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The encapsulation efficiency (EE%) reflects the amount of bioactive components that can be loaded into nanoliposomes. Obtaining a suitable nanoliposome stabiliser may be the key to improving their EE%. In this study, three polyphenols were screened as stabilisers of nanoliposomes with high nisin EE%, with curcumin nanoliposomes (Cu-NLs) exhibiting the best performance (EE% = 95.94%). Characterizations of particle size, PDI and zeta potential indicate that the Cu-NLs had good uniformity and stability. TEM found that nisin accumulated at the edges of the Cu-NLs' phospholipid layer. DSC and FT-IR revealed that curcumin was involved in the formation of the phospholipid layer and altered its structure. FT-IR and molecular docking simulations indicate that the interactions between curcumin and nisin are mainly hydrogen bonding and hydrophobic. In whole milk, Cu-NLs effectively protected nisin activity. This study provides an effective strategy for improving the EE% of nanoliposomes loaded with nisin and other bacteriocins., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Lipid nanoparticles (LNPs) are emerging nucleic acid delivery systems in the development of mRNA therapeutics such as the severe acute respiratory syndrome coronavirus 2 vaccines. However, a suitable analytical method for evaluating the encapsulation efficiency (EE) of the LNPs is required to ensure drug efficacy, as current analytical methods exhibit throughput issues and require long analysis times. Hence, we developed and validated an anion-exchange HPLC method using Analytical Quality by Design. Three critical method parameters (CMPs) were identified using risk assessment and Design of Experiments: column temperature, flow rate, and sodium perchlorate concentration. The CMPs were optimized using Face-Centered Central Composite Design. The discriminating power of the optimized HPLC method and RiboGreen assay was comparable. The main advantage of this method is that LNPs can be directly injected into the HPLC system without bursting the LNPs loaded with encapsulated poly(A). The optimized HPLC method was validated as robust, high-throughput, and sufficiently sensitive according to the ICH Q2 guidelines. We believe our findings could promote efficient LNPs-based drug development., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: All authors were employees of Eisai Co., Ltd. during the execution of this research project. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In recent years, the use of lipid nanoparticles (LNPs) for delivery of messenger RNA (mRNA)-based therapies has gained substantial attention in the field of drug development. In such an application, multiple LNP attributes have to be carefully characterized to ensure product safety and quality, whereas accurate and efficient characterization of these complex mRNA-LNP formulations remains a challenging endeavor. Here, we present the development and application of an online separation and characterization platform designed for the isolation and in-depth analysis of mRNAs and mRNA-loaded LNPs. Our asymmetrical flow field-flow fractionation with a multi-detector (MD-AF4) method has demonstrated exceptional resolution between mRNA-LNPs and mRNAs, delivering excellent recoveries (over 70%) for both analytes and exceptional repeatability. Notably, this platform allows for comprehensive and multi-attribute LNP characterization, including online particle sizing, morphology characterization, and determination of encapsulation efficiency, all within a single injection. Furthermore, real-time online sizing by synchronizing multi-angle light scattering (MALS) and dynamic light scattering (DLS) presented higher resolution over traditional batch-mode DLS, particularly in differentiating heterogeneous samples with a low abundance of large-sized particles. Additionally, our method proves to be a valuable tool for monitoring LNP stability under varying stress conditions. Our work introduces a robust and versatile analytical platform using MD-AF4 that not only efficiently provides multi-attribute characterizations of mRNA-LNPs but also holds promise in advancing studies related to formulation screening, quality control, and stability assessment in the evolving field of nanoparticle delivery systems for mRNAs., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

Iron is an important micronutrient that cannot be added directly into food products due to potential reactions with the food matrix, impact on color, and taste. Complexed biopolymeric nanocarriers can overcome these challenges particularly for oral delivery of iron, but selecting appropriate biopolymers, their ratio and pH of complexation is very important. In this study, whey protein concentrate (WPC)-pectin nanocomplexes were prepared at different concentrations (WPC 4, 6 and 8%; pectin 0.5, 0.75 and 1%), and pH (3, 6 and 9) to encapsulate iron. The smallest carriers were observed at pH 3; higher pH led to higher zeta potential (zero to -32.5 mV). Encapsulation efficiency of iron in nanocarriers formulated at pH = 3, 6 and 9 were 87.83, 75.92 and 20%, respectively. Scanning electron microscopy revealed the spherical particles at pH 3. To conclude, a WPC to pectin ratio of 4: 1 at pH 3 was the best conditions for loading iron., Competing Interests: Declaration of competing interest All authors declare that there is no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study focuses on the optimization of Hydrastis canadensis-based nanocarriers in environmental and microbial applications like antibacterial and dye degradation. Hydrastis canadensis (H. canadensis) is loaded into the nanocarrier using a gelation method. Characterization involves pH analysis, UV-VIS spectrophotometry, scanning electron microscopy, Fourier-transform infrared spectroscopy, dynamic light scattering, high-performance liquid chromatography, encapsulation efficiency. Further antimicrobial activity against Staphylococcus aureus and Escherichia coli were tested. Dye degradation was evaluated at concentrations of 1 % of high molecular (HM) and 1.5 % of low molecular (LM) chitosan nanoparticles with both 3C and 1000C concentrations of the drug. The obtained results confirm the presence of chitosan nanocarrier alongside the pure drug in 1 % HM and 1.5 % LM chitosan particles with a notable encapsulation efficiency activity in both 3C and 1000C concentrations. Antimicrobial studies were carried out using the agar well diffusion method and revealed a significant zone of inhibition of 20 mm and 25 mm for E. coli and S. aureus, respectively in chitosan nanocarrier-loaded samples compared to pure drug and chitosan nanocarriers samples. The dye degradation studies of four dyes methylene blue, methylene orange, methylene red, and safranin using both pure drugs and chitosan nanocarrier-loaded drugs showed the highest percentage of degradation (76 %) against methylene blue in the chitosan nanocarrier-drug loaded formulation. These findings cumulatively underscore chitosan nanoparticles can be used as an effective carrier for Hydrastis Canadensis, with enhanced antimicrobial and dye degradation capabilities. Varied concentrations and molecular weights highlight the versatility of the ionotropic gelation method in optimizing drug delivery. Enhanced efficacy of the nanocarrier was evident in the observed zone of inhibition in antimicrobial testing. The substantial degradation percentage in methylene blue emphasizes the formulation's applicability in environmental dye removal processes, with potential avenues for improvement explored through interactions between the chitosan nanocarrier and H. canadensis characteristics. Future investigations may focus on scaling up the optimized formulation for large-scale applications and exploring release kinetics and comprehensive toxicity assessments for a holistic understanding of potential environmental and biomedical implications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Tanshinone compounds are secondary metabolites which their application in food and pharmaceutical industry is limited due to the low solubility in water and sensitivity to heat. This study aimed to develop a novel biopolymer nanocarriers system based on pectin/zein for the encapsulation of tanshinone compounds using the anti-solvent precipitation method. The concentration of pectin and mass ratio of tanshinone/zein in the final formulation of nanoparticles were optimized. According to the results, a pectin concentration of 1 g/L and a tanshinone/zein ratio of 0.1:1 g/g were considered the optimal nanoparticle formulation. The resulting nanoparticles exhibited a spherical core-shell structure, with approximate values for size, zeta potential, TSI, and encapsulation efficiency of 132 ± 0.002 nm, -38.6 ± 0.019 mV, 0.600 ± 0.084, and 79.41 ± 0.62 %, respectively. The FTIR test confirmed the presence of hydrophobic, hydrogen, and electrostatic interactions among the constituents within the nanoparticles. Additionally, XRD and DSC tests verified the amorphous nature of the nanoparticles. Morphological examination conducted through TEM, and SEM revealed the characteristics of the resulting nanoparticles. Furthermore, this carrier system significantly enhanced the solubility of tanshinone compounds in water., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

