Emulsion Method Research Articles

Cubosomes; An Approach to Sustain and Improve the Ocular Delivery for Glaucoma Treatment: Box Benhken Optimization, Formulation, In Vitro Characterization and Ex Vivo Permeation Study

Introduction: In glaucoma, acetazolamide (ACZ) is used to lower intraocular pressure (IOP). Low aqueous solubility and decreased corneal permeation are two characteristics of ACZ. This research intends to enhance the ocular delivery of ACZ. Materials and Methods: Using a Box-Behnken design, cubosomes loaded with acetazolamide were made using the Melt dispersion emulsification and sonication method. The independent variables included Glyceryl monoolein (GMO), polyvinyl alcohol (PVA) and Poloxamer 407 (P407. The prepared formulations underwent evaluation for polydispersity index (PDI), particle size and entrapment efficiency. Thermal analysis, in vitro characterisation and permeation were among the additional tests performed on the developed formulation. Results: Optimized formulation showed PDI of 0.23 ± 0.03, mean particle size of 243 ± 4.2 nm, zeta potential of -26.1 ± 0.6 mV, entrapment efficiency of 73.99% and cubic structure under TEM. Drug penetration through goat cornea has increased, according to an ex vivo permeation study. The prepared cubosome safety, stability and ability to be delivered through the cornea were confirmed by the ocular irritation test. Conclusion: The optimized cubosomal formulation has the potential to improve glaucoma treatment and be regarded as promising for ocular delivery of ACZ.

Morphological and Structural Characterization of Encapsulated Arginine Systems for Dietary Inclusion in Ruminants

This research evaluated two methods of arginine encapsulation, melt emulsification and nanoprecipitation, using a lipid matrix of carnauba wax and commercial polymers (Eudragit®) as a protective material. The ratios of wax–arginine were 1:1, 2:1, 3:1, and 4:1, while those of Eudragit® RS:RL were 30:70 and 40:60 in proportions of 1:0.5 and 1:1 Eudragit®–arginine. The microcapsules were morphostructurally characterized by scanning electron microscopy, and a microelement analysis was performed via energy-dispersive X-ray spectroscopy and Fourier transform infrared spectroscopy. Additionally, in vitro digestibility was used to determine the protection efficiency. Both encapsulated systems presented regular (crystals) and spherical (microcapsules) polyhedral morphologies. Qualitative nitrogen decreased significantly as the wax ratio increased in the wax–arginine formulations. The formulations with a 1:1 Eudragit:–arginine ratio (1000 mg arginine) produced a higher nitrogen content in the encapsulated systems than the formulations containing 500 mg of arginine. The 2:1 and 3:1 wax–arginine formulations had the lowest degradability after 5 h of rumen fluid exposure (40.7 and 21.26%, respectively) in comparison with 100% unencapsulated arginine. The 3:1 wax–arginine formulation is an efficient encapsulating system which protects against rumen degradation. The more intense absorption bands at 1738 cm−1 and 1468 cm−1 associated with the C=O and C-H groups in carnauba wax indicate that arginine was more protected than in the other systems.

Green preparation of force-sensitive micro-capsules and their application in self-healing of cement sheath

Micro-capsule has been considered the most promising self-healing material, while the complex and high temperature polymerization process, and environmental unfriendly raw materials limits its wide application. Herein, the green micro-capsule composed of epoxy resin and calcium alginate were prepared at room temperature via needle drop method，O/W/O emulsion method and spraying method. The effects of preparation methods on the water absorption, encapsulation rate of micro-capsules were revealed, and the differences in the self-healing efficiency of cement containing these green micro-capsules were also studied. Among all samples, the micro-capsules prepared by O/W/O emulsion method (MCE-2) owned the highest water absorption 54.71 % and the highest epoxy resin content 48.1 %. Compared with the blank samples, adding micro-capsules improved the liquidity of cement slurry and facilitated the hydration process, related to the release of extra water. While, the micro-capsules caused additional defects in the cement matrix and weakened the compressive strength of cement. In the self-healing measurement, the composites with MCE-2 showed the biggest mass transfer pressure and least permeability before and after repair. The higher water absorption of MCE-2 significantly increased greater mass transfer resistance to block the erosion of external fluids before repair, and the higher epoxy resin content could ensure stronger sealing effect after repair, achieving the best self-healing performance.

