oral treatment, psoriasis-related changes persist (including irregular acanthosis, subcorneal pustules, hyperkeratosis, and granular layer loss). Topical treatment shows slight improvement, with noticeable hyperkeratosis and inflammation. However, in ITP group, symptoms are significantly alleviated, and only mild hyperkeratosis remains. The findings are also supported by the images captured of both ill and treated mice (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eb). The epidermal thickness was assessed using imageJ software\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/sup\u003e (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003ec). The findings revealed that psoriasis induction led to an increase in epidermal thickness, whereas treatment with MTX resulted in a decrease. The thickness values for the Oral, Top, and ITP groups were obtained about 123, 108, and 20 \u0026micro;m, respectively. Based on the results obtained, the epidermal thickness returned to normal in the patch group, while the epidermis was thinner in the topical group compared to the Oral group. Consequently, the transdermal form is superior to the topical and oral forms.\u003c/p\u003e\n \u003cp\u003eMicroscopic images of the spleens in psoriatic mice (Control group) showed an increase in red pulps size, elevated white pulps counts, and their close proximity to each other, confirming spleen enlargement.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e55\u003c/span\u003e\u003c/sup\u003e Additionally, extramedullary hematopoiesis was initiated within the spleens.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e56\u003c/span\u003e\u003c/sup\u003e The Stimuli group continued to exhibit signs of psoriasis. The Oral group displayed a more pronounced degree of extramedullary hematopoiesis compared to the Control psoriasis group. This could be ascribed to the influence of MTX. Conversely, the Top group showed a reduction in both extramedullary hematopoiesis and the enlargement of red pulps. Meanwhile, the ITP group demonstrated a slight separation of white pulps, a lesser expansion of red pulps, and low hemorrhage. Upon examining the liver\u0026apos;s histological images, it was noted that psoriasis induced lobular inflammation within the liver\u0026apos;s tissues.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e57\u003c/span\u003e\u003c/sup\u003e The Oral group experienced an intensification of inflammation from the effect of MTX,\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e58\u003c/span\u003e\u003c/sup\u003e while the Top and ITP groups showed the accumulation of fat in the liver cells. The histological analysis of lung tissues indicated that psoriasis led to scant necrosis, an increase in inflammatory cells, and bleeding within the lung parenchyma. The findings align with the outcomes reported by the researchers, which stem from the correlation between psoriasis and respiratory diseases.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e59\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e60\u003c/span\u003e\u003c/sup\u003e The group receiving oral treatment showed symptoms of bronchitis, further bleeding, and air space ruptures due to destructive effect of MTX.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e61\u003c/span\u003e\u003c/sup\u003e The Top group retained signs of psoriasis, including tissue congestion and bleeding. However, the group treated with the iontophoresis patch displayed normal lung tissue, suggesting successful treatment in the mice. The histological examination of the kidneys indicates that psoriasis triggers inflammatory cells to infiltrate the tubules and interstitium, revealing preliminary symptoms of nephrotoxicity.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e57\u003c/span\u003e\u003c/sup\u003e The Oral group exhibited initial kidney injury and extravasation of lymphoplasma cells into the kidney parenchyma. This could be ascribed to the influence of MTX.\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e62\u003c/span\u003e\u003c/sup\u003e The Top group displayed minor kidney damage and nephrotoxicity, albeit less severe than that observed in the psoriasis group. In contrast, the ITP group demonstrated a resurgence of the kidneys\u0026apos; typical operational capacity (Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e"},{"header":"3. Discussion","content":"\u003cp\u003eIontophoresis patches are safe and non-invasive devices for delivering therapeutic agents through the skin, minimizing skin irritation compared to other transdermal strategies like microneedling or electroporation. One of the main advantages of iontophoresis is its ability to significantly promote the release of various therapeutic agents, including high molecular weight drugs, and provide better control over their release through electrical voltage. This method ensures a high local concentration of the drug while keeping its systemic concentration minimal, thereby reducing side effects. To achieve effective therapeutic concentrations, materials with high loading capacity should be used, and portable iontophoresis patches are preferable as they are easier to handle and can be used without specialist assistance.