Formulation, Characterization, and Cytotoxicity Evaluation of Lactoferrin Functionalized Lipid Nanoparticles for Riluzole Delivery to the Brain

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with a very poor prognosis. Its treatment is hindered by a lack of new therapeutic alternatives and the existence of the blood–brain barrier (BBB), which restricts the access of drugs commonly used in ALS, such as riluzole, to the brain. To overcome these limitations and increase brain targeting, riluzole-loaded nanostructured lipid carriers (NLC) were prepared and functionalized with lactoferrin (Lf), facilitating transport across the BBB by interacting with Lf receptors expressed in the brain endothelium. NLC were characterized with respect to their physicochemical properties (size, zeta potential, polydispersity index) as well as their stability, encapsulation efficiency, morphology, in vitro release profile, and biocompatibility. Moreover, crystallinity and melting behavior were assessed by DSC and PXRD. Nanoparticles exhibited initial mean diameters between 180 and 220 nm and a polydispersity index below 0.3, indicating a narrow size distribution. NLC remained stable over at least 3 months. Riluzole encapsulation efficiency was very high, around 94–98%. FTIR and protein quantification studies confirmed the conjugation of Lf on the surface of the nanocarriers, with TEM images showing that the functionalized NLC presented a smooth surface and uniform spherical shape. An MTT assay revealed that the nanocarriers developed in this study did not cause a substantial reduction in the viability of NSC-34 and hCMEC/D3 cells at a riluzole concentration up to 10 μM, being therefore biocompatible. The results suggest that Lf-functionalized NLC are a suitable and promising delivery system to target riluzole to the brain.

Polyethylene Glycol-Stabilized Zein Nanoparticles Containing Gallic Acid

Research background. Gallic acid is a polyphenol presenting antioxidant and antitumor activities, however its use as a nutraceutical or drug is hindered by its low bioavailability. Zein is a natural protein found in corn and has been applied as nanoparticle for drug carrier. In this study, zein nanoparticles were obtained and stabilized with polyethylene glycol (PEG) as gallic acid carriers. Experimental approach. Nanoparticles were obtained by the liquid-liquid method and characterized in terms of mean size, polydispersity index, zeta potential, morphology, solid-state interactions, and encapsulation efficiency/drug loading. The stability of nanoparticles was evaluated in simulated gastrointestinal fluids and food simulants, and the antioxidant activity was determined by the scavenging of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. Results and conclusions. Zein nanoparticles containing gallic acid were obtained and stabilized only in the presence of PEG. The optimal conditions originated nanoparticles with mean size <200 nm, low polydispersity index (<0.25) and negative zeta potential (20 mV). The gallic acid encapsulation efficiency was about 40 %, drug loading about 5 %, and the compound was encapsulated in an amorphous state. FTIR did not identify chemical interactions after gallic acid nanoencapsulation. Zein nanoparticles were more susceptible to release the gallic acid in gastric than intestinal simulated medium, however more than 50 % of drug content was protected from premature release. In food simulants, the gallic acid release from nanoparticles was prolonged and sustained. Moreover, the nanoencapsulation did not reduce the antioxidant activity of gallic acid. Novelty and scientific contribution. The results show the importance of PEG on the formation and properties of zein nanoparticles obtained by the liquid-liquid dispersion method. This study indicates PEG-stabilized zein nanoparticles are potential carriers for gallic acid delivery by the oral route to take advantage of its antioxidant properties and be applied both in the pharmaceutical and food industry.

Application of Box Behnken Design and Response Surface Methodology for Optimization Sonication Conditions of Nanostructured Lipid Carrier from Mixtures of Palm Stearin and Palm Olein

The study aimed to determine the optimum conditions of the sonication in the manufacture of nanostructured lipid carriers (NLC). Nanostructured lipid carriers were developed using a mixture of palm stearin and palm olein, water, and tween as surfactants. Optimization was carried out to obtain nanostructured lipid carriers with a size below 200nm, zeta potential + 30 mV, and polydispersity index below 0.5. Optimization of NLC using response surface methodology and Box-Behnken experimental design. The independent variables were amplitude (A, kHz), pulse on pulse off (B, minute), and time of sonication (C, minute) each at three levels, while dependent variables were zeta potential (Y1, mV), particle size (Y2, nm), and polydispersity index (Y3). Measurement for dependent variables using the Zetasizer Nano ZS particle size analyzer utilized with Malvern software (Malvern, UK). The optimum formulation was obtained at a combination of amplitude (35 kHz), pulse on pulse off (pulse on 9 pulses off 3), and time of sonication (3 minutes 25 seconds ). This resulted in NLC having particle size 127.9 nm, polydispersity index 0.191, the zeta potential of -27.3 mV.

