Malvern Zetasizer Research Articles

Berberine-Loaded Bovine Serum Albumin Nanoparticles: Fabrication, Characterization, and Drug Release Evaluation

Abstract Introduction/Objective Introduction: Numerous studies have highlighted the diverse biochemical and pharmacological properties of berberine (BER), including its anti-inflammatory, antioxidant, anticancer, and hypolipidemic effects. Nanoparticles play a crucial role in enhancing drug delivery by providing a larger surface area, facilitating quicker dissolution in the bloodstream. Methods/Case Report Methods: Blank bovine serum albumin nanoparticles (BSA NPs) were prepared using a desolvation process, followed by the fabrication of berberine-loaded BSA nanoparticles (BER-NP). Physicochemical characterization, including particle size and zeta potential, was conducted using a Malvern Zetasizer. The mean particle size and distribution were determined using photon correlation spectroscopy (PCS), and transmission electron microscope (TEM) images were obtained for visualization. Fourier transform infrared (FTIR) spectra and differential scanning calorimetry (DSC) thermograms were recorded to assess drug encapsulation efficiency (EE). Berberine release from BER-NPs was evaluated using dialysis membranes. Results (if a Case Study enter NA) Results: FTIR spectra and DSC thermograms confirmed successful drug loading into the nanoparticles. Encapsulation efficiency was determined by UV absorption spectroscopy, revealing high efficiency in the encapsulation process. Dialysis membranes demonstrated controlled release kinetics of berberine from BER-NPs, indicating their potential for targeted drug delivery. Conclusion Conclusion: Berberine-loaded bovine serum albumin nanoparticles exhibit favorable characteristics for drug delivery applications, including enhanced permeation across biological barriers. These nanoparticles hold promise for targeted therapy and combination drug regimens, offering potential benefits for various medical interventions.

Preparation and Characterization of Berberine-Loaded Bovine Serum Albumin Nanoparticles: Encapsulation Efficiency and Release Rate Determination

Abstract Introduction/Objective Introduction: Berberine (BER) has garnered attention in various studies for its diverse biochemical and pharmacological properties, including anti-inflammatory, antioxidant, antidepressant, anticancer, antihyperglycemic, hypolipidemic, antihypertensive, and hepatoprotective effects. Nanoparticles serve as efficient carriers for drugs, offering increased surface area and enhanced dissolution in the bloodstream, thereby improving drug delivery. Methods/Case Report Methods: Blank Bovine Serum Albumin Nanoparticles (BSA NPs) were synthesized using a desolvation process, followed by the fabrication of Berberine-loaded Bovine Serum Albumin Nanoparticles (BER-NP). Physicochemical characterization, including particle size and zeta potential, was conducted using a Malvern Zetasizer and dynamic light scattering (DLS). Transmission electron microscopy (TEM) was employed to visualize the morphology of both blank BSA NPs and BER-loaded nanocarriers. Results (if a Case Study enter NA) Results: Fourier-transform infrared (FTIR) spectra and Differential Scanning Calorimetry (DSC) thermograms were obtained for free berberine, BSA NPs, and BER-NPs. Encapsulation efficiency (EE) was determined by measuring the concentration of free BER in the supernatant after centrifugation, with EE calculated based on the difference between total added berberine and the measured supernatant concentration. Dialysis membranes were utilized to assess BER release from BER-NPs in vitro, with initial BER content in nanoparticles determining the release profile. Conclusion Conclusion: The prepared nanoparticles have demonstrated the potential to enhance drug permeation across epithelial and endothelial barriers. This approach holds promise for targeted drug delivery to specific sites, facilitating combined therapy with distinct modalities or drugs.

Preparation and evaluation of liposomal gel containing Neolamarckia cadamba leaves extract for anti-inflammatory activity

