Modified Ethanol Injection Method Research Articles

Evaluation of the antifungal properties of nanoliposomes containing rhinacanthin-C isolated from the leaves of Rhinacanthus nasutus.

Fungal infections represent a challenging threat tothe human health. Microsporum gypseum and Trichophyton rubrum are pathogenic fungi causing various topical mycoses in humans. The globally emerging issue of resistance to fungi demands the development of novel therapeutic strategies. In this context, the application of nanoliposomes as vehicles for carrying active therapeutic agents can be a suitable alternative. In this study, rhinacanthin-C was isolated from Rhinacanthus naustus and encapsulated in nano-liposomal formulations, which were prepared by the modified ethanol injection method. The two best formulations composed of soybean phosphatidylcholine (SPC), cholesterol (CHL), and tween 80 (T80) in a molar ratio of 1:1:0 (F1) and 1:1:0.5 (F2) were proceeded for experimentation. The physical characteristics and antifungal activities were performed and compared with solutions of rhinacanthin-C. The rhinacanthin-C encapsulating efficiencies in F1 and F2 were 94.69 ± 1.20% and 84.94 ± 1.32%, respectively. The particle sizes were found to be about 221.4 ± 13.76 nm (F1) and 115.8 ± 23.33 nm (F2), and zeta potential values of -38.16 mV (F1) and -40.98 mV (F2). Similarly, the stability studies of rhinacanthin-C in liposomes demonstrated that rhinacanthin-C in both formulations was more stable in mediums with pH of 4.0 and 6.6 than pure rhinacanthin-C when stored at the same conditions. Rhinacanthin-C in F1 was slightly more stable than F2 when stored in mediums with a pH of 10.0 after three months of storage. However, rhinacanthin-C in both formulations was less stable than pure rhinacanthin-C in a basic medium of pH 10.0. The antifungal potential was evaluated against M. gypsum and T. rubrum. The findings revealed a comparatively higher zone of inhibition for F1. In the MIC study, SPC: CHL: T80 showed higher inhibition against M. gypseum and a slightly higher inhibition against T. rubrum compared to free rhinacanthin-C solution. Moreover, rhinacanthin-C showed significant interaction against 14α-demethylase in in silico study. Overall, this study demonstrates that nanoliposomes containing rhinacanthin-C can improve the stability and antifungal potential of rhinacanthin-C with sustained and prolonged duration of action and could be a promising vehicle for delivery of active ingredients for targeting various fungal infections.

Novel itraconazole nano-spanlastics gel with enhanced penetration to treat topical fungal infection

Candida infections of the skin are the most common that can be treated topically. Herein, an attempt was made to enhance low penetration of itraconazole through nano-spanlastic vesicles. Nano-spanlastics of itraconazole were prepared and optimized using a response surface method and further transformed into gel. A custom ‘response surface method’ was applied to optimize independent variables (Span 60 and Tween 80) at two levels to study effect on entrapment efficiency. Nano-spanlastics were prepared by modified ethanol injection method followed by ultrasonication using Tween 80 as an edge activator.Optimized batch of nano-spanlastics was found at the nano range 160.2nm with poly-dispersity index of 0.337, zeta potential −42.3 mV and entrapment efficiency 97.93 ± 0.51 %. Thus, zeta potential and polydispersity index indicated a stable formulation. The nano-spanlastic formulation was further converted into gel and was found to be sufficiently viscous and spreadable. Ex-vivo permeation study of vesicular spanlastics revealed higher flux (0.00650 μg/cm2/min) than marketed (0.00229 μg/cm2/min) and conventional gel (0.00073 μg/cm2/min). Further, antifungal activity of nano-spanlastics loaded gel against Candida albicans depicted greater zone of inhibition (28 mm) in comparison to the marketed formulation (22 mm). Thus, to conclude, nano-spanlastics can be a potential nanocarrier for improved penetration and for targeting topical fungal infections, thus providing new opportunities for dermal treatment.