Investigating the potential of Quercetin enthused nano lipoidal system for the management of dermatitis

Objective: The present research work was undertaken to develop quercetin enthused nanolipoidal systems and its characterization. The objective was to investigate potential of prepared system in the management of DNCB induced dermatitis. Method: Nanolipoidal system was prepared in different combinations with quercetin, L-α phosphatidylcholine (SPC) and ethanol and characterized for particle size, polydispersity index (PDI), zeta potential, drug entrapment efficiency, percentage drug release, skin retention and skin permeation. Selected batches were further incorporated into Carbopol 934 base gel. The vesicles were in size range 324.19-359 nm while polydispersity index (PDI) ranges from 0.241-0.554 and for zeta potential, it was from -26.33 to -39.3 nm. Entrapment efficiency was from 23.77-94.68 %. Confocal laser scanning microscopy showed penetration depth of rhodamine enthused ethosome across rat skin up to 45.23 µm which was significantly higher than the rhodamine solution (10 µm). In dinitrochlorobenzene (DNCB) induced mice dermatitis model histopathology study showed a marked decrease in amount of inflammatory cell nucleus in mice treated with quercetin loaded ethosomal gel followed by 76.13% decrease in-ear swelling and ear mass respectively in morphology study. The conventional marketed formulation showed a nominal decrease in epidermal thickness. Further Primary irritation index was less than 0.4 indicating negligible irritation in all the groups. Results: The optimized formulation F6 with SPC and ethanol in the ratio of 20:80 displayed the highest drug content and entrapment efficiency of 94.68±1.14%. PDI was 0.241±0.11 and skin retention 7.7%. Batch F6 with vesicle size and zeta potential of 324.9±19 nm and -26.33 mV, respectively, was incorporated in Carbopol 934 base gel and the prepared gel was evaluated for morphology, spreadability, in vitro, ex vivo release study, and kinetics study and in vivo studies. Conclusion: The present study revealed that the developed ethosomal gel can be used for enhanced delivery of Quercetin via skin. The in vitro studies indicated that the gel serves as an efficient carrier for Quercetin. It showed its effectiveness in the management of dermatitis. Further, Quercetin loaded nanoethosomal gel formulation can be viewed as a promising drug delivery system for the management of dermatitis.

Preparation, Evaluation & Optimization of Nanoparticles Composed of Pyridostigmine

The purpose of this study was to prepare Pyridostigmine nanoparticles for control release of Pyridostigmine to improve the oral bioavailability, enhance the solubility and dissolution rate by decreasing particle size of drug. Infrared spectroscopic studies confirmed that there was no interaction between drug and polymers. The controlled release Pyridostigmine nanoparticles were prepared by Solvent evaporation by using Ethyl cellulose, Chitosan & HPMC K100 at different ratios. The production yield of the formulated controlled release nanoparticles (F1 to F16) in the range of 76.11 % to 83.58 %. The drug content of the formulated controlled release nanoparticles (F1 to F16) in the range of 82.56 %to 98.20%. The Theoretical loading of the formulated controlled release nanoparticles (F1- F16) in the range of 24.43 % to 64.24%. The entrapment efficiency increased with increasing the concentration of polymers and the formulations containing chitosan nanoparticles F6 (1:2) showed better entrapment (90.94%) among all formulation. The solubility of selected formulation (F6) in 0.2 M Phosphate buffer pH 6.8 increased when compared to pure drug. Particle size distribution was determined by Malvern zeta size, the size range for produced nanoparticles in the range of 200 nm to 400 nm. The Polydispersity index of selected nanoparticle formulation (F6) was indicated a narrow range and a homogeneous size distribution of particles. The in vitro dissolution study was carried out in 0. 2N PBS for 2 hours and phosphate buffer pH 6.8 for 10 hours. The formulations shows controlled release of drug up to 12 hrs and all formulations showed more than 75% of drug release. The release kinetics showed that the formulations were complies with Zero order kinetics followed by diffusion controlled mechanism. The best formulation F6 was evaluated by infrared spectroscopy, particle size, Polydispersity index & zeta potential and Scanning Electron microscopy. Best formulation of nanoparticles shown the extent of drug release was found to be F6 (96.93%) in 12 hrs. SEM studies confirmed the morphology of the nanoparticle formulation. Keywords: Polydispersity index, Zeta potential, Scanning Electron microscopy, Pyridostigmine