High-Efficiency High-Temperature-Resistant Nanosilica Viscosity Reducer for Extra-Heavy Oil Viscosity Reduction.

Due to the fact that more conventional heavy oil recovery methods (heating, emulsification, dilution, and other methods) have many shortcomings, they cannot meet the demand of heavy oil exploitation. Therefore, there is a need for new recovery methods. In this paper, the surface of nano SiO2 was modified with a silane coupling agent, KH-560, to prepare a nanoviscosity reducer (NRV), which has high-temperature resistance (300 °C), calcium and magnesium resistance (Ca2+ + Mg2+ > 900 mg/L) and high viscosity reduction rate (>99%). FTIR and SEM measurements showed that KH560 has been successfully connected to the surface of SiO2. The particle size distribution of NRV is mainly distributed in 50-80 nm, which matches the results of SEM. The experimental results showed that the viscosity reduction rates of 1 wt % NRV on M-1 heavy oil before and after aging were 99.73% and 99.71%, respectively. The viscosity reduction effect of 1% NRV on M-1 heavy oil and the bleeding rate of emulsion formation were investigated when the oil-water ratio ranged from 9:1 to 1:9. The results showed that when the oil-water ratio was between 7:3 and 1:9, the viscosity reduction rate was greater than 99%. Besides, the bleeding rate of emulsion increases with the decrease of the oil-water ratio. What's more, static and dynamic adsorption experiments showed that the adsorption capacity of 1 wt % NRV was 1.746 mg/g and 1.668 mg/g sand, respectively. The interfacial tension experiment showed that the interfacial tension (IFT) between 1 wt % NRV and M-1 heavy oil was 0.052 mN/m, and low interfacial tension was beneficial to displace the oil in the formation pores. At the same time, the displacement effect of NRV on M-1 heavy oil at different concentrations (0.5, 1.0, and 1.5 wt %) and temperatures (200, 250, and 300 °C) was investigated by core flooding experiments. The results showed that the recovery rate increases with the increase of NRV concentration, and 1 wt % NRV at 300 °C will improve the recovery rate of M-1 heavy oil by 27.3% compared to steam flooding. NRV could reduce the viscosity of crude oil, which provides technical guidelines for the exploitation of heavy oil and extra heavy oil.

High efficiency azo dye removal via a combination of adsorption and photocatalytic processes using heterojunction Titanium dioxide nanoparticles on hierarchical porous carbon

Amidst the rapid development of the textile industry, wastewater problems also arise. High-performance materials for reactive black 5 (RB5) dye treatment by adsorption and photocatalysis were evolved using Titanium dioxide (TiO2) nanoparticles on carbon media. Herein, the synthesis of spherical carbon via the water–in–oil emulsion method alongside a sol–gel process and the production of TiO2 nanoparticles using the precipitation procedure of Titanium isopropoxide and carbonization at 700–900 °C for 2 h are a novel approach in this work. The characterization of these materials indicates that different temperatures result in distinct properties, for instance, raised pores on the surface of the media and changes in the crystal structure of TiO2. The results show that the as-synthesized material carbonized at 900 °C had distinguished dye adsorption, up to 430 ppm in 1 h, due to their high surface area and pore volume. On the contrary, the calcined 700 °C condition had the prominent photocatalytic efficiency on account of the heterojunction band gap between anatase and rutile crystal structure. A mixed phase minimizes the charge recombination, subsequently increasing the photocatalytic capability.