Evaluation of physicochemical, in vivo analgesic and antiinflammatory activities of Brachystegia eurycoma gum-based naproxen loaded niosomal gels

Protracted oral Naproxen administration for acute and chronic conditions, such as rheumatoid arthritis treatment, may cause peptic ulcers. The topical application of Naproxen will circumvent this effect. This research aims to assess the physicochemical, anti-inflammatory and analgesic properties of Naproxen-loaded niosomal gels formulated using Brachystegia eurycoma gum (BEG) as the gelling polymer. BEG was isolated by acetone precipitation of filtrate produced by cold maceration of Brachystegia eurycoma seeds powder. Naproxen and BEG compatibility was evaluated using Fourier transform infrared FTIR) spectroscopy. Naproxen-loaded niosomes were produced with cholesterol, surfactant (tween 80) and water using a modified ethanol injection method. Theformulated niosomes were incorporated into either BEG (5, 7.5 or 10%) or HPMC (1.5, 5%) gel base to produce the respective Naproxen-loaded niosomal gels. The niosomal gels were assessed for their physicochemical, antiinflammatory and analgesic properties using a commercially available gel as a positive control. There was no incompatibility between BEG and Naproxen. The formulated gels’ pH ranged from 6.4 to 7.3, while the viscosity ranged from 1476.2 to 2980 mPas. The formulated gels had good extrudability and spreadability compared to the commercially available gel (1%). In-vivo studies using white albino rats showed that gels from the optimized formulation (F1) had comparable anti-inflammatory (26.5% inhibition) and analgesic properties to the commercially available gel (31.3% inhibition). Naproxen-loaded niosomal gels having good physicochemical, analgesic and anti-inflammatory properties were successfully formulated using BEG as a gelling agent.

Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method.

Messenger RNA (mRNA)-based therapies are a novel class of therapeutics used in vaccination and protein replacement therapies for monogenic diseases. Previously, we developed a modified ethanol injection (MEI) method for small interfering RNA (siRNA) transfection, in which cationic liposome/siRNA complexes (siRNA lipoplexes) were prepared by mixing a lipid-ethanol solution with a siRNA solution. In this study, we applied the MEI method to prepare mRNA lipoplexes and evaluated the in vitro and in vivo protein expression efficiencies. We selected six cationic lipids and three neutral helper lipids to generate 18 mRNA lipoplexes. These were composed of cationic lipids, neutral helper lipids, and polyethylene glycol-cholesteryl ether (PEG-Chol). Among them, mRNA lipoplexes containing N-hexadecyl-N,N-dimethylhexadecan-1-aminium bromide (DC-1-16) or 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl) propan-2-yl) amino)-N,N,N-trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12) with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and PEG-Chol exhibited high protein expression in cells. Furthermore, mRNA lipoplexes composed of DC-1-16, DOPE, and PEG-Chol exhibited high protein expression in the lungs and spleen of mice after systemic injection and induced high antigen-specific IgG1 levels upon immunization. These results suggest that the MEI method can potentially increase the efficiency of mRNA transfection, both in vitro and in vivo.

Abstract 5071: Novel approach to intra- and peri-tumoral delivery of sustained-release chemotherapeutics