\u003c/p\u003e \u003cp\u003eIn this study, a portable, battery-powered, wearable iontophoresis transdermal paper patch was developed for treating psoriasis. The flexible paper patch features an electrode coated with polypyrrole, which contains the anti-inflammatory drug methotrexate. This patch offers several advantages for better-controlled and effective treatment of psoriasis: it uses polypyrrole to achieve high treatment efficiency by adjusting the electrical variable, releases an appropriate amount of drug during consecutive electrical stimulations, benefits from the flexibility and biodegradability of paper, increases drug amount while reducing passive leakage, and is easy to apply due to its portable battery.\u003c/p\u003e \u003cp\u003eThe therapeutic efficacy of the designed patch was tested \u003cem\u003ein-vivo\u003c/em\u003e on mice with psoriasis, showing that after five days of treatment, the symptoms of psoriasis, such as redness, dryness, folds, and thickening of the skin, disappeared, and the skin returned to normal. The side effects on various organs were minimal compared to other treatment methods, indicating that this method has fewer unwanted complications. The designed paper patch can assist patients with personal care in their treatment regimen.\u003c/p\u003e"},{"header":"4. Conclusion","content":"\u003cp\u003eThis study has successfully demonstrated the potential of a battery-powered iontophoresis paper patch for the effective delivery of MTX for the treatment of psoriasis. The patch, which uses an electrical signal as a stimulus for drug release, showed a significant enhancement in skin permeation. The study also highlighted the patch\u0026rsquo;s ability to control the timing and dosage of the drug, leading to a more efficient and targeted treatment approach. Furthermore, the patch demonstrated a significant reduction in passive drug leakage, a common issue with traditional drug delivery systems. Histological examinations of various tissues from treated psoriatic mice showed promising results, with significant improvements observed in the skin, spleen, liver, lungs, and kidneys. The ITP group displayed normal lung tissue and a resurgence of the kidneys\u0026rsquo; typical operational capacity, suggesting successful treatment of the psoriatic mice. Overall, the findings of this study underscore the potential of simultaneous voltage triggered drug release and iontophoresis as a more effective method for enhancing released drug permeation through the skin, paving the way for further research and development in this field. The designed iontophoresis patch, due to its compact size and battery-powered operation, is portable and can be used to treat diseases without the need for specialized personnel, making it a promising candidate for future transdermal drug delivery systems.\u003c/p\u003e"},{"header":"4. Methods","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e4.1. Materials\u003c/h2\u003e \u003cp\u003ePyrrole (98%), lactic acid (85%), sodium chloride (NaCl), potassium chloride (KCl), disodium hydrogen phosphate (Na\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e), sodium dihydrogen phosphate (NaH\u003csub\u003e2\u003c/sub\u003ePO\u003csub\u003e4\u003c/sub\u003e), urea, and graphite powder were purchased from Sigma-Aldrich. MTX injectable solution (100 mg mL\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), Ringer's injection serum, imiquimod cream (5%), and Formalin (10%) were purchased from Nanoalvand Co., Iran, Daru-Pakhsh Co., Iran, and Kimia Kala Razi Pharmaceutical Co., Iran, Alzahra hospital, Isfahan, Iran, respectively. Conductive silver ink was purchased from Electroninks Incorporation. All chemicals were selected to be of analytical grade or better. Throughout the process, deionized water was utilized using a Millipore Q water purification system.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e4.2. Design and fabrication of the drug delivery patch\u003c/h2\u003e \u003cp\u003eThe MTX-loaded drug delivery patch is constructed using paper-based electrodes. These electrodes, which include working, counter, and reference electrodes, are made by applying carbon and silver inks onto a paper substrate, forming a paper-based screen-printed electrode (P-SPE). The first layer of PPy is deposited on the working electrode by applying a solution (200 \u0026micro;L, 0.2 M pyrrole, 0.2 M sodium chloride) under a steady potential of 0.9 V for 250 seconds. Following this, a second layer of the drug-loaded PPy film is deposited by using a mixture (200 \u0026micro;L, 0.1 M pyrrole, 0.1 M sodium chloride, and 0.005 M MTX). When a constant voltage of 0.75 V is applied for 1200 seconds, the monomer (pyrrole) undergoes oxidation to form polypyrrole, and MTX is loaded into the film. Subsequently, the electrodes are rinsed to eliminate any MTX that might have been physically absorbed onto the electrode surface. Finally, the electrode is preserved at a temperature of 4\u0026deg;C for storage.