The utilization of drug delivery systems, such as lipid nanoparticles and polyplexes/micelleplexes, has shown promise in intracellularly delivering nucleic acids for addressing various diseases. Accurate quantification of the nucleic acid cargo within nanoparticles is essential for the development of safe and effective nanomedicines. Currently, the RiboGreen and SYBR Gold methods are regarded as standard techniques for the precise quantification of RNA in lipid nanoparticles and polyplexes/micelleplexes, respectively. In this chapter, we present a comprehensive protocol for the precise evaluation of the encapsulation efficiency in such formulations using these methods. Additionally, we offer detailed instructions for nanoparticle preparation, characterization, and a comparative analysis of the sensitivity of both methods in quantifying unencapsulated siRNA., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

This current research explored the application of cold plasma (CP) treatment to modify zein-alginate oligosaccharide (zein-AOS) composites in an ethanol-water solution. Anti-solvent method was used to prepare zein-AOS nanoparticles (NPs), and the objective was to investigate the mechanism by which CP promotes interaction between protein and saccharides. Characterization results indicated that CP treatment improved hydrogen bonding and electrostatic interaction between zein and AOS. The CP zein-AOS NPs underwent dispersion and rearrangement, resulting in smaller aggregates with better dispersibility. Among the various induction conditions tested, the zein-AOS85 NPs (induced at 85 W for 2 min) exhibited superior performance as delivery wall materials, with smaller particle size (234.67 nm), larger specific surface area (9.443 m 2 /g), and higher surface charge (-35.43 mV). In addition, zein-AOS85 showed high stability when used as delivery wall material, providing more binding sites and self-assembly dynamics for nutrients. Curcumin was used as the nutrient model in this study, and CP was found to enhance hydrogen bonding, electrostatic interaction, and hydrophobic interaction between zein, AOS, and nutrients, resulting in increased encapsulation efficiency (EE) from 63.80 % to 85.17 %. The delivery system also exhibited good pH, ionic strength, storage, and dispersion stability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The quantification of the drug associated to nanoparticle carriers, often expressed in terms of encapsulation efficiency, is a regulatory requirement. The establishment of independent methods to evaluate this parameter provides a means for measurement validation, which is critical in providing confidence in the methods and enabling the robust characterization of nanomedicines. Chromatography is traditionally used to measure drug encapsulation into nanoparticles. Here, we describe an additional independent strategy based on analytical centrifugation. The encapsulation of diclofenac into nanocarriers was quantified based on the mass difference between placebo (i.e. unloaded) and loaded nanoparticles. This difference was estimated using particle densities measured by differential centrifugal sedimentation (DCS) and size and concentration values measured by particle tracking analysis (PTA). The proposed strategy was applied to two types of formulations, namely poly(lactic-co-glycolic acid) (PLGA) nanoparticles and nanostructured lipid carriers, which were analysed by DCS operated in sedimentation and flotation modes, respectively. The results were compared to those from high performance liquid chromatography (HPLC) measurements. Additionally, X-ray photoelectron spectroscopy analysis was used to elucidate the surface chemical composition of the placebo and loaded nanoparticles. The proposed approach enables the monitoring of batch-to-batch consistency and the quantification of diclofenac association to PLGA nanoparticles from 0.7 ng to 5 ng of drug per 1 μg of PLGA, with good linear correlation between DCS and HPLC results (R 2  = 0.975). Using the same approach, similar quantification in lipid nanocarriers was possible for a loading of diclofenac ≥1.1 ng per 1 μg of lipids, with results in agreement with the HPLC method (R 2  = 0.971). Hence, the strategy proposed here expands the analytical tools available for evaluating nanoparticles encapsulation efficiency, being thus significant for increasing the robustness of drug-delivery nanocarriers characterization., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Conjugate drugs are evolving into potent techniques in the drug development process for enhancing the biopharmaceutical, physicochemical, and pharmacokinetic properties. Atorvastatin (AT) is the first line of treatment for coronary atherosclerosis; however its therapeutic efficacy is limited because of its poor solubility and fast pass metabolism. Curcumin (CU) is evidenced in several crucial signaling pathways linked to lipid regulation and inflammation. To enhance the therapeutic efficacy and physical properties of AT and CU, a new conjugate derivative (AT-CU) was synthesized and assessed by in silico, in vitro characterizations, and in vivo efficacy through mice model. Although the biocompatibility and biodegradability of Polylactic-co-Glycolic Acid (PLGA) in nanoparticles are well documented, burst release is a common issue with this polymer. Hence the current work used chitosan as a drug release modifier to the PLGA nanoparticles. The chitosan-modified PLGA AT-CU nanoparticles were prepaid by single emulsion and solvent evaporation technique. With raising the concentration of chitosan the particle size grew from 139.2 nm to 197.7 nm, the zeta potential rose from -20.57 mV to 28.32 mV, and the drug encapsulation efficiency improved from 71.81% to 90.57%. At 18 h, the burst release of AT-CU from PLGA nanoparticles was seen, hitting abruptly 70.8%. For chitosan-modified PLGA nanoparticles, the burst release pattern was significantly reduced which could be due to the adsorption of the drug on the surface of chitosan. The efficiency of the ideal formulation i.e F4 (chitosan/PLGA = 0.4) in treating atherosclerosis was further strongly evidenced by in vivo investigation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Curcumin (CUR) was encapsulated into yeast cells (YCs) through a pH-driven method with a 5.04-fold increase in loading capacity and a 43.63-fold reduction in incubation time compared to the conventional diffusion method. Optimal encapsulation was obtained when the mass ratio of CUR to YCs was 0.1, and the loading capacity and encapsulation efficiency were 8.07% and 80.66%, respectively. Encapsulation of CUR into YCs was confirmed by fluorescence microscopy, differential scanning calorimetry, and thermogravimetric analysis. Fourier transform infrared spectroscopy and X-ray diffraction further demonstrated that the encapsulated CUR was interacted with mannoprotein and β-glucan of the cell wall network through hydrophobic interaction and hydrogen bond in amorphous state. The in vitro bioaccessibility of YCs-loaded CUR was significantly increased by 6.05-fold. The enhanced encapsulation efficiency and rapid encapsulation process proposed in this study could facilitate YCs-based microcarriers to encapsulate bioactive substances with higher bioaccessibility., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Hepatitis B virus core protein (HBc) has become a hot spot in drug carrier protein research due to its natural particle self-assembly ability and ease of modification. The truncation of the C-terminal polyarginine domain (CTD, aa 151-183) of HBc does not affect the self-assembly of the particles. However, it does affect the internal and external charges of the particles, which may subsequently affect drug encapsulation. Thus, the truncated C-terminal polyarginine domain (CTD) of HBc and the inserted RGD peptide were selected to construct and express three HBc variants (RH) encapsulated with ICG (RH/ICG) with different C-terminal lengths to compare the stability and drug activity of their nanoformulations. RH160/ICG was found to have a great advantages in encapsulation efficiency and biological imaging. Compared with other HBc variants, RH160/ICG significantly improved encapsulation efficiency, up to 32.77%±1.23%. Cytotoxicity and hemolysis assays further demonstrated the good biocompatibility of RH160/ICG. Cell uptake and in vivo imaging experiments in mice showed that RH160/ICG could efficiently deliver ICG in tumor cells and tumor sites with good imaging effect. This research provides a new direction for further expanding the diagnosis and treatment application of ICG and development of HBc-based nanoparticle drug carrier platform.