Abstract INTRODUCTION: Intratumoral phenylephrine (α1 vasoconstrictor) combined with chemotherapy has been used for cutaneous malignancies with modest local response. The purpose of this study was to improve the efficacy of intratumoral melphalan using sustained-release, liposomal nanoparticles containing phenylephrine in a mouse model of melanoma. METHODS: A modified ethanol injection method was used to develop the liposomal phenylephrine nanoparticles. These were injected peri-tumorally to generate a local vasoconstrictive effect intended to “entrap” intratumorally injected melphalan. Drug exposure/dwell time was measured with fluorescent imaging. Mice bearing B16 (subclone F10) melanoma tumors were treated with intratumoral melphalan (20 ug/50 ul) with or without peri-tumoral liposomal phenylephrine (10 ug/50-100 ul x 4-5 injections around the tumor periphery) twice weekly x 3 weeks. RESULTS: UV-Absorbance spectroscopy confirmed encapsulation of phenylephrine within the liposomal nanoparticles with an expected absorbance of 275 nm. The modified ethanol injection method produced liposomal phenylephrine encapsulation of approximately 25%. Peri-tumoral, liposomal phenylephrine caused vasoconstriction of normal surrounding vessels with an approximate 10% decrease in vessel diameter. Our liposomal phenylephrine nanoparticles also led to increased retention of intratumorally injected fluorescently-labeled (FITC) liposomes 24-48 hours after inoculation, using both melanoma (B16) and breast (TUBO/4T1) cell lines orthotopically grown within mice (C57Bl/6 and BALB/c, respectively). The combination of intratumoral melphalan with peri-tumoral phenylephrine resulted in more effective control of tumor growth and a complete tumor response rate of 44.4%, compared to phenylephrine/melphalan only controls (p < 0.01, n=18 per group). However, there was significant adverse reaction with the combination therapy, resulting in ulceration of the injection/tumor site. Ulcers healed about 14 days after completion of treatment. Immunohistochemistry demonstrated increased infiltration of CD11b+ cells into the tumors, suggesting an immune-mediated response in addition to direct chemotoxicity. CONCLUSIONS: Liposomal nanoparticles containing phenylephrine increased retention/dwell time of intratumorally administered melphalan and resulted in superior anti-tumor response. Our preclinical model holds innovative, translational promise for the clinical advancement of intralesional therapies in cutaneous/subcutaneous malignancies. Citation Format: Emmanuel M. Gabriel, Vijay Madamsetty, Deborah Bahr, Jamie Kaplan, Kristopher Attwood, Sanjay Bagaria, Keith Knutson, Debabrata Mukhopadhyay. Novel approach to intra- and peri-tumoral delivery of sustained-release chemotherapeutics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5071.

Astragalus Polysaccharides/PVA Nanofiber Membranes Containing Astragaloside IV-Loaded Liposomes and Their Potential Use for Wound Healing.

Delayed wound healing is a common and serious complication in diabetic patients, especially the slow healing of foot ulcers, which seriously affects the quality of life of patients and is also the most important risk factor for lower limb amputation. The multifunctional novel dressing prepared by loading the polymer nanofibers with anti-inflammatory and prohealing plant extracts can promote the wound repair of these ulcers by electrospinning technology. Liposomes are nanoparticles prepared from phospholipids and have been widely used as drug delivery systems. Liposomes can be combined with electrospun nanofibrous webs to facilitate local and sustained delivery of loaded bioactive substances. In this study, liposomes were prepared with astragaloside IV (AS) by employing a modified ethanol injection method and conducting the physical and chemical characterization (e.g., the particle size, polydispersity index, zeta potential, and entrapment efficiency). Astragalus polysaccharides were extracted from Astragalus membranaceus. Subsequently, we prepared the electrospun polyvinyl alcohol (PVA)/astragalus polysaccharide (APS)/astragaloside IV (AS) nanofibers. The morphology of the produced ASL/APS/PVA, APS/PVA, and PVA nanofibers were analyzed by scanning electron microscopy (SEM), and it turns out that the addition of astragalus extract made the fiber diameter smaller and the fibers arranged neatly with no dripping. An induced diabetic rat model was built, and a diabetic ulcer model was built by total cortical resection to assess the prorepair ability of the prepared nanofibers. According to in vivo animal experiments, the nanofibrous membrane loaded with APS and ASL was reported to inhibit the occurrence of wound inflammation, enhance the deposition of collagen fibers (P < 0.05) and the repair of regenerated epithelium (P < 0.05), and effectively strengthen the wound healing of diabetic rats (P < 0.05). In brief, PVA-loaded APS/ASL nanofibrous membranes refer to a prominent wound healing dressing material, which can effectively facilitate the healing of diabetic wounds, and they are demonstrated to be highly promising for application in diabetic wound dressings and tissue engineering.

