Reverse-phase Evaporation Research Articles

Effect of Solvent and Cholesterol on Liposome Production by the Reverse-Phase Evaporation (RPE) Method.

Liposomal drug delivery is a promising approach for delivering therapeutics effectively. While most liposomes are designed to be nanometer-sized for efficient cellular uptake, micron-sized liposomes are gaining interest due to their larger drug-loading capacity. When combined with macroscale structures, such as implants and hydrogels, they offer prolonged therapeutic delivery. This study investigates how solvents affect the production of micron-sized liposomes, with or without cholesterol, using the reverse-phase evaporation (RPE) method. Although the RPE method is established for producing micron-sized liposomes, the influence of solvents on liposome size and uniformity is not well understood. The study explores whether controlling the size of inverse micelles, an intermediate product, through the use of different organic solvents affects the final liposome size. Three solvents─diethyl ether, methanol, and acetone─were tested for their effect on the formation of inverse micelles and liposomes and their sizes. Results showed that without cholesterol, diethyl ether produced uniform inverse micelles, leading to mostly nanosized liposomes. Methanol and acetone resulted in phase separation, preventing uniform liposome formation, although some micron-sized liposomes appeared. The acetone sample yielded mostly oil droplets. The results showed that forming inverse micelles lead to nanosized liposomes. With cholesterol, phase separation was dominant in methanol, but micron-sized liposomes still formed. Across all cases, cholesterol reduced the liposome size. The findings reveal that inverse micelles are not always reliable predictors of the final liposome size and that the RPE method is highly sensitive to solvent polarity and lipid-solvent interactions. Overall, the findings of this study provide valuable insights into how the choice of solvent and lipid composition can influence the production of liposomes via the RPE method. These insights are critical for optimizing liposome production and influencing future designs of liposomal drug delivery systems.

Enhancement of oral bioavailability of ibrutinib using a liposil nanohybrid delivery system.

Liposils, synthesized via the liposome templating method, offer a promising strategy for enhancing liposome stability by employing a silica coating. This study focuses on the development of nanocarriers utilizing silica-coated nanoliposomes for encapsulating the poorly water-soluble drug, ibrutinib. Ibrutinib-loaded nanoliposomes were meticulously formulated using the reverse-phase evaporation technique, serving as templates for silica coating, resulting in spherical liposils with an average size of approximately 240 nanometers. Comprehensive characterization of the liposil's physical and chemical properties was conducted using various analytical methods, including dynamic light scattering, transmission electron microscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction analysis. Liposils demonstrated superior performance compared to ibrutinib-loaded nanoliposomes, showing sustained drug release profiles in simulated intestinal fluids and resistance to simulated gastric fluid, as confirmed by dissolution studies. Moreover, ibrutinib liposils exhibited a significant increase in half-life (4.08-fold) and notable improvement in bioavailability (3.12-fold) compared to ibrutinib suspensions, as determined by pharmacokinetic studies in rats. These findings underscore the potential of liposils as nanocarriers for orally delivering poorly water-soluble drugs, offering enhanced stability and controlled release profiles, thereby improving bioavailability prospects and therapeutic efficacy. This approach holds promise for addressing challenges associated with the oral administration of drugs with limited solubility, thereby advancing drug delivery technologies and clinical outcomes in pharmaceutical research and development.

Liposome preparation of alpha-arbutin: stability and toxicity assessment using mouse B16F10 melanoma cells

ABSTRACT Melanoma is the most aggressive type of skin cancer, with few therapeutic alternatives following metastasis development. In recent years, drug delivery-associated nanotechnology has shown promising targeted results with diminished adverse effects compared to conventional treatments. This study aimed to (1) examine the effects of plant-derived α-arbutin, a natural compound and (2) compare these findings with bioactively developed liposomes containing α-arbutin utilizing the B16-F10 murine melanoma cell line as a model. Liposomes were obtained through reversed-phase evaporation by applying a spray dryer to assess their stability. The following biologic assays were measured cytotoxicity/antiproliferative (MTT, Neutral Red, and dsDNA PicoGreen). In addition, the levels of melanin and purinergic enzymes were also measured. The production of reactive oxygen species (ROS) and nitric oxide (NO) was determined as a measure of oxidative state. Treatment with nano-liposome containing alpha-arbutin induced a significant 68.4% cytotoxicity, similar to the positive control, in the B16-F10 murine melanoma cell line at 72 hr. Further, arbutin and liposomes containing alpha-arbutin increased levels of ROS and nitrite formation at 72 hr at the highest concentration (100 and 300 µg/ml) of treatments. Arbutin and liposomes containing alpha-arbutin reduced melanin levels at all tested concentrations. In addition, arbutin and alpha-arbutin containing liposomes lowered nucleotides (AMP, ADP, and ATP) and nucleoside (adenosine) levels in melanoma cells. Evidence suggests that α-arbutin containing liposome can be considered as an alternative immunosuppressive agent stimulated in melanoma treatment.