In the present study, nanoliposomes with tuneable structure elasticity were prepared by reverse-phase evaporation. Both Fluorescence Polarization and Fluorescence Resonance Energy Transfer was employed to characterize the structural elasticity of resultant nanoliposomes. Temozolomide, a kind of hydrophilic drug as the first-line treatment choice for glioblastoma, was encapsulated into nanoliposomes. The results showed that the flexibility of nanoliposomes gradually increased with sodium cholate, while decreasing with cholesterol, Labrafac CC and Labrafac PG adding. Furthermore, the structural flexibility of nanoliposomes was positively correlated with the encapsulation efficiency and release rate and cellular uptake. Our research reveals the structural flexibility of nanoliposomes could affect in vitro characteristics and thereafter in vivo behaviors of nanoliposomes.

Recent findings have proved the benefits of Pioglitazone (PGZ) against atherosclerosis and type 2 diabetes. Since the systematic and controllable release of this drug is of significant importance, encapsulation of this drug in nanoparticles (NPs) can minimize uncontrolled issues. In this context, drug delivery approaches based on several poly(lactic-co-glycolic acid) (PLGA) nanoparticles have been rising in popularity due to their promising capabilities. However, a fully reliable and reproducible synthetic methodology is still lacking. In this work, we present a rational optimization of the most critical formulation parameters for the production of PGZ-loaded PLGA NPs by the single emulsification-solvent evaporation or nanoprecipitation methods. We examined the influence of several variables (e.g., component concentrations, phases ratio, injection flux rate) on the synthesis of the PGZ-NPs. In addition, a comparison of these synthetic methodologies in terms of nanoparticle size, polydispersity index (PDI), zeta potential (ζ p ), drug loading (DL%), entrapment efficiency (EE%), and stability is offered. According to the higher entrapment efficiency content, enhanced storage time and suitable particle size, the nanoprecipitation approach appears to be the simplest, most rapid and most reliable synthetic pathway for these drug nanocarriers, and we demonstrated a very slow drug release in PBS for the best formulation obtained by this synthesis.

Liposomes are an attractive drug delivery platform for a wide variety of pharmaceutical molecules. Encapsulation efficiency, which refers to the amount of drug contained inside liposomes compared with the total amount of drug, is a critical quality attribute of liposome products, as the free drug in a liposomal formulation may cause toxicity or undesired biodistribution. The determination of encapsulation efficiency requires the measurement of at least two of the three drug populations: total drug, encapsulated drug and free drug. However, direct measurement of the encapsulated drug and free drug remains a challenging analytical task. Nanoparticle exclusion chromatography (nPEC), an emerging high-performance liquid chromatography (HPLC) technique, has shown great potential in separating and quantifying the free drug in liposomal formulations. In this study, nPEC was systematically evaluated for two representative liposomal formulations containing either hydrophilic or hydrophobic small molecule drugs. It is reported for the first time that the insoluble free drug suspended in the aqueous formulation can be directly measured by nPEC. This free drug in the suspension sample was quantified with excellent accuracy and precision. On the other hand, the total drug measurement from dissociated liposomes was confirmed by the benchmark methodology of reversed phase liquid chromatography (RPLC). The facile quantitation of free and total drug in the liposome formulation enables the fast and accurate determination of the encapsulation efficiency, which can be used to guide the formulation development and characterize the product quality., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Shellac can be used as an ideal delivery vehicle to deliver and protect the hydrophobic quercetagetin; the barriers such as low acid stability and encapsulation efficiency, however, heavily impede the application of shellac. The purpose of this work is to prepare quercetagetin-loaded shellac-quaternized chitosan nanoparticles (Que-Sh-QCS NPs) to overcome these challenges. Herein, quaternized chitosan, with 14% degree of substitution, was successfully synthesized via a quaternization modification. The concentration of quaternized chitosan over 0.05% can prevent the aggregation of shellac nanoparticles at the acid. The encapsulation efficiency of quercetagetin obviously increased from 37.92 to 65.48% with the concentration of QCS varying from 0 to 0.05%. Meanwhile, Que-Sh-QCS 0.05 NPs possessed good storage stability, antioxidant property, biocompatibility, and controlled release. Therefore, quaternized chitosan can improve the encapsulation efficiency and acid and storage stabilities of nutraceutical-loaded shellac nanoparticles, providing a new insight into the application of shellac in cosmetics, pharmaceuticals, and food.

In this study, we investigated the design and construct of a chitosan (CA)-based targeted gene delivery system and evaluated its function. To this end, CA-folic acid/pDNA (CA-FA/pDNA) nanoparticles were prepared in different formulations using the ion gelation method. All the synthesized nanoparticles were characterized using FTIR, TEM, SEM and DLS. Moreover, the effects of molecular weight (MW) of CA, DNA, and CA concentration were inspected on encapsulation efficiency (EE). The results showed that the EE of pDNA was directly proportional with MW of CA and CA concentration but was in an inverse proportion with DNA concentration. In addition, high MW of CA and low MW of CA nanoparticles showed lower and higher pDNA release in all pH ranges, respectively. It is concluded that the N/P ratio increase can cause controlled pDNA release., (© 2020 International Union of Biochemistry and Molecular Biology, Inc.)

The evaluation of encapsulation efficiency is a regulatory requirement for the characterization of drug delivery systems. However, the difficulties in efficiently separating nanomedicines from the free drug may compromise the achievement of accurate determinations. Herein, ultrafiltration was exploited as a separative strategy towards the evaluation of methotrexate (MTX) encapsulation efficiency in nanostructured lipid carriers and polymeric nanoparticles. The effect of experimental conditions such as pH and the amount of surfactant present in the ultrafiltration media was addressed aiming at the selection of suitable conditions for the effective purification of nanocarriers. MTX-loaded nanoparticles were then submitted to ultrafiltration and the portions remaining in the upper compartment of the filtering device and in the ultrafiltrate were collected and analyzed by HPLC-UV using a reversed-phase (C18) monolithic column. A short centrifugation time (5 min) was suitable for establishing the amount of encapsulated MTX in nanostructured lipid carriers, based on the assumption that the free MTX concentration was the same in the upper compartment and in the ultrafiltrate. The defined conditions allowed the efficient separation of nanocarriers from the free drug, with recoveries of >85% even when nanoparticles were present in cell culture media and in pig skin surrogate from permeation assays.

A phase inversion based cold water dilution method was developed to encapsulate Vitamin D 3 (Vit D) in nano-structured lipid carrier (NLC) by blending caprylic-/capric triglyceride, Leciva S70 and Kolliphor HS®15, Vit D and sodium chloride. To optimize the process; a total of forty one formulations prepared by varying in their composition were tested for presence of NLC. Out of forty one formulations, only thirteen formulations resulted in NLC formation which were further evaluated for their physico-chemical attributes (particle size, zeta potential, transmittance, encapsulation efficiency and Vit D release). During principal component analysis using XLstats it was found that NLC-19, fabricated with 20% (v/v) Kolliphor, 20% (v/v) CCTG and 60% (v/v) water, 2.5% (w/v) Leciva, 2% (w/v) Vit D and 5% (w/v) sodium chloride was the most suitable for purpose of encapsulating Vitamin D. Hence, NLC-19 formulation was further taken up for stability studies under the following environmental stress conditions: (a) Temperature and humidity: accelerated condition: 45 ± 2 °C and RH 75 ± 5%, ambient condition: 25 ± 3 °C and RH 65 ± 5% and refrigerated condition: 6 ± 2 °C and RH 55 ± 5%, (b) pH: 3, 4, 5, 6, and 7, and (c) Ionic strength (NaCl concentration): 0 mM, 250 mM, 500 mM and 750 mM. The sensory evaluation of 'Lassi' (fortified with NLC-19) and its acceptability further confirmed the suitability of NLC-19 for the purpose of fortification of Vitamin D 3 in 'Lassi' (A milk based beverage)., (Copyright © 2019. Published by Elsevier Ltd.)

