{"paper_id":"4b077e63-1530-4309-a1b7-eb60df40a7c7","body_text":"Application of 32 Factorial Design of Loratadine-Loaded Nanosponge in Topical Gel system: In Vitro, Ex Vivo, and In Vivo Assessments | 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 Application of 32 Factorial Design of Loratadine-Loaded Nanosponge in Topical Gel system: In Vitro, Ex Vivo, and In Vivo Assessments Durgaramani Sivadasan, Krishnaraju Venkatesan, Jamal Moideen Muthu Mohamed, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3852896/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 16 Mar, 2024 Read the published version in Scientific Reports → Version 1 posted 10 You are reading this latest preprint version Abstract Loratadine (LoR) is a highly lipophilic and practically insoluble in water, hence having a low oral bioavailability. As it is formulated as topical gel, it competitively binds with the receptors, thus reducing the side-effects. The objective of this study was to prepare LoR loaded nanosponge (LoR-NS) in gel for topical delivery. Nine different formulations of emulsion were prepared by solvent evaporation method with polyvinyl alcohol (PVA), ethyl cellulose (EC), and dichloromethane (DCM). Based on 3 2 Full Factorial Design (FFD), optimization was carried out by varying the concentration of LOR:EC ratio and stirring rate. The preparations were subjected for the evaluation of particle size (PS), in vitro release, zeta potential (ZP) and entrapment efficiency (EE). The results revealed that the NS dispersion was nanosized with sustained release profiles and significant PS. The optimized formulation was formulated and incorporated into carbopol 934P hydrogel. The formulation was then examined to surface morphological characterizations using scanning electron microscopy (SEM) which depicted spherical NS. Stability studies, undertaken for 2 months at 40 ± 2 ℃ and 75 ± 5% RH, concluded to the stability of the formulation. The formulation did not cause skin irritation. Therefore, the prepared NS hydrogel proved to be a promising applicant for LoR as a novel drug delivery system (NDDS) for safe, sustained and controlled topical application. Nanosponge Loratadine Ethyl cellulose Hydrogel NDDS Sustained release Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Allergies arise from an immune system response to a foreign body. Allergic reaction is common and is characterized by the release of histamine, cytokines, and other mediators that may induce mast-cell degranulation. Antihistamines provide symptomatic relief to allergic conditions. Anti-histaminic drugs include loratadine, cetirizine, meclizine, bromopheniramine [ 1 ]. Tricyclic piperidine derivative loratadine (LoR) is a member of the second generation of H 1 -antihistamines and is used to treat urticaria, angioedema, and other allergic skin symptoms. It prevents the symptoms associated with histamines on gastrointestinal smooth muscle, bronchial smooth muscle, capillaries including increased capillary permeability, vasodilation. It is administrated by oral route for the treatment of skin conditions categorized by localized allergic reaction [ 2 ]. The cutaneous route should be a better option for drug delivery because the oral route has a limited bioavailability and causes side effects. Because of its high lipophilicity (log p = 5.2) and low molecular weight (382.88 Da), LoR is an excellent option for cutaneous distribution. However, LoR's limited topical applicability is due to its low water solubility [ 3 ]. Nanosponges (NS) are a new class of nanoparticles (NPs) with a small mesh-like 3D structure, smaller than 100 nm, with wide nanometric cavities that encapsulate a wide range of hydrophilic and lipophilic substances. Because of the inner hydrophobic cavities and external hydrophilic branching, they can easily load the particles [ 4 ]. The backbone is represented by a long chain of polyesters. The polymers are bound together by small molecules named as cross-linkers. NS can be used to carry water insoluble drugs which mainly belongs to BCS class II and IV [ 5 ]. NS are non-irritating, non-mutagenic, non-toxic, non-allergenic and biodegradable which are advantageous compared to other nanoparticulate systems; also, their release is predictable, are stable up to 130 ℃ and at pH of about 1–11; they can entrap a wide variety of ingredients, protecting the drug from degradation, and offering reduced side effects; they have overall better physical, chemical and thermal stability; they allow extended release for up to 12 h; eventually, these preparations are cost effective [ 6 ]. Hydrogels are cross-linked water-soluble polymers with 3D structure. If the molecular entanglement and/or secondary forces like hydrogen bond, ionic bond are responsible for the formation of linkage, then the hydrogel can be termed as reversible or physical gels. The hydrogels can be prepared by various physical forms like microparticles, slab, coating, films. The loading of the drug in the matrix and the drug release rate also depends upon the porosity of the gel matrix [ 7 , 8 ]. The novelty of NS hydrogel lies in its unique properties as a highly porous material with nanoscale dimensions that can absorb and retain large quantities of substances such as drugs, toxins, or heavy metals. It can also release these substances in a controlled manner, making it a promising candidate for drug delivery, wound healing, and environmental remediation applications. Additionally, the biocompatibility and biodegradability of NS hydrogels make them a safer and more sustainable alternative to other materials currently used in similar applications. Pharmaceutical formulations including NPs, NS, liposomes are prepared by employing specific processes that consider a number of different variables and aspects. These independent factors interact to generate efficacy, utility, stability, and safety. As a result, in order to get the intended result, it is frequently required to adjust the formulation processing settings. The quantitative methods intricacy in building the design represents the real link between the contributing elements and reactions, and the detailed for one property isn't necessarily the best for the others [ 9 ]. It is common knowledge that traditional experimentation takes a significant amount of time and effort, particularly when evaluating complicated systems. A full factorial experiment includes every possible level for every component. There are total of 2k experiments in order to analyse k components at 2 levels. The 2k full factorial design is quite useful in the early stages of experimental work, particularly in cases when there are less than or equal to 4 process parameters, design parameters, or other components [ 10 ]. For factors at 2-levels, it assumes that the response is approximately linear over the range of the chosen factor setting. There are merely two variables in the first design of the 2k series, A and B, which will each be investigated twice. The current study used the emulsion solvent evaporation approach to create LoR-loaded NS (LoR-NS) in gel. To optimise a 3 2 Full Factorial Design (FFD), the concentration of the LoR:EC ratio and the stirring rate were changed. The chosen formula was also examined for cutaneous irritation and histopathology. 2. Materials and Methods 2.1. Materials Loratadine (LoR) was a generous gift sample from Apotex Research Pvt Ltd, Bengaluru. Ethyl cellulose was purchased from Yarrow Chem (Mumbai, India), Dichloromethane was purchased from KFC (Bengaluru, India), PVA was purchased from SDFCL (Bengaluru, India), Carbopol 934P was purchased from Rolex Chemicals (Bengaluru, India), HPLC water was purchased from SDFCL (Mumbai, India). The remaining chemicals and reagents were all analytical grade. 2.2. Optimization and preparation of LoR-NS LoR-NS formulations were prepared by emulsion solvent evaporation method. The preparation of the NS involves 2 different phases namely- aqueous phase and dispersed phase (Table 1 ). The dispersed phase consisting100 mg of LoR and ethyl cellulose being dispersed in 20 mL DCM [ 11 ]. The aqueous phase includes 1% PVA solution. The dispersed phase was dropped slowly to the aqueous phase which was stirred at a constant rate for 2 h using mechanical stirrer (IKA RW 20 digital, Germany). Table 1 Formulation chart of LoR-NS. Formulation Code MEM concentration (mg) PVA DCM Water Stirring rate S1 1:1 1 20 100 1000 S2 1:2 1000 S3 1:3 1000 S4 1:1 1500 S5 1:2 1500 S6 1:3 1500 S7 1:1 2000 S8 1:2 2000 S9 1:3 2000 The prepared LoR-NS were fitted into the statistical factorial designs. A 3 2 FFD was used to examine the impact of the LoR:EC ratio (X 1 ) and stirring rate (X 2 ) as independent variables on the prepared formulations in vitro properties. LoR:EC ratios of 1:1, 1:2 and 1:3 (w/w), and 1000, 1500, 2000 rpm stirring rates were tested. Nine formulations were produced by the combination of the three levels of each variable. PS of the NS (Y 1 ) release rate (Y 2 ) and ZP (Y 3 ) were dependent variables analysed using software from Design-Expert 12 (Stat-Ease) (Table 2 ). The formulations were optimized based on the factors discussed above. The optimized formulation was selected and formulated, based on the higher desirability factor. The rationale of selecting control factor was to obtain a desired PS with a long-acting drug release in the NS preparation. To ascertain the degree of significance of the independent factors on the response variables and their interaction, an analysis of variance, or ANOVA, was utilised. Table 2 Levels and factors used in 32 FFD. Factors Levels -1 0 1 LoR:EC ratio, w/w (X 1 ) 1 : 1 1: 2 1: 3 Stirring rate, rpm (X 2 ) 1000 1500 2000 2.2.1. Preparation of LoR-NS gel Hydrogel was prepared by using 1% Carbopol 934P (gelling agent). Specified quantity of carbopol 934P was permitted to swell in 100 mL of double distilled water. Approximately 15 mL of gel was added to the optimized NS dispersion by 1%. About 0.02 g of methyl paraben was added for preservative action, the pH of the preparation was adjusted by using triethanolamine. It was continuously stirred at a constant rate for 10 min to allow the formation of carbopol hydrogel integrating LoR-NS [ 12 ]. In order to release any trapped air, the produced hydrogel was left undisturbed for 15 minutes before being stored in a tightly-sealed wide-mouth container for additional research. 2.3. Particle size (PS) polydispersibility index (PDI) and zeta potential (ZP) The mean PS, ZP of NS dispersion was determined by Dynamic light scattering (DLS) technique using Malvern Zeta sizer Nano S-90 (UK). The dilutions were done using HPLC water. Each measurement was done 3 times based to the technique explained by Mohamed et al. (2020) [ 13 ]. 2.4. In vitro release study $$\\left(\\%\\right)\\text{E}\\text{E} = \\frac{\\text{q}\\text{u}\\text{a}\\text{n}\\text{t}\\text{i}\\text{t}\\text{y} \\text{o}\\text{f} \\text{M}\\text{E}\\text{M} \\text{i}\\text{n} \\text{N}\\text{C}\\text{s}}{\\text{q}\\text{u}\\text{a}\\text{n}\\text{t}\\text{i}\\text{t}\\text{y} \\text{o}\\text{f} \\text{M}\\text{E}\\text{M} \\text{i}\\text{n} \\text{t}\\text{h}\\text{e} \\text{p}\\text{r}\\text{e}\\text{p}\\text{a}\\text{r}\\text{a}\\text{t}\\text{i}\\text{o}\\text{n}} \\times 100$$ 1 The in vitro diffusion of LoR-NS was studied using the dialysis bag method. The dialysis membrane-50 (molecular weight- 12 kDa) was saturated overnight in the pH 7.4 phosphate buffer solution (PBS). The donor compartment consists of 5 mL of the NS dispersion being filled in the dialysis bag. The receptor compartment consists of 200 mL of pH 7.4 PBS taken in 250 mL beaker. The beaker was positioned over a magnetic stirrer, and 37 ± 0.5°C and 100 rpm were maintained as the study state constant [ 14 ]. At predefined intervals (i.e., 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7 and 8 h), samples (1 mL) were removed and replaced with equivalent volumes of new PBS. The samples were examined for drug concentration using a UV-Vis spectrophotometer set to 250 nm after being suitably diluted. Each and every experiment was run in triplicate. 2.5. Scanning electron microscopy The optimized NS dispersion was investigated for surface morphology using SEM (Tescan VEGA3) at different magnifications at room temperature (RT) according to our previous studies [ 13 ]. 2.6. Spreadability study $$S = \\frac{\\text{M}\\times \\text{L}}{\\text{T}}$$ The spreadability was determined by method of horizontal plate glass. A similar weight (2 g) was attached to the upper glass plate, and between two horizontal glass plates about 1 g of the optimized LoR-NS gel was mounted. It had been held upright. It was noticed the duration it took for the top glass plate to separate from the bottom glass plate. The formula below was used to determine the spreadability [ 15 ]: Where, S = spreadability; L = length of glass slide (cm), M = weight of the upper slide (g), and T = time taken (sec). 