Zein–resin composite nanoparticles with coencapsulated carvacrol

<b><sc>Abstract.</sc></b> Curcumin has diverse pharmacological applications as anti-inflammatory, anti-cancer, anti-fungal, and antibacterial. However, high hydrophobicity, rapid metabolism, and low bioavailability limit its application. Encapsulation improves its application by enhancing the bioavailability, stability, and solubility of this active compound. The study's objective was to investigate the ultrasonication effect and the compositional parameters on the physical attributes and encapsulation efficiency (EE) of the curcumin nanoparticles (NP). Through the emulsion evaporation method, (CN) were loaded into (poly-lactic acid -PLA). The selected parameters: curcumin concentration, surfactant concentration, sonication amplitude, and time were performed to achieve the highest nanoparticle attributes. Curcumin concentration of 0.2, 0.4, and 0.6 mg/mL at sonication amplitude of 80 and 100 µm with varying sonication time was used to prepare CN. The CN encapsulated in PLA (CNPLA) was characterized by a differential scanning calorimeter. A dynamic light scattering instrument was used to determine the Poly-dispersity Index (PDI), average hydrodynamic diameter (HD), and the zeta potential (ZP) and EE determined by UV vis-spectrophotometer. The results show that an increase in amplitude and 5 minutes sonication time resulted in significant (p < 0.05) decreased nanoparticle length. Concentrations of 0.2 mg/mL and 400 mg of dimethylamine boren at 100 µm exhibited optimum PDI, HD, ZP, and EE values. At 5min sonication time, PDI of 0.221±0.048, mean HD was 458.6±58.6 nm, and ZP was -57.3 mV. The EE was highest at 0.2 mg/mL concentration of Curcumin. Additionally, longer sonication time resulted in improved dispersion of the solvent. The results show that ultrasonication and compositional parameters can affect the processing of curcumin nanoparticles.

This study was focused on synthesizing, characterizing, and evaluating the antimicrobial effect of polymer nanoparticles (NPs) loaded with ampicillin. For this, the NPs were produced through polymeric self-assembly in aqueous media assisted by high-intensity sonication, using anionic polymers corresponding to the sodium salts of poly(maleic acid-co-vinylpyrrolidone) and poly(maleic acid-co-vinylpyrrolidone) modified with decyl-amine, here named as PMA-VP and PMA-VP-N10, respectively. The polymeric NPs were analyzed and characterized through the formation of polymeric pseudo-phases utilizing pyrene as fluorescent probe, as well as by measurements of particle size, zeta potential, polydispersity index, and encapsulation efficiency. The antimicrobial effect was evaluated by means of the broth microdilution method employing ampicillin sensitive and resistant Staphylococcus aureus strains. The results showed that PMA-VP and PMA-VP-N10 polymers can self-assemble, forming several types of hydrophobic pseudo-phases with respect to the medium pH and polymer concentration. Likewise, the results described that zeta potential, particle size, polydispersity index, and encapsulation efficiency are extremely dependent on the medium pH, whereas the antimicrobial activity displayed an interesting recovery of antibiotic activity when ampicillin is loaded in the polymeric NPs.