Permeation enhancer nanovesicles mediated topical delivery of curcumin for the treatment of hyperpigmentation

The main aim of the present study was to develop curcumin (CUR) loaded permeation enhancer-lipid vesicles for the treatment of hyperpigmentation. Hyperpigmentation is an acquired skin disorder characterized by uneven skin coloration, mainly in the regions of the facial skin, affecting millions of people worldwide. It often occurs in visible areas, hence causing significant negative psychological and social impacts. In the present study, curcumin-loaded permeation enhancer nanovesicles (PE-NVs) were developed by modified ethanol injection method and dimethyl sulfoxide was added as a penetration enhancer. PE-NVs were subjected to various physicochemical characterizations and drug permeation studies across the skin. The PE-NVs were tested for their efficacy in a sunlight-induced hyperpigmented rabbit skin model. Topical application of PE-NVs reduced symptoms of hyperpigmentation as compared with CUR methanolic solution because of higher accumulation because of better permeation into skin layers. Histopathological studies also confirmed the effectiveness of PE-NVs, since they reduced hyperpigmentation-induced lesions. Results confirmed that PE-NVs is a potential drug delivery system for topical administration drugs to treat skin-associated inflammatory disorders.

Novel nano spanlastic carrier system for buccal delivery of lacidipine

Lacidipine, a calcium channel antagonist with its highly lipophilic and poorly water-soluble nature (BCS Class II Drug), when administered via the conventional oral route undergoes extensive hepatic metabolism by Cytochrome P450 3A4 (CYP3A4), results in low bioavailability (∼10%) and summons a challenge to anti-hypertensive therapy. This present research study aimed to override this drug delivery obstacle by formulating lacidipine loaded spanlastic orally dissolving films. A 23 factorial design model was employed in the spanlastics development process to examine the influence of the formulation variables over the response variables. Spanlastics composed of Span 60 and Tween 80 were prepared by modified ethanol injection method and developed into an orally dissolving film using hypromellose E5, polyvinyl alcohol and polyethylene glycol 400 by the solvent casting technique. The characterization results of the formulation showed that the spanlastics possessed nano-sized vesicles and good entrapment efficiency (86.84%). Optical microscopy and high resolution transmission electron microscopy displayed lamellar, circularly shaped, and uniformly dispersed nanovesicles. Evaluation studies revealed that the oral film possessed considerable flexibility, good mechanical strength, disintegrated within 35 s and revealed a drug release of 92.96% within 5 min. The scanning electron microscopy displayed a smooth, uniform surface of the film with pores. Stability studies of the films at 25 ± 2 °C and 60 ± 5% RH for 3 months showed no alteration in physical appearance or mechanical properties. The developed spanlastic vesicles, owing to their nano size have an enhanced barrier permeation profile while the oral films offer better bioavailability of the drug by absorption through the oro-mucosal tissue. In combination with the increased permeability, the circumvention of hepatic metabolism, improved dissolution and fast onset of action, this new drug delivery approach makes lacidipine efficiently available for antihypertensive therapy.

Enhancement of in vitro antitumour activity of epirubicin in HER2+ breast cancer cells using immunoliposome formulation.