2.7. pH determination and Viscosity A Digital pH metre 335 was used to measure the pH of LoR-NS gel. Prior to analysis, standard buffer solutions with pH values of 4.0, 7.0, and 9.2 were used to calibrate the pH metre. Following calibration, 50 g of gel was submerged in the glass electrode, and the pH was recorded [ 16 ]. Using a Brookfield viscometer, the optimised LoR-NS gel's viscosity was determined. The gel was kept at a temperature of 25°C. After being fastened to the viscometer, Helipath T-bar Spindle No. 95F was submerged in the 50 g gel. Viscosity was measured in centipoises (cps) using a viscometer run at different rpms. 2.8. Drug content 1 g of the gel, precisely weighed, was added to a 100 mL volumetric flask holding 20 mL of PBS (pH 7.4). After 30 min of shaking, 100 mL of PBS pH 7.4 solution was added to the volumetric flask. The sample was examined using an Agilent Technologies Cary 60 UV-Vis spectrophotometer set to 250 nm after an appropriate dilution. 2.9. Ex vivo permeation study of optimized LoR-NS gel Male Wistar albino rats in good health, weighing between 200 and 250 g, were purchased from an animal shelter in Chennai, India, and housed at 25°C with a 12-hour light/dark cycle. Ad libitum water and regular laboratory pellet diet were provided to the animals. The institutional animal committee (ABMRCP/IAEC /8/2020–2021) gave its approval to this study methodology. The ex vivo permeation study of the optimised LoR-NS (test) and the pure LoR (control) plain gel were studied. The rats were sacrificed by cervical dislocation (generally approved as minimal suffering for small rodents) and the dorsal skin was shaved and removed. The skin was divided for test and control. The skin was then attached to the Franz diffusion cell. The stratum corneum faced the donor compartment, while the dermis faced the receptor compartment, due to the manner it was linked. To the donor compartment, about 5 g of the gel for test and control was applied. The receptor compartment was filled with PBS (pH 7.4, mimicking the pH of blood). Subsequently, the beaker was kept on the magnetic stirrer (100 rpm) with temperature of 37 ± 0.5 ℃. At predefined intervals (i.e., 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7 and 8 hours), the samples (1 mL) were removed and replaced with an equivalent amount of freshly prepared buffer. The samples were diluted appropriately, and then their drug concentration was measured using a UV spectrophotometer set at 250 nm. The data were fit into different kinetic models [ 17 ]. The permeation of LoR-NS gel across the skin was calculated using the following formula: $$J = \\frac{\\text{V}}{\\text{A}} \\times \\frac{\\text{d}\\text{c}}{\\text{d}\\text{t}}$$ Where, J = flux; A = surface area; V = receptor volume; C = concentration; t = time. 2.10. Histopathological study After performing the ex vivo permeation study, the excised skin was stored in 10% formalin. Then, the skin was dehydrated using ethanol and fixed in paraffin. Hematoxylin and eosin (H&E) was used for further staining, and the results were examined using a light microscope [ 18 ]. 2.11. Skin irritation study A study on skin irritation was conducted on six rats (N = 6) of either sex and 200–250 g in weight. The animals were kept in cages made of polypropylene and had free access to water and a conventional meal. There were two groups of animals (n = 3 per group). The dorsal region's hair follicles were extracted with an operating blade and around 4 cm of scissors. Gel was used and wrapped with a cotton bandage after the hair was removed [ 19 ]. The following number method was used to score the reaction at the application site after it was examined: a) Formation of eschars and erythema: 0 indicates no erythema, 1 indicates very little erythema, 2 indicates well-defined erythema, and 3 indicates moderate to severe erythema. b) Formation of edoema: 0 indicates no edoema; 1 indicates extremely minor edoema (barely noticeable); 2 indicates slight edoema (area margins well raised); 3 indicates moderate edoema (raised about 1 mm); 4 indicates severe edoema (increased more than 1 mm). c) Initial cutaneous irritation score: This is how the main skin irritation was rated: One (1) is non-irritating (0.0); two (2) is inconsequential (0.1–0.4); three (3) is minor (0.41–1.9); and four (4) is severe (5.0–8.0) irritant. 2.12. Stability study Stability studies were performed for the optimized LoR-NS gel. It was stored in a wide mouth screw capped container and was placed in the stability chamber (Thermolab, Scientific equipment Ltd, India) maintained at temperature 40 ± 2 ℃/ 75 ± 5% RH for 2 months [ 12 ]. 2.12. Statistical analysis The means ± standard deviations were used to present the quantitative data. One-way analysis of variance (ANOVA) was used to conduct statistical comparisons using SPSS 13.0 for Windows software (SPSS, USA). p -values were deemed statistically significant if they were less than 0.05. 3. Results and Discussion Information on NS drug delivery systems from the past research was carried out using FFD. The researchers reported that factor design plays a very important role in optimizing drug dosage forms; by using the design of optimization, the researchers can carry out the formulations with minimum number of runs and obtain optimizing results [ 20 ]. So FFD was employed in the current research. The two independent variable was controlled at the same time and the effects were analyzed either individually or together in the design. 3.1. PS, drug release and optimization Table 3 shows the average PS of the prepared LoR-NS. R 2 = 0.8714, the mean PS varied between 284 ± 7.1 and 368 ± 74.97 nm. Every formulation exhibited PS in the nanoscale (< 1 µm) range. ANOVA was used for statistical analysis, and the results revealed a linear relationship, showing that the interaction between the two independent variables on the PS of NS was not statistically significant (p > 0.01). At higher stirring rates (> 1000 rpm), the reduced PS could be attributed to the increased mechanical shear forming smaller NS [ 21 ]. Accordingly, the emulsion globules aggregate and lead to increased PS at lower stirring rates. Collectively, the increase of PS values (formulations S1 and S2) by emulsion globules is most likely due to increase of the LoR:EC concentration along with slow stirring rate (1000 rpm). Table 3 PS and ZP of LoR-NS in gel and LoR drug release from NS gel. Formulation PS (nm) ZP Percentage release after 8h (%) S1 357.5 ± 14.6 -12.22 ± 1.6 90.76 ± 55.5 S2 328.6 ± 25.4 -15.18 ± 1.08 91.75 ± 7.62 S3 368.7 ± 74.97 -11.45 ± 1.12 90.46 ± 7.54 S4 233.9 ± 10.5 -18.81 ± 1.62 95.22 ± 6.2 S5 274.7 ± 28.6 -17.14 ± 1.81 86.2 ± 7.26 S6 284.23 ± 15.1 -17.22 ± 1.04 92.74 ± 8.55 S7 298.90 `± 14.2 -16.09 ± 1.06 95.99 ± 8.31 S8 287.9 ± 23.23 -16.18 ± 1.77 95.13 ± 9.23 S9 285.2 ± 25.2 -15.82 ± 1.92 95.18 ± 9.47 All the values are expressed as Mean ± SD, n = 3 The in vitro release profiles of LoR-NS (percentage of cumulative drug release) are illustrated graphically in Fig. 1 a. According to Table 4 , the proportion of LoR released after eight hours varied from 86.2 ± 0.26% (S5) to 95.99 ± 0.31% (S7). ANOVA was used in the statistical study to compare the release of LoR from various formulations, and the results showed that the release was significant (R 2 = 0.9976). The increase in the wall thickness of NS may be the cause of the decrease in the LoR % released after 8 hours with the rise in the polymer ratio ( p > 0.001). This can result in a longer diffusional path which reduces the release of LoR-NS [ 22 ]. It was evident that, the stirring rate increases and subsequently the NS size decreases. This increase could be the result of increased LoR entrapment in the hydrophobic matrix due to decreased porosity, which could have lowered release rates and the similar result was reported by Li et al (2023) [ 23 ]. Likewise, Costa et al (2022) investigated that the larger particles with higher porosity would allow hydrogel to leak the drug, resulting in faster release rates. Because of ECs hydrophobic and plastic nature, the particles adjacent to the surface matrix may initially be released resulting in an initial burst effect [ 24 ]. Table 4 Viscosity, pH, Drug content, and Spreadability of LoR-NS gel. Formulation Viscosity (cps) pH Drug content (%) Spreadability (g.cm/sec) LoR-NS gel 75.9 ± 5.63 6.98 ± 0.07 94.6 ± 66.15 0.65 ± 0.03 *All the values are expressed as Mean ± SD, n = 3 Factors that influence NS quality included PS with long-acting drug release. Combination of optimal level factor which influenced PS with long-acting drug release are LoR:EC ratio at level 1 (1:2), stirring rate at level 2 (1350 rpm). There are three factors has been selected considering the various factor and level. There are LoR:EC ratio, X 1 , and stirring rate, X 2 . The effect of studied variables (LoR:EC ratio, X 1 ; stirring rate, X 2 ) on the response variables (PS of the NS, Y 1 ; release rate, Y 2 ; ZP, Y 3 ) was statistically analyzed using Design-Expert 12 (Stat-Ease) software. To determine the significance and the interaction between the independent factors of the response variables, the ANOVA was performed. The relationship between independent variable and the response was described by means of polynomial equations. The equation was based on the selected model [ 25 ]. The responses such as ZP and drug release have been found to be insignificant (Fig. 1 b). The formula with highest desirability (0.895) was selected as optimized formula and was prepared for further studies. 3.2. Surface morphology The average diameter was 298 ± 4.45 nm and was determined by counting the more than 450 number of NPs visible in several SEM pictures, and by collecting SEM images of native LoR using the same dispersion and staining methodology as for the LoR dispersion (Fig. 2 a). The surface morphology of the NS was determined using SEM. The images revealed that the particles were spherical in shape, nanometric in size and was found to be porous in nature. It is most likely the result of dichloromethane diffusing from the surface of the NPs during preparation because there are tiny pores on the NS's surface (Fig. 2 b). To measure the average diameter of LoR-NS, which had previously been sonicated into a dispersion, SEM images were acquired [ 26 ]. 3.3. Viscosity, pH, Drug content, and Spreadability The spreadability of the optimized LoR-NS gel formulation was found to be 0.65 ± 0.03 g.cm/sec. Its pH was found to be 6.98 ± 0.01, which is near to skin pH. Viscosity is an important parameter of the gel because as the viscosity decreases the spreadability also decreases. Viscosity was found to be 75.9 ± 0.63 cps by using T-bar spindle (95F). Drug content was found to be 94.6 ± 0.15% (Table 4 ) which in good agreement with the previously published literature [ 27 ]. NS was considered to have a strongly spreadable because of its limited time spread. The therapeutic efficacy of gels is influenced by their distribution. To help ensure that the gel is applied to the skin uniformly, the produced gels must be easily spreadable and satisfy the ideal standards for topical application. Furthermore, this is believed to be a crucial component of patient adherence to treatment [ 28 ]. Given that anti-inflammatory and topical analgesic formulations are applied to the thin layers of the skin, the consistency of the material is one of the most crucial components. The gels viscosity is crucial in regulating the penetration of the medication. 3.4. Ex vivo permeation of LoR-NS gel Ex vivo permeability of LoR-NS gel across the skin was found to be 2.266 mg/cm2/min. Different models were adopted for fitting the drug release data in kinetic modelling for optimized formulation and revealed that ex vivo permeation study follows zero order kinetics (R2 = 0.9752), drug diffused from the matrix framework (Fig. 3 ). The 'n' value refers to the non-Fickian diffusion of all formulations (n = 0.6618), that indicates that there was absence of limit that splits the medium and the drug as described the Yadav et al (2022) [ 29 ]. Consequently, the amount of LoR increases that permeates the skin. EC included in DCM increase skin permeability by changing or disorganizing the ordered alkyl chains of phospholipids, which causes lipid fluidization [ 30 ]. Lipid fluidization increases the LoR ability to penetrate the skin. The lipid bilayers in the stratum corneum are able to penetrate using DCM, which lowers the stratum corneums barrier resistance and increases intracellular transport by dekeratinizing corneocytes. 