Liposomal Drug Delivery against Helicobacter pylori Using Furazolidone and N-Acetyl Cysteine in Augmented Therapy.

Helicobacter pylori (H. pylori) infection is a significant global health concern, affecting approximately 50% of the world's population and leading to gastric ulcers, gastritis, and gastric cancer. The increase in antibiotic resistance has compromised the efficacy of existing therapeutic regimens, necessitating novel approaches for effective eradication. This study aimed to develop a targeted liposomal drug delivery system incorporating furazolidone and N-acetylcysteine (NAC) to enhance mucopenetration and improve Helicobacter pylori eradication. Liposomes were formulated with furazolidone, NAC, and Pluronic F-127 using a modified reverse-phase evaporation technique. The formulations were categorized based on charge as neutral, negative, and positive and tested for mucopenetration using a modified silicon tube method with coumarin-6 as a fluorescent marker. The encapsulation efficiency and particle size were analyzed using HPLC and an Izon q-nano particle size analyzer. The results indicated that charged liposomes showed a higher encapsulation efficiency than neutral liposomes with Pluronic F-127. Notably, combining furazolidone with 1% NAC achieved complete eradication of H. pylori in 2.5 h, compared to six hours without NAC. The findings of this study suggest that incorporating NAC and Pluronic F-127 into liposomal formulations significantly enhances mucopenetration and antimicrobial efficacy.

Optimizing gefitinib nanoliposomes by Box-Behnken design and coating with chitosan: A sequential approach for enhanced drug delivery.

This study aimed to improve the stability and prolonged gefitinib release from the nanoliposomes. Nanoliposomes were prepared by reverse-phase evaporation and optimized using Box-Behnken design to investigate the influence of sonication time (X 1), tween 80 / soya phosphatidylcholine ratio (X 2), and cholesterol/soya phosphatidylcholine ratio (X 3) on nanoliposomes. Optimized nanoliposomes were quasi-spherical shaped, with a mean dimension of 93.2 nm and an encapsulation efficiency of 87.56±0.17 %. Surface decoration of the optimized batch was done using different concentrations of chitosan. The optimal chitosan concentration required to adorn the nanoliposome surface was 0.01 %. In comparison to unadorned nanoliposomes (82.16±0.65 %), adorned nanoliposomes (78.04±0.35 %) released the drug consistently over 24 h via Fickian diffusion. The IC50 values for surface-adorned nanoliposomes in A549 and H1299 cells were 6.53±0.75 and 4.73±0.46 μM, respectively. Cytotoxicity of the surface-decorated nanoliposomes may be due to their higher zeta potential and prolonged drug release. At the end of the sixth month, the samples stored at 4 °C were more stable than those stored at 25 °C and 45 °C. The stability of plain nanoliposomes has increased after chitosan coating. Thus, by using different concentrations of chitosan solution as coating material, we can develop a suitable sustained drug-release surface-adorned nanoliposomal formulation. The developed nanoliposomes may offer a new path for melanoma clinics.

Liposomes as versatile agents for the management of traumatic and nontraumatic central nervous system disorders: drug stability, targeting efficiency, and safety.