The purpose of this study was to optimize the preparation conditions of podophyllotoxin liposomes (PPT-Lips), and to investigate their effects on PC3 cells. PPT-Lips were prepared by using a thin-film dispersion method. In order to achieve maximum drug encapsulation efficiency (EE), the process and formulation variables were optimized by response surface methodology (RSM). The optimum preparation conditions were cholesterol to lecithin ratio of 3.6:40 (w/w), lipid to drug ratio of 15.8:1 (w/w), and the ultrasonic intensity of 35% (total power of 400 W). The experimental EE of PPT-Lips was 90.425%, which was consistent with the theoretically predicted value. The characterization studies showed that PPT-Lips were well-dispersible spherical particles with an average size of 106 nm and a zeta potential of -10.1 mV. A gradual and time-dependent pattern of PPT from liposomes was found in in vitro drug release with a cumulative release amount up to 70.3% in 24 h. Results of cell viability experiments on PC3 cells demonstrated that PPT-Lips exhibited more effective anticancer activity in comparison with free PPT. Therefore, PPT-Lips represent an efficient and promising drug delivery system for PPT.

Objective: To develop an oral sustained release formulation of mycophenolate mofetil (MMF) for once-daily dosing, using chitosan-coated polylactic acid (PLA) or poly(lactic-co-glycolic) acid (PLGA) nanoparticles. The role of polymer molecular weight (MW) and drug to polymer ratio in encapsulation efficiency (EE) and release from the nanoparticles was explored in vitro., Methods: Nanoparticles were prepared by a single emulsion solvent evaporation method where MMF was encapsulated with PLGA or PLA at various polymer MW and drug: polymer ratios. Subsequently, chitosan was added to create coated cationic particles, also at several chitosan MW grades and drug: polymer ratios. All the formulations were evaluated for mean diameter and polydispersity, EE as well as in vitro drug release. Differential scanning calorimetry (DSC), surface morphology, and in vitro mucin binding of the nanoparticles were performed for further characterization., Results: Two lead formulations comprise MMF: high MW, PLA: medium MW chitosan 1:7:7 (w/w/w), and MMF: high MW, PLGA: high MW chitosan 1:7:7 (w/w/w), which had high EE (94.34% and 75.44%, respectively) and sustained drug release over 12 h with a minimal burst phase. DSC experiments revealed an amorphous form of MMF in the nanoparticle formulations. The surface morphology of the MMF NP showed spherical nanoparticles with minimal visible porosity. The potential for mucoadhesiveness was assessed by changes in zeta potential after incubation of the nanoparticles in mucin., Conclusion: Two chitosan-coated nanoparticles formulations of MMF had high EE and a desirable sustained drug release profile in the effort to design a once-daily dosage form for MMF.

Entrapment efficiency (EE) is a crucial parameter for the evaluation of nanocarriers. The accurate measurement of EE demands clear separation of nanocarriers from free drugs, which so far has not been clearly validated due to a lack of functional tools to identify nanocarriers. Herein, an environment-responsive water-quenching fluorophore was employed to label and identify model nanocarriers, polycaprolactone nanoparticles (PN), methoxy polyethylene glycol-poly(d,l-lactic acid) polymeric micelles (PM) and solid lipid nanoparticles (SLN). The separation process of three commonly used methods (centrifugation, ultrafiltration and gel permeation chromatography) was visualized by live imaging. The separation efficiency of the centrifugation method is very poor, especially for PM (40 nm), SLN (100 nm) and PN (100 nm); only PN (200 nm) can be efficiently separated but at a consumption of enormous energy. The ultrafiltration method shows the best separation efficiency with only 0.32-0.93% of leakage of the nanocarriers. Gel permeation chromatography exhibits good separation as well but suffers from low recovery, a potential factor that might compromise the accuracy of EE measurement. In conclusion, the ultrafiltration method is the method of choice for efficient separation and accurate measurement of EE., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Context: Size, encapsulation efficiency and stability affect the sustained release from nanoparticles containing protein-type drugs., Objectives: Insulin was used to evaluate effects of formulation parameters on minimizing diameter, maximizing encapsulation efficiency and preserving blood glucose control following intraperitoneal (IP) administration., Methods: Homogenization or sonication was used to incorporate insulin into poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles with increasing poly(ethylene glycol) (PEG) content. Effects of polymer type, insulin/polymer loading ratio and stabilizer in the internal aqueous phase on physicochemical characteristics of NP, in vitro release and stability of encapsulated insulin were investigated. Entrapment efficiency and release were assessed by radioimmunoassay and bicinconnic acid protein assay, and stability was evaluated using SDS-PAGE. Bioactivity of insulin was assessed in streptozotocin-induced, insulin-deficient Type I diabetic mice., Results: Increasing polymeric PEG increased encapsulation efficiency, while the absence of internal stabilizer improved encapsulation and minimized burst release kinetics. Homogenization was shown to be superior to sonication, with NP fabricated from 10% PEG-PLGA having higher insulin encapsulation, lower burst release and better stability. Insulin-loaded NP maintained normoglycaemia for 24 h in diabetic mice following a single bolus, with no evidence of hypoglycemia., Conclusions: Insulin-loaded NP prepared from 10% PEG-PLGA possessed therapeutically useful encapsulation and release kinetics when delivered by the IP route.

Background and Objectives: The use of milk proteins for drug delivery is a new trend in functional foods and pharmaceutical. Recently, researchers have focused on the utilization of whey proteins in the preparation of nanoparticle and carrier for drugs and micronutrients. The objectives of this paper were to use whey proteins isolate (WPI) nanoparticles for the encapsulation of zinc citrate micronutrients and characterization of the prepared nanoparticles., Materials and Methods: Nanoparticles were prepared from WPI with pH cycling and used for the encapsulation and sustained release of zinc citrate with three ratios (7, 14 and 28 mM) of zinc citrate per gram WPI. The particle size of the prepared nanoparticles was characterization and examined by transmission electron microscopy. The release of Zinc from the prepared nanoparticles was carried out using simulated gastric fluid at pH 1.2 using dialysis membranes, the amount of zinc citrate loaded whey protein (14.36 mg Zinc in 1 g WPI) within range of daily dose of zinc for healthy adults., Results: The WPI nanoparticles were able to encapsulate efficiently zinc, with encapsulation efficiency that ranged between 99.79 and 96.31%. Zinc was highly released from the prepared nanoparticles in acidic media (pH 1.2)., Conclusion: It can be concluded that WPI can be used as an effective vehicle for the protection and sustained release of zinc in food and pharmaceutical preparations.