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e4.3. Patch Characterization\u003c/h2\u003e \u003cp\u003eThe surface structure of the paper (Pap), and the prepared films (Pap/C/PPy, Pap/C/PPy/MTX) were examined using field emission scanning electron microscopy (FESEM) (MIRA3 TESCAN, Czech Republic) at an accelerating voltage of 25 KV. The presence of PPy and MTX was confirmed using attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) with a Jasco FTIR-6300 spectrometer in the wavenumber range of 650\u0026ndash;4000 cm\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003e4.4. In-Vitro Release Studies\u003c/h2\u003e \u003cp\u003eMTX release in response to an electrical stimulus was investigated in a laboratory environment. This involved placing Ringer\u0026rsquo;s serum (200 \u0026micro;L) on the P-SPE to cover all three electrodes. The released MTX concentration was calculated using absorbance measurements at 303 nm and the MTX calibration curve. The study also explored the effect of loading parameters, such as Py monomer, dopant (NaCl), and MTX concentrations, on the amount of released MTX in Ringer\u0026rsquo;s serum (200 \u0026micro;L) at an applied potential of -1 V. Furthermore, the release of the drug under optimal conditions was assessed by applying different potentials (-0.5, -1, -1.2, and \u0026minus;\u0026thinsp;1.5 V) for a duration of 5 minutes. The passive leakage of MTX was measured under the same conditions, but without the application of an electrical stimulus. The release profiles of MTX at various time intervals were examined by adding Ringer\u0026rsquo;s serum (200 \u0026micro;L) to the P-SPE to cover all three electrodes, followed by the application of an appropriate potential (-1.2 V) for a specific duration.\u003c/p\u003e \u003cp\u003eThe release of MTX was examined under various pH levels of artificial sweat at the optimal potential of -1.2 V for a duration of 5 minutes. To create artificial sweat, we dissolved specific weight/volume ratios of materials (0.5% NaCl, 0.1% KCl, 0.1% lactic acid, and 0.1% urea) in deionized water.\u003csup\u003e\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e\u003c/sup\u003e We then adjusted the pH by adding ammonium hydroxide (0.001 M) using a laboratory pH meter (827 pH lab, Metrohm, Switzerland). In our study, we tested the impact of four different pH values (4.4, 5.4, 6.4, and 7.4) on drug release. The choice of sweat over other fluids was made to optimize the patch for transdermal drug delivery. Following the release, the amount of released MTX was measured for 200 \u0026micro;L of the prepared solution placed on the P-SPE. The absorbance of the resulting solutions was then recorded.\u003c/p\u003e \u003cp\u003ePulsed release was studied by adding sweat (300 \u0026micro;L, pH\u0026thinsp;=\u0026thinsp;5.4) to the P-SPE. Four consecutive electrical stimulations were applied at -1.2 V in 1.5-minute intervals. The potentiostat was turned off for 1.5 minutes between each stimulation. After each stimulation, sample of the stimulated solution (200 \u0026micro;L) was taken and its absorbance was recorded. Then fresh sweat solution (200 \u0026micro;L) was added to the remaining solution on the P-SPE, and the potentiostat was turned off for 1.5 minutes. The absorbances of the solutions were recorded at 1.5-minute intervals.\u003c/p\u003e \u003cp\u003eLastly, the performance of the prepared P-SPE was evaluated under different electrical actuator conditions (potentiostat and battery) by applying a voltage of -1.2 V. For tests using a battery as an electrical driving source, a 9 V battery equipped with a voltage-reducing module was used to apply a voltage of -1.2 V. The release was carried out under the same conditions (200 \u0026micro;L of sweat at pH 5.4 for 5 min), and the absorbance of the resulting solution was measured at 303 nm.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e4.5. In-vitro skin Permeation Test\u003c/h2\u003e \u003cp\u003eA cell resembling the Franz diffusion cell (with a penetration area of 1.54 cm\u0026sup2; and a receptor volume of 10 mL) was created and utilized for skin permeation experiments using normal hairless rat skin. Male Wistar rats (weighing 180\u0026ndash;230 g) were anesthetized with a mixed isopentane gas (1\u0026ndash;3%). The dorsal skin of the rat was removed, and then the subcutaneous fat was separated. The skin was subsequently divided into appropriate segments. To keep it hydrated, the rat skin was placed on a Franz cell and left for 30 minutes. The temperature within the cell containing PBS (pH\u0026thinsp;=\u0026thinsp;7.4) was maintained at 37\u0026deg;C, with normal rat skin secured on top. Three groups of skin permeability tests were conducted (The experiment was repeated three times in each group). In the first group (ITP), an iontophoresis patch containing MTX was applied to the skin (donor cell) and the drug was released