3.5. histopathological outcome The potential histopathological changes were examined on the excised skin of rat. The histopathology of the skin exposed to either LoR-NS loaded gel (Fig. 4 ) or LoR gel (Fig. 4 b) revealed intact epidermal layers and dermal structure. The histoarchitectural presentation of this skin presented a few autolytic changes in a diffused manner. No signs of infiltration of inflammatory cells or degenerative changes was observed in the given tissue section. The histoarchitectural presentation of skin revealed intact epidermis and dermal structures (Fig. 4 c). No inflammatory infiltrates were observed, nor any degenerative changes were observed in the given tissue section [ 31 ]. The histoarchitectural presentation of skin presented with intact epidermal layers as well as intact dermal structures. No signs of infiltration of inflammatory cells or degenerative changes was observed in the given tissue section (Fig. 4 d). 3.6. Skin irritation The skin irritation studies of control and test group up to day 7 illustrates in Fig. 5 . The erythemal scores were recorded in control and test group for all the animals. The mean erythemal scores were found to be 0.00, which means that there was no edema or erythema on the skin of the shaved rats in the optimized formulation [ 32 ]. 3.7. Stability of LoR-NS gel The formulations were stable based on drug EE, and cumulative % drug release (CDR %) after their storage for 2 months at 40 ± 2 ℃, 75 ± 5% RH. Also, the spreadability, viscosity, and pH of LoR-NS gel were stable in the same experimental conditions. The data obtained with LoR-NS suspension corroborate previously published data [33]. There was no significant difference ( p > 0.05) in stability after 2 months, as given in Table 5 and Fig. 6 , indicating that the optimized formulation should remain stable when topically applied in humans. Table 5 Stability profile of LoR-NS gel at 40 ± 2 ℃, 75 ± 5% RH for 60 days. Days LoR-NS suspension LoR-NS gel Drug EE (%) CDR (%) Spreadability (g.cm/sec) Viscosity (cps) pH CDR (%) 0 97.51 ± 3.63 86.42 ± 5.08 0.65 ± 0.03 75.9 ± 3.63 6.98 ± 0.34 79.47 ± 5.13 15 95.12 ± 4.35 87.78 ± 6.06 0.68 ± 0.02 74.5 ± 4.14 6.98 ± 0.67 80.87 ± 4.17 30 96.60 ± 5.1 86.88 ± 7.05 0.63 ± 0.02 74.8 ± 4.21 6.98 ± 0.88 80.90 ± 4.20 60 98.85 ± 5.03 87.1 ± 5.06 0.67 ± 0.02 71.6 ± 3.14 6.98 ± 0.47 81.87 ± 3.09 90 96.29 ± 8.18 85.37 ± 10.6 0.61 ± 0.45 69.4 ± 5.59 6.48 ± 0.18 78.31 ± 7.16 *All the values are expressed as mean ± SD, n = 3 4. Conclusion In conclusion, the comprehensive exploration of the Loratadine-loaded nanosponge incorporated into a topical gel through a Complete Factorial Design has yielded promising results through a series of rigorous assessments. The in vitro studies provided valuable insights into thepreparations physicochemical attributes, drug release kinetics, and stability. The ex-vivo assessments, utilizing skin permeation studies, further demonstrated the efficacy of the nanosponge-based gel in facilitating the transdermal delivery of Loratadine. These collective findings highlight the potential of the Loratadine-loaded nanosponge in topical gel as a promising candidate for transdermal drug delivery, with implications for improved therapeutic outcomes and patient compliance. Further studies and clinical trials will be crucial to validate and expand upon these results, paving the way for the potential translation of this innovative formulation into clinical practice. Declarations Acknowledgements The authors are thankful to the Faculty of pharmacy & BioMedical Sciences, MAHSA University, Bandar Saujana Putra, 42610 Jenjarom, Selangor. Malaysia for providing the necessary lab facilities during the experimental study. Author contributions D.S. and J.M.M.M. conceived and designed research, K.V and S.A. conducted experiments, J.M.M.M., Y.I.A, M.F., and A.E. analyzed data and wrote the manuscript, S.E and F.M. supervision of the work. All authors read and approved the manuscript. Funding The authors extend their sincere appreciation to the Deanship of Scientific Research at King Khalid University for funding this study through the Large Research Group Project under grant number \"RGP 2/109/44\". Competing interests The authors declare no competing interests. Additional information Correspondence and requests for materials should be addressed to S.E. or F.M. Data Availability The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request. References Nur Husna SM, Tan HT, Md Shukri N, Mohd Ashari NS, Wong KK. Allergic Rhinitis: A Clinical and Pathophysiological Overview. Front Med (Lausanne). 2022 Apr 7;9:874114. doi: 10.3389/fmed.2022.874114. Sidhu G, Akhondi H. Loratadine. [Updated 2023 Mar 13]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK542278/ Yu YQ, Yang X, Wu XF, Fan YB. Enhancing Permeation of Drug Molecules Across the Skin via Delivery in Nanocarriers: Novel Strategies for Effective Transdermal Applications. Front Bioeng Biotechnol. 2021 Mar 29;9:646554. doi: 10.3389/fbioe.2021.646554. Akash Garg a, Wen-Cheng Lai b, Himansu Chopra a, Rutvi Agrawal a, Talever Singh a, Ramkumar Chaudhary a, Braj Nandan Dubey a. Nanosponge: A promising and intriguing strategy in medical and pharmaceutical Science. Heliyon Volume 10, Issue 1, 15 January 2024, e23303. https://doi.org/10.1016/j.heliyon.2023.e23303 Moideen, J.M.M.; Alqahtani, A.; Venkatesan, K.; Ahmad, F.; Krisharaju, K.; Gayasuddin, M.; Shaik, R.A. Application of the Box-Behnken design for the production of soluble curcumin: Skimmed milk powder inclusion complex for improving the treatment of colorectal cancer. Food Sci. Nutr . 2020 , 8, 6643–6659. Himangshu Bhowmik, D. Nagasamy Venkatesh * , Anuttam Kuila, Kammari Harish Kumar. Nanosponges: A Review. Int J App Pharm, Vol 10, Issue 4, 2018, 1-5. Bashir S, Hina M, Iqbal J, Rajpar AH, Mujtaba MA, Alghamdi NA, Wageh S, Ramesh K, Ramesh S. Fundamental Concepts of Hydrogels: Synthesis, Properties, and Their Applications. Polymers (Basel). 2020 Nov 16;12(11):2702. doi: 10.3390/polym12112702. Ahmad Z, Salman S, Khan SA, Amin A, Rahman ZU, Al-Ghamdi YO, Akhtar K, Bakhsh EM, Khan SB. Versatility of Hydrogels: From Synthetic Strategies, Classification, and Properties to Biomedical Applications. Gels. 2022 Mar 7;8(3):167. doi: 10.3390/gels8030167. Mohamed, J.M.; Alqahtani, A.; Ahmad, F.; Krishnaraju, V.; Kalpana, K. Stoichiometrically governed curcumin solid dispersion and its cytotoxic evaluation on colorectal adenocarcinoma cells. Drug Des. Deliv. Ther . 2020 , 14 , 4639–4658. Antony, Jiju. (2003). 6-Full factorial designs. Design of Experiments for Engineers and Scientists. 54-72. 10.1016/B978-075064709-0/50007-7. Kashif Maroof, Ronald F. S. Lee, Lee Fong Siow & Siew Hua Gan. (2022) Microencapsulation of propolis by spray drying: A review. Drying Technology 40:6, pages 1083-1102. Mahmood A, Mahmood A, Ibrahim MA, Hussain Z, Ashraf MU, Salem-Bekhit MM, Elbagory I. Development and Evaluation of Sodium Alginate/Carbopol 934P-Co-Poly (Methacrylate) Hydrogels for Localized Drug Delivery. Polymers . 2023; 15(2):311. https://doi.org/10.3390/polym15020311 Mohamed, J.M.; Alqahtani, A.; Ahmad, F.; Krishnaraju, V.; Kalpana, K. Pectin co-functionalized dual layered solid lipid nanoparticle made by soluble curcumin for the targeted potential treatment of colorectal cancer. Carbohydr. Polym . 2020 , 252, 117180. Al Fatease, A., Alqahtani, A., Khan, B.A., Mohamed, J.MM, Farhana SA. Preparation and characterization of a curcumin nanoemulsion gel for the effective treatment of mycoses. Sci Rep 13 , 22730 (2023). https://doi.org/10.1038/s41598-023-49328-2. Nnamani PO, Ugwu AA, Nnadi OH, Kenechukwu FC, Ofokansi KC, Attama AA, Lehr CM. Formulation and evaluation of transdermal nanogel for delivery of artemether. Drug Deliv Transl Res. 2021 Aug;11(4):1655-1674. doi: 10.1007/s13346-021-00951-4. Kulasekaran, A. & Gopal, Andal & Lakshimipathy, R. & Alexander, J.. (2015). Modification in pH measurements for getting accurate pH values with different pH meters irrespective of aging and drifts in the meters. International Journal of ChemTech Research. 8. 16-24. Das MK, Ahmed AB. Formulation and ex vivo evaluation of rofecoxib gel for topical application. Acta Pol Pharm. 2007 Sep-Oct;64(5):461-7. Goindi S, Kumar G, Kumar N, Kaur A. Development of novel elastic vesicle-based topical formulation of cetirizine dihydrochloride for treatment of atopic dermatitis. AAPS PharmSciTech. 2013 Dec;14(4):1284-93. doi: 10.1208/s12249-013-0017-3. Prajapati DP, Patel M, Dharamsi A. Beneficial effect of polyherbal formulation in letrozole induced Polycystic ovarian syndrome (PCOS). J Tradit Complement Med. 2022 Aug 10;12(6):575-583. doi: 10.1016/j.jtcme.2022.08.003. Singh, Bhupinder & Dahiya, Manju & Kharb, Vandana & Ahuja, Naveen. (2005). Optimizing Drug Delivery Systems Using Systematic \"Design of Experiments.\" Part II: Retrospect and Prospects. Critical reviews in therapeutic drug carrier systems. 22. 215-94. 10.1615/CritRevTherDrugCarrierSyst.v22.i3.10. Nazief AM, Hassaan PS, Khalifa HM, Sokar MS, El-Kamel AH. Lipid-Based Gliclazide Nanoparticles for Treatment of Diabetes: Formulation, Pharmacokinetics, Pharmacodynamics and Subacute Toxicity Study. Int J Nanomedicine. 2020 Feb 18;15:1129-1148. doi: 10.2147/IJN.S235290. Senthivel CK, Karuppaiyan K, Ahamad F, Mohamed JMM*, El-Sherbiny M, Alotaibi AT, Abed SY, Ibraheem KM, Salama M. Terminalia chebula loaded polymeric nanoparticles: a preliminary approach for status epilepticus. Chemical Papers 2023, DOI: 10.1007/s11696-023-03164-w. Zhimin Li, Xianjing Feng, Shixing Luo, Yanfeng Ding, Zhi Zhang, Yifeng Shang, Doudou Lei, Jinhong Cai, Jinmin Zhao, Li Zheng, Ming Gao, High drug loading hydrophobic cross-linked dextran microspheres as novel drug delivery systems for the treatment of osteoarthritis, Asian Journal of Pharmaceutical Sciences, Volume 18, Issue 4, 2023, 100830, https://doi.org/10.1016/j.ajps.2023.100830. Ana Letícia Rodrigues Costa, Stephanie M. Willerth, Lucimara Gaziola de la Torre, Sang Won Han, Trends in hydrogel-based encapsulation technologies for advanced cell therapies applied to limb ischemia, Materials Today Bio, Volume 13, 2022, 100221, https://doi.org/10.1016/j.mtbio.2022.100221. Mohamed JMM, Alqahtani A, Khan BA, Al Fatease A, Alqahtani T, Venkatesan K, Ahmad F, Alzghoul BI, Alamri A. Preparation of Soluble Complex of Curcumin for the Potential Antagonistic Effects on Human Colorectal Adenocarcinoma Cells. Pharmaceuticals (Basel). 2021 Sep 19;14(9):939. doi: 10.3390/ph14090939. Ahmad F, Al-Subaie AM, Gayasuddin M, Mohamed JM, Krishnaraju V. Review on the Medicinal uses and Pharmacological aspects of Plectranthus tenuiflorus from the Labiatae Family of Saudi Arabia. International Journal of Pharmaceutical Sciences Review and Research, 2020, vol. 64, no.2, pp. 43-48. Mohamed JMM, Khan BA, Rajendran V, El-Sherbiny M, Othman G, Hussamuldin, ABA, Al-Serwi, RH. Polymeric Ethosomal Gel Loaded with Nimodipine: Optimisation, Pharmacokinetic and Histopathological Analysis. Saudi Pharmaceutical Journal, Volume 30, Issue 11, November 2022, Pages 1603-1611. https://doi.org/10.1016/j.jsps.2022.09.003 Yadav K, Yadav D, Kumar S, Narra N, Mohamed JMM* et al. Natural Biodegradable and Biomass Polymeric Nanoparticles for the Delivery of Noscapine for cancer treatment. Biomass Conversion and Biorefinery, DOI: 10.1007/s13399-022-03334-y Kurmi BD, Tekchandani P, Paliwal R, Paliwal SR. Transdermal Drug Delivery: Opportunities and Challenges for Controlled Delivery of Therapeutic Agents Using Nanocarriers. Curr Drug Metab. 2017;18(5):481-495. doi: 10.2174/1389200218666170222150555. Landén NX, Li D, Ståhle M. Transition from inflammation to proliferation: a critical step during wound healing. Cell Mol Life Sci. 2016 Oct;73(20):3861-85. doi: 10.1007/s00018-016-2268-0. Wang J, Li Z, Sun F, Tang S, Zhang S, Lv P, Li J, Cao X. Evaluation of dermal irritation and skin sensitization due to vitacoxib. Toxicol Rep. 2017 Jun 10;4:287-290. doi: 10.1016/j.toxrep.2017.06.003. Elhassan GO, Mohamed JMM. Development And In Vitro Evaluation Of Valsartan-Loaded Resealed Erythrocytes, International Journal of Applied Pharmaceutics, Vol. 14, 2022, 201-205, 10.22159/ijap.2022.v14ti.57 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 16 Mar, 2024 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 23 Jan, 2024 Reviews received at journal 21 Jan, 2024 Reviews received at journal 18 Jan, 2024 Reviewers agreed at journal 17 Jan, 2024 Reviewers agreed at journal 17 Jan, 2024 Reviewers invited by journal 17 Jan, 2024 Editor assigned by journal 17 Jan, 2024 Editor invited by journal 17 Jan, 2024 Submission checks completed at journal 17 Jan, 2024 First submitted to journal 11 Jan, 2024 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. 