This study evaluates the effect of modified lecithin (ML) and sodium caseinate (SC) on the formulation, stability and bioaccessibility of astaxanthin (AXT) loaded oil-in-water (O/W) nanoemulsions. These nanoemulsions were formulated using high-pressure homogenization in four passes at 100MPa. The volume mean diameter (d 4,3 ) of nanoemulsions produced by ML and SC were 163±5 and 144±12 nm, respectively. The physiochemical stability of nanoemulsions was recorded at 25°C. The nanoemulsions prepared by ML were stable for 30 minutes against a wide range of pH and heating temperatures (60-120 °C). However, ML-stabilized nanoemulsions showed droplet growth when treated at high NaCl concentrations. In comparison, droplet growth was observed in SC-stabilized nanoemulsions at pH4 and at high temperature treatment. However, SC-stabilized nanoemulsions were stable at high NaCl concentration (500 mM). The SC-stabilized nanoemulsions showed good physical and chemical stability (>70%) after 30 days of storage. The bioaccessibility of AXT in nanoemulsions was significantly higher in ML (33%) than in SC-stabilized nanoemulsions (6%), indicating a strong influence of emulsifier on bioaccessibility. These findings provide valuable information in designing nutritional products such as aqueous based AXT fortified beverages., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The antibacterial property of thyme essential oil due to different volatile compounds, has been well documented in the literature. To overcome the high volatility of essential oil components, encapsulation has emerged as a new alternative. In this work, chitosan and thyme essential oil-loaded chitosan nanoparticles (TEO-CSNPs) and nanocapsules (TEO-CSNCs) were prepared by nanoprecipitation and nanoencapsulation, respectively. The morphology, encapsulation efficiency, release kinetics, and inhibitory activity were evaluated. Average size of nanocapsules (9.1±1.6nm) was slightly higher than nanoparticles (6.4±0.5nm). The percentage encapsulation of thymol and carvacrol, more than 68%, was similar for nanoparticles and nanocapsules. However, thymol and carvacrol release time from TEO-CSNPs was faster compared to TEO-CSNCs. The release kinetics data were fitted to three analytical kinetic models with no statistical differences among them. The inhibitory activity was higher for nanoparticles than for nanocapsules when tested against six foodborne bacteria. The inhibitory effect of TEO-CSNPs was the highest against Staphylococcus aureus (inhibition halo 4.3cm) and for TEO-CSNCs it was against Bacillus cereus (inhibition halo 1.9cm)., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objective: The prime objective of current work was to develop a strategy for preparation of combinational nano-formulation for reversal of drug resistance., Methods: As a model system, doxorubicin (DOX)-resistant COLO205 cells were developed and validated. From co-treatment studies with DOX, curcumin was selected as it reversed DOX-resistance at lowest concentration. In an attempt to increase its solubility, curcumin was encapsulated into hydroxypropyl-β-cyclodextrin (HP-β-CD). Here, we propose that presence of stabilizer overcomes its low encapsulation efficiency. Thus, we evaluated curcumin encapsulation in HP-β-CD in presence of different stabilizers and organic solvents. Finally, the effect of nanocurcumin with liposomal DOX was studied for reversal of resistance in COLO205 cells., Key Findings: In the process encapsulation, selective optimization of organic solvent by freeze-drying was found to be appropriate among other methods. From optimization studies with different organic solvent (acetone and dichloromethane) and stabilizer [polyvinyl alcohol (PVA) and Pluronics], HP-β-CD-encapsulated curcumin prepared using acetone in PVA-stabilized dispersion increased encapsulation (60%) with size of ~40 nm. Prepared nano-curcumin reversed the DOX resistance effectively in combination with liposomal DOX., Conclusions: Curcumin reversed DOX resistance in COLO205 cells at low concentration and enhanced curcumin encapsulation in HP-β-CD was noted in presence of PVA. Further, it was observed that prepared HP-β-CD-encapsulated curcumin is equi-efficacious to nano-dispersed curcumin., (© 2017 Royal Pharmaceutical Society.)

The influence of mPEG-PCL and mPEG-PLGA on encapsulation efficiency and drug-loading of nanoparticles was very important. SN-38 NPs were prepared from a series of diblock copolymers: mPEG1000-PLGA2000, mPEG2000-PCLs, mPEG5000-PCLs, mPEG2000-PLGAs, and mPEG5000-PLGAs by the thin film-hydration method. The prepared nanoparticles were characterized by morphology, size, encapsulation efficiency, drug-loading, and in vitro release behavior. This experiment suggested that the encapsulation efficiency and drug-loading of SN-38 NPs were attained the maximum values when the ratio of hydrophilic to hydrophobic block was between 1:2 and 1:3.

Nanoscale delivery systems have been widely investigated to overcome the penetration barrier of stratum corneum for effective transcutaneous application. The aim of this study is the development of effective vesicular formulations of ovalbumin and saponin which are able to promote penetration through the skin layers. Three kinds of vesicular formulations have been investigated as carriers, including liposomes, transfersomes and ethosomes, in which cholesterol and/or cationic lipid stearylamine are incorporated. The impact of membrane composition variations on the protein entrapment has been evaluated for each vesicle type. Formulations were characterized for particle size, polydispersity and encapsulation efficiency. The best formulations for each type of vesicle were subjected to in vivo transdermal immunization in mice. Among the three kinds of vesicular carrier, ethosomal nano carrier not only showed the best stability over a two months' storage, but also enabled the highest increase in the titer of serum antibody. In this regard, cationic nano-ethosomes can be considered as a promising vesicular carrier for transdermal vaccines. Meanwhile, we have developed a simple method to determine encapsulation efficiency of vesicular systems, which has potential application as a high throughput screening for vesicular formulations., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Effective encapsulation of drugs into the delivery systems could increase the efficiency of nanoparticles in prevention and treatment of diseases., Objective: The purpose of this study was to compare the different methods for determination of encapsulation efficiency of a model protein in the PLGA nanoparticles., Methods: The various direct methods include dichloromethane, acetonitrile, modified acetonitrile and NaOH based extraction and radioactive methods were used to directly calculate the encapsulation efficiency of the loaded protein in the PLGA nanoparticles. Furthermore, indirect methods include BCA, Fluorescent and radioactive methods were compared., Results: The encapsulation efficiencies determined by indirect methods include dichloromethane, acetonitrile, modified acetonitrile, NaOH based extraction and radioactive methods were 12.62% ± 1.97, 17.43% ± 2.51, 64.69% ± 4.31, 86.36% ± 2.25 and 90.15% ± 1.78, respectively. Moreover, the encapsulation efficiencies determined by indirect methods include BCA, fluorescent and radioactive methods were 81.46% ± 1.92, 88.23% ± 1.15 and 89.6% ± 1.9, respectively., Conclusions: Among the results obtained by indirect methods, radioactive and fluorescent methods showed more reliable. Moreover, NaOH and radioactive methods were the most reliable methods among the direct methods.

To design a useful lipid drug carrier having a high encapsulation efficiency (EE%) for the antiprostate cancer drugs flutamide (FT) and nilutamide (NT), a lipid nano-emulsion (LNE) was prepared with soybean oil (SO), phosphatidylcholine (PC), and sodium palmitate, and the partition coefficients (K p s) of the drugs for the LNE were determined by 19 F nuclear magnetic resonance (NMR) spectrometry. The 19 F NMR signal of the trifluoromethyl group of both drugs showed a downfield shift from an internal standard (trifluoroethanol) and broadening according to the increase in the lipid concentration due to their interaction with LNE particles. The difference in the chemical shift (Δδ) of each drug caused by the addition of LNE was measured under different amounts of LNE, and the K p values were calculated from the Δδ values. The results showed that FT has higher lipophilicity than NT. The total lipid concentration (SO + PC) required to encapsulate each drug into LNE with an EE% of more than 95% was calculated from the K p values as 93.3 and 189.9 mmol/L for FT and NT, respectively. For an LNE prepared with the total lipid concentration of 215 mmol/L, the predicted EE% values were 98 and 96% for FT and NT, respectively, while the experimental EE% values determined by a centrifugation method were approximately 99% for both drugs. Thus, the 19 F NMR spectrometric method is a useful technique to obtain the K p values of fluorinated drugs and thereby predict the theoretical lipid concentrations and prepare LNEs with high EE% values.