for 5 minutes. In the second group (Stimuli-Top), the designed patch without MTX was initially placed on the skin, followed by the application of voltage (-1.2 V) using artificial sweat (pH\u0026thinsp;=\u0026thinsp;5.4) for 5 minutes. Subsequently, MTX solution (200 \u0026micro;L, 160 \u0026micro;g mL\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e) dissolved in artificial sweat were applied to the skin to assess its permeability. In the third group (Top), MTX solution (200 \u0026micro;L, 200 \u0026micro;g mL\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e) was applied topically to the skin, and the drug\u0026rsquo;s penetration rate was assessed. In each group, we collected samples (200 \u0026micro;L) at various time intervals (0.25, 0.5, 0.75, 1, and 2 hours) from receptor cells and measured their absorption. Same volume of buffer solution (200 \u0026micro;L) was periodically added to the receptor during sampling intervals to maintain volume. Detection was performed using UV absorbance at 303 nm and a graph illustrating the cumulative drug permeation per unit area over time was created.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003e4.6. Animal Tests (In-Vivo Experiment)\u003c/h2\u003e \u003cp\u003eEighteen BALB/c mice, aged 5\u0026ndash;6 weeks and of random gender, were obtained from the animal house at the School of Pharmacy, Isfahan University of Medical Sciences. The mice were kept in groups of three per cage, with libitum access to food and water, in a room maintained at 22\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C and 50% humidity. A 12-hour light-dark cycle was followed. Ethical guidelines from the Ethical Committee of University of Isfahan (IR.UI.REC.1402.132) were strictly adhered to during the animal studies. All procedures were performed under anesthesia, with efforts made to minimize pain. The mice\u0026rsquo;s dorsal hair was shaved (area: 2\u0026times;2.5 cm), and psoriasis was induced by applying a 5% IMQ cream at a dose of 65 mg cm\u003csup\u003e-\u003c/sup\u003e\u0026sup2; for 6 consecutive days. Throughout the treatment, the cream application was continued to sustain the psoriasis.). Six groups of mice randomly were established, each containing three mice: Normal group (healthy mice), Control group (Consisting of psoriatic mice without any treatment), Stimuli group (Psoriatic mice subjected to electrical stimulation (-1.2 V) using a specially designed drug-free patch for 5 minutes daily), Oral group (Psoriatic mice that received MTX tablets via gavage. The oral MTX tablet dosage was 5.143 mg kg\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e week\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e (equivalent to a clinical dose of 25 mg for a 60 kg human).\u003csup\u003e\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e\u003c/sup\u003e The MTX solution (300 \u0026micro;L, 0.5 mg mL\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e) was administered once during the treatment period using gavage tubes), Top group (Psoriatic mice treated with topical MTX solution (150 \u0026micro;g mL\u003csup\u003e-\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e) once a day), and ITP group (Psoriatic mice treated with a battery-powered iontophoresis patch using electrical stimulation (-1.2 V) for five minutes once a day). Lesion images were captured daily prior to treatment. On the fifth day, the mice were euthanized, and samples of treated skin, spleen, and kidney, liver, and lung tissues were collected for immunohistochemical analysis. These tissues were preserved in formalin solution (10%) at 4\u0026deg;C separately. for the purpose of analysis and comparison, tissues were stained using hematoxylin and eosin colors (H\u0026amp;E). subsequently, microscopic images were captured using a microscope equipped with a digital camera.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003e4.7. Statistics and reproducibility\u003c/h2\u003e \u003cp\u003eThe data presented in this study underwent statistical analysis using Tukey\u003csup\u003e,\u003c/sup\u003es multiple-comparison post hoc test and One-way analysis of variance (ANOVA) in GraphPad Prism software (version 8). The data in the figures were marked by * for **P\u0026thinsp;\u0026lt;\u0026thinsp;0.01, ***P\u0026thinsp;\u0026lt;\u0026thinsp;0.001, ****p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001, ns for no significances. The results were expressed as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD). No animals were excluded from the analysis. No data were excluded from the analyses.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003eThe Ethical Committee of the University of Isfahan approved the study. All animal studies conducted in this research strictly adhered to the ethical guidelines set forth by the Ethical Committee of the University of Isfahan (IR.UI.REC.1402.132). The welfare of the animals was a primary concern, and all procedures were designed to minimize suffering and ensure humane treatment throughout the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eReporting summary\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFurther information on research design is available in the Nature Portfolio Reporting Summary linked to this article.