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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-3852896\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Article\",\"associatedPublications\":[],\"authors\":[{\"id\":267792835,\"identity\":\"b8ca5bb1-64c0-4ad1-8add-087da7756a30\",\"order_by\":0,\"name\":\"Durgaramani Sivadasan\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Jizan University\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Durgaramani\",\"middleName\":\"\",\"lastName\":\"Sivadasan\",\"suffix\":\"\"},{\"id\":267792836,\"identity\":\"b57f2d9d-3e7a-4e60-a583-51ff7d9fb2d8\",\"order_by\":1,\"name\":\"Krishnaraju Venkatesan\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"King Khalid University\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Krishnaraju\",\"middleName\":\"\",\"lastName\":\"Venkatesan\",\"suffix\":\"\"},{\"id\":267792837,\"identity\":\"4790ddeb-c6b9-4ccf-9005-be52e7e7ab41\",\"order_by\":2,\"name\":\"Jamal Moideen Muthu Mohamed\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"MAHSA University\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Jamal\",\"middleName\":\"Moideen Muthu\",\"lastName\":\"Mohamed\",\"suffix\":\"\"},{\"id\":267792838,\"identity\":\"29b6f5cf-45a4-4f79-a7ef-f1ad08eee8c5\",\"order_by\":3,\"name\":\"Saud Alqahtani\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"King Khalid University\",\"correspondingAuthor\":false,\"submittingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Saud\",\"middleName\":\"\",\"lastName\":\"Alqahtani\",\"suffix\":\"\"},{\"id\":267792839,\"identity\":\"5f34e6ef-ec82-42ca-afba-4a1841b17b96\",\"order_by\":4,\"name\":\"Yahya I. 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The significance variance associated with control (LoR gel) are denoted by **\\u003cem\\u003ep\\u003c/em\\u003e \\u0026gt;0.05 as estimated by ANOVA text.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3852896/v1/b4f95d50b63fc16f4c425bb2.png\"},{\"id\":49810792,\"identity\":\"436e9ef0-d5e8-4c65-8bea-b3b1dac28b68\",\"added_by\":\"auto\",\"created_at\":\"2024-01-18 11:58:35\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":774194,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eHistopathological reports of (a) normal skin (Untreated-control), (b) LoR solution, (b) LoR-NS dispersion, and (d) LoR-NS gel.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3852896/v1/31a0374a028758f943e84b86.png\"},{\"id\":49811468,\"identity\":\"343b951e-ddac-438c-9259-bfd5528fbcf0\",\"added_by\":\"auto\",\"created_at\":\"2024-01-18 12:06:35\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":344468,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e(a) control group, (b) test group at day 0, (c) test group at day 1, (d) test group at day 2, (e) test group at day 3, (f) test group at day 5, (g) test group at day 6, and (h) test group at day 7.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3852896/v1/76f097686001dec81813e0ab.png\"},{\"id\":49810790,\"identity\":\"170a0b58-f462-4b31-b592-916b2e8d1cde\",\"added_by\":\"auto\",\"created_at\":\"2024-01-18 11:58:35\",\"extension\":\"png\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":127230,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003e\\u003cem\\u003eIn vitro\\u003c/em\\u003e drug release profile during 8h in which medium (PBS, pH 7.4?) at 40 ± 2 ℃, 75±5 % RH of (a) LoR-NS suspension and (b) LoR-NS gel. No significant differences were observed overtime and between (a) and (b).\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3852896/v1/1b19e3981669c138a7b52c04.png\"},{\"id\":52907941,\"identity\":\"da2b1a35-8081-434c-b961-39019115bb4f\",\"added_by\":\"auto\",\"created_at\":\"2024-03-18 15:14:38\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":3072159,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-3852896/v1/271335a0-d3ec-4142-8eff-53512ebda3db.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Application of 32 Factorial Design of Loratadine-Loaded Nanosponge in Topical Gel system: In Vitro, Ex Vivo, and In Vivo Assessments\",\"fulltext\":[{\"header\":\"1. Introduction\",\"content\":\"\\u003cp\\u003eAllergies arise from an immune system response to a foreign body. Allergic reaction is common and is characterized by the release of histamine, cytokines, and other mediators that may induce mast-cell degranulation. Antihistamines provide symptomatic relief to allergic conditions. Anti-histaminic drugs include loratadine, cetirizine, meclizine, bromopheniramine [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eTricyclic piperidine derivative loratadine (LoR) is a member of the second generation of H\\u003csub\\u003e1\\u003c/sub\\u003e-antihistamines and is used to treat urticaria, angioedema, and other allergic skin symptoms. It prevents the symptoms associated with histamines on gastrointestinal smooth muscle, bronchial smooth muscle, capillaries including increased capillary permeability, vasodilation. It is administrated by oral route for the treatment of skin conditions categorized by localized allergic reaction [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e]. The cutaneous route should be a better option for drug delivery because the oral route has a limited bioavailability and causes side effects. Because of its high lipophilicity (log p\\u0026thinsp;=\\u0026thinsp;5.2) and low molecular weight (382.88 Da), LoR is an excellent option for cutaneous distribution. However, LoR's limited topical applicability is due to its low water solubility [\\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eNanosponges (NS) are a new class of nanoparticles (NPs) with a small mesh-like 3D structure, smaller than 100 nm, with wide nanometric cavities that encapsulate a wide range of hydrophilic and lipophilic substances. Because of the inner hydrophobic cavities and external hydrophilic branching, they can easily load the particles [\\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e]. The backbone is represented by a long chain of polyesters. The polymers are bound together by small molecules named as cross-linkers. NS can be used to carry water insoluble drugs which mainly belongs to BCS class II and IV [\\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e]. NS are non-irritating, non-mutagenic, non-toxic, non-allergenic and biodegradable which are advantageous compared to other nanoparticulate systems; also, their release is predictable, are stable up to 130 ℃ and at pH of about 1\\u0026ndash;11; they can entrap a wide variety of ingredients, protecting the drug from degradation, and offering reduced side effects; they have overall better physical, chemical and thermal stability; they allow extended release for up to 12 h; eventually, these preparations are cost effective [\\u003cspan citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eHydrogels are cross-linked water-soluble polymers with 3D structure. If the molecular entanglement and/or secondary forces like hydrogen bond, ionic bond are responsible for the formation of linkage, then the hydrogel can be termed as reversible or physical gels. The hydrogels can be prepared by various physical forms like microparticles, slab, coating, films. The loading of the drug in the matrix and the drug release rate also depends upon the porosity of the gel matrix [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eThe novelty of NS hydrogel lies in its unique properties as a highly porous material with nanoscale dimensions that can absorb and retain large quantities of substances such as drugs, toxins, or heavy metals. It can also release these substances in a controlled manner, making it a promising candidate for drug delivery, wound healing, and environmental remediation applications. Additionally, the biocompatibility and biodegradability of NS hydrogels make them a safer and more sustainable alternative to other materials currently used in similar applications.\\u003c/p\\u003e \\u003cp\\u003ePharmaceutical formulations including NPs, NS, liposomes are prepared by employing specific processes that consider a number of different variables and aspects. These independent factors interact to generate efficacy, utility, stability, and safety. As a result, in order to get the intended result, it is frequently required to adjust the formulation processing settings. The quantitative methods intricacy in building the design represents the real link between the contributing elements and reactions, and the detailed for one property isn't necessarily the best for the others [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. It is common knowledge that traditional experimentation takes a significant amount of time and effort, particularly when evaluating complicated systems.\\u003c/p\\u003e \\u003cp\\u003eA full factorial experiment includes every possible level for every component. There are total of 2k experiments in order to analyse k components at 2 levels. The 2k full factorial design is quite useful in the early stages of experimental work, particularly in cases when there are less than or equal to 4 process parameters, design parameters, or other components [\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e]. For factors at 2-levels, it assumes that the response is approximately linear over the range of the chosen factor setting. There are merely two variables in the first design of the 2k series, A and B, which will each be investigated twice.\\u003c/p\\u003e \\u003cp\\u003eThe current study used the emulsion solvent evaporation approach to create LoR-loaded NS (LoR-NS) in gel. To optimise a 3\\u003csup\\u003e2\\u003c/sup\\u003e Full Factorial Design (FFD), the concentration of the LoR:EC ratio and the stirring rate were changed. The chosen formula was also examined for cutaneous irritation and histopathology.\\u003c/p\\u003e\"},{\"header\":\"2. Materials and Methods\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.1. Materials\\u003c/h2\\u003e \\u003cp\\u003eLoratadine (LoR) was a generous gift sample from Apotex Research Pvt Ltd, Bengaluru. Ethyl cellulose was purchased from Yarrow Chem (Mumbai, India), Dichloromethane was purchased from KFC (Bengaluru, India), PVA was purchased from SDFCL (Bengaluru, India), Carbopol 934P was purchased from Rolex Chemicals (Bengaluru, India), HPLC water was purchased from SDFCL (Mumbai, India). The remaining chemicals and reagents were all analytical grade.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec4\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.2. Optimization and preparation of LoR-NS\\u003c/h2\\u003e \\u003cp\\u003eLoR-NS formulations were prepared by emulsion solvent evaporation method. The preparation of the NS involves 2 different phases namely- aqueous phase and dispersed phase (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). The dispersed phase consisting100 mg of LoR and ethyl cellulose being dispersed in 20 mL DCM [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e]. The aqueous phase includes 1% PVA solution. The dispersed phase was dropped slowly to the aqueous phase which was stirred at a constant rate for 2 h using mechanical stirrer (IKA RW 20 digital, Germany).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab1\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 1\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eFormulation chart of LoR-NS.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"6\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eFormulation\\u003c/p\\u003e \\u003cp\\u003eCode\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eMEM\\u003c/p\\u003e \\u003cp\\u003econcentration (mg)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003ePVA\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eDCM\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eWater\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003eStirring rate\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\" morerows=\\\"8\\\" rowspan=\\\"9\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\" morerows=\\\"8\\\" rowspan=\\\"9\\\"\\u003e \\u003cp\\u003e20\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\" morerows=\\\"8\\\" rowspan=\\\"9\\\"\\u003e \\u003cp\\u003e100\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e1000\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e1000\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e1000\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e1500\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e1500\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e1500\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS7\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2000\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2000\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1:3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2000\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe prepared LoR-NS were fitted into the statistical factorial designs. A 3\\u003csup\\u003e2\\u003c/sup\\u003e FFD was used to examine the impact of the LoR:EC ratio (X\\u003csub\\u003e1\\u003c/sub\\u003e) and stirring rate (X\\u003csub\\u003e2\\u003c/sub\\u003e) as independent variables on the prepared formulations \\u003cem\\u003ein vitro\\u003c/em\\u003e properties. LoR:EC ratios of 1:1, 1:2 and 1:3 (w/w), and 1000, 1500, 2000 rpm stirring rates were tested. Nine formulations were produced by the combination of the three levels of each variable. PS of the NS (Y\\u003csub\\u003e1\\u003c/sub\\u003e) release rate (Y\\u003csub\\u003e2\\u003c/sub\\u003e) and ZP (Y\\u003csub\\u003e3\\u003c/sub\\u003e) were dependent variables analysed using software from Design-Expert 12 (Stat-Ease) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e). The formulations were optimized based on the factors discussed above. The optimized formulation was selected and formulated, based on the higher desirability factor. The rationale of selecting control factor was to obtain a desired PS with a long-acting drug release in the NS preparation. To ascertain the degree of significance of the independent factors on the response variables and their interaction, an analysis of variance, or ANOVA, was utilised.