In vitro evaluation and in situ intestinal absorption characterisation of paeoniflorin nanoparticles in a rat model.

Purpose: the aim of this study was to improve the stability and bioavailability of paeoniflorin (PF) by using nanoparticle encapsulation technology. Methods: paeoniflorin nanoparticles (PF NPs) were prepared with PLGA as the carrier using the compound emulsion method. The nanoparticles were characterised by using a Malvern laser particle sizer, transmission electron microscope (TEM), X-ray diffraction (XRD) analyser, and Fourier-transform infrared (FT-IR) spectrometry. The PF NPs were subjected to a series of stability investigations (such as for 4 °C storage stability, pH stability, and thermal stability), lyophilisation protection technology investigations, and in vitro release studies. Finally, the intestinal absorption properties of PF and PF NPs were studied by the in situ single-pass intestinal perfusion (SPIP) rat model, using the effective permeability coefficient (P eff) and the absorption rate constant (K a) as relevant indexes. Results: the prepared nanoparticles had a particle size of 105.0 nm with blue opalescent, rounded morphology, uniform size, good stability and slow release. We found that 4% alginate was the best lyoprotectant for the PF NPs. In the intestinal absorption experiments, P eff was higher for the PF NPs group compared with the original PF material drug group in all intestinal segments (P < 0.05), and the absorption rate constant K a increased with the increase in the drug concentration. Conclusion: the nanoparticles produced by this method have good stability and a slow-release effect; they can thus improve the absorption of PF in rat intestines, helping improve the stability and bioavailability of PF and enhancing its pharmacological effects.

Simple-by-design approach for production of stabilizer-free cubosomes from phosphatidylglycerol and docosahexaenoic acid monoacylglycerol

Docosahexaenoic acid monoacylglycerol represents a promising lipid constituent in the development of drug nanocarriers owing to its amphiphilicity and the beneficial health effects of this docosahexaenoic acid precursor in various disorders including cancer and inflammatory diseases. Here, we describe the formation and characterization of simple-by-design and stabilizer-free lamellar and non-lamellar crystalline nanoparticles (vesicles and cubosomes, respectively) from binary mixtures of docosahexaenoic acid monoacylglycerol and phosphatidylglycerol, which is a ubiquitous amphiphilic component present in biological systems. At the physiological temperature of 37 °C, these single amphiphilic components tend to exhibit inverse hexagonal and lamellar liquid crystalline phases, respectively, on exposure to excess water. They can also be combined and dispersed in excess water by employing a high-energy emulsification method (by means of ultrasonication) to produce through an electrostatic stabilization mechanism colloidally stable nanodispersions. A colloidal transformation from vesicles to cubosomes was detected with increasing MAG-DHA content. Through use of synchrotron small-angle X-ray scattering, cryo-transmission electron microscopy, and nanoparticle tracking analysis, we report on the structural and morphological features, and size characteristics of these nanodispersions. Depending on the lipid composition, their internal liquid crystalline architectures were spanning from a lamellar (Lα) phase to biphasic features of coexisting inverse bicontinuous (Q2) cubic Pn3m and Im3m phases. Thus, a direct colloidal vesicle-cubosome transformation was detected by augmenting the concentration of docosahexaenoic acid monoacylglycerol. The produced cubosomes were thermally stable within the investigated temperature range of 5–60 °C. Collectively, our findings contribute to understanding of the imperative steps for production of stabilizer-free cubosomes from biocompatible lipids through a simple-by-design approach. We also discuss the potential therapeutic use and future implications for development of next-generation of multifunctional vesicles and cubosomes for co-delivery of docosahexaenoic acid and drugs in treatment of diseases.

Poly Lactic-co-Glycolic Acid (PLGA) Loaded with a Squaraine Dye as Photosensitizer for Antimicrobial Photodynamic Therapy.

