Ethanol Injection Method Research Articles

Transfersomes: An Innovative Vesicular Carrier for Boosted Transdermal Delivery System

One may find difficulties with oral and parenteral drug delivery systems in a routine of clinical practice because they do not have sufficient compliance and bioavailability for patients. So, nowadays transdermal route is a greater area of interest of pharmaceutical research for delivering drug. But skin is the most challenging area to cross in transdermal delivery of drug as the stratum corneum & the outer layer of the skin have tight intracellular junctions. Researchers have developed various approaches like micro needle, sonophoresis, electrophoresis, and iontophoresis etc to overcome those complications for the transdermal delivery of drugs. Chemical permeation enhancers are needed in vesicular drug delivery system such as niosomes, liposomes, elastic liposomes (transfersomes and ethosomes) to improve their penetration property. Transferosomes can be prepared by a number of methods like vortexing, sonication method, freeze–thaw method, ethanol injection method, Reverse-phase evaporation method, etc. Transfersomes can carry wide ranges of drugs having a wide range of solubility within it as they are constructed of hydrophobic as well as hydrophilic moieties. The main property of transferosome is deformability. This flexible nature of the vesicle membrane helps transfersome to go across the narrow pores with a maximum amount of drugs present within it. They have high deformable capacity which exhibits advanced penetration capability of intact vesicles. Both high and low molecular weight drugs like albumin, insulin, corticosteroids, sex hormones, anesthetic, anticancer, analgesic can be fused within transfersome.

CD73 downregulation by EGFR-targeted liposomal CD73 siRNA potentiates antitumor effect of liposomal doxorubicin in 4T1 tumor-bearing mice

Blocking CD73 ectonucleotidase has been proposed as a potential therapeutic approach for cancer treatment. The present study aimed to investigate the antitumor effect of a novel EGFR-Targeted liposomal CD73 siRNA formulation in combination therapy with liposomal doxorubicin in the 4T1 mouse model. CD73 siRNA was encapsulated into nanoliposomes by the ethanol injection method. After preparation, characterization, morphology, and stability evaluation of formulations, the toxicity was measured by MTT assay. Uptake assay and efficiency of the liposomal formulations were investigated on the 4T1 cell line. The liposomal formulation containing CD73 siRNA was targeted with GE11 peptide for in vivo evaluations. Following biodistribution analysis, the antitumor activity of prepared formulations in combination with liposomal doxorubicin was studied in mice bearing 4T1 metastatic breast cancer cells. Finally, the induction of immune response of formulations in concomitant treatment with liposomal doxorubicin was evaluated in the tumor microenvironment of a mouse model of breast cancer. The size of prepared liposomal formulations at N/P = 16 for the liposomal CD73 siRNA and GE11-liposomal CD73 siRNA groups were 89 nm ± 4.4 and 95 nm ± 6.6, respectively. The nanoparticle’s PDI was less than 0.3 and their surface charge was below 10 mV. The results demonstrated that N/P = 16 yielded the best encapsulation efficiency which was 94% ± 3.3. AFM results showed that the liposomes were spherical in shape and were less than 100 nm in size. The results of the MTT assay showed significant toxicity of the liposomes containing CD73 siRNA during the 48-h cell culture. Real-time PCR and flow cytometry results showed that liposomes containing CD73 siRNA could effectively downregulate CD73 expression. Liposomal formulations were able to significantly downregulate CD73 gene expression, in vivo. However, CD73 downregulation efficiency was significantly higher for the targeted form compared to the non-targeted formulation (P value < 0.01). The combination showed maximum tumor growth delay with remarkable survival improvement compared to the control group. Studying the immune responses in the treatment groups which received doxorubicin, showed decreased number of lymphocytes in the tumor environment. However, this decrease was lower in the combination therapy group. Finally, our results clearly showed that CD73 downregulation increases the activity of CD8+ lymphocytes (IFN-ℽ production) and also significantly decreases the Foxp3 in the CD25+ lymphocytes compared to the control group. GE11-Lipo CD73 siRNA formulation can efficiently knockdown CD73 ectonucleotidase. Also, the efficacy of liposomal doxorubicin is significantly enhanced via the downregulation of CD73 ectonucleotidase.