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab2\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 2\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eLevels and factors used in 32 FFD.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"4\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eFactors\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c4\\\" namest=\\\"c2\\\"\\u003e \\u003cp\\u003eLevels\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e-1\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eLoR:EC ratio, w/w (X\\u003csub\\u003e1\\u003c/sub\\u003e)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1 : 1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1: 2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1: 3\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eStirring rate, rpm (X\\u003csub\\u003e2\\u003c/sub\\u003e)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e1000\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1500\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e2000\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cdiv id=\\\"Sec5\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.2.1. Preparation of LoR-NS gel\\u003c/h2\\u003e \\u003cp\\u003eHydrogel was prepared by using 1% Carbopol 934P (gelling agent). Specified quantity of carbopol 934P was permitted to swell in 100 mL of double distilled water. Approximately 15 mL of gel was added to the optimized NS dispersion by 1%. About 0.02 g of methyl paraben was added for preservative action, the pH of the preparation was adjusted by using triethanolamine. It was continuously stirred at a constant rate for 10 min to allow the formation of carbopol hydrogel integrating LoR-NS [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e]. In order to release any trapped air, the produced hydrogel was left undisturbed for 15 minutes before being stored in a tightly-sealed wide-mouth container for additional research.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec6\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.3. Particle size (PS) polydispersibility index (PDI) and zeta potential (ZP)\\u003c/h2\\u003e \\u003cp\\u003eThe mean PS, ZP of NS dispersion was determined by Dynamic light scattering (DLS) technique using Malvern Zeta sizer Nano S-90 (UK). The dilutions were done using HPLC water. Each measurement was done 3 times based to the technique explained by Mohamed et al. (2020) [\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec7\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.4. In vitro release study\\u003c/h2\\u003e \\u003cp\\u003e \\u003cdiv id=\\\"Equ1\\\" class=\\\"Equation\\\"\\u003e \\u003cdiv format=\\\"TEX\\\" class=\\\"mathdisplay\\\" id=\\\"FileID_Equ1\\\" name=\\\"EquationSource\\\"\\u003e\\n$$\\\\left(\\\\%\\\\right)\\\\text{E}\\\\text{E} = \\\\frac{\\\\text{q}\\\\text{u}\\\\text{a}\\\\text{n}\\\\text{t}\\\\text{i}\\\\text{t}\\\\text{y} \\\\text{o}\\\\text{f} \\\\text{M}\\\\text{E}\\\\text{M} \\\\text{i}\\\\text{n} \\\\text{N}\\\\text{C}\\\\text{s}}{\\\\text{q}\\\\text{u}\\\\text{a}\\\\text{n}\\\\text{t}\\\\text{i}\\\\text{t}\\\\text{y} \\\\text{o}\\\\text{f} \\\\text{M}\\\\text{E}\\\\text{M} \\\\text{i}\\\\text{n} \\\\text{t}\\\\text{h}\\\\text{e} \\\\text{p}\\\\text{r}\\\\text{e}\\\\text{p}\\\\text{a}\\\\text{r}\\\\text{a}\\\\text{t}\\\\text{i}\\\\text{o}\\\\text{n}} \\\\times 100$$\\u003c/div\\u003e \\u003cdiv class=\\\"EquationNumber\\\"\\u003e1\\u003c/div\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe \\u003cem\\u003ein vitro\\u003c/em\\u003e diffusion of LoR-NS was studied using the dialysis bag method. The dialysis membrane-50 (molecular weight- 12 kDa) was saturated overnight in the pH 7.4 phosphate buffer solution (PBS). The donor compartment consists of 5 mL of the NS dispersion being filled in the dialysis bag. The receptor compartment consists of 200 mL of pH 7.4 PBS taken in 250 mL beaker. The beaker was positioned over a magnetic stirrer, and 37\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.5\\u0026deg;C and 100 rpm were maintained as the study state constant [\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e]. At predefined intervals (i.e., 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7 and 8 h), samples (1 mL) were removed and replaced with equivalent volumes of new PBS. The samples were examined for drug concentration using a UV-Vis spectrophotometer set to 250 nm after being suitably diluted. Each and every experiment was run in triplicate.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.5. Scanning electron microscopy\\u003c/h2\\u003e \\u003cp\\u003eThe optimized NS dispersion was investigated for surface morphology using SEM (Tescan VEGA3) at different magnifications at room temperature (RT) according to our previous studies [\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec9\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.6. Spreadability study\\u003c/h2\\u003e \\u003cp\\u003e \\u003cdiv id=\\\"Equa\\\" class=\\\"Equation\\\"\\u003e \\u003cdiv format=\\\"TEX\\\" class=\\\"mathdisplay\\\" id=\\\"FileID_Equa\\\" name=\\\"EquationSource\\\"\\u003e\\n$$S = \\\\frac{\\\\text{M}\\\\times \\\\text{L}}{\\\\text{T}}$$\\u003c/div\\u003e \\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe spreadability was determined by method of horizontal plate glass. A similar weight (2 g) was attached to the upper glass plate, and between two horizontal glass plates about 1 g of the optimized LoR-NS gel was mounted. It had been held upright. It was noticed the duration it took for the top glass plate to separate from the bottom glass plate. The formula below was used to determine the spreadability [\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e]:\\u003c/p\\u003e \\u003cp\\u003eWhere, S\\u0026thinsp;=\\u0026thinsp;spreadability; L\\u0026thinsp;=\\u0026thinsp;length of glass slide (cm), M\\u0026thinsp;=\\u0026thinsp;weight of the upper slide (g), and T\\u0026thinsp;=\\u0026thinsp;time taken (sec).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec10\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.7. pH determination and Viscosity\\u003c/h2\\u003e \\u003cp\\u003eA Digital pH metre 335 was used to measure the pH of LoR-NS gel. Prior to analysis, standard buffer solutions with pH values of 4.0, 7.0, and 9.2 were used to calibrate the pH metre. Following calibration, 50 g of gel was submerged in the glass electrode, and the pH was recorded [\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e]. Using a Brookfield viscometer, the optimised LoR-NS gel's viscosity was determined. The gel was kept at a temperature of 25\\u0026deg;C. After being fastened to the viscometer, Helipath T-bar Spindle No. 95F was submerged in the 50 g gel. Viscosity was measured in centipoises (cps) using a viscometer run at different rpms.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.8. Drug content\\u003c/h2\\u003e \\u003cp\\u003e1 g of the gel, precisely weighed, was added to a 100 mL volumetric flask holding 20 mL of PBS (pH 7.4). After 30 min of shaking, 100 mL of PBS pH 7.4 solution was added to the volumetric flask. The sample was examined using an Agilent Technologies Cary 60 UV-Vis spectrophotometer set to 250 nm after an appropriate dilution.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.9. Ex vivo permeation study of optimized LoR-NS gel\\u003c/h2\\u003e \\u003cp\\u003eMale Wistar albino rats in good health, weighing between 200 and 250 g, were purchased from an animal shelter in Chennai, India, and housed at 25\\u0026deg;C with a 12-hour light/dark cycle. Ad libitum water and regular laboratory pellet diet were provided to the animals. The institutional animal committee (ABMRCP/IAEC /8/2020\\u0026ndash;2021) gave its approval to this study methodology. The \\u003cem\\u003eex vivo\\u003c/em\\u003e permeation study of the optimised LoR-NS (test) and the pure LoR (control) plain gel were studied. The rats were sacrificed by cervical dislocation (generally approved as minimal suffering for small rodents) and the dorsal skin was shaved and removed. The skin was divided for test and control. The skin was then attached to the Franz diffusion cell. The stratum corneum faced the donor compartment, while the dermis faced the receptor compartment, due to the manner it was linked. To the donor compartment, about 5 g of the gel for test and control was applied. The receptor compartment was filled with PBS (pH 7.4, mimicking the pH of blood). Subsequently, the beaker was kept on the magnetic stirrer (100 rpm) with temperature of 37\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.5 ℃. At predefined intervals (i.e., 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7 and 8 hours), the samples (1 mL) were removed and replaced with an equivalent amount of freshly prepared buffer. The samples were diluted appropriately, and then their drug concentration was measured using a UV spectrophotometer set at 250 nm. The data were fit into different kinetic models [\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e]. The permeation of LoR-NS gel across the skin was calculated using the following formula:\\u003cdiv id=\\\"Equb\\\" class=\\\"Equation\\\"\\u003e\\u003cdiv format=\\\"TEX\\\" class=\\\"mathdisplay\\\" id=\\\"FileID_Equb\\\" name=\\\"EquationSource\\\"\\u003e\\n$$J = \\\\frac{\\\\text{V}}{\\\\text{A}} \\\\times \\\\frac{\\\\text{d}\\\\text{c}}{\\\\text{d}\\\\text{t}}$$\\u003c/div\\u003e\\u003c/div\\u003e\\u003c/p\\u003e \\u003cp\\u003eWhere, J\\u0026thinsp;=\\u0026thinsp;flux; A\\u0026thinsp;=\\u0026thinsp;surface area; V\\u0026thinsp;=\\u0026thinsp;receptor volume; C\\u0026thinsp;=\\u0026thinsp;concentration; t\\u0026thinsp;=\\u0026thinsp;time.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.10. Histopathological study\\u003c/h2\\u003e \\u003cp\\u003eAfter performing the ex vivo permeation study, the excised skin was stored in 10% formalin. Then, the skin was dehydrated using ethanol and fixed in paraffin. Hematoxylin and eosin (H\\u0026amp;E) was used for further staining, and the results were examined using a light microscope [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.11. Skin irritation study\\u003c/h2\\u003e \\u003cp\\u003eA study on skin irritation was conducted on six rats (N\\u0026thinsp;=\\u0026thinsp;6) of either sex and 200\\u0026ndash;250 g in weight. The animals were kept in cages made of polypropylene and had free access to water and a conventional meal. There were two groups of animals (n\\u0026thinsp;=\\u0026thinsp;3 per group). The dorsal region's hair follicles were extracted with an operating blade and around 4 cm of scissors. Gel was used and wrapped with a cotton bandage after the hair was removed [\\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e]. The following number method was used to score the reaction at the application site after it was examined:\\u003c/p\\u003e \\u003cp\\u003e\\u003cspan\\u003ea) Formation of eschars and erythema: 0 indicates no erythema, 1 indicates very little erythema, 2 indicates well-defined erythema, and 3 indicates moderate to severe erythema.\\u003cbr\\u003e\\u003c/span\\u003e\\u003cspan\\u003eb) Formation of edoema: 0 indicates no edoema; 1 indicates extremely minor edoema (barely noticeable); 2 indicates slight edoema (area margins well raised); 3 indicates moderate edoema (raised about 1 mm); 4 indicates severe edoema (increased more than 1 mm).\\u003cbr\\u003e\\u003c/span\\u003e\\u003cspan\\u003ec) Initial cutaneous irritation score: This is how the main skin irritation was rated: One (1) is non-irritating (0.0); two (2) is inconsequential (0.1\\u0026ndash;0.4); three (3) is minor (0.41\\u0026ndash;1.9); and four (4) is severe (5.0\\u0026ndash;8.0) irritant.\\u003cbr\\u003e\\u003c/span\\u003e\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec15\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.12. Stability study\\u003c/h2\\u003e \\u003cp\\u003eStability studies were performed for the optimized LoR-NS gel. It was stored in a wide mouth screw capped container and was placed in the stability chamber (Thermolab, Scientific equipment Ltd, India) maintained at temperature 40\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2 ℃/ 75\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5% RH for 2 months [\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e].