Antimicrobial Photodynamic Therapy (aPDT) is an innovative and promising method for combating infections, reducing the risk of antimicrobial resistance compared to traditional antibiotics. Squaraine (SQ) dyes can be considered promising photosensitizers (PSs) but are generally hydrophobic molecules that can self-aggregate under physiological conditions. To overcome these drawbacks, a possible solution is to incorporate SQs inside nanoparticles (NPs). The present work deals with the design and development of innovative nanophotosensitizers based on poly lactic-co-glycolic acid (PLGA) NPs incorporating a brominated squaraine (BrSQ) with potential application in aPDT. Two designs of experiments (DoEs) based on the single emulsion and nanoprecipitation methods were set up to investigate how different variables (type of solvent, solvent ratio, concentration of PLGA, stabilizer and dye, sonication power and time) can affect the size, zeta (ζ)-potential, yield, entrapment efficiency, and drug loading capacity of the SQ-PLGA NPs. SQ-PLGA NPs were characterized by NTA, FE-SEM, and UV-Vis spectroscopy and the ability to produce reactive oxygen species (ROS) was evaluated, proving that ROS generation ability is preserved in SQ-PLGA. In vitro antimicrobial activity against Gram-positive bacteria in planktonic state using Staphylococcus aureus was conducted in different conditions and pH to evaluate the potential of these nanophotosensitizers for aPDT in the local treatment of infections.

Innovative dodecane-hexadecane/expanded graphite composite phase change material for sub-zero cold storage applications

Utilization of phase change materials (PCMs) for cold energy storage has garnered increasing attention. This study focuses on the development of a composite PCM by integrating a novel eutectic dodecane-hexadecane (DD-HD) PCM into expanded graphite (EG), to cater to the demands of sub-zero cold energy storage applications, such as cold chain logistics. The study identified the optimal mass ratio of DD-HD as 80:20. A series of DD-HD/EG composites were fabricated through a combination of high-speed emulsification and physical adsorption methods. The adsorption capacity of EG for the eutectic PCM mixture was evaluated by examining the impact of EG fraction on the leakage of the composite PCM, revealing that the optimum EG fraction is 10 wt%. The composite PCM displayed a sub-zero phase change temperature of -12.09 ℃, a high melting enthalpy of 178.75 kJ/kg, and a high thermal conductivity of 5.18 W/(m•K). The thermal conductivity of the composite was enhanced by 22.52 times compared to the base PCM. Thermogravimetric analysis indicated that the decomposition temperature of DD-HD/EG was significantly higher than that of the base PCM. The composite demonstrated outstanding thermal reliability, as evidenced by thermal cycle tests. With its impressive thermal characteristics, this composite shows great potential for various cold energy storage applications.

A novel nanodrug for the sensitization of photothermal chemotherapy for breast cancer in vitro.

Owing to the complexity of tumor treatment, clinical tumor treatment has evolved from a single treatment mode to multiple combined treatment modes. Reducing the tolerance of tumors to heat and the toxicity of chemotherapy drugs to the body, as well as increasing the sensitivity of tumors to photothermal therapy and chemotherapy drugs, are key issues that urgently need to be addressed in the current cancer treatment. In this work, polylactic acid-based drug nanoparticles (PLA@DOX/GA/ICG) were synthesized with good photothermal conversion ability by encapsulating the water-soluble anticancer drug doxorubicin (DOX), photothermal conversion agent indocyanine green (ICG) and liposoluble drug gambogic acid (GA) using a double emulsion method. The preparation process of PLA@DOX/GA/ICG was examined. Gambogic acid entrapped in PLA@DOX/GA/ICG nanoparticles could act as an HSP90 protein inhibitor to achieve bidirectional sensitization to chemotherapy and photothermal therapy under 808 nm laser irradiation for the first time, effectively ablating breast cancer cells in vitro. This nanodrug was expected to be used for the efficient treatment of tumors.