Various nanoparticle-based drug delivery systems for the treatment of neurological disorders have been widely studied. However, their inability to cross the blood-brain barrier hampers the clinical translation of these therapeutic strategies. Liposomes are nanoparticles composed of lipid bilayers, which can effectively encapsulate drugs and improve drug delivery across the blood-brain barrier and into brain tissue through their targeting and permeability. Therefore, they can potentially treat traumatic and nontraumatic central nervous system diseases. In this review, we outlined the common properties and preparation methods of liposomes, including thin-film hydration, reverse-phase evaporation, solvent injection techniques, detergent removal methods, and microfluidics techniques. Afterwards, we comprehensively discussed the current applications of liposomes in central nervous system diseases, such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, traumatic brain injury, spinal cord injury, and brain tumors. Most studies related to liposomes are still in the laboratory stage and have not yet entered clinical trials. Additionally, their application as drug delivery systems in clinical practice faces challenges such as drug stability, targeting efficiency, and safety. Therefore, we proposed development strategies related to liposomes to further promote their development in neurological disease research.

Comparative analysis of lipid-peptide nanoparticles prepared via microfluidics, reverse phase evaporation, and ouzo techniques for efficient plasmid DNA delivery

In the current “era of lipid carriers,” numerous strategies have been developed to manufacture lipid nanoparticles (LNPs). Nevertheless, the potential impact of various preparation methods on the characteristics, use, and/or stability of these LNPs remains unclear. In this work, we attempted to compare the effects of three different preparation methods: microfluidics (MF), reverse phase evaporation (RV), and ouzo (OZ) on lipid-peptide NPs (LPNPs) as plasmid DNA delivery carriers. These LPNPs had the same components, namely DOTMA cationic lipid, DSPC, cholesterol, and protamine. Subsequently, we compared the LPNPs in terms of their physicochemical features, functionality as gene delivery vehicles in two distinct cell lines (NT2 and D1-MSCs), and finally, their storage stability over a six-month period.It was clear that all three LPNP formulations worked to deliver EGFP-pDNA while keeping cells alive, and their physicochemical stability was high for 6 months. However, the preparation technique had a significant impact on their physicochemical characteristics. The MF produced LPNPs with a lesser size, polydispersity index, and zeta potential than the other synthesis methods. Additionally, their DNA entrapment efficiency, cell viability, and functional stability profiles were generally superior. These findings provide new insights for comparing different manufacturing methods to create LPNPs with the desired characteristics for effective and safe gene delivery.

Exploring Niosomes: A Comprehensive Review of their Structure, Formulation, and Biomedical Applications

Niosomes are a novel class of non-ionic surfactant vesicles. They have emerged as promising vesicles for the delivery of a diverse range of therapeutic agents. Niosomes are implied for the improvement of stability and solubility of molecules used in pharmaceutical components. The current study delved into the formulation methods, applications, and recent advancements in niosomal technology. They offer various benefits over other drug delivery systems due to their biocompatibility, biodegradability, and versatility. The first section of this study outlined the background, detailed structure, and characteristics of niosomes. Niosomes are quite distinct in their characteristics as they have the ability to encapsulate both hydrophilic and lipophilic drugs. The second section of the current study highlighted various methods employed for niosome preparation, such as film hydration, reverse-phase evaporation, and microfluidic techniques. By modifying the lipid composition and surfactants, specific physicochemical properties, such as size, lamellarity, and surface charge can be achieved.The final section discussed applications of niosomes across pharmaceutical, cosmetic, and biotechnological fields. These include targeted drug delivery, gene delivery, cosmetic ingredient encapsulation, and vaccine adjuvant systems. Additionally, the recent advancements in niosome research have been discussed as well.

N‐Acetyl cysteine‐loaded liposomes to reduce iron overload‐induced toxicity in human kidney cells

AbstractBACKGROUNDIron overload in the body generates free radicals through the Fenton reaction, leading to adverse effects on various organs. Several antioxidants have been recommended to treat iron‐related diseases effectively. N‐Acetyl cysteine (NAC), a potent antioxidant, was investigated in this study for its potential to mitigate iron overload‐induced cell death. A liposomal formulation was developed to encapsulate NAC, employing the reverse phase evaporation method, to enhance its delivery and effectiveness. The liposomal NAC was characterized by evaluating its size, zeta potential, morphology and release profiles.RESULTSThe protective effect of liposomal NAC against iron overload‐induced toxicity was assessed in human kidney 2 (HK‐2) cells. Cell viability assays (Cell Counting Kit 8) and reactive oxygen species assays (DCFDA assay) confirmed the effectiveness of liposomal NAC in inhibiting ferroptosis induced by slow iron uptake (1 mmol L−1 ferric ammonium citrate). Additionally, liposomal NAC protected cells from toxicity resulting from rapid iron uptake (50 μmol L−1 ferric ammonium citrate, 20 μmol L−1 8‐hydroxyquinoline). The liposomal formulation demonstrated an enhanced protective effect compared to free NAC.CONCLUSIONThese findings suggest that liposomal NAC more effectively protects human kidney cells from toxicity caused by different iron uptake models than free NAC, highlighting its potential as a superior treatment option for iron overload‐induced cellular damage. © 2024 The Author(s). Journal of Chemical Technology and Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry (SCI).