\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec16\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.12. Statistical analysis\\u003c/h2\\u003e \\u003cp\\u003eThe means\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;standard deviations were used to present the quantitative data. One-way analysis of variance (ANOVA) was used to conduct statistical comparisons using SPSS 13.0 for Windows software (SPSS, USA). \\u003cem\\u003ep\\u003c/em\\u003e-values were deemed statistically significant if they were less than 0.05.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"3. Results and Discussion\",\"content\":\"\\u003cp\\u003eInformation on NS drug delivery systems from the past research was carried out using FFD. The researchers reported that factor design plays a very important role in optimizing drug dosage forms; by using the design of optimization, the researchers can carry out the formulations with minimum number of runs and obtain optimizing results [\\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e20\\u003c/span\\u003e]. So FFD was employed in the current research. The two independent variable was controlled at the same time and the effects were analyzed either individually or together in the design.\\u003c/p\\u003e \\u003cdiv id=\\\"Sec18\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.1. PS, drug release and optimization\\u003c/h2\\u003e \\u003cp\\u003eTable\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e shows the average PS of the prepared LoR-NS. R\\u003csup\\u003e2\\u003c/sup\\u003e\\u0026thinsp;=\\u0026thinsp;0.8714, the mean PS varied between 284\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;7.1 and 368\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;74.97 nm. Every formulation exhibited PS in the nanoscale (\\u0026lt;\\u0026thinsp;1 \\u0026micro;m) range. ANOVA was used for statistical analysis, and the results revealed a linear relationship, showing that the interaction between the two independent variables on the PS of NS was not statistically significant (p\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.01). At higher stirring rates (\\u0026gt;\\u0026thinsp;1000 rpm), the reduced PS could be attributed to the increased mechanical shear forming smaller NS [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e]. Accordingly, the emulsion globules aggregate and lead to increased PS at lower stirring rates. Collectively, the increase of PS values (formulations S1 and S2) by emulsion globules is most likely due to increase of the LoR:EC concentration along with slow stirring rate (1000 rpm).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab3\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 3\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003ePS and ZP of LoR-NS in gel and LoR drug release from NS gel.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"4\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eFormulation\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003ePS (nm)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eZP\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003ePercentage\\u003c/p\\u003e \\u003cp\\u003erelease after 8h (%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e357.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;14.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-12.22\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e90.76\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;55.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e328.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;25.4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-15.18\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.08\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e91.75\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;7.62\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS3\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e368.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;74.97\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-11.45\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.12\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e90.46\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;7.54\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS4\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e233.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;10.5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-18.81\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.62\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e95.22\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;6.2\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS5\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e274.7\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;28.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-17.14\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.81\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e86.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;7.26\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e284.23\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;15.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-17.22\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.04\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e92.74\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;8.55\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS7\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e298.90 `\\u0026plusmn; 14.2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-16.09\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.06\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e95.99\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;8.31\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS8\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e287.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;23.23\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-16.18\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.77\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e95.13\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;9.23\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eS9\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e285.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;25.2\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e-15.82\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;1.92\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e95.18\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;9.47\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"4\\\"\\u003eAll the values are expressed as Mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD, n\\u0026thinsp;=\\u0026thinsp;3\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eThe \\u003cem\\u003ein vitro\\u003c/em\\u003e release profiles of LoR-NS (percentage of cumulative drug release) are illustrated graphically in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ea. According to Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e, the proportion of LoR released after eight hours varied from 86.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.26% (S5) to 95.99\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.31% (S7). ANOVA was used in the statistical study to compare the release of LoR from various formulations, and the results showed that the release was significant (R\\u003csup\\u003e2\\u003c/sup\\u003e\\u0026thinsp;=\\u0026thinsp;0.9976). The increase in the wall thickness of NS may be the cause of the decrease in the LoR % released after 8 hours with the rise in the polymer ratio (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.001). This can result in a longer diffusional path which reduces the release of LoR-NS [\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e]. It was evident that, the stirring rate increases and subsequently the NS size decreases. This increase could be the result of increased LoR entrapment in the hydrophobic matrix due to decreased porosity, which could have lowered release rates and the similar result was reported by Li et al (2023) [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. Likewise, Costa et al (2022) investigated that the larger particles with higher porosity would allow hydrogel to leak the drug, resulting in faster release rates. Because of ECs hydrophobic and plastic nature, the particles adjacent to the surface matrix may initially be released resulting in an initial burst effect [\\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab4\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 4\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eViscosity, pH, Drug content, and Spreadability of LoR-NS gel.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"5\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eFormulation\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eViscosity (cps)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003epH\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eDrug content (%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eSpreadability (g.cm/sec)\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eLoR-NS gel\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e75.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.63\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.98\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.07\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e94.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;66.15\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.65\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"5\\\"\\u003e*All the values are expressed as Mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD, n\\u0026thinsp;=\\u0026thinsp;3\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003eFactors that influence NS quality included PS with long-acting drug release. Combination of optimal level factor which influenced PS with long-acting drug release are LoR:EC ratio at level 1 (1:2), stirring rate at level 2 (1350 rpm).\\u003c/p\\u003e \\u003cp\\u003eThere are three factors has been selected considering the various factor and level. There are LoR:EC ratio, X\\u003csub\\u003e1\\u003c/sub\\u003e, and stirring rate, X\\u003csub\\u003e2\\u003c/sub\\u003e. The effect of studied variables (LoR:EC ratio, X\\u003csub\\u003e1\\u003c/sub\\u003e; stirring rate, X\\u003csub\\u003e2\\u003c/sub\\u003e) on the response variables (PS of the NS, Y\\u003csub\\u003e1\\u003c/sub\\u003e; release rate, Y\\u003csub\\u003e2\\u003c/sub\\u003e; ZP, Y\\u003csub\\u003e3\\u003c/sub\\u003e) was statistically analyzed using Design-Expert 12 (Stat-Ease) software. To determine the significance and the interaction between the independent factors of the response variables, the ANOVA was performed. The relationship between independent variable and the response was described by means of polynomial equations. The equation was based on the selected model [\\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e]. The responses such as ZP and drug release have been found to be insignificant (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eb). The formula with highest desirability (0.895) was selected as optimized formula and was prepared for further studies.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec19\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.2. Surface morphology\\u003c/h2\\u003e \\u003cp\\u003eThe average diameter was 298\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;4.45 nm and was determined by counting the more than 450 number of NPs visible in several SEM pictures, and by collecting SEM images of native LoR using the same dispersion and staining methodology as for the LoR dispersion (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ea). The surface morphology of the NS was determined using SEM. The images revealed that the particles were spherical in shape, nanometric in size and was found to be porous in nature. It is most likely the result of dichloromethane diffusing from the surface of the NPs during preparation because there are tiny pores on the NS's surface (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003eb). To measure the average diameter of LoR-NS, which had previously been sonicated into a dispersion, SEM images were acquired [\\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec20\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.3. Viscosity, pH, Drug content, and Spreadability\\u003c/h2\\u003e \\u003cp\\u003eThe spreadability of the optimized LoR-NS gel formulation was found to be 0.65\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03 g.cm/sec. Its pH was found to be 6.98\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.01, which is near to skin pH. Viscosity is an important parameter of the gel because as the viscosity decreases the spreadability also decreases. Viscosity was found to be 75.