Afatinib liposomal dry powder inhaler: Targeted pulmonary delivery of EGFR inhibitor for the management of lung cancer

The present research work aims to prepare a liposomal dry powder inhaler (LDPI) of afatinib dimaleate for the treatment of non-small cell lung cancer through pulmonary delivery. The influence of formulation independent variables, lipid concentration (X1, mol%) and organic phase (X2, mL), on dependent variables, entrapment efficiency (%) and particle size (nm), were investigated using a three-level factorial design. The optimized drug encapsulated PEGylated liposomal batch of 2 mL prepared by ethanol injection method was further scaled up to 25 mL, to enable lyophilization used to convert liposomal dispersion into LDPI. Scale-up batch of liposomes was found to have an average particle size of 181.2 ± 5.0 nm, % entrapment efficiency of 75.31 ± 2.70%, and zeta potential of 9.65 ± 0.31 mV. LDPI manufactured with trehalose as a cryoprotectant and l-leucine as an antiadherent offered porous particles with smooth morphologies that improved aerodynamic efficiency. The prepared LDPI formulation exhibited 9.639 ± 1.369 μm of particle size distribution, 72.56 ± 1.04% drug content, and good flow properties. In vitro lung deposition study implied that particles could reach the deeper region of the lung with more than 85% dose emission from an inhaler device. In vitro drug release study revealed that the drug release profile in pH 7.4 buffer was lower than in pH 5.5 and pH 6.5 buffers, demonstrating the stability of LDPI in lung fluid. Cytotoxicity study utilizing A549 adenocarcinoma cell line uncovered the greater potential of liposomal formulation in inhibiting tumor cell growth as compared to plain drug solution. The cellular uptake across adenocarcinoma cells exhibited more intracellular uptake of fluorescein isothiocyanate (FITC) encapsulated afatinib liposomal formulation than plain FITC dye. Thus, the optimized LDPI can offer an excellent approach for managing lung cancer through site-specific delivery.

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 &amp;lt; 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.

Preparation, Characterization, and In Vitro Performance of Gambogic Acid-Layered Double Hydroxide/Liposome Nanocomposites

Gambogic acid (GA) refers to a xanthonoid that exhibits significant antitumor activity due to its poor solubility and low bioavailability. For this reason, its shortcoming should be overcome using novel approaches to improve its practical effectiveness. In this study, with the use of ion exchange method, GA was encapsulated in layered double hydroxide (LDH). In the GA-LDH nanohybrid, GA was distributed and stabilized in the interlamellar region of LDH through intermolecular interactions. GA-LDH was further modified by liposome (LS) through ethanol injection method. The drug encapsulation efficiency of GA-LDH/LS was obtained as 56.28%. The chemical structures and physicochemical properties exhibited by GA-LDH/LS were characterized and confirmed using different instruments, and drug release showed that GA-LDH/LS had significantly sustained release due to the combined effect of the matrix LDH and the phospholipid bilayer. Furthermore, GA-LDH/LS displayed lower hemolysis percentage than GA-LDH during the hemolysis test. This study suggested that GA-LDH/LS nanocomposite could be a promising antitumor drug delivery system due to its outstanding performance in biomedical research.

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.

Merging Experimental Design and Nanotechnology for the Development of Optimized Simvastatin Spanlastics: A Promising Combined Strategy for Augmenting the Suppression of Various Human Cancer Cells.