Development and pharmacological evaluation of liposomes and nanocapsules containing paroxetine hydrochloride

Depression is one of the most common psychiatric disorders. Nanotechnology has emerged to optimize the pharmacological response. Therefore, the aim of this work was to develop and characterize liposomes and nanocapsules containing paroxetine hydrochloride and evaluate their antidepressant-like effect using the open field and tail suspension tests in mice. Liposomes and nanocapsules were prepared using the reverse-phase evaporation and nanoprecipitation methods, respectively. The particle size of the formulation ranged from 121.81 to 310.73 nm, the polydispersity index from 0.096 to 0.303, the zeta potential from −11.94 to −34.50 mV, the pH from 5.31 to 7.38, the drug content from 80.82 to 94.36 %, and the association efficiency was 98 %. Paroxetine hydrochloride showed slower release when associated with liposomes (43.82 %) compared to nanocapsules (95.59 %) after 10 h. In Vero cells, in vitro toxicity showed a concentration-dependent effect for paroxetine hydrochloride nanostructures. Both nanostructures decreased the immobility time in the TST at 2.5 mg/kg without affecting the number of crossings in the open field test, suggesting the antidepressant-like effect of paroxetine. In addition, the nanocapsules decreased the number of groomings, reinforcing the anxiolytic effect of this drug. These results suggest that the nanostructures were effective in preserving the antidepressant-like effect of paroxetine hydrochloride even at low doses.

An Overview of Transferosomal Technology

Abstract: Ever since the invention of liposomes by Bangham in 1963, researchers have been fascinated by the vesicular carriers. Liposomes and niosomes have been used extensively by researchers for various routes such as oral and nasal. However, lately, it has been understood that traditional liposomes are not very significant when it comes to penetration. The use of nanovesicles in transdermal drug delivery systems has been enhanced exponentially ever since the discovery of ultra- deformable liposomes known as transfersomes or transferosomes. Transferosomes have numerous advantages, such as biocompatibility, biodegradability, flexibility, and deformability, so that they can pass through narrow constrictions. They have good entrapment efficiency and can act as a depot to sustain the release of drugs. The methods of preparation include the rotary film evaporation method, reverse phase evaporation method, vortexing sonication method, ethanol injection method, and freeze-thaw method. Transfersomes are characterized by particle size, zeta potential, polydispersity index, surface morphology, and encapsulation efficiency. Transferosomes have been successfully exploited for the enhancement of efficacy of many drugs like Hydroquinone, Itraconazole, Ivabradine, lornoxicam, minoxidil etc., via transdermal and nasal routes. The technology is easy to scale up. Consequently, it can be inferred that transfersomes are the future of transdermal drug delivery systems.

Nanoliposomes Encapsulated Rapamycin/Resveratrol to Induce Apoptosis and Ferroptosis for Enhanced Colorectal Cancer Therapy

ObjectivesMonotherapy is often ineffective for treating colorectal cancer. In this study, we developed PEG-modified liposomes loaded with rapamycin (Rapa) and resveratrol (Res) (Rapa/Res liposomes, or RRL) to investigate their therapeutic potential in colorectal cancer. MethodsRRL were constructed using the reversed-phase evaporation method. We assessed the cytotoxicity, apoptosis, and ferroptotic effects of RRL on colorectal cancer HCT116 cells. The anti-tumor efficacy of RRL was evaluated in HCT116 xenograft mice. ResultsRRL had a particle size of 86.67 ± 1.10 nm and a zeta potential of -33.13 ± 0.49 mV. The coloaded formulation demonstrated satisfactory performance both in vitro and in vivo, resulting in increased cytotoxicity to HCT116 cells and significant suppression of HCT116 xenografts tumor growth. Mechanically, RRL significantly increased the apoptosis rate of HCT116 cells, induced ROS accumulation in tumor cells, and effectively downregulated the expression of the ferroptosis-associated proteins GPX4 and SLC7A11, demonstrating its superior efficacy compared to that of Rapa liposomes (Rapa/Lps) or Res liposomes (Res/Lps) alone. ConclusionColoading Rapa and Res into liposomes to promote apoptosis and ferroptosis in tumor cells represents a promising strategy for the treatment of colorectal cancer.