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.63 cps by using T-bar spindle (95F). Drug content was found to be 94.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.15% (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e) which in good agreement with the previously published literature [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eNS was considered to have a strongly spreadable because of its limited time spread. The therapeutic efficacy of gels is influenced by their distribution. To help ensure that the gel is applied to the skin uniformly, the produced gels must be easily spreadable and satisfy the ideal standards for topical application. Furthermore, this is believed to be a crucial component of patient adherence to treatment [\\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e]. Given that anti-inflammatory and topical analgesic formulations are applied to the thin layers of the skin, the consistency of the material is one of the most crucial components. The gels viscosity is crucial in regulating the penetration of the medication.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec21\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.4. Ex vivo permeation of LoR-NS gel\\u003c/h2\\u003e \\u003cp\\u003e \\u003cem\\u003eEx vivo\\u003c/em\\u003e permeability of LoR-NS gel across the skin was found to be 2.266 mg/cm2/min. Different models were adopted for fitting the drug release data in kinetic modelling for optimized formulation and revealed that ex vivo permeation study follows zero order kinetics (R2\\u0026thinsp;=\\u0026thinsp;0.9752), drug diffused from the matrix framework (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e). The 'n' value refers to the non-Fickian diffusion of all formulations (n\\u0026thinsp;=\\u0026thinsp;0.6618), that indicates that there was absence of limit that splits the medium and the drug as described the Yadav et al (2022) [\\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e29\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eConsequently, the amount of LoR increases that permeates the skin. EC included in DCM increase skin permeability by changing or disorganizing the ordered alkyl chains of phospholipids, which causes lipid fluidization [\\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e30\\u003c/span\\u003e]. Lipid fluidization increases the LoR ability to penetrate the skin. The lipid bilayers in the stratum corneum are able to penetrate using DCM, which lowers the stratum corneums barrier resistance and increases intracellular transport by dekeratinizing corneocytes.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec22\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.5. histopathological outcome\\u003c/h2\\u003e \\u003cp\\u003eThe potential histopathological changes were examined on the excised skin of rat. The histopathology of the skin exposed to either LoR-NS loaded gel (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e) or LoR gel (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003eb) revealed intact epidermal layers and dermal structure. The histoarchitectural presentation of this skin presented a few autolytic changes in a diffused manner. No signs of infiltration of inflammatory cells or degenerative changes was observed in the given tissue section. The histoarchitectural presentation of skin revealed intact epidermis and dermal structures (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003ec). No inflammatory infiltrates were observed, nor any degenerative changes were observed in the given tissue section [\\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e31\\u003c/span\\u003e]. The histoarchitectural presentation of skin presented with intact epidermal layers as well as intact dermal structures. No signs of infiltration of inflammatory cells or degenerative changes was observed in the given tissue section (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003ed).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec23\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.6. Skin irritation\\u003c/h2\\u003e \\u003cp\\u003eThe skin irritation studies of control and test group up to day 7 illustrates in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e. The erythemal scores were recorded in control and test group for all the animals. The mean erythemal scores were found to be 0.00, which means that there was no edema or erythema on the skin of the shaved rats in the optimized formulation [\\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e32\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec24\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.7. Stability of LoR-NS gel\\u003c/h2\\u003e \\u003cp\\u003eThe formulations were stable based on drug EE, and cumulative % drug release (CDR %) after their storage for 2 months at 40\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2 ℃, 75\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5% RH. Also, the spreadability, viscosity, and pH of LoR-NS gel were stable in the same experimental conditions. The data obtained with LoR-NS suspension corroborate previously published data [33]. There was no significant difference (\\u003cem\\u003ep\\u003c/em\\u003e\\u0026thinsp;\\u0026gt;\\u0026thinsp;0.05) in stability after 2 months, as given in Table\\u0026nbsp;\\u003cspan refid=\\\"Tab5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e and Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e, indicating that the optimized formulation should remain stable when topically applied in humans.\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab5\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 5\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eStability profile of LoR-NS gel at 40\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2 ℃, 75\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5% RH for 60 days.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"7\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eDays\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c3\\\" namest=\\\"c2\\\"\\u003e \\u003cp\\u003eLoR-NS suspension\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"4\\\" nameend=\\\"c7\\\" namest=\\\"c4\\\"\\u003e \\u003cp\\u003eLoR-NS gel\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eDrug EE (%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003eCDR (%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003eSpreadability (g.cm/sec)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003eViscosity (cps)\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003epH\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003eCDR (%)\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e97.51\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.63\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e86.42\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.08\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.65\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e75.9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.63\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.98\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.34\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e79.47\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.13\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e15\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e95.12\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;4.35\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e87.78\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;6.06\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.68\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e74.5\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;4.14\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.98\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.67\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e80.87\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;4.17\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e30\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e96.60\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.1\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e86.88\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;7.05\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.63\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e74.8\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;4.21\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.98\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.88\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e80.90\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;4.20\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e60\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e98.85\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.03\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e87.1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.06\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.67\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.02\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e71.6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.14\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.98\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.47\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e81.87\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;3.09\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003e90\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e96.29\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;8.18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e85.37\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;10.6\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.61\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.45\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e69.4\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5.59\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.48\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.18\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"char\\\" char=\\\"\\u0026plusmn;\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e78.31\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;7.16\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003ctfoot\\u003e \\u003ctr\\u003e\\u003ctd colspan=\\\"7\\\"\\u003e*All the values are expressed as mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD, n\\u0026thinsp;=\\u0026thinsp;3\\u003c/td\\u003e\\u003c/tr\\u003e \\u003c/tfoot\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"4. Conclusion\",\"content\":\"\\u003cp\\u003eIn conclusion, the comprehensive exploration of the Loratadine-loaded nanosponge incorporated into a topical gel through a Complete Factorial Design has yielded promising results through a series of rigorous assessments. The \\u003cem\\u003ein vitro\\u003c/em\\u003e studies provided valuable insights into thepreparations physicochemical attributes, drug release kinetics, and stability. The \\u003cem\\u003eex-vivo\\u003c/em\\u003e assessments, utilizing skin permeation studies, further demonstrated the efficacy of the nanosponge-based gel in facilitating the transdermal delivery of Loratadine. These collective findings highlight the potential of the Loratadine-loaded nanosponge in topical gel as a promising candidate for transdermal drug delivery, with implications for improved therapeutic outcomes and patient compliance. Further studies and clinical trials will be crucial to validate and expand upon these results, paving the way for the potential translation of this innovative formulation into clinical practice.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgements\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors are thankful to the Faculty of pharmacy \\u0026amp; BioMedical Sciences, MAHSA University, Bandar Saujana Putra, 42610 Jenjarom, Selangor. Malaysia for providing the necessary lab facilities during the experimental study.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthor contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eD.S. and J.M.M.M. conceived and designed research, K.V and S.A. conducted experiments, J.M.M.M., Y.I.A, M.F., and A.E. analyzed data and wrote the manuscript, S.E and F.M. supervision of the work. All authors read and approved the manuscript.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunding\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors extend their sincere appreciation to the Deanship of Scientific Research at King Khalid University for funding this study through the Large Research Group Project under grant number \\u0026quot;RGP 2/109/44\\u0026quot;.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCompeting interests\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors declare no competing interests.