Epirubicin (EPI) is one of the potent breast cancer (BC) chemotherapeutic agents, but its adverse effects limit its efficacy. Herein, EPI was selected to be loaded in liposomal carrier, which has been targeted by a monoclonal antibody, Herceptin. The preparation process of liposomes was a modified ethanol injection method followed by Herceptin conjugation. The in vitro cell toxicity and cellular uptake of optimum formulation against HER2+ and HER2− cancer cell lines were evaluated. The results showed that the drug loading (DL%) and encapsulation efficiency (EE%) of liposome preparation method yielded 30.62% ± 0.49% and 62.39% ± 8.75%, respectively. The average size of naked liposomes (EPI‐Lipo) and immunoliposomes (EPI‐Lipo‐mAb) was 234 ± 9.86 and 257.26 ± 6.25 nm, with a relatively monodisperse distribution, which was confirmed by SEM micrographs. The release kinetic followed Higuchi model for both naked and immunoliposomes. In vitro cytotoxicity study on three different BC cell lines including BT‐20, MDA‐MB‐453 and MCF‐7 demonstrated higher toxicity of EPI in the Herceptin conjugated form (EPI‐Lipo‐mAb) in comparison with the free EPI and EPI‐Lipo in HER2 overexpressing cell line. In addition, the cellular uptake study showed a higher uptake of immunoliposomes by MCF‐7 cells in comparison with naked liposomes. In conclusion, these data show that the targeted delivery of EPI to breast cancer cells can be achieved by EPI‐Lipo‐mAb in vitro, and this strategy could be used for breast cancer therapy with further studies.

Statistical optimization of hyaluronic acid enriched ultradeformable elastosomes for ocular delivery of voriconazole via Box-Behnken design: in vitro characterization and in vivo evaluation

Voriconazole (VCZ) is a well-known broad spectrum triazole antifungal, mainly used orally and intravenously. The study aimed to formulate VCZ into ultradeformable elastosomes for the topical treatment of ocular fungal keratitis. Different formulae were prepared using a modified ethanol injection method, employing a 33 Box-Behnken design. They were characterized by measuring their entrapment efficiency (EE%), particle size (PS), polydispersity index (PDI) and zeta potential (ZP). The optimized formula was subjected to further in vitro investigations and in vivo evaluation studies. The prepared vesicles had satisfactory EE%, PS, PDI and ZP values. The numerical optimization process suggested an optimal elastosomal formula (OE) composed of phosphatidyl choline and brij S100 at the weight ratio of 3.62: 1, 0.25%w/v hyaluronic acid and 5% (percentage from phosphatidyl choline/brij mixture) polyvinyl alcohol. It had high EE (72.6%), acceptable PS and PDI (362.4 nm and 0.25, respectively) and highly negative ZP of −41.7 mV. OE exhibited higher elasticity than conventional liposomes, with acceptable stability for three months. Transmission electron microscopy demonstrated the spherical morphology of vesicles with an external transparent coat of Hyaluronic acid. OE was expected to cause no ocular irritation or blurring in vision as reflected by pH and refractive index measurements. The histopathological study revealed the safety of OE for ocular use. The fungal susceptibility testing using Candida albicans demonstrated the superiority of OE to VCZ suspension, with greater and more durable growth inhibition. Therefore, OE can be regarded as a promising formula, achieving both safety and efficacy.

Oleosomes Encapsulating Sildenafil Citrate as Potential Topical Nanotherapy for Palmar Plantar Erythrodysesthesia with High Ex vivo Permeation and Deposition.

Palmar plantar erythrodysesthesia (PPE) is a commonly reported skin toxicity of chemotherapeutic agents that significantly affects patients' quality of life. PPE is described as inflammation, swelling, and even cracks and ulcers in the skin of palms and soles of the feet. Conventional treatment includes topical creams, analgesics, or corticosteroids. However, serious cases are not responding to these medications. PPE has been reported to cause drug cessation or dose reduction if not properly treated. Sildenafil citrate (SC) has a well-documented activity in wound healing through improving blood supply to the affected area. However, SC has poor physicochemical properties limiting its transdermal permeation and deposition. This research endeavored to elaborate novel vesicular system with natural components, phospholipids and oleic acid, loaded with sildenafil citrate for topical management of PPE. Sildenafil-loaded oleosomes were prepared using modified ethanol injection method. Optimized oleosomes had nanometric particle size (157.6nm), negative zeta potential (- 85.2 mv), and high entrapment efficiency (95.56%). Ex vivo studies on human skin revealed that oleosomes displayed 2.3-folds higher permeation and 4.5-folds more deposition through the human skin compared to drug suspension. Results endorsed SC oleosomes as suitable topical treatment of PPE providing ameliorated sildenafil permeability in addition to acting as a reservoir for gradual release of the drug. Graphical abstract.