The objective of present research work is to develop liposomes as a carrier system for methanolic extract, its incorporation in to gel formulations and to characterize the prepared and develop liposomal gel formulation. Formulation was carried out by thin film technique. Scanning electron micrograph of the prepared nanosponges at 50.00 kx magnification showed that the liposome was porous with a smooth surface morphology and spherical shape. The porous nature of liposome was clearly observed in the SEM images. Particle size and zeta potential was determined by Malvern Zeta sizer. The particle size analysis confirmed that the prepared sample were in the nanometer range. Average particle size obtained for the formulations F1 to F5 were 162.7 nm to 195.6 nm. Zeta potential values of liposome indicated that the formulated liposomes are stable. The amount of drug being entrapped in liposome was calculated and all the prepared liposome was found to possess very high entrapment efficiency. The viscosity of liposome loaded gel is found to 6842±0.32cps. The pH of liposome loaded gel is 6.3 and spreadability is 12.11, indicating that liposome loaded gel has high release and permeability. The in vitro anti-inflammatory effect of Neolamarckia cadamba extract was evaluated against denaturation of egg albumin. The present findings exhibited a concentration dependent inhibition of protein (albumin) denaturation by Neolamarckia cadamba throughout the concentration range of 20-100μg/ml. Diclofenac sodium (at the concentration range of 20-100 μg/ml) was used as reference drug which also exhibited concentration dependent inhibition of protein denaturation; however, the effect of diclofenac sodium was found to be less active when compared with extract name. Moreover, the results demonstrated that the liposome-based drug delivery approach could be a valuable tool to improve the therapeutic efficacy of phytochemicals by improving their absorption, and bioavailability via altering their physicochemical and release properties. Keywords: Liposomes, Neolamarckia cadamba, Qualitative phytochemical screening, Thin film technique, In vitro anti-inflammatory activity.

Developing fish oil emulsion gel enriched with Lentinula edodes single cell protein and its effect on controlling the growth of Acinetobacter baumannii

In this study, the characterization of fish oil (FO) emulsion gel (EGEL) containing single cell protein (SCP) produced from Lentinula edodes (L. edodes) and its potential inhibition against Acinetobacter baumannii (A. baumannii) were investigated. Oil extracted from the fish liver was emulsified with tween 80 and water, and then gelled using gelatin with the assistance of an ultrasonic homogenizer. The characteristics and surface analysis of SCP-EGEL were examined using FTIR (Fourier-transform infrared spectroscopy) and SEM (Scanning electron microscope). The particle size distribution and zeta potential of SCP-EGEL were measured using a Malvern Zetasizer. When SCP-EGEL was applied to the surface of the medium inoculated with A. baumannii, the inhibition zone (IZ) was 8.2 mm. An expansion of the IZ was observed (10.2 mm) when SCP-EGEL was applied to a fish skin (FS) surface prepared in the shape of a 6-mm diameter disc. In the SEM images, when SCP was added to lipo gel, the gel structure appeared flattened or swollen in some areas. The appearance of SCP cells being covered with gel gave the impression that they have a secondary wall. Therefore, the resulting complex can potentially be used as an additive in animal and human nutrition, in functional food coatings to suppress A. baumannii, and in fish feed to enrich it with protein.

Effect of surfactant's charge properties on behavior, physiology, and biochemistry and the release of microcystins of Microcystis aeruginosa

Surfactant pollution is escalatitheng in eutrophic waters, but the effect of surfactant charge properties on the physiological and biochemical properties of toxin-producing microalgae remains inadequately explored. To address this gap, this study explores the effects and mechanisms of three common surfactants―cetyltrimethylammonium bromide (CTAB, cationic), sodium dodecyl sulfate (SDS, anionic), and Triton X-100 (nonionic)―found in surface waters, on the agglomeration behavior, physiological indicators, and Microcystin-LR (MC-LR) release of Microcystis aeruginosa (M. aeruginosa) by using UV–visible spectroscope, Malvern Zetasizer, fluorescence spectrometer, etc. Results suggest that charge properties significantly affect cyanobacterial aggregation and cellular metabolism. The CTAB-treated group demonstrates a ∼5.74 and ∼9.74 times higher aggregation effect compared to Triton X-100 and SDS (300 mg/L for 180 min) due to strong electrostatic attraction. Triton X-100 outperforms CTAB and SDS in polysaccharide extraction, attributed to its higher water solubility and lower critical micelle concentration. CTAB stimulates cyanobacteria to secrete proteins, xanthohumic acid, and humic acids to maintain normal physiological cells. Additionally, the results of SEM and ion content showed that CTAB damages the cell membrane, resulting in a ∼90% increase in the release of intracellular MC-LR without cell disintegration. Ionic analyses confirm that all three surfactants alter cell membrane permeability and disrupt ionic metabolic pathways in microalgae. This study highlights the relationship between the surface charge properties of typical surfactants and the dispersion/agglomeration behavior of cyanobacteria. It provides insights into the impact mechanism of exogenous surfactants on toxic algae production in eutrophic water bodies, offering theoretical references for managing surfactant pollution and treating algae blooms.