Simvastatin (SMV) is an antihyperlipidemic agent that has been investigated as a possible anti-cancer agent. An obstacle to malignant tumor therapy using drugs is the delivery of adequate levels to the cancer cells while minimizing side effects following their systemic administration. To circumvent this challenge, the researchers directed towards the field of nanotechnology to benefit from the nano-size of the formulation in passively targeting the tumor cells. Thus, our study aimed at investigating the potential of a combined mixture–process variable design for optimization of SMV spanlastics (SMV-SPNs) with minimized particle size and maximized zeta potential to enhance the anticancer activity of the drug. The study investigated the effects of Span® 20 and Tween® 80 as mixture components and sonication time as a process variable on particle size, polydispersity index, and zeta potential as responses. SPNs were prepared using an ethanol injection method. Combining the predicted optimized variables’ levels is supposed to achieve the set goals with a desirability of 0.821. The optimized spanlastics exhibited a measured globule size of 128.50 nm, PDI of 0.329, and ZP of −29.11 mV. The percentage relative error between predicted responses and the observed ones were less than 5% for the three responses, indicating the optimization technique credibility. A significant improvement in the cytotoxicity of the optimized formulation against three different cancerous cell lines was observed in comparison with SMV. The inhibitory concentration (IC50) values of MCF-7, HCT-116, and HEPG2 were found to be 0.89, 0.39, and 0.06 μM at 24 h incubation. The enhanced cytotoxicity could be assigned to the possible improved permeation and preferential build-up within the cancerous cells by virtue of the minimized size. These findings imply that SMV-SPNs could be an ideal strategy to combat cancer.

Extraction, Encapsulation into Lipid Vesicular Systems, and Biological Activity of Rosa canina L. Bioactive Compounds for Dermocosmetic Use.

Valorization of wild plants to obtain botanical ingredients could be a strategy for sustainable production of cosmetics. This study aimed to select the rosehip extract containing the greatest amounts of bioactive compounds and to encapsulate it in vesicular systems capable of protecting their own antioxidant activity. Chemical analysis of Rosa canina L. extracts was performed by LC-DAD-MS/MS and 1H-NMR and vitamins, phenolic compounds, sugars, and organic acids were detected as the main compounds of the extracts. Liposomes, prepared by the film hydration method, together with hyalurosomes and ethosomes, obtained by the ethanol injection method, were characterized in terms of vesicle size, polydispersity index, entrapment efficiency, zeta potential, in vitro release and biocompatibility on WS1 fibroblasts. Among all types of vesicular systems, ethosomes proved to be the most promising nanocarriers showing nanometric size (196 ± 1 nm), narrow polydispersity (0.20 ± 0.02), good entrapment efficiency (92.30 ± 0.02%), and negative zeta potential (−37.36 ± 0.55 mV). Moreover, ethosomes showed good stability over time, a slow release of polyphenols compared with free extract, and they were not cytotoxic. In conclusion, ethosomes could be innovative carriers for the encapsulation of rosehip extract.

QbD-based optimization of raloxifene-loaded cubosomal formulation for transdemal delivery: ex vivo permeability and in vivo pharmacokinetic studies.

Raloxifene (RLX) is a drug that is commonly recommended to postmenopausal women at high risk of invasive breast cancer and to prevent osteoporosis. However, limited water solubility (0.000512mg/ml) and low oral bioavailability (2%) of RLX limit its therapeutic utility. The objective of the present study was to develop an alternative transdermal delivery of RLX to improve its absorption, bypass first pass metabolism, and subsequently improve bioavailability. RLX-loaded cubosomes were prepared using the ethanol injection method followed by microfluidization technique and optimized using the QbD-based 23 factorial design. The average particle size, entrapment efficiency, and zeta potential of the optimized formulation were found to be 110.6nm, 98.23%, and 26.2mV, respectively. In vitro dissolution study indicated that the RLX-loaded cubosomes released 98.26% of the drug compared to pure RLX dispersion (58.6%). Histopathological examination revealed no sign of inflammation, indicating the safety of the developed formulation. Accelerated stability study as per ICH guidelines displayed no significant change in the formulation characteristics and drug-related performance of the developed formulation. Ex vivo permeability studies demonstrated a prolonged release from cubosomal formulation. In vivo pharmacokinetic studies revealed that the relative bioavailability of the optimized transdermal RLX-loaded cubosomes increased by 2.33-fold and 1.22-fold when compared with the oral RLX dispersion and transdermal RLX hydro-ethanolic solution respectively. IVIVC showed level C correlation with linear regression. Thus, the developed RLX-loaded cubosomes may have potential to overcome the problems associated with the existing marketed oral dosage forms of RLX.

Formulation and Evaluation of Transdermal Gel Containing Tacrolimus-Loaded Spanlastics: In Vitro, Ex Vivo and In Vivo Studies.