Andrographolide-loaded ethosomal gel for transdermal application: Formulation and in vitro penetration study

Background: Andrographolide is a phytoconstituent with anti-inflammatory activity, however, the compound’s poor oral bioavailability has hindered its effective formulation for oral administration. This study, therefore, aims to develop an ethosome for improving andrographolide penetration through the transdermal delivery system. Methods: This study developed 3 ethosome formulas with different andrographolide-phospholipid weight ratios (1:8, 1:9; 1:10), using the thin-layer dispersion-sonication method. Subsequently, the ethosomes were evaluated for particle size, polydispersity index, zeta potential, morphology, as well as entrapment efficiency, and incorporated into a gel dosage form. Subsequently, an in vitro penetration study was performed using Franz diffusion cells for 24 hours and the stability of the gels at 5 ± 2°C, 30 ± 2°C, and 40 ± 2°C, were studied for 3 months. Results: The results showed the optimal formula was E2, a 1:9 weight ratio formula of andrographolide and phospholipid. Based on the transmission electron micrograph, E2 possessed unilamellar, as well as spherical-shaped vesicles, and exhibited superior characteristics for transdermal delivery, with a particle size of 89.95 ± 0.75 nm, polydispersity index of 0.254 ± 0.020, a zeta potential of -39.3 ± 0.82 mV, and entrapment efficiency of 97.89 ± 0.02%. Furthermore, the cumulative andrographolide penetration and transdermal flux for the ethosomal gel of E2 (EG2) were 129.25 ± 4.66 µg/cm2 and 5.16 ± 0.10 µg/cm2/hours, respectively. All the ethosomal gel formulations exhibited improved penetration enhancement of andrographolide, compared to the nonethosomal formulations. Also, the andrographolide levels in the ethosomal and nonethosomal gels after 3 months ranged from 98.13 to 104.19%, 97.93 to 104.01%, and 97.23 to 102.26% at storage temperatures of 5 ± 2°C, 30 ± 2°C/RH 65% ± 5%, and 40 ± 2°C/RH 75% ± 5%, respectively. Conclusions: This study concluded that encapsulation into ethosome enhances andrographolide delivery through the skin.

Nanodispersions of TiO2 in Water for Removing Acrylic Films Used in Conservation

The influence of a nanodispersion of TiO2 in water (nanoparticle size: 40 nm, polydispersity index: 0.25), brushed on a Paraloid film and subjected to UV–Vis irradiation was evaluated. The TiO2 nanodispersions showed a tendency to reduce the molecular weight of Paraloid due to its photocatalytic properties. FTIR and GPC analyses and SEM images suggested the degradation of the polymer, while chromatic variations of the films were scarcely detected. This study is very remarkable in the perspective of using this material for the removal of polymeric films used in conservation.