IntroductionBreast cancer treatment is plagued by systemic toxicity and drug resistance, prompting the search for better drug delivery systems, with oryzanol, a natural compound with anti-tumor potential but poor water solubility, emerging as a candidate. PLGA nanoparticles, a biodegradable and FDA-approved platform, are designed to encapsulate oryzanol, addressing its solubility issues and enabling targeted, controlled release to enhance anti-breast cancer efficacy. This study focuses on developing and characterizing γ-oryzanol-loaded PLGA (γ-oryzanol@PLGA) nanoparticles, evaluating their formulation, cellular effects, and mechanisms, intending to lay a preclinical foundation for oryzanol as a safe adjuvant therapy for breast cancer.MethodsTo address this unmet need, this study developed γ-oryzanol@PLGA nanoparticles (NPs) as a potential therapeutic strategy. Transmission electron microscopy (TEM) was used to characterize the morphology of the NPs. The colloidal stability and uniformity of nanoparticles were evaluated by measuring the polydispersity index (PDI) and zeta potential. Encapsulation efficiency and loading capacity were determined through UV-visible spectrophotometry. Flow cytometry was employed to assess the cellular uptake of the NPs compared to the free drug, and cytotoxicity assays were conducted to measure the effective concentration. Transcriptomic analysis was performed to identify differentially expressed genes and enriched cancer-related pathways.ResultsTEM results showed that the NPs were spherical with uniform morphology, with blank NPs having a size of 232.50 ± 1.27 nm and drug-loaded NPs being 241.60 ± 7.89 nm. The NPs exhibited excellent colloidal stability (PDI <0.03, zeta potential: −20 to −26 mV). Effective package load (86.22% ± 3.43%) and loading capacity (7.89% ± 0.31%) were achieved. Flow cytometry indicated a 3.2-fold enhanced cellular uptake compared to the free drug at 4 H (p < 0.05), and cytotoxicity assays showed a 3-fold reduction in the effective concentration. Transcriptomic analysis identified 576 differentially expressed genes and enriched cancer-related pathways, revealing the molecular mechanisms of the enhanced antitumor effects.ConclusionCollectively, these findings demonstrate that γ-oryzanol@PLGA NPs significantly improve drug delivery efficiency and therapeutic potency while maintaining excellent biocompatibility. This presents a promising nanoplatform for breast cancer treatment, warranting further preclinical development. Future studies should focus on in vivo validation and the exploration of combination therapies to fully realize the potential of this novel approach.

Lipid-PLGA hybrid nanoparticles of paclitaxel: Preparation, characterization, in vitro and in vivo evaluation.

Comparative study of lipid nanoparticle-based mRNA vaccine bioprocess with machine learning and combinatorial artificial neural network-design of experiment approach

Objective The objective of this study was to prepare the liver targeting drug delivery system (TDDS) of artesunate (ART)-loaded polyethylene glycol (PEG)-poly(d,l-lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) modified by glycyrrhetinic acid (GA), and evaluate its in vitro cytotoxicity. Significance The GA-PEG-PLGA-ART NPs enhanced the in vitro cytotoxicity on HCC cell lines. The development of GA-PEG-PLGA NPs will greatly push the clinical applications of ART as a novel anticancer drug. Methods The NPs were prepared using solvent evaporation method, and the formulation was optimized through an orthogonal design. In addition, physical properties were determined, including particle size, polydispersity index (PDI), zeta potential (ZP), morphology, drug loading capacity (LC) and encapsulation efficiency (EE), and in vitro drug release. Moreover, the in vitro cytotoxicity of NPs with three human cancer cell lines viz. HepG2, Hep3B, and SMCC-7721 was conducted using the SRB assay. Additionally, lyophilization was conducted to improve the long-term physical stability. Results The GA-PEG-PLGA-ART NPs have spherical shape, small particle size (around 88 nm) with a narrow size distribution (PDI < 0.3), high drug LC (up to 59.3 ± 1.65%), and high EE (up to 73.13 ± 5.17%). In vitro drug release behavior showed that drugs were released from NPs in a sustained and controlled release pattern. Cytotoxicity study indicated the NPs achieved lower cancer cell survival fraction. The GA-PEG-PLGA NPs freeze-dried with 3% (w/v) of mannitol showed better effect on long-term physical stability. Conclusion The GA-PEG-PLGA-ART NPs appear as a potential liver targeted intracellular delivery platform for ART.