Preparation and Characterization of Ivabradine HCl Transfersomes for Enhanced Transdermal Delivery

Ivabradine HCl (IVB) is used in treatment of stable angina pectoris. The aim of the work was to prepare a once daily transdermal film of transfersomes loaded IVB. Transfersomes were prepared by modified ethanol injection method using three formulation factors namely: surfactant type, drug: phospholipid ratio and surfactant: phospholipid ratio. The transfersomes were characterized for entrapment efficiency, particle size, zeta potential, in vitro drug release, optical and transmission electron microscopy imaging, differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FTIR). The best transfersomes showed optimal properties as well as pure drug were further formulated in transdermal film and characterized for content uniformity, folding endurance, weight and thickness, moisture content, moisture uptake, tensile strength and permeation through excised ear rabbit skin. Pharmacokinetic parameters of transfersomes incorporated film were compared with oral solution of the same drug dose. A maximum of about 82.03% entrapment efficiency, size of 206.7 nm, zeta potential of −88.3 Mv and 66.7% of drug release after 24 h were obtained from sodium lauryl sulphate based transfersomes at drug: phospholipid and surfactant: phospholipid ratio of 1:20 and 75:25 w/w, respectively. Films contained IVB transfersomes exhibited improved permeability and skin retention when compared with pure drug as well as enhanced pharmacokinetic parameters.

Identification of Abies sibirica L. Polyprenols and Characterisation of Polyprenol-Containing Liposomes.

The needles of conifer trees are one of the richest sources of natural polyprenols. Polyprenol homologs from Abies sibirica L. lipophilic 80% purified extract were analyzed and quantified. In total, 10 peaks (Prenol-11 to Prenol-20) were observed in the ultra-high-performance liquid chromatography–diode array detector (UHPLC-DAD) chromatogram of Siberian fir with the most abundant compound being Prenol-15 (relative amount 37.23 + 0.56% of the total polyprenol yield). Abies sibirica L. polyprenol solubility and incorporation efficiency into liposomes were studied in various commercially available lecithin mixtures (Phosal IP40, Phosal 75SA, and Lipoid P45). The resulting multilamellar polyprenol liposomes were morphologically characterized by Light and Transmission Electron Microscopy, and the liposome size was discovered to be polymodal with the main peak at 1360 nm (90% of the volume). As polyprenols are fully soluble only in lipids, a liposomal formulation based upon co-solubilization and a modified ethanol injection method of polyprenols into the ethanol-phospholipid system was developed for the entrapment and delivery of polyprenols for potential commercial applications in food supplement and cosmetic industries.

Preparation and In Vitro/Vivo Evaluation of Nano-Liposomal Form of Febrifugine Hydrochloride.

Febrifugine hydrochloride (FFH) has strong pharmacological antimalarial effect. However, compared with oral administration, the efficacy of intravenous administration is significantly reduced. In this study, we prepared conventional liposomes and PEGylated liposomes to improve the efficacy of its intravenous injection. Both liposome formulations were prepared using a modified ethanol injection method. Their mean particle sizes were 126.23 and 114.93 nm, mean zeta potentials were -6.25 and -26.33 mV, and entrapment efficiencies (EE) were 89.43 and 96.42%, respectively. The in vitro release profile indicated that the release of FFH from PEGylated liposomes and conventional liposomes was slower than free FFH, with sustained-release effect of PEGylated liposomes being more significant. PEGylated liposomes demonstrated excellent antimalarial activities in vitro superior to free FFH and conventional FFH-loaded liposomes. In addition, the PEGylated liposomes resulted in enhanced antimalarial effect in P. berghei infected mice in vivo with delayed recrudescence and prolonged survival time, compared with free FFH and conventional FFH-loaded liposomes administration. Based on these exciting experimental results, PEGylated liposomes could be a potential drug delivery system for FFH, with enhanced pharmacodynamics of intravenous injection.