Exploring the Antitumor Efficacy of PEGylated Liposomes Loaded with Licorice Extract for Cancer Therapy.

Glycyrrhizic Acid (GA), a compound derived from licorice, has exhibited promising anticancer properties against several cancer types, including Prostate Cancer (PCa) and Gastric Cancer (GCa). This study has introduced a novel approach involving the encapsulation of GA and Licorice extract (Lic) into Polyethylene Glycol Liposomes (PEG-Lip) and assessed their efficacy against AGS (human gastric cancer) and PC-3 (human prostate cancer) cells, marking the first report of this endeavor. We synthesized GA-loaded PEG-Lip (GA PEG-Lip) and Lic-loaded PEG-Lip (Lic PEG-Lip) through the reverse-phase evaporation method. Characterization of these liposomal formulations revealed their size, drug encapsulation, and loading efficiencies to be 110 ± 2.05 nm, 117 ± 1.24 nm; 61 ± 0.81%, 34 ± 0.47%; and 8 ± 0.41% and 4.6 ± 0.21%, respectively. Importantly, the process has retained the chemical structure of both GA and Lic. Furthermore, GA and Lic have been released from the PEG-Lip formulations in a controlled manner. In our experiments, both nanoformulations exhibited enhanced cytotoxic effects against AGS and PC-3 cells. Notably, GA PEG-Lip outperformed Lic PEG-Lip, reducing the viability of PC-3 and AGS cells by 12.5% and 15.9%, respectively. These results have been corroborated by apoptosis assays, which have demonstrated GA PEG-Lip and Lic PEG-Lip to induce stronger apoptotic effects compared to free GA and Lic on both PC-3 and AGS cells. This study has underscored the potential of encapsulating GA and Lic in PEG-Lip as a promising strategy to augment their anticancer efficacy against prostate and gastric cancers.

Improving Cancer Therapy: Design, Synthesis, and Evaluation of Carboplatin-Based Nanoliposomes against Breast Cancer Cell Lines

Objective: Cancer poses a significant challenge to modern medicine, with breast carcinoma being one of the most prevalent forms of the disease. Carboplatin is a commonly used chemotherapy drug for treating breast cancer, but its efficacy can be limited due to its poor water solubility and associated side effects. This study intended to enhance the therapeutic potency of carboplatin by encapsulating it within liposomal nanocarriers. Methods: We fabricated nanoscale liposomes loaded with carboplatin via a technique called reverse phase evaporation, and examined their physical attributes. We also studied the toxicity of these nanoliposomes on MDA-MB 231 breast cancer cells. Results: Our findings indicated that the liposomal nanoparticles possessed a negative zeta potential of -18.2 mV and an average size of 278 nm. The drug loading level was 2.2%, while the efficiency of drug encapsulation reached 58.5%. Of note, the cytotoxicity of carboplatin in its nanoliposomal form was markedly more potent against the MDA-MB 231 breast cancer cell line than the free drug (p-value less than 0.05). Conclusion: Our findings suggest that carboplatin-loaded liposomal nanocarriers could potentially serve as an advanced chemotherapeutic approach for the treatment of breast cancer, offering enhanced efficacy and reduced side effects compared to conventional carboplatin therapy. Further research is warranted to explore this novel delivery system’s benefits fully.

Methods for obtaining phospholipid vesicles to improve the bioavailability of sparingly soluble active pharmaceutical ingredients

Low bioavailability of practically insoluble pharmaceutical substances is a serious issue in the field of drug development. The article provides an overview of the main methods for preparing liposomal drugs, such as the lipid film hydration method, ultrasonication method, extrusion method, high pressure homogenization method, solvent injection method, detergent removal method, and the reversephase evaporation method.