In vitro and ex vivo evaluation of chitosan gel containing metformin-loaded polymeric nanoparticles for topical treatment of melanoma

Objective The purpose of this study was to prepare and evaluate chitosan (CS) gel containing metformin hydrochloride (MET)-loaded polycaprolactone (PCL) nanoparticles (NPs) for topical treatment of melanoma. Significance Topical administration of MET-PCL NPs-CS gel improves penetration of drug, decreases side effects, and increases efficacy of treatment. Methods MET-PCL NPs were prepared by double emulsion method. Particle size, charge, encapsulation efficiency (EE), release, and morphology were evaluated. MET-PCL NPs-CS gel formulation was characterized in terms of organoleptic properties, pH, gelling time, viscosity, spreadability, release, and morphology. Cytotoxicity was performed on B16F10 cells. Ex vivo permeability was done with pig skin. Results The size, charge, and EE were found to be 180 ± 10 nm, −11.4 mV, and 93%. SEM images showed that NPs were spherical and smooth. An initial burst release followed by a slower release was observed. MET-PCL NPs-CS gel was found to be transparent. The pH was 4.9 ± 0.05. The gelation time was 1.6 ± 0.2 min. The viscosity results confirm pseudoplastic behavior of gel. The spreadability by % area was 392 ± 6.4 cm. The images showed that gelling network of CS gel was composed of suspended NPs. The viscosity was between 554 and 3503 cP. MET-PCL NPs-CS gel showed prolonged release up to 72 h. On B16F10 cells, gel showed higher cytotoxicity compared to MET solution. MET-PCL NPs-CS gel had twofold higher permeability in pig skin compared with MET-CS gel. Conclusion Topical administration of MET-PCL NPs-CS gel into the skin resulted in improved dermal penetration and this promising approach may be of value in effective treatment of melanoma and other skin cancers.

Innovative insights into the preparation of tamarind gum base film loaded with pomelo essential oil via a low-energy emulsion method combined with melatonin: Maximizing the dual functions of tamarind gum

Pomelo essential oil has good antioxidant and antimicrobial properties. The objective of this study was to modify the active packaging loaded with essential oil through low-energy emulsification, in order to achieve both film-forming and emulsifying properties of tamarind gum, and to investigate the surface activity of melatonin in the emulsion. When α:β mixing ratio of 60:40, the resulting film solution exhibited favorable apparent viscosity and reduced emulsion flocculation and precipitation phenomena. Particle size distribution analysis and optical microscopy results demonstrated that the obtained emulsion was uniformly dispersed at an α:β mixing ratio of 60:40, with droplet diameters mainly concentrated within the range of 1–100 μm. After undergoing film-forming treatment, E-3-F (α:β mixing ratio of 60:40) exhibited a smoother and more compact microstructure, leading to significant improvements in mechanical properties and barrier performance. Laser confocal microscopy imaging revealed that the addition of melatonin further reduced interfacial tension between oil and water, while simultaneously weakening hydrophobic interaction forces among oil droplets, thereby effectively inhibiting droplet aggregation behavior. Moreover, incorporation of melatonin also enhanced material compactness and imparted excellent antioxidant effects to the packaging material. Consequently, this study ultimately unveils potential for low-energy emulsification combined with melatonin application in bio-based packaging field while providing novel insights into developing multifunctional bio-based packaging.

Determination of Chlorogenic Acid in Solid-lipid Nanoparticles: Validation by UV-spectroscopy