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated during the current study are available from the corresponding author (Masoud A. Mehrgardi) on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was funded by Isfahan University Research Council and Stanford University.The authors express their deep sincere thanks to\u0026nbsp;Azar Baradaran for her constructive comments and guidance throughout this project.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eE.M.Conceptualization, Investigation, Validation, Methodology, Visualization, Formal analysis, Data Curation, Writing - Original Draft; M A.M. Conceptualization, Validation, Formal analysis, Resources, Writing - Review \u0026amp; Editing, Supervision, Project administration, Funding acquisition.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing financial interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAdditional Information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSupplementary information\u003c/strong\u003e. The supplementary information contains supplementary figures available at http://www.nature.com.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCorresponding author\u003c/strong\u003e. correspondence to Masoud A. Mehrgardi.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eQu F, Geng R, Liu Y, Zhu J (2022) Advanced nanocarrier-and microneedle-based transdermal drug delivery strategies for skin diseases treatment. 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New J Chem 42:8512\u0026ndash;8523\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGemeiner P et al (2015) Polypyrrole-coated multi-walled carbon nanotubes for the simple preparation of counter electrodes in dye-sensitized solar cells. Synth Met 210:323\u0026ndash;331\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCai Z et al (2017) Electrochemical synthesis of graphene/polypyrrole nanotube composites for multifunctional applications. Synth Met 227:100\u0026ndash;105\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLiu S et al (2010) Green electrochemical synthesis of Pt/graphene sheet nanocomposite film and its electrocatalytic property. J Power Sources 195:4628\u0026ndash;4633\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlizadeh N, Shamaeli E (2014) Electrochemically controlled release of anticancer drug methotrexate using nanostructured polypyrrole modified with cetylpyridinium: Release kinetics investigation. 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Biology (Basel) 10:1121\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Voltage-triggered drug release, Transdermal, Psoriasis, Iontophoresis, Paper patch","lastPublishedDoi":"10.21203/rs.3.rs-4877240/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4877240/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study introduces a wearable iontophoresis transdermal paper patch for delivering methotrexate, a potent anti-inflammatory drug, to treat imiquimod (IMQ)-induced psoriasis-like inflammations. This patch features a paper-based electrode that has been coated with polypyrrole and loaded with the anti-inflammatory medication methotrexate. Incorporating paper into the design enhanced the amount of drug that could be loaded and reduced its unintended release. The most effective release of the drug, was achieved with a voltage of -1.2 V. The specially designed iontophoresis patch ensured that the methotrexate optimally penetrated the mouse skin (19.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 \u0026micro;g cm\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e after 2 h). The patch's ability to alleviate psoriasis, which was experimentally induced in BALB/c mice (gender random), was confirmed through successful testing. Histological analysis of the skin and internal organs such as the spleen, lungs, liver, and kidneys showed that methotrexate is highly effective and has minimal adverse effects.\u003c/p\u003e","manuscriptTitle":"Could Wearable Voltage-Triggered Paper Patch Revolutionize Psoriasis Treatment by Enhancing Skin Permeation of Methotrexate?","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-20 13:32:40","doi":"10.21203/rs.3.rs-4877240/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c1d01a89-b10c-4e40-9a39-c2dad651d96a","owner":[],"postedDate":"August 20th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":36083433,"name":"Physical sciences/Chemistry/Chemical biology/Drug delivery"},{"id":36083434,"name":"Biological sciences/Chemical biology"}],"tags":[],"updatedAt":"2024-08-20T13:32:42+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-20 13:32:40","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4877240","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4877240","identity":"rs-4877240","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}