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAdditional information\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eCorrespondence and requests for materials should be addressed to S.E. or F.M.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eData Availability\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eNur Husna SM, Tan HT, Md Shukri N, Mohd Ashari NS, Wong KK. Allergic Rhinitis: A Clinical and Pathophysiological Overview. Front Med (Lausanne). 2022 Apr 7;9:874114. doi: 10.3389/fmed.2022.874114.\\u003c/li\\u003e\\n\\u003cli\\u003eSidhu G, Akhondi H. Loratadine. [Updated 2023 Mar 13]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK542278/\\u003c/li\\u003e\\n\\u003cli\\u003eYu YQ, Yang X, Wu XF, Fan YB. Enhancing Permeation of Drug Molecules Across the Skin via Delivery in Nanocarriers: Novel Strategies for Effective Transdermal Applications. Front Bioeng Biotechnol. 2021 Mar 29;9:646554. doi: 10.3389/fbioe.2021.646554.\\u003c/li\\u003e\\n\\u003cli\\u003eAkash Garg a, Wen-Cheng Lai b, Himansu Chopra a, Rutvi Agrawal a, Talever Singh a, Ramkumar Chaudhary a, Braj Nandan Dubey a. Nanosponge: A promising and intriguing strategy in medical and pharmaceutical Science. Heliyon Volume 10, Issue 1, 15 January 2024, e23303. https://doi.org/10.1016/j.heliyon.2023.e23303\\u003c/li\\u003e\\n\\u003cli\\u003eMoideen, J.M.M.; Alqahtani, A.; Venkatesan, K.; Ahmad, F.; Krisharaju, K.; Gayasuddin, M.; Shaik, R.A. Application of the Box-Behnken design for the production of soluble curcumin: Skimmed milk powder inclusion complex for improving the treatment of colorectal cancer. \\u003cem\\u003eFood Sci. Nutr\\u003c/em\\u003e. \\u003cstrong\\u003e2020\\u003c/strong\\u003e, 8, 6643\\u0026ndash;6659.\\u003c/li\\u003e\\n\\u003cli\\u003eHimangshu Bhowmik, D. Nagasamy Venkatesh\\u003csup\\u003e*\\u003c/sup\\u003e, Anuttam Kuila, Kammari Harish Kumar. Nanosponges: A Review. Int J App Pharm, Vol 10, Issue 4, 2018, 1-5.\\u003c/li\\u003e\\n\\u003cli\\u003eBashir S, Hina M, Iqbal J, Rajpar AH, Mujtaba MA, Alghamdi NA, Wageh S, Ramesh K, Ramesh S. Fundamental Concepts of Hydrogels: Synthesis, Properties, and Their Applications. 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Lee, Lee Fong Siow \\u0026amp; Siew Hua Gan. (2022) Microencapsulation of propolis by spray drying: A review. \\u003cem\\u003eDrying Technology\\u003c/em\\u003e 40:6, pages 1083-1102.\\u003c/li\\u003e\\n\\u003cli\\u003eMahmood A, Mahmood A, Ibrahim MA, Hussain Z, Ashraf MU, Salem-Bekhit MM, Elbagory I. Development and Evaluation of Sodium Alginate/Carbopol 934P-Co-Poly (Methacrylate) Hydrogels for Localized Drug Delivery. \\u003cem\\u003ePolymers\\u003c/em\\u003e. 2023; 15(2):311. https://doi.org/10.3390/polym15020311\\u003c/li\\u003e\\n\\u003cli\\u003eMohamed, J.M.; Alqahtani, A.; Ahmad, F.; Krishnaraju, V.; Kalpana, K. Pectin co-functionalized dual layered solid lipid nanoparticle made by soluble curcumin for the targeted potential treatment of colorectal cancer. \\u003cem\\u003eCarbohydr. Polym\\u003c/em\\u003e. \\u003cstrong\\u003e2020\\u003c/strong\\u003e, 252, 117180. \\u003c/li\\u003e\\n\\u003cli\\u003eAl Fatease, A., Alqahtani, A., Khan, B.A., Mohamed, J.MM, Farhana SA. Preparation and characterization of a curcumin nanoemulsion gel for the effective treatment of mycoses. \\u003cem\\u003eSci Rep\\u003c/em\\u003e\\u003cstrong\\u003e13\\u003c/strong\\u003e, 22730 (2023). https://doi.org/10.1038/s41598-023-49328-2.\\u003c/li\\u003e\\n\\u003cli\\u003eNnamani PO, Ugwu AA, Nnadi OH, Kenechukwu FC, Ofokansi KC, Attama AA, Lehr CM. Formulation and evaluation of transdermal nanogel for delivery of artemether. Drug Deliv Transl Res. 2021 Aug;11(4):1655-1674. doi: 10.1007/s13346-021-00951-4.\\u003c/li\\u003e\\n\\u003cli\\u003eKulasekaran, A. \\u0026amp; Gopal, Andal \\u0026amp; Lakshimipathy, R. \\u0026amp; Alexander, J.. (2015). Modification in pH measurements for getting accurate pH values with different pH meters irrespective of aging and drifts in the meters. International Journal of ChemTech Research. 8. 16-24.\\u003c/li\\u003e\\n\\u003cli\\u003eDas MK, Ahmed AB. Formulation and ex vivo evaluation of rofecoxib gel for topical application. 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Critical reviews in therapeutic drug carrier systems. 22. 215-94. 10.1615/CritRevTherDrugCarrierSyst.v22.i3.10.\\u003c/li\\u003e\\n\\u003cli\\u003eNazief AM, Hassaan PS, Khalifa HM, Sokar MS, El-Kamel AH. Lipid-Based Gliclazide Nanoparticles for Treatment of Diabetes: Formulation, Pharmacokinetics, Pharmacodynamics and Subacute Toxicity Study. Int J Nanomedicine. 2020 Feb 18;15:1129-1148. doi: 10.2147/IJN.S235290.\\u003c/li\\u003e\\n\\u003cli\\u003eSenthivel CK, Karuppaiyan K, Ahamad F, Mohamed JMM*, El-Sherbiny M, Alotaibi AT, Abed SY, Ibraheem KM, Salama M. Terminalia chebula loaded polymeric nanoparticles: a preliminary approach for status epilepticus. Chemical Papers 2023, DOI: 10.1007/s11696-023-03164-w.\\u003c/li\\u003e\\n\\u003cli\\u003eZhimin Li, Xianjing Feng, Shixing Luo, Yanfeng Ding, Zhi Zhang, Yifeng Shang, Doudou Lei, Jinhong Cai, Jinmin Zhao, Li Zheng, Ming Gao, High drug loading hydrophobic cross-linked dextran microspheres as novel drug delivery systems for the treatment of osteoarthritis, Asian Journal of Pharmaceutical Sciences, Volume 18, Issue 4, 2023, 100830, https://doi.org/10.1016/j.ajps.2023.100830.\\u003c/li\\u003e\\n\\u003cli\\u003eAna Let\\u0026iacute;cia Rodrigues Costa, Stephanie M. Willerth, Lucimara Gaziola de la Torre, Sang Won Han, Trends in hydrogel-based encapsulation technologies for advanced cell therapies applied to limb ischemia, Materials Today Bio, Volume 13, 2022, 100221, https://doi.org/10.1016/j.mtbio.2022.100221.\\u003c/li\\u003e\\n\\u003cli\\u003eMohamed JMM, Alqahtani A, Khan BA, Al Fatease A, Alqahtani T, Venkatesan K, Ahmad F, Alzghoul BI, Alamri A. Preparation of Soluble Complex of Curcumin for the Potential Antagonistic Effects on Human Colorectal Adenocarcinoma Cells. Pharmaceuticals (Basel). 2021 Sep 19;14(9):939. doi: 10.3390/ph14090939.\\u003c/li\\u003e\\n\\u003cli\\u003eAhmad F, Al-Subaie AM, Gayasuddin M, Mohamed JM, Krishnaraju V. Review on the Medicinal uses and Pharmacological aspects of Plectranthus tenuiflorus from the Labiatae Family of Saudi Arabia. International Journal of Pharmaceutical Sciences Review and Research, 2020, vol. 64, no.2, pp. 43-48.\\u003c/li\\u003e\\n\\u003cli\\u003eMohamed JMM, Khan BA, Rajendran V, El-Sherbiny M, Othman G, Hussamuldin, ABA, Al-Serwi, RH. Polymeric Ethosomal Gel Loaded with Nimodipine: Optimisation, Pharmacokinetic and Histopathological Analysis. Saudi Pharmaceutical Journal, Volume 30, Issue 11, November 2022, Pages 1603-1611. https://doi.org/10.1016/j.jsps.2022.09.003\\u003c/li\\u003e\\n\\u003cli\\u003eYadav K, Yadav D, Kumar S, Narra N, Mohamed JMM* et al. Natural Biodegradable and Biomass Polymeric Nanoparticles for the Delivery of Noscapine for cancer treatment. Biomass Conversion and Biorefinery, DOI: 10.1007/s13399-022-03334-y\\u003c/li\\u003e\\n\\u003cli\\u003eKurmi BD, Tekchandani P, Paliwal R, Paliwal SR. Transdermal Drug Delivery: Opportunities and Challenges for Controlled Delivery of Therapeutic Agents Using Nanocarriers. Curr Drug Metab. 2017;18(5):481-495. doi: 10.2174/1389200218666170222150555.\\u003c/li\\u003e\\n\\u003cli\\u003eLand\\u0026eacute;n NX, Li D, St\\u0026aring;hle M. Transition from inflammation to proliferation: a critical step during wound healing. Cell Mol Life Sci. 2016 Oct;73(20):3861-85. doi: 10.1007/s00018-016-2268-0.\\u003c/li\\u003e\\n\\u003cli\\u003eWang J, Li Z, Sun F, Tang S, Zhang S, Lv P, Li J, Cao X. Evaluation of dermal irritation and skin sensitization due to vitacoxib. Toxicol Rep. 2017 Jun 10;4:287-290. doi: 10.1016/j.toxrep.2017.06.003.\\u003c/li\\u003e\\n\\u003cli\\u003eElhassan GO, Mohamed JMM. Development And In Vitro Evaluation Of Valsartan-Loaded Resealed Erythrocytes, International Journal of Applied Pharmaceutics, Vol. 14, 2022, 201-205, 10.22159/ijap.2022.v14ti.57\\u003cstrong\\u003e\\u003c/strong\\u003e\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"scientific-reports\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"scirep\",\"sideBox\":\"Learn more about [Scientific Reports](http://www.nature.com/srep/)\",\"snPcode\":\"\",\"submissionUrl\":\"\",\"title\":\"Scientific Reports\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Scientific Reports\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true},\"keywords\":\"Nanosponge, Loratadine, Ethyl cellulose, Hydrogel, NDDS, Sustained release\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-3852896/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-3852896/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eLoratadine (LoR) is a highly lipophilic and practically insoluble in water, hence having a low oral bioavailability. As it is formulated as topical gel, it competitively binds with the receptors, thus reducing the side-effects. The objective of this study was to prepare LoR loaded nanosponge (LoR-NS) in gel for topical delivery. Nine different formulations of emulsion were prepared by solvent evaporation method with polyvinyl alcohol (PVA), ethyl cellulose (EC), and dichloromethane (DCM). Based on 3\\u003csup\\u003e2\\u003c/sup\\u003e Full Factorial Design (FFD), optimization was carried out by varying the concentration of LOR:EC ratio and stirring rate. The preparations were subjected for the evaluation of particle size (PS), \\u003cem\\u003ein vitro\\u003c/em\\u003e release, zeta potential (ZP) and entrapment efficiency (EE). The results revealed that the NS dispersion was nanosized with sustained release profiles and significant PS. The optimized formulation was formulated and incorporated into carbopol 934P hydrogel. The formulation was then examined to surface morphological characterizations using scanning electron microscopy (SEM) which depicted spherical NS. Stability studies, undertaken for 2 months at 40\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;2 ℃ and 75\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5% RH, concluded to the stability of the formulation. The formulation did not cause skin irritation. Therefore, the prepared NS hydrogel proved to be a promising applicant for LoR as a novel drug delivery system (NDDS) for safe, sustained and controlled topical application.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Application of 32 Factorial Design of Loratadine-Loaded Nanosponge in Topical Gel system: In Vitro, Ex Vivo, and In Vivo Assessments\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-01-18 11:58:31\",\"doi\":\"10.21203/rs.3.rs-3852896/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"decision\",\"content\":\"Revision requested\",\"date\":\"2024-01-23T07:48:08+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-01-21T23:05:36+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-01-18T20:53:46+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"19fbdf4d-f338-4be3-ac88-42c8e4ed9bb5\",\"date\":\"2024-01-17T14:24:02+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"54eaf3be-42a3-452f-8cf1-4d9c239aa025\",\"date\":\"2024-01-17T13:25:55+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2024-01-17T10:06:47+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2024-01-17T09:47:11+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvited\",\"content\":\"\",\"date\":\"2024-01-17T09:40:11+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2024-01-17T09:39:29+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Scientific Reports\",\"date\":\"2024-01-11T09:00:53+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"scientific-reports\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"scirep\",\"sideBox\":\"Learn more about [Scientific Reports](http://www.nature.com/srep/)\",\"snPcode\":\"\",\"submissionUrl\":\"\",\"title\":\"Scientific Reports\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Scientific Reports\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"255170c5-4ac7-49fe-a87b-cfec58001e22\",\"owner\":[],\"postedDate\":\"January 18th, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-03-18T15:12:25+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-3852896\",\"link\":\"https://doi.org/10.1038/s41598-024-55953-2\",\"journal\":{\"identity\":\"scientific-reports\",\"isVorOnly\":false,\"title\":\"Scientific Reports\"},\"publishedOn\":\"2024-03-16 15:01:32\",\"publishedOnDateReadable\":\"March 16th, 2024\"},\"versionCreatedAt\":\"2024-01-18 11:58:31\",\"video\":\"\",\"vorDoi\":\"10.1038/s41598-024-55953-2\",\"vorDoiUrl\":\"https://doi.org/10.1038/s41598-024-55953-2\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-3852896\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-3852896\",\"identity\":\"rs-3852896\",\"version\":[\"v1\"]},\"buildId\":\"FbvkV6FR0MCFSLy54lSbu\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}