Abstract 5750: Development of prostate specific membrane antigen-targeted liposomal zinc for the treatment of prostate cancer

Abstract Introduction: Treatment choices for early-stage prostate cancer include mostly watchful waiting, radical prostatectomy, or radiation therapy, whereas for advanced cases hormone therapy followed by chemo/radiotherapy are the preferred options, despite their notable side effects. There is a need for more effective therapy with lower side effects, especially for advanced scenarios. Unlike normal epithelial prostate cells, cancerous ones show reduced zinc (Zn) levels due to ZIP1 zinc transporter downregulation. Zn deficiency allows m-aconitase to remain active, making prostate cancer cells energy efficient and grow. High levels of Zn are needed to inhibit m-aconitase, the enzyme responsible for the first reaction in the Krebs cycle. This study aims to induce energy starvation and cell death of prostate cancer cells by inhibiting m-aconitase using the prostate-specific membrane antigen (PSMA)-targeting liposomal zinc. Methods: Liposomes were prepared by standard reverse phase evaporation method with DSPC, Cholesterol and DSPE-mPEG(2000) lipids. Zinc chloride was used to load Zn+2 into liposome following passive loading method. PSMA-expressing (PC3-PIP) and PSMA-null (PC3-LUC) human prostate cancer cells were used for in vitro studies. In the cell uptake study, cells were incubated with Dil-dye labeled liposomes for 2 hours before analyzing them by microscopy and flow cytometry. For the uptake inhibition study, cells were pre-incubated with PSMA-specific peptide for 2 hours before incubation with liposomes. Cytotoxicity studies were performed with PSMA-targeting Zn-loaded liposomes in PC3-PIP cells. Results: We conjugated the PSMA-specific peptide and control peptide (ASP3) with DSPE-PEG(3400)-NHS, and used them at various amounts (1 to 4 mol% of total lipid) to prepare Zn-loaded PSMA-targeting liposomes (TL) and control liposomes (CL), respectively. The average hydrodynamic diameter of all the liposomes was 120 to 130 nm, and zeta potential ranged from -16 to -12 mV. The encapsulation efficiency and loading capacity of Zn+2 were 0.6% and 8.5%, respectively. Cellular uptake studies showed a higher accumulation of PSMA-targeting liposome (TL) compared to non-targeting liposome (CL) in PSMA-expressing PC3 cells, whereas both liposomes showed low uptake in PSMA non-expressing cell. Furthermore, pre-treatment of PSMA-expressing cells with PSMA-specific peptide reduced the uptake of TL, indicating that the uptake of TL liposome is PSMA dependent. Finally, cytotoxicity studies showed that Zn-loaded TL, induces cell death in the PSMA-expressing cells. Overall, we developed PSMA-targeting Zn-containing liposomes that selectively target and kill PSMA-expressing prostate cancer cells. Future work includes testing the effect of Zn-loaded TL on m-aconitase activity and testing of in vivo efficacy of the formulation in prostate cancer models. Citation Format: Sujan K. Mondal, Alexander L. Klibanov, Anna Moore. Development of prostate specific membrane antigen-targeted liposomal zinc for the treatment of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5750.

DESIGN AND IN VIVO EVALUATION OF NAPROXEN-LOADED TRANSFEROSOMAL GEL FOR TRANSDERMAL DELIVERY

Objective: The main objective of the present study was to formulate, optimize, and evaluate naproxen transfersomal gels. Methods: Reverse phase evaporation was used to create thirteen different formulations of naproxen-loaded transfersomes. Using Response Surface Methodology (RSM) and Central Composite Designs (CCD), the influence of independent process variables, such as soy lecithin, cholesterol content and surfactant concentration, on dependent variables, such as entrapment effectiveness and vesicle size of naproxen transfersomes, was assessed. In vitro, ex-vivo, and in vivo drug release of formulations were also studied. Results: It was discovered that the NTG7 formulation of transfersomes had the maximum entrapment effectiveness and ideal vesicle diameter. The optimized NTG7 formulation displayed a maximum drug content of 97.4% and a maximum drug release of 88.03%. The release of naproxen from the final gel adhered to the Korsmeyer-Peppas release model. The ex-vivo drug release of the optimized formulation was found to be 85.91% for 24 h. The maximum drug concentration after oral administration was 843.54±7.67ng/ml, and Tmax was 3.0±0.08h. The improved formulation's AUC0-∞ was greater than the commercial formulation's. A higher drug concentration in the blood compared to the marketed formulation suggested better systemic absorption of naproxen from the gel formulation. After three months, at a temperature range of 2 to 8 °C, the formulation demonstrated correct semisolid consistency and good stability and there was also no appreciable change in the initial values of appearance, pH, and % drug content. Conclusion: The above findings imply that the gel created using naproxen-loaded transfersomes may be a potentially valuable new formulation.