PREPARATION OF LIPID NANOCARRIER FORMULATIONS AND CYTOTOXICITY STUDIES OF DONEPEZIL

Objective: Our research endeavors to discover innovative formulations for the pharmaceutical component of radiopharmaceuticals, which are used to diagnose Alzheimer's disease. Our approach involves the incorporation of Donepezil, a proven active ingredient, into lipid-based nanocarrier systems. Additionally, we have conducted a comprehensive study on the cytotoxicity of Donepezil as a vital aspect of our research. Material and Method: Two distinct techniques were employed in creating nanocarrier formulations: emulsion and sonication. Malvern Zeta Sizer measurements were conducted to assess the properties of the prepared formulations. In addition, the cell proliferation kit II (XTT) was used to evaluate the cytotoxicity of the active ingredient Donepezil. Result and Discussion: Formulations with particle sizes ranging from 100-200 nm have been selected based on the results of characterization studies. Cytotoxicity assays have shown that amounts of Donepezil (50, 100, 500, 1000, 2000, and 5000 µg/ml) are biocompatible. These findings confirm the optimal formulation parameters for producing high-quality Donepezil-based pharmaceutical products. The characterization studies of the prepared formulations have shown that they have the potential to be used in the diagnosis of Alzheimer's disease.

Capture of fully assembled secretory immunoglobulin A by affinity chromatography with nanobodies as ligands

Secretory immunoglobulin A (sIgA) has strongpotential for passive immunization and other therapeutic applications. This complex molecule with a molecular mass of 410 kDa consists of two IgA monomers linked by a connecting chain (J) and a secretory component (SC). Fully assembled sIgA was overexpressed in CHO cells. In order to exploit the therapeutic potential of recombinant sIgA expressed in CHO cells, a platform process for capture of the antibody directly from the clarified culture supernatant using available affinity chromatography material was investigated. The hydrodynamic radius was 10.2 ± 0.9 nm determined by dynamic light scattering and the isoelectric point 5.5 by electrophoretic mobility using the Malvern Zetasizer. Capture Select IgA and Capture Select IgA-CH1 chromatography media with camelid-derived single-domain antibody fragments comprising the 3 CDR domain antibody fragment with a 13 kD or 14 kDa as ligand were used to purify the antibody. sIgA was directly captured from the clarified culture supernatant and a highly purified, full-length, correctly assembled sIgA was obtained. The residence time must be in the range of 5–10 min to achieve a high binding capacity. The effective diffusivity of sIgA was in the range of 10−9 cm2 s−1 and the dynamic binding capacity was in the range of 10 mg/ml packed bed. We conclude that the current version of the resin has a pore size that is too small for this type of molecule to achieve substantial resin utilization and productivity, although affinity ligand-based purification led to high degree of purity suited for an industrial capture process.

IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8+ T, and CD4+ T Cells.

The drug resistance and low response rates of immunotherapy limit its application. This study aimed to construct a new nanoparticle (CaCO3-polydopamine-polyethylenimine, CPP) to effectively deliver interleukin-12 (IL-12) and suppress cancer progress through immunotherapy. The size distribution of CPP and its zeta potential were measured using a Malvern Zetasizer Nano-ZS90. The morphology and electrophoresis tentative delay of CPP were analyzed using a JEM-1400 transmission electron microscope and an ultraviolet spectrophotometer, respectively. Cell proliferation was analyzed by MTT assay. Proteins were analyzed by Western blot. IL-12 and HMGB1 levels were estimated by ELISA kits. Live/dead staining assay was performed using a Calcein-AM/PI kit. ATP production was detected using an ATP assay kit. The xenografts in vivo were estimated in C57BL/6 mice. The levels of CD80+/CD86+, CD3+/CD4+ and CD3+/CD8+ were analyzed by flow cytometry. CPP could effectively express EGFP or IL-12 and increase ROS levels. Laser treatment promoted CPP-IL-12 induced the number of dead or apoptotic cell. CPP-IL-12 and laser could further enhance CALR levels and extracellular HMGB1 levels and decrease intracellular HMGB1 and ATP levels, indicating that it may induce immunogenic cell death (ICD). The tumors and weights of xenografts in CPP-IL-12 or laser-treated mice were significantly reduced than in controls. The IL-12 expression, the CD80+/CD86+ expression of DC from lymph glands, and the number of CD3+/CD8+T or CD3+/CD4+T cells from the spleen increased in CPP-IL-12-treated or laser-treated xenografts compared with controls. The levels of granzyme B, IFN-γ, and TNF-α in the serum of CPP-IL-12-treated mice increased. Interestingly, CPP-IL-12 treatment in local xenografts in the back of mice could effectively inhibit the growth of the distant untreated tumor. The novel CPP-IL-12 could overexpress IL-12 in melanoma cells and achieve immunotherapy to melanoma through inducing ICD, activating CD4+ T cell, and enhancing the function of tumor-reactive CD8+ T cells.