Objective: The objective of the research was the development and validation of a simple, sensitive, accurate, robust, and precise UV-spectroscopic method for the quantitative determination of chlorogenic acid loaded in solid-lipid nanoparticles as per the guidelines of the International Conference on Harmonization. Methods: The solid-lipid nanoparticles of chlorogenic acid were prepared using the hot melt emulsification method and the high-speed homogenizer method. Glyceryl monostearate was used as a solid lipid, and Tween 80 was used as a surfactant for the preparation of chlorogenic acid-loaded solid lipid nanoparticles. The method was validated in terms of linearity, accuracy, precision, robustness, ruggedness, limit of detection, and limit of quantification. Results: The chlorogenic acid exhibited absorption maxima at the wavelength of 330 nm. The regression equation from the calibration curve was y=0.006x + 0.0193 with a correlation coefficient of 0.9989. The percentage recovery was found to be 99.92, 99.80, and 99.86, respectively (within the acceptable limit of 98-102%), which validated the accuracy of the method. Furthermore, the method exhibited precision, robustness, and ruggedness, as illustrated by a relative standard deviation (RSD) of less than 2%. The limit of detection and limit of quantification were found to be 6.97 and 21.13 μg/ml, respectively. Conclusion: It was concluded that the proposed spectrophotometer analytical method for the determination of Chlorogenic acid was found reliable, accurate, consistent, precise, accurate, and robust. Therefore, the proposed analytical technique could be an integral part of further evaluation and characterization of Chlorogenic acid-solid lipid nanoparticles.

Sustained release of inactivated H1N1 virus from degradable microparticles for extended vaccination

Influenza vaccines administered as intramuscularly injected inactivated viruses or intranasally administered live-attenuated viruses usually provide short-term protection against influenza infections. Biodegradable particles that provide sustained release of the antigen has been studied as an approach to extend vaccine protection. Here, we investigate sustained release of ultraviolet killed influenza A virus (A/PR/8/34(H1N1)) (kPR8) loaded into poly(D,L-lactic-co-glycolic acid) (PLGA) microparticles. Particles were prepared using the double emulsion method, and polymer molecular weight (MW), polymer hydrophobicity, polymer concentration in the organic phase, and the amount of killed virus were varied to obtain a range of particles. Formulations included PLGA 50:50 (2–6, 7–17 kDa), PLGA 75:25 (4–15 kDa), and 50/50 PLGA 75:25 (4–15 kDa)/PCL (14 kDa). Additionally, NaOH was co-encapsulated in some cases to enhance particle degradation. The structure of the particles was explored by size measurements and electron microscopy. The kPR8 release profiles were measured using hemagglutinin ELISA. The concentration of the polymer (PLGA) in the organic phase and polymer MW significantly influenced virus loading, while polymer MW and co-encapsulation of NaOH modulated the release profiles. Mice receiving a single intramuscular injection of NaOH microparticle-encapsulated kPR8 were partially protected against a lethal influenza challenge 32 weeks post immunization. Microparticle (MP) vaccination induced a gradual increase in PR8-specific IgGs dominated by IgG1 in contrast to the rapid IgG2a-biased response elicited by soluble kPR8 immunization. Our results indicate that vaccine-NaOH co-loaded PLGA particles show potential as a single dose vaccination strategy for extended protection against influenza virus infection.

Kojic Acid Containing Novel Drug Delivery System on Facial Dyschromia: Characterization and Their Evaluation

Stubborn skin pigmentation issues like post-inflammatory hyperpigmentation (PIH) and melasma are the primary reasons people seek cosmetic consultations. Treating these conditions topically is challenging, as it involves inhibiting various stages of production of the pigment process. A powerful tyrosinase inhibitor like, kojic acid (KA) is employed as a formulation to regulate pigmentation production by suppressing the melanogenesis process. It’s important to note that the application of KA has been approved by the Food and Drug Administration (FDA), US, for dermatological treatments. The goal of this investigation was to formulate a nanoemulsion containing kojic acid for skin delivery using an emulsification method. The characteristics of the KA nanoemulsion were thoroughly examined through techniques like fourier transform infrared spectroscopy, (FTIR) particle size analysis and transmission electron microscopy (TME). In addition, the formulation’s performance was evaluated through both ex-vivo permeation study and in-vitro release study. Analysis of the FTIR, X-ray diffraction (XRD), and differential scanning calorimetry (DSC) results revealed that kojic acid with other ingredients in the formulation did not exhibit any chemical interactions. The kojic acid nanoparticles that were produced exhibited a spherical shape and were uniformly distributed, with an average size diameter of 184 nm. In in-vitro tests, it was observed that 87.67% of the drug was released within 12 hours. Moreover, ex-vivo permeation evaluation demonstrated that 81.24% of the drug permeated the skin within 8 hours of application. The thermal stability studies confirmed the stability of the kojic acid nanoemulsion, with no signs of creaming, cracking, or phase separation in the formulation. In conclusion, the findings of this study suggest that the kojic acid nanoemulsion holds great promise as an effective means for delivering kojic acid within the upper layers of the skin for treating of facial dyschromia.