Bilosomes: An Overview of Advancements in Preparation, Characterization, and Applications

Bilosomes, also known as bile vesicles, are bilayer vesicles that are formed by the hydration of bile salts. They have gained significant attention in recent years due to their potential use in various applications, including drug delivery and nutraceuticals. In this review, we discuss the various techniques used for the preparation of Bilosomes, including the film hydration, reverse phase evaporation, and ether injection methods. We also discuss the characterization techniques used to study the properties of Bilosomes, such as transmission electron microscopy (TEM), dynamic light scattering (DLS), and small angle X-ray scattering (SAXS). Additionally, we discuss the potential applications of Bilosomes, including their use as drug delivery systems for hydrophobic drugs, as well as their use in the food and nutraceutical industries. Finally, we discuss the current state of the Bilosomes market, including the currently available commercial products. Overall, this review provides a comprehensive overview of the recent advancements in Bilosomes research and highlights the potential of Bilosomes in various fields.

A novel strategy for partial purification of alkane hydroxylase from P. chrysogenum SNP5 through reconstituting its native membrane into liposome

Integral proteins or enzymes are still challenging to purify into their native state because of their need for an amphipathic environment and cofactors. Alkane hydroxylase (AlkB) is a membrane-bound enzyme that catalyzes the hydroxylation of a range of alkanes that have a broad spectrum of applications. In the current study, a novel approach has been explored for partial purification of alkane hydroxylase (AlkB) in its native state through restructuring the lipid bilayer of Penicilliumchrysogenum SNP5 into a liposome to extend the native and protective environment to AlkB enzyme. Three different methods i.e., reverse-phase evaporation method (RPEM), detergent-based method (DBM), and ethanol injection method (EIM) have been used for reconstituting its native membrane into liposome. On characterizing liposomes through fluorescence imaging, AFM, and particle size analysis, the reverse-phase evaporation method gave the best results based on the size distribution (i.e., 100–300 nm), the morphology of liposomes, and maximum AlkB specific activity (i.e., 140.68 U/mg). The maximum reconstitution efficiency of 29.48% was observed in RPEM followed by 17.3% in DBM and 12.3% in EIM. On the characterization of the purified AlkB, the molecular weight was measured of 44.6 KDa and the thermostability of liposomes synthesized with the RPEM method was obtained maximum at 55 °C. This approach may open a new strategy for the purification of integral enzymes/proteins in their native state in the field of protein purification and its applications in diversified industries.

LIPOSOMAL DELIVERY OF IMMUNOMODULATORY ANTIGEN OF B. MALAYI: ISOLATION, PREPARATION, CHARACTERIZATION, AND IMMUNE RESPONSES ASSESSMENT

Objective: The present study was aimed on developing and characterizing liposomal delivery system loaded with antigen of filaria parasite Brugia malayi extracted protein for assessment of humoral immune responses of antigen. Methods: Liposomes were prepared by reverse-phase evaporation method with slight modification using molar ratio of Soya PC: PE:Cholesterol in different molar concentrations. Results: The levels of F6 specific immunoglobulin (Ig) G1, IgG2a and IgG2b antibodies were found to be elevated in immunized animals over non-immunized controls. Analysis of IgG-subclasses revealed that all the subclasses at (1:25dilution) increased several folds over the controls with IgG1 showing the greatest increase (25.0-fold) followed by IgG2b (3.0-fold). Antibodies titers showed the many fold increment of titers on liposomized antigen groups (Gr.I; without booster dose and Gr.IV; with booster dose). IgG showed about 2.2 fold increment in Gr.IV than control group (Gr.V). IgG1 after booster dose showed about 25-fold increment followed by IgG2b than IgG2a. Conclusion: These results suggest that the liposomal antigen delivery system shows 25-fold IG-G responses in comparison to plain administrated antigen.