An effective treatment approach of liposomally encapsulated metformin in colon cancer.

Metformin is a drug that is widely used in the treatment of type-2 diabetes and its anticarcinogenic effect has been detected in many studies since the 2000s. Metformin has a short half-life and poor biocompatibility, which limits the activity of the drug. As a solution to this situation, our study aimed to increase the anticarcinogenic effects and reduce the side effects of metformin in colon cancer by liposomal encapsulation. For this purpose, in our study, liposome production was carried out using the thin film hydration method. The amount of metformin loaded in liposomes was determined by a standard absorbance curve at 237nm. Size distributions and membrane zeta potentials of the liposomes were evaluated with Malvern Zetasizer ZS90. Transmission electron microscopy was performed by staining the liposomes negatively with uranyl acetate. Cultured HT-29 cells were treated with liposomal metformin or free metformin at concentrations of 0, 10, 20, and 40mM for 24 and 48h. At the end of the treatment period, cell viability was evaluated by CellTiter-Glo luminescent cell viability test. The anticarcinogenic effects of liposomal and free metformin on HT-29 cells were compared. As a result, liposome encapsulated metformin treatment for 24h was more effective on HT-29 cells at 20- and 40-mM concentrations causing significantly greater decrease in the IC-50 dose compared to the free metformin. The result suggests that liposomal encapsulated metformin may offer a promising approach to increase the efficacy of the drug in the treatment of colon cancer.

Optimization of the filtration and centrifugation steps for the isolation of exosome-like nanoparticles (ELNs) from grapes

ABSTRACT Exosome-like nanoparticles (ELNs) are a class of nanosized extracellular vesicles from plants that can deliver drugs, siRNA, DNA, and proteins to various types of cells. Purification of ELNs from plants are tricky when compared to that from body fluids or cell culture medium because the initial suspension of the former contains high amounts of sugar, fiber, fat, etc. In this study, we have optimized the different steps for the isolation of ELNs from grapes. After preparing the juice, the bulk density was measured by Brix analysis and followed the same bulk density whenever doing the isolation. Differential centrifugation at various speeds and times and membrane filtration using different pore-size membranes were also optimized before the final step of ELN separation by ultracentrifugation. The size, zeta potential, and PDI value of the isolated ELNs were measured using a Malvern Zeta Sizer. SEM analysis showed that the grape ELN has an average size of 140.8 ± 10.75 nm. The results indicated that filtration and centrifugation at different levels are essential before the ultracentrifugation for isolating homogenous grape ELNs with desired size, zeta potential and PDI value. From the optimized method, we have obtained 4.8 ± 0.509 mg protein equivalents of ELNs from 100 g of grapes.

Bovine serum albumin and folic acid-modified aurum nanoparticles loaded with paclitaxel and curcumin enhance radiotherapy sensitization for esophageal cancer

Background Nanocarrier systems have been used in the study of esophageal cancer (EC) and other diseases, with significant advantages in improving the non-targeted and nonspecific toxicity of traditional formulations. Some chemotherapeutic drugs and high atomic number nanomaterials have sensitization effects on ionizing radiation and can be used as chemoradiation sensitizers. Methods Aurum (Au) nanoparticles were modified by bovine serum albumin (BSA) and folic acid (FA), and were core-loaded with paclitaxel (PTX) and curcumin (CUR). The basic characteristics of FA-BSA-Au@PTX/CUR nanomedicines were evaluated by transmission electron microscopy, Fourier transform infrared spectroscopy, and Malvern Zetasizer. The encapsulation and release of drugs were monitored by ultraviolet–visible spectrophotometry (UV–Vis). The biological toxicity and radiotherapy sensitization effect of FA-BSA-Au@PTX/CUR were observed by cell viability, colony formation, cell apoptosis, cell cycle distribution, and γ-H2AX analysis experiments. Results The prepared nanomedicines showed good stability and spherical morphology. The results of cell uptake and cell viability detection revealed that FA-BSA-Au@PTX/CUR could specifically target EC cell KYSE150 and exert a certain inhibitory effect on proliferation, with no obvious toxicity on healthy cells Het-1A. In addition, the results of the colony formation experiment, cell apoptosis detection, cell cycle distribution, and γ-H2AX analysis showed that compared with X-rays alone, FA-BSA-Au@PTX/CUR combined with X-rays exhibited relatively stronger radiotherapy sensitization and anti-tumor activity. Conclusions FA-BSA-Au@PTX/CUR could target EC cancer cells and act as a safe and effective radiotherapy sensitizer to improve the radiotherapy efficacy of EC.