Experimental Study on the Resistance of Synthetic Penicillin Solid Lipid Nanoparticles to Clinically Resistant Staphylococcus aureus

Background. With the increasing resistance of antibiotics to bacteria, new and effective methods are needed to transform existing antibiotics to solve the problem of long development cycles for new drugs. The antibiotic nanodelivery system has proven to be a promising strategy. Aim. The purpose of this study is to synthesize penicillin solid lipid nanoparticles (penicillin SLNs) to enhance the antibacterial activity of penicillin against drug-resistant Staphylococcus aureus. Materials and Methods. Penicillin SLNs were synthesized. And particle size, the polydispersity index (PI), and zeta potential (ZP) of penicillin SLNs were measured. The surface morphology of penicillin SLNs was observed using a transmission electron microscope. Results. The particle size of penicillin SLNs is 112.3 ± 11.9 nm , the polydispersity index (PI) and zeta potential (ZP) of penicillin SLNs are 0.212 ± 0.03 and − 27.6 ± 5.5 mV . The encapsulation efficiency and drug loading were 98.31 ± 1.2 % and 4.98 ± 0.05 ( % w / w ), respectively. Penicillin SLNs had a more significant inhibitory effect on the growth of methicillin-sensitive Staphylococcus aureus (MSSA) after the drug and the bacteria were incubated for 12 hours. The number of MRSA colonies in the penicillin group increased after 12 hours, while the number of MRSA colonies in the penicillin SLNs group did not change significantly. Conclusion. Penicillin SLNs enhance the ability of penicillin to enter cells and increase the concentration of penicillin in the cell and also extend the residence time of penicillin in the cell. Our findings indicated that penicillin SLNs enhance the inhibitory effect of penicillin on drug-resistant Staphylococcus aureus.

USING A SIMPLEX CENTROID DESIGN AND FATTY ACIDS TO OPTIMIZE FLUCONAZOLE-LOADED SOLID LIPID NANOPARTICLES (SLNs)

Objective: This study aimed to prepare fluconazole (FZ)-loaded solid lipid nanoparticles (SLNs) using a simplex centroid design and fatty acids to optimize the SLNs to get small-sized nanoparticles with a narrow distribution. Methods: Hot emulsification was used to prepare the FZ-loaded SLNs. Stearic acid (Sa) (X1), palmitic acid (Pa) (X2), and myristic acid (Ma) (X3) were the solid lipids. The effect of various types and amounts of fatty acids on the particle size, polydispersity index, zeta potential, and pH of the SLNs was studied using the simplex centroid design. Results: The particle size of all formulations ranged between 16.49 nm and 56.65 nm, and the polydispersity index (PDI) ranged between 0.258 and 0.676, indicating a relatively narrow size distribution. The zeta potential ranged from–7.47 to–12.2 mV. The pH was around 4.63–4.77, indicating that the SLN system was a weak acid. Design-Expert® software was used to design the responses of all model formulations and to select the optimized formulation. The optimal formulation comprised 0.190 g Sa, 0.048 g Pa, and 0.002 g Ma. The experimental values of the particle size and PDI of the optimal formulation did not differ significantly from the predicted values and lay within a 95% confidence interval (CI). Conclusion: Therefore, the simplex centroid design using fatty acids could efficiently formulate and optimize FZ-loaded SLNs.

Application of Plackett-Burman design for screening of factors affecting pitavastatin nanoparticle formulation development

Introduction: Nanoparticle formulation of pitavastatin calcium is a potential alternative to solve the solubility related problem. However, the formulation of nanoparticle involves various parameters that affect product quality. Plackett-Burman design could facilitate an economical experimental plan that focuses on determining the relative significance of many. Aim: The objective of this study was to screen the variables which could significantly affect the pitavastatin nanoparticle formulation. Materials and methods: The pitavastatin nanoparticles were formulated by preparing nanosuspension using the emulsion solvent evaporation technique followed by freeze-drying. A Plackett-Burman screening design methodology was employed in which seven factors at two levels were tested at 12 runs to study the effect of formulation and process variables on particle size and polydispersity index of nanoparticles. The surface morphology and crystalline nature of nanoparticle were also evaluated. Results: The particle size and polydispersity index of nanosuspension was found in the range of 113.1 to 768.5 nm and 0.068 to 0.508, respectively. Statistical analysis of various variables revealed that stabilizer concentration, injection flow rate, and stirring rate were the most influential factors affecting the particle size and polydispersity index of the formulation. X-ray diffraction (XRD) and scanning electron microscopy (SEM) study suggested the amorphous nature of nanoparticles. Conclusions: This study concluded that the Plackett-Burman design was an efficient tool for screening the process and formulation variables affecting the properties of pitavastatin nanoparticles and also for the identification of the most prominent factor.