Our goal was to prepare Span 60-based elastic nanovesicles (spanlastics (SPLs)) of tacrolimus (TCR) using the adapted ethanol injection method, characterize them, and evaluate their ability to improve the transdermal permeation of the active substance. The impact of two different concentrations of penetration enhancers, namely, propylene glycol and oleic acid, on the entrapment efficiency, vesicle size, and zeta potential was assessed. Moreover, in vitro release through a semipermeable membrane and ex vivo penetration through hairless rat skin were performed. Morphological examination and pharmacokinetics were performed for one selected formulation (F3OA1). TCR-loaded SPLs were effectively formulated with two different concentrations of permeation enhancers, and the effect of these enhancers on their physicochemical properties differed in accordance with the concentration and kind of enhancer used. The results of in vitro release displayed a considerable (p < 0.05) enhancement compared to the suspension of the pure drug, and there was a correlation between the in vitro and ex vivo results. The selected TCR-loaded nanovesicles incorporated into a gel base showed appreciable advantages over the oral drug suspension and the TCR-loaded gel. Additionally, the pharmacokinetic parameters were significantly (p < 0.05) improved based on our findings. Moreover, the AUC0–7 ng·h/mL form F3 OA1 was 3.36-fold higher than that after the administration of the TCR oral suspension.

Irinotecan-loaded ROS-responsive liposomes containing thioether phosphatidylcholine for improving anticancer activity

In this work, irinotecan (IR)-loaded reactive oxygen species (ROS)-responsive liposomes were developed for improving anticancer activity. First of all, thioether phosphatidylcholine (S-PC) based liposomes (S-LP) were prepared via ethanol injection method. After that, IR was actively loaded into liposomes (IR/S-LP) by using sucrose octasulfate triethylamine (TEA8-SOS) with encapsulation efficiency of 97.0 ± 1.0% and loading efficiency of 30.6 ± 0.1%. The IR/S-LP liposomal formulation was further characterized through dynamic light scattering (DLS) and transmission electron microscopy (TEM), confirming regular spherical structure with particle size of 100.2 ± 5.2 nm and zeta potential of −25.1 ± 2.2 mV. Moreover, the IR/S-LP formulation exhibited improved drug release profiles in the presence of H2O2. Cellular uptake behavior of IR/S-LP was observed by using confocal laser scanning microscopy (CLSM). Pharmacokinetic analyses further confirmed that IR/S-LP could lead to extending SN38 exposure. Ultimately, in vivo antitumor test in 4T-1 tumor-bearing mice revealed that IR/S-LP had dose-dependent tumor growth inhibition and superior anticancer activity compared to free IR and convenient IR-liposomes (Onivyde®-like) ascribed to ROS-responsiveness and raised bioavailability. Taken together, IR loaded S-PC based liposomes have ROS responsiveness leading to enhanced anticancer activity, which might be an alternative of convenient IR-liposomes.

Fenticonazole nitrate loaded trans-novasomes for effective management of tinea corporis: design characterization, in silico study, and exploratory clinical appraisal

The current investigation aimed for loading fenticonazole nitrate (FTN), an antifungal agent with low aqueous solubility, into trans-novasomes (TNs) for management of tinea corporis topically. TNs contain Brij® as an edge activator besides the components of novasomes (cholesterol, Span 60, and oleic acid) owing to augment the topical delivery of FTN. TNs were fabricated applying ethanol injection method based on D-optimal experiment. TNs were evaluated with regard to entrapment efficiency percent (EE%), particle size (PS), polydispersity index (PDI), and zeta potential (ZP). Further explorations were conducted on the optimum formulation (F7). F7 showed spherical appearance with EE%, PS, PDI, and ZP of 100.00 ± 1.10%, 358.60 ± 10.76 nm, 0.51 ± 0.004, and −30.00 ± 0.80 mV, respectively. The in silico study revealed the ability of the FTN–cholesterol complex to maintain favorable interactions throughout the molecular dynamics simulation (MDS) study. Moreover, Trichophyton mentagrophytes growth was inhibited effectively by F7 than by FTN suspension applying 2,3-bis(2-methyloxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay. Furthermore, a clinical appraisal on patients with tinea corporis fungal lesions confirmed the superiority of F7 compared to Miconaz® cream in the magnitude of clinical cure of tinea corporis. Thereby, TNs could be considered as promising vesicles for enhancing the antifungal potential of FTN for the topical management of tinea corporis.