Emulsion Polymerisation of Poly(Methyl Methacrylate)-Grafted-Graphene Oxide (PMMA-GO): Effect of Surfactant Concentration on Colloidal Stabilization

Graphene oxide (GO) exhibits a wide range of outstanding mechanical, electrical, and physical characteristics, and it is of substantial interest to impart such qualities onto polymeric materials such as poly (methyl methacrylate) PMMA for wider specialized functionalization. The attention of this work is on the development of emulsion polymerization procedure to prepare PMMA-GO nanocomposite and the effect of sodium dodecyl sulphate (SDS) surfactant dosages incorporated during the polymerization, on the effect of PMMA surface interactions in oil. The grafting efficiency is quantified using the Fourier-transform infrared spectroscopy (FTIR), and the effect of surfactant concentration on PMMA-GO stability is examined using UV-Visible spectroscopy, zeta potential and particle size analyses using the Malvern Zetasizer. The surfactant free emulsion has a better stabilization in terms of zeta potential analysis compared to emulsion of PMMA-GO with 0.32, and 0.4 wt. % of SDS. The polymerized PMMA-GO can be used as a model system to alter wax crystallization at low temperatures in oil and gas industries.

Using a novel approach to characterize the surface reactivities of silica-rich ferrihydrite and biogenic cyanobacteria-ferrihydrite aggregates and the implications for Archean ocean geochemistry

Precambrian banded iron formations (BIF) are iron- and silica-rich chemical sedimentary rocks that are commonly used as paleo-redox proxies for Archean and Paleoproterozoic seawater geochemistry. At the onset of the Great Oxidation Event (herein GOE) around 2.4 Ga, cyanobacteria flourished with increasing nutrient fluxes due to oxidative weathering on land. In turn, this led to increased primary productivity that facilitated the permanent shift from a reducing Earth atmosphere to an oxidizing one. Interestingly, the duration of GOE also overlapped with one of the most prolific periods of BIF deposition.It is widely accepted that cyanobacteria were likely responsible for BIF formation during the GOE. Oxidation of dissolved Fe(II) by oxygen produced from cyanobacteria forms a metastable and amorphous mineral phase ferrihydrite, Fe(OH)3. As an essential component in both ancient BIF deposits and various modern ecosystems, the surface reactivity of ferrihydrite has been extensively studied under different conditions (i.e., pH and ionic strengths). Not only are the highly reactive surfaces of ferrihydrite particles important shuttles for trace element transport from the water column to the sediment pile, but previous studies have also demonstrated that cyanobacterial cells and ferrihydrite tend to aggregate at seawater pH. This means that ferrihydrite was also a vector for the transport of organic carbon to the seafloor. However, a complicating issue is how co-ions affect the surface reactivity of ferrihydrite, specifically dissolved silica which was abundant in ancient seawater. Although previous studies have demonstrated that silica can passivate the surface reactivity of ferrihydrite, what remains unclear is how silica impacts ferrihydrite-biomass aggregation. To fill this knowledge gap, we formed both silica-spiked ferrihydrite and cyanobacteria-ferrihydrite aggregates in situ and subsequently conducted empirical potentiometric acid-base titrations and Cd adsorption experiments on the fresh aggregate samples at three different ionic strengths (0.56 M, 0.1 M and 0.01 M). We minimized sample processing (i.e., drying and powdering) to a simple washing step, in which the aggregate pellets remained hydrated to avoid any mineral transformation thus altering their true surface reactivity in seawater. Experimental results were then fitted with non-electrostatic model to predict both surface charges and metal-adsorption behavior of ferrihydrite aggregates. Different from previous surface-complexation modelling studies, here we used a novel and more powerful modelling program called Phreefit. It utilizes the global optimization algorithms instead of more commonly used Newton-Raphson method in FITEQL program, which is often too limited for precisely modelling complex systems such as the two samples in this study. Furthermore, we also measured the surface charges of both samples over the pH range from 3 to 9 on a Malvern Zetasizer and characterized the surface functional groups through Fourier-Transform Infrared Spectroscopy to help with our interpretation of the experimental data.Preliminary results show that cyanobacteria-ferrihydrite aggregates formed primarily due to ionic bridging. Cyanobacterial cells likely facilitated the precipitation of dissolved silica. Findings from titration and Cd adsorption experiments indicate that the surface reactivity and capacity of both silica-rich ferrihydrite cyanobacteria-ferrihydrite aggregates to adsorb trace elements differ from their individual components, likely due to site blockage. This distinction is particularly prominent when considering the expected Archean seawater pH from 6 to 8. This disparity implies that the biogenic ferrihydrite aggregates do not exhibit an additive surface reactivity, which is in agreement with similar previous studies. Our combined results are crucial to accurately predict the adsorption of trace elements onto the aggregate surface and, ultimately, comprehend the archive of trace elements in sedimentary rocks used to reconstruct Precambrian ocean chemistry.