Phenylethyl Resorcinol Loaded in Liposomal Cream Formulation for Cosmeceutical Application

Phenylethyl Resorcinol (PR) is a cosmeceutical skin lightening agent and the purpose of this study was to enhance its stability by using liposomal cream formulation which increases local efficacy and safety. Liposome formulation was prepared by modified ethanol injection method, and it contained soy phosphatidyl choline (SPC), cholesterol (CHO), Tween 80 (TW80) and deoxycholic acid (DA) mixed with 2% PR. The physicochemical properties, skin permeation as well as cellular study were evaluated in order to obtain the optimized formulation. The optimized liposome formulation composed of SPC:TW80:DA (84:16:2.5) and exhibited vesicle size, polydispersity index (PDI) and zeta potential of 286.4±8.04 nm, 0.317±0.03 and -39.20±3.85 mV, respectively. Entrapment efficiency (EE) of liposome formulation was 93.55±0.05%. The vesicle was spherical in shape and showed good physicochemical stability for 4 months. The skin permeation study demonstrated that liposome with a negative charge could result in a high PR skin deposition value of 1732.76±216.24 µg/cm2 after 24 h. Cellular study showed that liposome formulation could inhibit melanin content in B16 melanoma cells and enhance cell viability in HaCaT keratinocyte cells. The optimized PR liposome was incorporated in cream and investigated physicochemical properties, stability and skin permeation. Liposomal PR cream showed a good stability and a superior result than PR cream in skin permeation parameters, as well as in tyrosinase inhibition.

Investigation of the Effect of Gammaoryzanol Addition on the Properties of the Nanoliposomes by using Response Surface Methodology: Preparation, Characterization and Optimization

Liposomes are the most important lipid-based nanocarriers, which are used for encapsulation of both hydrophilic and hydrophobic active compounds. The aim of this study was to investigate the effect of Gammaoryzanol (GO) addition to lipid bilayers on characteristics of liposomes prepared by a modified ethanol injection method. The GO bearing nanoliposomes were prepared with different molar ratios of phosphatidylcholine (PC): GO. PC: GO molar ratio was concerned as the independent variable while particle size (PS), encapsulation efficiency percentage (EE%) and drug release over 24 h (D24h) were considered as the dependent variables. The variables were analyzed and optimized by employing response surface methodology (RSM). Also, graphical response surfaces and contour plots were drawn to understand the interaction effects of different variables. Finally, optimum levels of the variables were obtained from the optimization plots. The mean PS, EE% and D24h values of Celecoxib-loaded nanoliposomes were found to be 102.6 ± 9.5 nm, 67.6 ± 11.2% and 53.97 ± 9.6 %, respectively. The results indicated that both PC: GO and PC: Drug ratios are important variables contributing to PS and EE% of the nanoliposomes. However, just PC: GO had an effect on D24h (P<0.05). According to the findings, optimized formulation of Celecoxib nanoliposomes is associated with PC: GO and PC: Drug ratios of 4.6 and 3, which provides PS = 102 nm, EE% = 74.2% and D24h = 59.9 %. In conclusion, addition of GO and using ethanol injection method, with respect to optimization of the parameters by RSM technique, presents a simple, rapid and beneficial approache for preparation of nanoliposomes with optimum characteristics.

Preparation and evaluation of vitamin C and folic acid-coloaded antioxidant liposomes

ABSTRACTVitamin C (VC) and folic acid (FA) are the important nutrients and antioxidants in human body. To protect them from pro-oxidant elements and reveal their co-operation effects, VC-loaded liposomes (VC-Lip), FA-loaded liposomes (FA-Lip), and their coloaded liposomes (VCFA-Lip) are prepared using the modified ethanol injection method in this study. The properties of the prepared liposomes including morphology, encapsulation efficiency, DPPH radical scavenging activity, hydrogen peroxide scavenging activity, and stability are determined and discussed in detail. The experimental results show that the vesicle size of the prepared liposomes are about 100–150 nm. The encapsulation efficiency of VC- and FA-coloaded liposomes is higher than that of the individual loaded ones. In vitro antioxidant activity study shows that the prepared liposomes have excellent antioxidant activity and the co-operation of VC and FA are better than the individual ones. Furthermore, the stability experiment reveals that the coloaded liposomes have a good stability in the relatively low temperature. These results indicate that VC- and FA-coloaded liposomes could be used as an effective antioxidant and it could be applied as a promising delivery system for the antioxidant defense system to food industry.