Solid lipid nanoparticles (SLN) have demonstrated good potential for oral peptide delivery. However, their hydrophobic nature generally accounts for low peptide entrapment efficiency (EE%). In this study, a new strategy was adopted to improve peptide EE% by incorporating a hydrophilic viscosity-enhancing agent (VA) within SLN cores to develop viscosity enhanced nanocarriers (VEN). Three agents namely, propylene glycol (PG), polyethylene glycol (PEG) 400 and PEG 600, were tested with human insulin serving as a model peptide drug. The effects of VA were both concentration- and type-dependent. 70% w/w PG had achieved the highest EE% (54.5%), versus the two PEGs, compared to only 20.4% in unmodified SLN. PG based VEN had demonstrated good dispersion stability at gastrointestinal (GI) pHs and preferential uptake by intestinal Caco2 cells while showing low cytotoxicity. Additionally, they preserved the integrity of insulin and significantly protected it against GI enzymatic degradation. Freeze dried VEN had shown good stability upon storage at -20°C. Orally administered insulin-VEN had achieved good hypoglycemic effect in fasted rats with relative bioavailability of 5.1%. To conclude, an easily implementable technique to improve peptide entrapment within SLN has been validated, and the resulting VEN had proved promising efficacy for oral peptide delivery., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Encapsulation of hydrophilic compounds for drug delivery systems with high loading efficiency is not easily feasible and remains a challenge, mainly due to the leaking of the drug to the outer aqueous phase during nanoparticle production. Usually, encapsulation of hydrophilic drugs is achieved by using double emulsion or inverse miniemulsion systems that often require the use of organic solvents, which may generate toxicological issues arising from solvent residues. Herein, we present the preparation of solid lipid nanoparticles loaded with a hydrophilic compound by a novel organic solvent free double emulsion/melt dispersion technique. The main objective of this study was to investigate the influence of important process and formulation variables, such as lipid composition, surfactant type, sonication parameters and lipid solidification conditions over physicochemical characteristics of SLN dispersion. Particle size and dispersity, as well as dispersion stability were used as responses. SLN dispersions with average size ranging from 277 to 550 nm were obtained, showing stability for over 60 days at 4 °C depending on the chosen emulsifying system. Entrapment efficiency of fluorescent dyes used as model markers was assessed by fluorescence microscopy and UV-vis spectrophotometry and results suggest that the obtained lipid based nanoparticles could be potentially applied as a delivery system of water soluble drugs., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In this study, PLGA or PLGA-PEG blend nanoparticles were developed loading amphotericin B (AmB), an antifungal agent broadly used in therapy. A 2(2) × 3(1) factorial experimental design was conducted to indicate an optimal formulation of nanoparticles containing AmB and demonstrate the influence of the interactions of components on the mean particle size and drug encapsulation efficiency. The independent variables analyzed were polymer amount (two levels) and organic phase (three factors in one level). The parameters methanol as cosolvent and higher polymer amount originated from the higher AmB encapsulation, but with the larger particle size. The selected optimized parameters were set as the lower polymer amount and ethyl acetate as cosolvent in organic phase, for both PLGA and PLGA-PEG nanoparticles. These parameters originated from nanoparticles with the size of 189.5 ± 90 nm and 169 ± 6.9 nm and AmB encapsulation efficiency of 94.0 ± 1.3% and 92.8 ± 2.9% for PLGA and PLGA-PEG nanoparticles, respectively. Additionally, these formulations showed a narrow size distribution indicating homogeneity in the particle size. PLGA and PLGA-PEG nanoparticles are potential carrier for AmB delivery and the factorial design presented an important tool in optimizing nanoparticles formulations.

Carboxymethyl-β-cyclodextrin (CMβ-CD)-modified glycol chitosan (GCS) nanoparticles (GCS-CMβ-CD NPs) were synthesized, and their pH-sensitive drug-release properties were investigated. GCS-CMβ-CD NPs could encapsulate doxorubicin hydrochloride (DOX), and the encapsulation efficiency and loading capacity increased with the amount of CMβ-CD. Drug-release studies indicate that DOX released was greater in acidic medium (pH 5.0) than in weakly basic medium (pH 7.4). The mechanism underlying the pH-sensitive properties of the carrier was analyzed. Finally, the MCF-7 (human breast cancer) and SW480 cell lines (human colon cancer) were used to evaluate the cytotoxicity of the NPs. The drug-loaded carriers show good inhibition of the growth of cancer cells compared with free DOX, and the carriers have good biocompatibility. In addition, the drug-loaded NPs have sustained drug-release properties. All these properties of the newly synthesized GCS-CMβ-CD NPs suggest a promising potential as an effective anticancer drug-delivery system for controlled drug release.

Ultrasmall polymer nanoparticles are rapidly gaining importance as nanocarriers for drugs and contrast agents. Here, a straightforward modular approach to efficiently loaded and stable sub-20-nm polymer particles is developed. In order to obtain ultrasmall polymer nanoparticles, we investigated the influence of one to two charged groups per polymer chain on the size of particles obtained by nanoprecipitation. Negatively charged carboxylate and sulfonate or positively charged trimethylammonium groups were introduced into the polymers poly(d,l-lactide-co-glycolide) (PLGA), polycaprolactone (PCL), and poly(methyl methacrylate) (PMMA). According to dynamic light scattering, atomic force and electron microscopy, the presence of one to two charged groups per polymer chain can strongly reduce the size of polymer nanoparticles made by nanoprecipitation. The particle size can be further decreased to less than 15 nm by decreasing the concentration of polymer in the solvent used for nanoprecipitation. We then show that even very small nanocarriers of 15 nm size preserve the capacity to encapsulate large amounts of ionic dyes with bulky counterions at efficiencies >90%, which generates polymer nanoparticles 10-fold brighter than quantum dots of the same size. Postmodification of their surface with the PEG containing amphiphiles Tween 80 and pluronic F-127 led to particles that were stable under physiological conditions and in the presence of 10% fetal bovine serum. This modular route could become a general method for the preparation of ultrasmall polymer nanoparticles as nanocarriers of contrast agents and drugs.

In the field of keratoconus treatment, a lipid-based liquid crystal nanoparticles system has been developed to improve the preocular retention and ocular bioavailability of riboflavin, a water-soluble drug. The formulation of this ophthalmic drug delivery system was optimized by a simplex lattice experimental design. The delivery system is composed of three main components that are mono acyl glycerol (monoolein), poloxamer 407 and water and two secondary components that are riboflavin and glycerol (added to adjust the osmotic pressure). The amounts of these three main components were selected as the factors to systematically optimize the dependent variables that are the encapsulation efficiency and the particle size. In this way, 12 formulas describing experimental domain of interest were prepared. Results obtained using small angle X-rays scattering (SAXS) and cryo-transmission electron microscopy (cryo-TEM) evidenced the presence of nano-objects with either sponge or hexagonal inverted structure. In the zone of interest, the percentage of each component was determined to obtain both high encapsulation efficiency and small size of particles. Two optimized formulations were found: F7 and F1. They are very close in the ternary phase diagram as they contain 6.83% of poloxamer 407; 44.18% and 42.03% of monoolein; 46.29% and 48.44% of water for F7 and F11, respectively. These formulations displayed a good compromise between inputs and outputs investigated.