Microbial reduction of synthetic Biogenic Iron Oxides containing various amounts of Organic Carbon and Silica

Iron oxides formed in close association with bacteria are referred to as biogenic minerals (BIOS). Both the organic part of BIOS and the iron oxide particles affect the net surface charge of those iron-organic carbon aggregates and offer reactive sites that can immobilize many soluble contaminants (Warren and Haack 2001) making BIOS a contender in bioremediation technologies. However, before using BIOS in bioremediation, it is essential to understand the interactions of impurities such as organic matter and other minor components (including silica) (Dyer et al. 2010). This project involves the synthesis of Biogenic Iron Oxides (BIOS) using various silica contents and different soluble alginate concentrations (as an analogue for bacterial exopolysaccharides) close to natural environmental conditions. The mineralogical, chemical and physical composition of the synthesized samples was determined by X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM), Field Emission Scanning Electron Microscopy (FESEM), Fourier-transform Infrared Spectroscopy (FTIR), and with a Malvern Zetasizer Nano instrument. The various samples (mainly ferrihydrite) were then reduced in the presence of Shewanella putrefaciens CN32, a well-known iron reducing bacterium. All microbial reduction experiments (see Table 1) with different types of BIOS were performed under anoxic conditions. Results indicate that the ratio of organic matter and silica to Fe (III) in BIOS affects the reduction rate. It is proposed that alginate binds to iron oxide particles and protects them from reduction. However, samples in presence of high concentration of silica (i.e.,> 0.05) showed structural disorder which likely prevented nucleation of well ordered ferrihydrite, which in return increased their rate of reduction. In addition, higher reduction rates of ferrihydrite were reported at higher concentrations of silica in BIOS, even in the presence of alginate.

Design-of-Experiment-Assisted Fabrication of Biodegradable Polymeric Nanoparticles: In Vitro Characterization, Biological Activity, and In Vivo Assessment.

Berberine (BER) is an alkaloid obtained from berberis plant having broad biological activities including anticancer. BER-encapsulated alginate (ALG)/chitosan (CHS) nanoparticles (BER-ALG/CHS-NPs) were developed for long-acting improved treatment in breast cancer. The surface of the NPs was activated by a conjugation reaction, and thereafter, the BER-ALG/CHS-NP surface was grafted with folic acid (BER-ALG/CHS-NPs-F) for specific targeting in breast cancer. BER-ALG/CHS-NPs-F was optimized by applying the Box-Behnken design using Expert design software. Moreover, formulations are extensively evaluated in vitro for biopharmaceutical performances and tested for cell viability, cellular uptake, and antioxidant activity. The comparative pharmacokinetic study of formulation and free BER was carried out in animals for estimation of bioavailability. The particle size recorded for the diluted sample using a Malvern Zetasizer was 240 ± 5.6 nm. The ζ-potential and the predicted % entrapment efficiency versus (vs) observed were +18 mV and 83.25 ± 2.3% vs 85 ± 3.5%. The high % drug release from the NPs was recorded. The analytical studies executed using infrared spectroscopy, differential scanning calorimetry, and X-ray diffraction expressed safe combinations of the components in the formulation and physical state of the drug revealed to be amorphous in the formulation. Cytotoxicity testing demonstrated that the formulation effectively lowered the cell viability and IC50 of the tested cell line in comparison to a raw drug. The cellular uptake of BER-ALG/CHS-NPs-F was 5.5-fold higher than that of BER-suspension. The antioxidant capacities of BER-ALG/CHS-NPs-F vs BER-suspension by the DPPH assay were measured to be 62.3 ± 2.5% vs 30 ± 6%, indicating good radical scavenging power of folate-conjugated NPs. The developed formulation showed a 4.4-fold improved oral bioavailability compared to BER-suspension. The hemolytic assay intimated <2% destruction of erythrocytes by the developed formulation. The observed experimental characterization results such as cytotoxicity, cellular uptake, antioxidant activity, and improved absorption suggested the effectiveness of BER-ALG/CHS-NPs-F toward breast cancer.