Targeted delivery of SN38 to breast cancer using amphiphilic diblock copolymers PHPMA-b-PBAEM as micellar carriers with AS1411 aptamer

Breast cancer treatment can be challenging, but a targeted drug delivery system (DDS) has the potential to make it more effective and reduce side effects. This study presents a novel nanotherapeutic targeted DDS developed through the self-assembly of an amphiphilic di-block copolymer to deliver the chemotherapy drug SN38 specifically to breast cancer cells. The vehicle was constructed from the PHPMA-b-PEAMA diblock copolymer synthesized via RAFT polymerization. A single emulsion method was then used to encapsulate SN38 within nanoparticles (NPs) formed from the PHPMA-b-PEAMA copolymer. The AS1411 DNA aptamer was covalently bonded to the surface of the micellar NPs, producing a targeted DDS. Molecular dynamics (MD) simulation studies were also performed on the di block polymeric system, demonstrating that SN38 interacted well with the di block. The in vitro results demonstrated that AS1411- decorated SN38-loaded HPMA NPs were highly toxic to breast cancer cells while having a minimal effect on non-cancerous cells. Remarkably, in vivo studies elucidated the ability of the targeted DDS to enhance the antitumor effect of SN38, suppressing tumor growth and improving survival rates compared to free SN38.

Oral administration of recombinant outer membrane protein A-based nanovaccine affords protection against Aeromonas hydrophila in zebrafish.

Aeromonas hydrophila, an opportunistic warm water pathogen, has always been a threat to aquaculture, leading to substantial economic losses. Vaccination of the cultured fish would effectively prevent Aeromoniasis, and recent advancements in nanotechnology show promise for efficacious vaccines. Oral delivery would be the most practical and convenient method of vaccine delivery in a grow-out pond. This study studied the immunogenicity and protective efficacy of a nanoparticle-loaded outer membrane protein A from A. hydrophila in the zebrafish model. The protein was over-expressed, purified, and encapsulated using poly lactic-co-glycolic acid (PLGA)nanoparticles via the double emulsion method. The PLGA nanoparticles loaded with recombinant OmpA (rOmpA) exhibited a size of 295 ± 15.1nm, an encapsulation efficiency of 72.52%, and a polydispersity index of 0.292 ± 0.07. Scanning electron microscopy confirmed the spherical and isolated nature of the PLGA-rOmpA nanoparticles. The protective efficacy in A. hydrophila-infected zebrafish after oral administration of the nanovaccine resulted in relative percentage survival of 77.7. Gene expression studies showed significant upregulation of immune genes in the vaccinated fish. The results demonstrate the usefulness of oral administration of nanovaccine-loaded rOmpA as a potential vaccine since it induced a robust immune response and conferred adequate protection against A. hydrophila in zebrafish, Danio rerio.

NLC Delivery of EGFP Plasmid to TM4 Cell Nuclei for Targeted Gene Therapy.