Monophosphoryl lipid A (MPL) and Quil A are two immunological adjuvants commonly used in vaccines. At present no simple, validated methods for the quantification of Quil A and MPL have been previously reported therefore the aim of the current study was to develop a simple, fast and validated method to quantify MPL and Quil A using high performance liquid chromatography evaporative light scattering detection (HPLC-ELSD). The HPLC-ELSD technique was carried out using a ZORBAX Eclipse XDB-C8 column (2.1×50 mm; particle size, 3.5 μm) in an isocratic elution mode at 25 °C. MPL was eluted at a retention time of 1.8 min with methanol-water as the mobile phase and a detector temperature of 75 °C. Quil A was resolved as three peaks with retention times of 4.1, 5.5 and 6.4 min with a detector temperature of 30 °C and with water-acetonitrile and 0.01% formic acid as the mobile phase. The nebulizer pressure and gain were set at 3.5 bar and 10, respectively. Calibration curves plotted for both the adjuvants had an R(2)>0.997. Accuracy, intra- and inter-day precision were within the accepted limits. The limit of detection for MPL and Quil A were calculated as 1.343 and 2.06 μg/mL, respectively. The limit of quantification was 2.445 for MPL and 8.97 μg/mL for Quil A. This analytical method was used to quantify the entrapment and in vitro release of MPL and Quil A in a poly lactic-co-glycolic acid (PLGA) nanoparticle vaccine., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Context: Vitamins have been shown to reduce chemotherapy-related fatigue (CRF) by conserving energy loss both during and after cancer treatment. However, it remains unknown whether this reduction of fatigue interferes with the cancer drugs or alters the effectiveness of these agents., Objectives: The objective was to synthesize vitamin-cisplatin-loaded chitosan nano-particles for chemoprevention and cancer fatigue., Materials and Methods: Multi-vitamin (C, D3, and B12)-cisplatin composite nano-formulation called NanoCisVital (NCV) to overcome CRF. The interactions between vitamins and NCV were characterized using scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, and a particle size analyser. The chemo-preventive activity was performed by in vitro bio assays., Results: SEM analysis showed spherical shape and the size is < 225 nm. NCV inhibited the proliferating yeast cells as well as denaturation of bovine serum albumin, and it also reduced the sprouting of new blood vessels in dose-dependent manner., Conclusion: Collectively, these results demonstrate that the NCV particles can be used to reduce CRF without much affecting the anti-cancer properties of cisplatin.

To develop self-assembled liquid crystalline nanoparticles as a drug delivery system for keratoconus treatment, a formulation containing riboflavin a water-soluble drug, two surfactants (poloxamer 407 and mono acyl glycerol - monoolein-) and water was optimized and prepared by emulsification and a homogenization process. A fractional factorial design was applied to estimate the main effects and interaction effects of five parameters on two responses, namely particle size and encapsulation efficiency. The five parameters are the temperature of the two phases, the duration of emulsification, the presence of heating during homogenization, the number of passes and pressure. The most influent parameters are the presence of heating during the homogenization and the pressure that led to the production of nanoparticles with an average size of 145 nm and an average encapsulation efficiency of 46%.

Hypothesis: Solid lipid nanoparticles (SLNs) produced by conventional microemulsion techniques using thermal heat have specific limitations (e.g. high polydispersity, instability and low encapsulation). Replacing thermal heat with microwave heat may produce SLNs which overcome some of these limitations., Experiments: Stearic acid-based SLNs prepared with Tween® 20 as the emulsifier were chosen as the optimum formulation to encapsulate and potentially deliver the antibacterial drug tetracycline. All formulations were characterized for their particle size, zeta potential, encapsulation efficiency, loading capacity, thermal and X-ray diffraction analyses. Short-term stability and in vitro drug studies were also performed., Findings: Microwave heating helps to overcome several disadvantages associated with thermal heating (nonuniform, inefficient and slow) and results in improved particle characteristics. There is thus the potential for new opportunities in the development of colloidal carriers. The particle sizes of microwave-produced SLNs were in the desired nanometer range (200-250 nm) with both lower size and lower polydispersity than the conventional SLNs. We take this as an indication of improved stability; however zeta potential measurements were not different, indicating similar stability. True stability testing (visual observation with time) did show that the microwave-induced SLNs were found to be more stable, particularly when refrigerated. The microwave-produced SLNs also demonstrated improved encapsulation efficiency and loading capacity. Thermal and diffraction analysis confirmed a lowered crystallinity of stearic acid with successful incorporation of tetracycline into the SLNs. In vitro release studies indicated that, after an initial burst release, SLNs could provide prolonged release of tetracycline. The presence of tetracycline and non-toxicity of carriers towards microbes was confirmed by antimicrobial susceptibility tests., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Ion pairing of a fatty acid with an antibiotic may be an effective strategy for formulation optimization of a nanoantibiotic system. The aim of this study was therefore to explore the potential of linoleic acid (LA) as an ion pairing agent to simultaneously enhance encapsulation efficiency and antibacterial activity of triethylamine neutralized vancomycin (VCM) in solid lipid nanoparticles (SLNs). The prepared VCM-LA2 conjugate was characterized by Fourier transform-infrared (FT-IR) spectroscopy, logP and binding energy calculations. The shifts in the FT-IR frequencies of COOH, NH2 and CO functionalities, an increase in logP value (1.37) and a lower interaction energy between LA and VCM (-125.54 kcal/mol) confirmed the formation of the conjugate. SLNs were prepared by a hot homogenization and ultrasonication method, and characterized for size, polydispersity index (PI), zeta potential (ZP), entrapment efficiency (%EE), surface morphology and physical stability. In vitro antibacterial activity studies against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) were conducted. Size, PI and ZP for VCM-LA2&#95;SLNs were 102.7±1.01, 0.225±0.02 and -38.8±2.1 (mV) respectively. SLNs were also stable at 4 °C for 3 months. %EE for VCM-HCl&#95;SLNs and VCM-LA2&#95;SLNs were 16.81±3.64 and 70.73±5.96 respectively, indicating a significant improvement in encapsulation of the drug through ion pairing with LA. Transmission electron microscopy images showed spherical nanoparticles with sizes in the range of 95-100 nm. After 36 h, VCM-HCl showed no activity against MRSA. However, the minimum inhibitory concentration for VCM-HCl&#95;SLNs and VCM-LA2&#95;SLNs were 250 and 31.25 μg/ml respectively against S. aureus, while against MRSA it was 500 and 15.62 μg/ml respectively. This confirms the enhanced antibacterial activity of VCM-LA2&#95;SLNs over VCM-HCl&#95;SLNs. These findings therefore suggest that VCM-LA2&#95;SLNs is a promising nanoantibiotic system for effective treatment against both sensitive and resistant S. aureus infections., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Nanoparticles of Mentha × piperita essential oil were prepared by melt-dispersion method. The nanoparticles prepared at varying oil doses (5-10%, w/v) showed an encapsulation efficiency of 78.2-83.4%, while the oil load was observed to range between 3.64 and 7.46%. The average particle size of the nanoparticles varied between 226 and 331 nm, while polydispersity index showed variation between 0.547 and 1.000. DSC analysis indicated endothermic reaction during formation of nanoparticles, while a 2-term exponential kinetic model was followed during oil release. Nanoparticles showed considerable mortality against housefly larvae in lab (100%) as well as simulated field condition after first week (93%) and 6th week (57%) of application. This was the first study utilizing controlled release property of nanoparticles to formulate a cost effective product for breeding site application against housefly., (Copyright © 2013 Elsevier B.V. All rights reserved.)