Thermostability and Immunostimulatory Activity of Cationic Liposome-based Newcastle Disease Virus Vaccine

Cold storage is a huge challenge in maintaining vaccine potency especially in low and middle income countries (LMIC). The aim of this research was to investigate the heat and freeze-thaw stability of Newcastle disease virus (NDV) vaccine encapsulated in 1,2-dioleoyl-3-trimethyl ammonium propane (DOTAP) cationic liposomes as well as its immunostimulatory activity. DOTAP cationic liposomes were prepared using the thin-film hydration technique and characterized for morphology, particle size, polydispersity index, zeta potential and encapsulation efficiency. Experimental birds were challenged with Herts 33 velogenic strain of NDV. Haemagglutination inhibition test was used to assess the formation of antibodies in the birds. The cationic liposomes were further stored at 28 oC or subjected to freeze-thaw cycles for 7 weeks and then assessed for stability using the same parameters of particle size, polydispersity index, zeta potential, morphology, antibody titre or differential white blood cell count. Using TEM, the morphology of the liposomes was found to be spherical, and using Malvern Zetasizer, particle size was in the nanosize range. Zetapotential and polydispersity indices were positive and below 0.1. The HI test showed that the DOTAP cationic liposomes stimulated a higher titre of antibodies than the commercial vaccine. Heat and freeze-thaw stability studies after 7 weeks of storage showed slight aggregation and elevated particle sizes and wider distribution. The HI titer was appreciably lower after secondary immunization with the DOTAP cationic liposomes stored at 28 oC but elevated for the product kept at freeze-thaw conditions. The immunized birds were 100 % protected from the virulent strain of NDV. In conclusion, the development of the DOTAP-cationic liposomes encapsulating NDV vaccine administered orally was found to be more stable under freeze-thaw conditions than when it was exposed to heat for 7 weeks. The liposome-based vaccine had more immunostimulatory effects than the commercial sample.

Green synthesis of graphene quantum dot- and its electrochemical, antioxidant and antimicrobial activities

The present work reports the synthesis of graphene quantum dots with a green approach. Graphene quantum dots (ssGQDs) were synthesized via Sarcopoterium spinosum (L.) Spach. fruit powder extract. The prepared ssGQDs were characterized by UV-Vis Spectroscopy, X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), X-ray Photoelectron Spectroscopy (XPS) and Malvern Zeta Sizer. The electrochemical activity of ssGQDs was investigated by cyclic voltammetry (CV) and differential puls voltammetry (DPV) methods. The synthesized ssGQDs were used to prepare electrosensor and tested for the electrochemical determination of dopamine. DPV studies showed a low detection limit of 0.107 μM over a wide linear concentration range of 10–800 μM. Antibacterial and antioxidant activities of synthesized ssGQDs were also investigated. The results showed that ssGQDs exhibited different antibacterial activity on Gram (+), Gram (-) bacterial strains and antifungal activity against fungal strains. 1,1-diphenyl-2-picryl-hydrazyl (DPPH) hydrogen peroxide (H2O2) scavenging methods were used to examine the antioxidant activity of ssGQDs and were found to be effective.

Self-assembly of Hyaluronic Acid-Cu-Quercetin flavonoid nanoparticles: synergistic chemotherapy to target tumors.