This study evaluated whether a nanostructured lipid carrier (NLC) delivery system could safely and accurately deliver nucleic acids to the cell nucleus using the enhanced green fluorescent protein (EGFP)-C1 plasmid model. The NLC was formulated using the emulsification method and equipped for cationic lipid-mediated transfection with 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), which interacts electrostatically with nucleic acid. The NLC attributes, including size, polydispersity index, and zeta potential, were assessed by dynamic light scattering (DLS). The morphological structure was analyzed using transmission electron microscopy. Entrapment efficiency was evaluated by a direct method. Cellular uptake mechanisms of pEGFP-C1-NLC and the ability of pEGFP-C1 to penetrate the nucleus of TM4 cells to express EGFP were observed using confocal microscopy. pEGFP-C1-NLC exhibited particle sizes in the range 56-88 nm with a particle charge range of -6.0 to+1.3 mV. The polydispersity index<0.5 showed good size uniformity, and entrapment efficiency of pEGFP-C1in the NLC was 92.06±2.295%. The NLC formulation was internalized predominantly via caveolae-mediated endocytosis, as indicated by EGFP expression following successful delivery of pEGFP by the NLC into the cells. NLC formulation could deliver genetic material to the nucleus and could be considered a gene therapy candidate for spermatogenesis.

Pickering emulsion stabilized by cellulose nanofibril from pineapple leaves for biofoam manufacture

Cellulose nano fibril (CNF) can be selected as an agent of pickering emulsion in biofoam manufacture. This study thoroughly investigates the pickering emulsion method for producing biofoam, the characterization of CNF from pineapple leaves, the production of biofoam with various concentration of CNF and surfactant, and the effect of these parameters on the properties of wet foam and dry foam. CNF was created by mechanical grinding then the stability and morphology were evaluated. The pickering emulsion mechanism in biofoam manufacture was investigated using foamability and foam stability with various CNF and surfactant concentration. As the results, CNF concentrations will create varying zeta potential values. The morphology of CNF shows that it has a structure-like entangled network. The stability test demonstrates that adding CNF improves the stability and foamability of biofoam by the pickering effect. Biofoam without CNF has an unstable structure and is easily collapse when dried in an oven. The concentration of CNF and the amount of surfactant utilized altered the qualities of both wet and dry foam. The lower concentration of CNF and the addition of surfactant could increases foamability, high porosity, water absorption, and biodegradability; as well as low density, contact angle and bending characteristics. In biofoam that consists of 2 % CNF, the stability of wet foam increases as the amount of surfactant added, resulting in a biofoam with low tensile strength.

Antiproliferative activity of syringic acid-loaded nanostructured lipid carriers against MCF-7 human breast carcinoma cells

Syringic acid (SA) is a phenolic herbal compound known to exert antiproliferative properties. However, its poor aqueous solubility and low bioavailability lead to limited therapeutic efficacy in treating breast cancer. Hence, to overcome the above problems, the current study aims to explore the quality by design (QbD) optimized syringic acid loaded nanostructured lipid carriers (SA-NLCs) to treat breast cancer. Here, SA-NLCs were formulated via the hot-melt emulsification method and optimized using the Box-Behnken design. The SA-NLCs exhibited spherical shapes with a particle size and zeta potential of 142.7 ± 4 nm and −14.5 ± 3.6 mV, respectively. The differential scanning calorimetry and powder XRD analyses showed the conversion of crystalline to amorphous form of SA after entrapping into NLCs. Also, the optimized SA-NLCs showed sustained drug release of 75.5 % and 97.6 % in pH 5.5 acetate buffer solution and pH 7.4 phosphate buffer saline respectively. The drug release kinetics study revealed that the optimized SA-NLCs exhibit the Weibull model with the Fickian drug release mechanism. Further, a significantly improved cytotoxicity of the optimized SA-NLCs (IC50: 3.8 ± 0.2 μg/mL) was observed as compared to free SA (IC50: 14.5 ± 1.2 μg/mL) on MCF-7 human breast carcinoma cell lines. Thus, the improved cytotoxicity of SA-NLCs on MCF-7 cell lines can be a suitable strategy for breast cancer treatment.