Synthesis and Assessment of Lipid Nanovesicles for Efficient Transdermal Delivery of Hydrophilic Molecules

The transdermal delivery of hydrophilic molecules through the dermis is a challenging task because of the various skin barriers. Thus, lipid nanovesicles, including liposomes, ethosomes and transethosomes, are advantageous to transdermal drug delivery. This study was designed to develop nanovesicles like liposomes (LF1), ethosomes (EF1) and transethosomes (TF1) containing hydrophilic agents such as Rhodamine B (RB) as a model compound. Nanovesicles loaded with RB were synthesized using an ethanol injection method followed by probe sonication (hot method). The nonsonicated nanovesicles had vesicle sizes ranging from [Formula: see text] to [Formula: see text][Formula: see text]nm, while sonicated nanovesicles sizes ranged from [Formula: see text] to [Formula: see text][Formula: see text]nm. In addition, the nonsonicated nanovesicles had polydispersity index (PdI) ranging from [Formula: see text] to [Formula: see text], whereas the PdI of the sonicated nanovesicles ranged from [Formula: see text] to [Formula: see text]. Transmission electron microscopy revealed the distinguishable features of the sonicated nanovesicles depending on the compositions. The entrapment efficiency of RB was observed in the order of [Formula: see text]. At the end of 10[Formula: see text]h, RB permeated through the Strat-M[Formula: see text] membrane at [Formula: see text], [Formula: see text] and [Formula: see text]g/cm2 from the LF1, EF1 and TF1 containing RB, respectively. The flux ([Formula: see text]) of TF1 was obtained 5.53 times higher than RB itself. Therefore, the TF1 nanovesicle allowed the highly efficient permeation and entrapment of hydrophilic RB compared to LF1 and EF1 nanovesicles. Hence, it can be concluded that transethosomes would be a better choice for hydrophilic compounds to cross the skin barriers via the transdermal route.

Improved stability and skin penetration through glycethosomes loaded with glycyrrhetinic acid.

In recent years, glycyrrhetinic acid (GA) has been popularly used in cosmetics because of its anti-inflammatory and anti-oxidant effects. However, due to the poor water solubility of GA and the barrier effect of human skin, the penetration of GA through the skin may be hindered. Liposomes are a common delivery system for functional compounds in cosmetics. Nonetheless, the stability and transdermal effect of traditional liposomes are limited. The aim of this work was to prepare a new liposome system that contained glycerol and ethanol to enhance the stability of the vesicles and promote the penetration of GA into the skin. The glycethosomes were prepared by ethanol injection and sonication method. The effects of different concentrations of glycerol and ethanol on the particle size, polydispersity (PDI), entrapment efficiency (EE), stability and rheological properties of vesicles were evaluated. Lipophilic and hydrophilic fluorescent probes were used to investigate the microviscosity of vesicles. In vitro permeation tests were performed with pig skin in Franz cells and the concentration of GA in different skin layers was determined by high-performance liquid chromatography (HPLC). The ability of different vesicles to induce lipid extraction and fluidization was analysed by using attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR). When glycerol was 50% and ethanol was 25%, the obtained glycethosomes had the smallest particle size and the best stability with a mean particle size of 94.5nm, PDI 0.216 and 99.8% EE. Fluorescence probe studies indicated that the microviscosity of glycethosomes was the largest when the concentration of glycerol and ethanol was 50% and 25%, which was consistent with the storage stability of glycethosomes. It was found that the glycethosomes had the best transdermal effect and the total skin permeation percentage of GA was 20.67%, while those of ethosomes, glycerosomes, liposomes and dispersion were 10.56%, 9.38%, 7.78% and 5.02%, respectively. And glycethosomes had effectively lipid extraction and fluidization effect on the skin stratum corneum. Compared with other traditional liposomes, glycethosomes can significantly improve the stability of vesicles and the transdermal effect of GA. Glycethosomes is promising vesicles for the delivery of GA.