In this study, a natural compound quercetin (Qu) was investigated for its various antitumor effects. However, due to its poor water solubility and low bioavailability, its clinical application is limited. To overcome this constraint, a modification was to Qu, which resulted in the creation of novel flavonoid self-assembling nanoparticles (HCQ NPs). HCQ NPs were synthesized by a self-assembly method and characterized using transmission electron microscopy, the Malvern Zetasizer instrument, X-ray photoelectron spectroscopy (XPS), the ultraviolet-visible spectrophotometric method (UV-vis), Fourier transform infrared (FITR) and inductively coupled plasma mass spectrometry. Extracellular, methylene blue spectrophotometric analysis was used to determine the ability of HCQ NPs to react with different concentrations of H2O2 to form hydroxyl radicals (•OH). Intracellular, DCFH-DA staining was used to detect the ability of HCQ NPs to react with H2O2 to generate reactive oxygen species. Flow cytometry was used to detect the uptake of HCQ NPs by MDA-MB-231 cells at different time points. The biocompatibility of HCQ NPs was evaluated using the Cell Counting Kit-8 (CCK-8) assay. Calcein AM/PI double staining and the CCK-8 assay were used to evaluate the synergistic antitumor effect of HCQ NPs and H2O2. HCQ NPs showed uniformly sized analogous spherical shapes with a hydrodynamic diameter of 55.36 ± 0.27 nm. XPS revealed that Cu was mainly present as Cu2+ in the HCQ NPs. UV-vis absorption spectrum of the characteristic peak of HCQ NPs was located at 296 nm. Similarly, FTIR spectroscopy revealed a complex formation of Qu and Cu2+ that substantially changed the wavenumber of the 4-position C = O characteristic absorption peak. Based on the proportion of Qu and Cu2+ (1:2), the total drug loading of Qu and Cu2+ in the HCQ NPs for therapeutic purposes was calculated to be 9%. Methylene blue spectrophotometric analysis of •OH indicated that Cu can lead to the generation of •OH by triggering Fenton-like reactions. HCQ NPs rapidly accumulated in MDA-MB-231 cells with the extension of time, and the maximum accumulation concentration was reached at about 0.5 h. Calcein AM/PI double staining and CCK-8 revealed synergistic antitumor effects of HCQ NPs including the chemotherapeutic effect of Qu and chemodynamic therapy by Cu2+ in a simulated tumor microenvironment. HCQ NPs demonstrated very low toxicity in LO2 cells in the biocompatibility experiment. This study show cases a new method of creating self-assembled flavonoid HCQ NPs that show great for fighting cancer.

Development, systematic optimisation and biofilm disruption activity of eugenol-based nanosized emulsions stabilised with Tween 80

The aims of this study were to systematically optimise a formula for eugenol emulsions via face-centered central composite design and to assess the activity against two-different bacterial strains (Staphylococcus aureus and Propionibacterium acnes) present at planktonic and biofilm forms. The molecular interaction of excipients, mean particle size (MPS) including zeta potential (ZP), drug entrapment efficiency (DEE) and in vitro drug release of optimised emulsions was done using FT-IR, Malvern Zetasizer, ultracentrifugation technique and membrane-free dissolution model, respectively. The emulsions consisted of 151.3 ± 1.45 nm MPS, −21.3 ± 1.25 mV ZP and 93.98 ± 1.41% DEE values. On storage of emulsions at 25 °C for 3 months, the value of DEE was found to be 72.12 ± 2.82%. The Tween 80 emulsifier film coverage onto the dispersed eugenol droplets of emulsions delayed significantly the drug release (12%–19%) compared to the drug release occurred from pure eugenol. The treatment of planktonic S. aureus and P. acnes with diluted eugenol emulsions showed the minimum inhibitory concentration and minimum bactericidal concentration values at 1.25–2.5 mg/ml whereas it occurred at 10 mg/ml for pure eugenol. Treating the biofilms with eugenol emulsions (1–2 mg/ml) yielded 59–70% minimum biofilm eradication concentration but 10 mg/ml pure eugenol showed 60%. Hence, the eugenol emulsions displayed antibacterial activity and could be projected as an antibiofilm or biofilm disruption agent.

Design and Characterisation of Rosuvastatin Calcium Nanosuspension Loaded Transdermal Patch

Objective: The present investigation highlights the formulation and evaluation of nanosuspension-loaded transdermal patches of poorly soluble antilipidemic drug (rosuvastatin) Methods: Dissolution characteristics of drug was improved by preparation of nanosuspension employing precipitation–ultrasonication technique using a different blend of water-soluble film forming polymers such as hydroxyl propyl methyl cellulose (HPMC K4M), Eudragit and were further characterized by fourier transformation infrared spectroscopy (FTIR), Malvern zetasizer and SEM. Propylene glycol and tween 80 were used as stabilizers. Based on the particle size of the formulation, the best one of NS4 was selected and fabricated into the transdermal patch by solvent casting method. The prepared formulations were evaluated for various parameters like drug excipient compatibility, appearance, morphology, thickness, tensile strength, folding endurance, % elongation, % moisture content, % moisture uptake, % drug content, in-vitro drug permeation, and stability studies were performed for the optimized formula according to the ICH Q1A (R2) guideline under 4°C and 25°C RH for three months. Results: Amongst all formulation, P4 transdermal patches were considered the optimized formulation. It has the highest in-vitro drug permeation (86.01±0.05%), thickness (0.86±0.09), weight uniformity (475.0±8.60 mg), folding endurance (279.3±9.39), moisture uptake (7.06±1.75%), moisture content (6.81±2.34 %) and stable formula Conclusion: Therefore, transdermal patches were considered to be potentially suitable for the sustained release of drugs to improve patient compliance.