Nanoethosomal formulation of gammaoryzanol for skin-aging protection and wrinkle improvement: a histopathological study

Background: Free radical scavengers and antioxidants, with the main focus on enhanced targeting to the skin layers, can provide protection against skin ageing.Objective: The aim of the present study was to prepare nanoethosomal formulation of gammaoryzanol (GO), a water insoluble antioxidant, for its dermal delivery to prevent skin aging.Methods: Nanoethosomal formulation was prepared by a modified ethanol injection method and characterized by using laser light scattering, scanning electronic microscope (SEM) and X-ray diffraction (XRD) techniques. The effects of formulation parameters on nanoparticle size, encapsulation efficiency percent (EE%) and loading capacity percent (LC%) were investigated. Antioxidant activity of GO-loaded formulation was investigated in vitro using normal African green monkey kidney fibroblast cells (Vero). The effect of control and GO-loaded nanoethosomal formulation on superoxide dismutase (SOD) and malondialdehyde (MDA) content of rat skin was also probed. Furthermore, the effect of GO-loaded nanoethosomes on skin wrinkle improvement was studied by dermoscopic and histological examination on healthy humans and UV-irradiated rats, respectively.Results: The optimized nanoethosomal formulation showed promising characteristics including narrow size distribution 0.17 ± 0.02, mean diameter of 98.9 ± 0.05 nm, EE% of 97.12 ± 3.62%, LC% of 13.87 ± 1.36% and zeta potential value of –15.1 ± 0.9 mV. The XRD results confirmed uniform drug dispersion in the nanoethosomes structure. In vitro and in vivo antioxidant studies confirmed the superior antioxidant effect of GO-loaded nanoethosomal formulation compared with control groups (blank nanoethosomes and GO suspension).Conclusions: Nanoethosomes was a promising carrier for dermal delivery of GO and consequently had superior anti-aging effect.

Using Factorial Experimental Design To Prepare Size-Tuned Nanovesicles

The aim of this work was to prepare size-tuned nanovesicles using a modified ethanol injection method (EIM) by applying factorial experimental design. Stable size-tuned nanovesicles (liposomes and niosomes) with controlled sizes and high EE values for hydrophobic compounds (Sudan Red 7B and vitamin D3) were achieved. Equations that were able to predict the mean particle sizes, in the ranges of 55–156 nm for liposomes and 224–362 nm for niosomes with PDI values between 0.032 and 0.378, were obtained. These customized soft nanoparticles could be suitable in food, cosmetic, pharmaceutical, or medical applications, such as diagnosis or therapy.

Iron-entrapped niosomes and their potential application for yogurt fortification

Several formulations were tested for iron-entrapped niosomes with high stability and encapsulation efficiency. A modified ethanol injection method was selected for niosomes preparation.Niosomes formulated with Glycerol monooleate, Polyglyceryl-3 dioleate or Sorbitan monooleate as food-grade surfactants and dodecanol as membrane stabilizer showed iron encapsulation efficiencies in the 72–84% range.Niosomes stability against simulated gastric fluid and simulated intestinal fluid was evaluated by measuring the entrapment efficiency and oxidation of iron.The best formulations were selected for preparation of iron-fortified yogurts. Sensory, rheological and stability properties of the control yogurt were only slightly affected by iron-entrapped niosomes. These results demonstrated that yogurt could be a suitable vehicle for the iron-entrapped niosomes formulated in this work.