Myricetin Loaded Nano-micelles Delivery System Reduces Bone Loss Induced by Ovariectomy in Rats Through Inhibition of Osteoclast Formation

In recent years, much attention has been paid to the therapeutic effects of phytochemicals on osteoporosis. Other studies have shown that myricetin (MY) could promote osteogenic activity and inhibit osteoclastic effect, albeit little is known about effect of MY micellar system on osteoporosis. Therefore, we sought to discuss the therapeutic effect and mechanism of MY-loaded bone-targeting micelles on osteoporosis induced by ovariectomy (OVA) in rats. The AL-P(LLA-CL)-PEG-P(LLA-CL)-MY micelles were prepared via ethanol injection method, while in vitro release study, bone targeting, pharmacokinetic studies, and the effect on proliferation of osteoblasts were investigated. Further, the therapeutic effect on osteoporosis was studied through ovariectomized rats. Compared with free MY, oral bioavailability of AL-P(LLA-CL)-PEG-P(LLA-CL)-MY micelles in rats was increased by 3.54 times. The AL-P(LLA-CL)-PEG-P(LLA-CL)-MY micelles exhibited bone targeting potential, and could significantly increase the activity of alkaline phosphatase and promote the proliferation of osteoblasts. Importantly, AL-P(LLA-CL)-PEG-P(LLA-CL)-MY micelles mainly regulated bone metabolism by inhibiting bone resorption, thereby improving the symptoms of osteoporosis in OVA rats. The AL-P(LLA-CL)-PEG-P(LLA-CL)-MY micelles substantially enhanced the oral bioavailability of MY and demonstrated good bone targeting capability, thereby suggesting its prospect as carrier for osteoporotic improvement in OVA rats.

Nanoliposome Use to Improve the Stability of Phenylethyl Resorcinol and Serve as a Skin Penetration Enhancer for Skin Whitening

Phenylethyl resorcinol (PR) is a potent tyrosinase inhibitor and a cosmeceutical skin lightening agent. However, the application of PR is limited by photoinstability and poor solubility. In this study, we formulated and optimized phenylethyl resorcinol loaded nanoliposomes (PR-NLPs) to improve the stability and effective delivery of PR. PR-NLPs were prepared by the ethanol injection method and optimized by a single factor experimental and Box–Behnken design. In addition, Diethylamino Hydroxybenzoyl Hexyl Benzoate (DHHB) as the UBA absorber was added to PR-NLPs, which significantly improved the photostability of PR. The mean size, polydispersity index (PDI), and zeta potential of the optimized PR-NLPs were 130.1 ± 3.54 nm, 0.225 ± 0.02, and −43.9 ± 3.44 mV, respectively. The drug encapsulation efficiency (EE) and loading efficiency (LC) of PR-NLPs were 96.81 ± 3.46% and 8.82 ± 0.6%, respectively. These PR-NLPs showed good physicochemical stability for 3 months at 4 °C and 25 °C in the dark. They showed typical sustained and prolonged drug-release behavior in vitro. The in vitro cytotoxicity assay and cellular uptake demonstrated that the PR-NLPs had excellent biocompatibility and cell transport ability. It significantly inhibited tyrosinase activity and reduced melanin production in B16F10 cells at concentrations of 20 or 30 μg/mL. Moreover, the PR-NLPs enhanced the PR into the skin. These results indicate that PR-NLPs can be used as a nanocarrier to improve the transdermal delivery of PR.

Donepezil HCl Liposomes: Development, Characterization, Cytotoxicity, and Pharmacokinetic Study.

The current research work aims to study the pharmacokinetic and nasal ciliotoxicity of donepezil liposome-based in situ gel to treat Alzheimer's disease. The physicochemical properties and first-pass metabolism of donepezil HCl result in low concentrations reaching the brain post oral administration. To overcome this problem, donepezil HCl-loaded liposomes were formulated using the ethanol injection method. The donepezil HCl-loaded liposomes were spherical with a size of 103 ± 6.2 nm, polydispersity index of 0.108 ± 0.008, and entrapment efficiency of 93 ± 5.33 %. The optimized in situ gel with donepezil HCl-loaded liposomes showed 80.11 ± 7.77 % drug permeation than donepezil HCl solution-based in situ gel (13.12 ± 4.84 %) across sheep nasal mucosa. The nasal ciliotoxicity study indicated the safety of developed formulation for administration via nasal route. The pharmacokinetics and biodistribution study of developed formulation showed higher drug concentration (1239.61 ± 123.60 pg/g) in the brain after nasal administration indicating its better potential via the nasal pathway. To treat Alzheimer's disease, the administration of liposome-based in situ gel through the nasal pathway can therefore be considered as an effective and promising mode of drug delivery.