This work investigated the suitability of lipid carriers as potential encapsulation method to improve the physical and chemical stability of microalgae oil high in docosahexaenoic acid (DHA). Lipid carriers with various oil contents were successfully prepared by a microfluidization method using stearic acid as solid lipid, microalgae oil as liquid lipid, and poloxamer 188 as surfactant. Results show that the mean particle diameter of the lipid carriers was in the range of 300 to 350 nm with the polydispersity index below 0.2. The lipid carriers were found to have spherical shape when examined under the transmission electron microscope. Data from the encapsulation efficiency and loading capacity indicate high distribution of microalgae oil throughout the lipid carriers and good physical stability as reflected by the particle size and size distribution during storage. Furthermore, the lower DPPH scavenging activity of lipid carriers compared with that of free microalgae oil suggests better chemical stability of microalgae oil encapsulated in lipid carriers. The addition of microalgae oil into lipid phase could disturb the crystalline order and form lattice defects to enable encapsulation of DHA as revealed by the results from differential scanning calorimetery. Current results suggest that this type of novel lipid carriers could be an efficient and promising carrier system for delivery of microalgae oil., (© 2014 Institute of Food Technologists®)

In the present work comparative evaluation of acetate and pH gradient techniques for effective drug loading in liposomes has been investigated. The acetazolamide (ACZ) loaded liposomes prepared by two methods were analyzed by vesicle size analysis, zeta potential, percent encapsulation efficiency, in vitro drug release studies and intraocular pressure lowering activity. ICH guidelines were followed for determining stability of the prepared liposomes. The superiority of acetate gradient method for active loading of acetazolamide has been established. The prepared acetate gradient positive liposomes showed extended hypotensive effect when compared to other liposomal formulations. Thus ACZ loaded liposomes prepared by acetate gradient technique may serve as promising ocular delivery system in the treatment of glaucoma. The current work emphasizes the fact that the techniques used for active drug loading into liposomes strongly influence the pharmaceutical performance of the final formulation., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background: Nanoparticles fabricated from the biodegradable and biocompatible polymer, polylactic-co-glycolic acid (PLGA), are the most intensively investigated polymers for drug delivery systems. The objective of this study was to explore fully the development of a PLGA nanoparticle drug delivery system for alternative preparation of a commercial formulation. In our nanoparticle fabrication, our purpose was to compare various preparation parameters., Methods: Docetaxel-loaded PLGA nanoparticles were prepared by a single emulsion technique and solvent evaporation. The nanoparticles were characterized by various techniques, including scanning electron microscopy for surface morphology, dynamic light scattering for size and zeta potential, x-ray photoelectron spectroscopy for surface chemistry, and high-performance liquid chromatography for in vitro drug release kinetics. To obtain a smaller particle, 0.2% polyvinyl alcohol, 0.03% D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS), 2% Poloxamer 188, a five-minute sonication time, 130 W sonication power, evaporation with magnetic stirring, and centrifugation at 8000 rpm were selected. To increase encapsulation efficiency in the nanoparticles, certain factors were varied, ie, 2-5 minutes of sonication time, 70-130 W sonication power, and 5-25 mg drug loading., Results: A five-minute sonication time, 130 W sonication power, and a 10 mg drug loading amount were selected. Under these conditions, the nanoparticles reached over 90% encapsulation efficiency. Release kinetics showed that 20.83%, 40.07%, and 51.5% of the docetaxel was released in 28 days from nanoparticles containing Poloxamer 188, TPGS, or polyvinyl alcohol, respectively. TPGS and Poloxamer 188 had slower release kinetics than polyvinyl alcohol. It was predicted that there was residual drug remaining on the surface from x-ray photoelectron spectroscopy., Conclusion: Our research shows that the choice of surfactant is important for controlled release of docetaxel.

Messenger RNA (mRNA) is the template for protein biosynthesis and is emerging as an essential active molecule to combat various diseases, including viral infection and cancer. Especially, mRNA-based vaccines, as a new type of vaccine, have played a leading role in fighting against the current global pandemic of COVID-19. However, the inherent drawbacks, including large size, negative charge, and instability, hinder its use as a therapeutic agent. Lipid carriers are distinguishable and promising vehicles for mRNA delivery, owning the capacity to encapsulate and deliver negatively charged drugs to the targeted tissues and release cargoes at the desired time. Here, we first summarized the structure and properties of different lipid carriers, such as liposomes, liposome-like nanoparticles, solid lipid nanoparticles, lipid-polymer hybrid nanoparticles, nanoemulsions, exosomes and lipoprotein particles, and their applications in delivering mRNA. Then, the development of lipid-based formulations as vaccine delivery systems was discussed and highlighted. Recent advancements in the mRNA vaccine of COVID-19 were emphasized. Finally, we described our future vision and perspectives in this field.

Messenger RNA (mRNA) is the template for protein biosynthesis and is emerging as an essential active molecule to combat various diseases, including viral infection and cancer. Especially, mRNA-based vaccines, as a new type of vaccine, have played a leading role in fighting against the current global pandemic of COVID-19. However, the inherent drawbacks, including large size, negative charge, and instability, hinder its use as a therapeutic agent. Lipid carriers are distinguishable and promising vehicles for mRNA delivery, owning the capacity to encapsulate and deliver negatively charged drugs to the targeted tissues and release cargoes at the desired time. Here, we first summarized the structure and properties of different lipid carriers, such as liposomes, liposome-like nanoparticles, solid lipid nanoparticles, lipid-polymer hybrid nanoparticles, nanoemulsions, exosomes and lipoprotein particles, and their applications in delivering mRNA. Then, the development of lipid-based formulations as vaccine delivery systems was discussed and highlighted. Recent advancements in the mRNA vaccine of COVID-19 were emphasized. Finally, we described our future vision and perspectives in this field. Lipid carriers with high biocompatibility have excellent mRNA encapsulation and delivery efficacy. Meanwhile, the emergence of lipid carriers provides an effective solution for delivering mRNA vaccines. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: The Box–Behnken design of experiments (BBD) is a statistical modelling technique that allows the determination of the significant factors in developing nanoparticles (NPs) using a limited number of runs. It also allows the prediction of the best levels of variables to obtain the desired characteristics (size, charge, and encapsulation efficiency) of the NPs. The aim of this study was to examine the effect of the independent variables (amount of polymer and drug, and surfactant concentration) and their interaction on the characteristics of the irinotecan hydrochloride (IRH)-loaded polycaprolactone (PCL) NPs and to determine the most optimum conditions for producing the desired NPs. Methods: The development of the NPs was carried out by a double emulsion solvent evaporation technique with yield enhancement. The NPs data were fitted in Minitab software to obtain the best fit model. Results: By using BBD, the most optimum conditions for producing the smallest size, highest magnitude of charge, and highest EE% of PCL NPs were predicted to be achieved by using 61.02 mg PCL, 9 mg IRH, and 4.82% PVA, which would yield 203.01 nm, −15.81 mV, and 82.35% EE. Conclusion: The analysis by BBD highlighted that the model was a good fit to the data, confirming the suitability of the design of the experiments. [ABSTRACT FROM AUTHOR]

The objective of this work is to investigate the nanoencapsulation of valuable bioactive compounds with the innovative electrohydrodynamic technology. More specifically, electrospraying, was applied for the encapsulation of three different active agents with lipolytic, anti-aging and antioxidant activity: Deoxycholic acid (DCA), olive leaves’ enzymatically modified extract and a hybrid of Deoxycholic acid-Hydroxytyrosol (DCA-HXT) in zein (ZN) biodegradable polymer. The resulting powders were evaluated regarding the system's: compositional and chemical characteristics, thermal stability and humidity effect, morphology, shape and size, encapsulation efficiency and release kinetics from the wall material. Encapsulation led to stable nanosystems, regarding the moisture and temperature effect, with high glass Transition Temperatures (Tg between 130-139 °C), and a mean particle size of 400-600 nm. Finally, high encapsulation efficiencies (DCA= 84.01%, DCA-HXT = 79.34% and modified extract =80.75%) and long-term release profiles (DCA=192 h, DCA-HXT=228 h and mod. extract=100 h) were measured for all three systems. Based on these results, we conclude that electrospraying process is an effective methodology for the development of bioactive agents’ nanosystems with excellent physicochemical properties and increased bioavailability; a crucial factor for the formation of high added-value products such as cosmetics.