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.

Decoquinate liposomes: highly effective clearance of Plasmodium parasites causing severe malaria

BackgroundSevere malaria caused by Plasmodium falciparum leads to most malaria-related deaths globally. Decoquinate (DQ) displays strong activity against multistage infection by Plasmodium parasites. However, the development of DQ as an oral dosage form for the treatment of malaria at the blood stage has not been successful. In this study, liposome formulations of DQ were created for intravenous (IV) injection to suppress Plasmodium berghei, a parasite that causes severe malaria in mice.MethodsDQ liposomes were prepared by conventional ethanol injection method with slight modifications and encapsulation efficiency evaluated by the well-established centrifugation method. Potency of the DQ liposomes against P. falciparum was assessed in vitro using freshly isolated human red blood cells. The efficacy of the DQ liposomes was examined in the mouse model of severe malaria.ResultsThe DQ liposomes were around 150 nm in size and had the encapsulation efficiency rates > 95%. The freshly prepared and lyophilized liposomes were stable after storage at − 20 °C for 6 months. The liposomes were shown to have excellent activity against P. falciparum in vitro with DQ IC50 0.91 ± 0.05 nM for 3D7 (chloroquine sensitive strain) and DQ IC50 1.33 ± 0.14 nM for Dd2 (multidrug resistant strain), which were 18- and 14-fold more potent than artemisinin, respectively. Mice did not have any signs of toxicity after receiving high dose of the liposomes (DQ 500 mg/kg per mouse) by IV injection. In the mouse model of severe malaria, the liposomes had impressive efficacy against P. berghei with DQ ED50 of 0.720 mg/kg.ConclusionThe DQ liposomes prepared in this study were stable for long term storage and safe for IV injection in mammalian animals. The newly created liposome formulations had excellent activity against Plasmodium infection at the blood-stage, which encourages their application in the treatment of severe malaria.

Fabrication and optimization of raloxifene loaded spanlastics vesicle for transdermal delivery

This research work is envisioned for the development and optimization of Raloxifene HCL loaded spanlastics. Raloxifene HCL, a selective estrogen receptor modulator (SERM), is active for the deterrence and management of postmenopausal osteoporosis. Spanlastics nanocarrier was prepared by ethanol injection method and optimized for independent variables by using Box Behnken design. The independent variables used in the experiment were concentration of Span 60, Tween 80 as an edge activator, and sonication time. The individual and joined impacts of independent variables were assessed on vesicle size, polydispersity index (PDI), and encapsulation efficiency (EE). Raloxifene HCL and formulation were analyzed by FT-IR, DSC and XRD. Raloxifene HCL loaded spanlastics were characterized for particle size, PDI, EE, and in-vitro drug release. Ex vivo skin permeation and stability studies were performed for the optimized formulation. The optimized spanlastics vesicle showed a particle size of 275.4 nm with PDI, 0.267 which was suitable for permeation through skin layers. TEM image illustrated well-dispersed nanocarrier with spherical shape. The % cumulative drug release and transdermal flux for the optimized spanlastics formulation were found to be 85% (Approx. after 24 h) and 4.24 μg/cm2/hr respectively. The enhanced permeation across the skin layers was proposed to be due to the presence of ethanol which will increase the lipid fluidity of the stratum corneum and will help to penetrate the drug. Later the skin penetration ability was determined by confocal laser scanning microscopy (CLSM) and showed that rhodamine loaded spanlastics formulation penetrated to the depth of 54.9 μm while rhodamine suspension limited to 20 μm in the skin. We can conclude that raloxifene HCL loaded spanlastics could serve as a better approach to deliver drug in deeper layers of the skin.