We prepared and characterized topical ophthalmic formulations containing brimonidine-loaded bioadhesive cationic chitosan or anionic alginate nanoparticles (NPs) for sustained release of brimonidine as once daily regimen for management of glaucoma. Nanoparticles were prepared using a spontaneous emulsification solvent diffusion method. Different concentrations of polymers, emulsifiers, and NPs stabilizers were used for formulation optimization. Nanoparticles were characterized regarding particle size, zeta potential, morphology, and drug content. Brimonidine-loaded NPs were incorporated into eye drops, a temperature-triggered in situ gelling system, and a preformed gel. They then were characterized regarding their pH, viscosity, uniformity of drug content, in vitro release characteristics, in vitro cytotoxicity, and in vivo intraocular pressure (IOP) lowering effects. Characteristics of optimized brimonidine-loaded chitosan and alginate NPs, respectively, are: particle size, 115.67 ± 3.58 and 157.67 ± 5.53 nm; zeta potential, +35.27 ± 3.39 and -37.8 ± 3.77 mV; encapsulation efficiency, 74.34% ± 2.05% and 70.40% ± 2.77%; drug loading, 11.81% ± 0.67% and 13.14% ± 0.90%; and yield, 87.91% ± 5.92% and 76.53% ± 3.32%. Transmission electron microscope (TEM) analyses revealed that NPs have spherical shapes with a dense core and distinct coat. Formulations possessed uniform drug content. Furthermore, pH and viscosity were compatible with the eye. Formulations showed a sustained release without any burst effect with a Higuchi non-Fickian diffusion mechanism. Cytotoxicity studies revealed that all formulations are biocompatible. Importantly, all formulations possessed a sustained IOP lowering effect compared to the marketed brimonidine tartrate eye drops. These formulations provide a great improvement in topical ocular brimonidine delivery. The application of a single drop is sufficient to provide extended IOP reduction, which should improve patient compliance. We have developed a novel biocompatible topical delivery system for brimonidine, a first line glaucoma medication. Once daily application should have positive effects on patient compliance and, therefore, preservation of vision.

Nanoencapsulation of ascorbic acid loaded in pluronic® F127 coated by chitosan-alginate polyelectrolyte complex and application of a direct quantification method to enhance its accuracy.

Pharmaceutical Nanotechnology Nanoparticle-Based Topical Ophthalmic Formulations for Sustained Celecoxib Release

Cytotoxicity of CuO nanoparticles (NPs) are an impediment in utilizing them as an effective nanocarriers of chemotherapeutic drugs for targeted drug delivery in nasopharyngeal cancer. In our current study, we have designed a two-step synthesis and coating of CuO NPs with different concentrations of PLGA (polylactide-co-glycolide) to reduce the cytotoxicity. This was further conjugated with folic acid to enhance targeting to specific tissue. The multiple drugs loaded in the NPs were two potent anticancer drugs doxorubicin and docetaxel. A complete characterization studies including micrographic analysis, zeta potential measurements, polydispersity index, Fourier transform infrared spectroscopy (FTIR), encapsulation and loading efficiencies, stability and in vitro release studies were done. Cytoxicity studies were done with MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, acridine orange/ethidium bromide and DAPI (4, 6-diamidino-2-phenylindole, dihydrochloride) staining procedures. Impediametric studies were also carried out to reinforce the reduction in cytotoxicity. Finally the cellular uptake of the NPs was seen. It was evident from the results that the multiple drugs loaded CuO NPs formed with PLGA coating were uniform, non-agglomerated in size ranging from 180 to 195 nm. The FTIR revealed no major changes in drug peaks. Encapsulation and loading efficiencies showed sufficient amount of drug being loaded into the NPs. The drug loaded NPs showed no change in size or zeta potential even after a period of 30 days. The cytotoxicity studies revealed significant reduction in toxicity after coating the surface treated with PLGA as evident from the microscopic analysis of cells. Hence the current study may be prioritized and further in vivo/in vitro studies may be carried out.

The genomic revolution has identified therapeutic targets for a plethora of diseases, creating a need to develop robust technologies for combination drug therapy. In the present work, we describe a self-assembled polymeric nanoparticle (NP) platform to target and control precisely the codelivery of drugs with varying physicochemical properties to cancer cells. As proof of concept, we codelivered cisplatin and docetaxel (Dtxl) to prostate cancer cells with synergistic cytotoxicity. A polylactide (PLA) derivative with pendant hydroxyl groups was prepared and conjugated to a platinum(IV) [Pt(IV)] prodrug, c,t,c-[Pt(NH(3))(2)(O(2)CCH(2)CH(2)COOH)(OH)Cl(2)] [PLA-Pt(IV)]. A blend of PLA-Pt(IV) functionalized polymer and carboxyl-terminated poly(D,L-lactic-co-glycolic acid)-block-poly(ethylene glycol) copolymer in the presence or absence of Dtxl, was converted, in microfluidic channels, to NPs with a diameter of ∼100 nm. This process resulted in excellent encapsulation efficiency (EE) and high loading of both hydrophilic platinum prodrug and hydrophobic Dtxl with reproducible EEs and loadings. The surface of the NPs was derivatized with the A10 aptamer, which binds to the prostate-specific membrane antigen (PSMA) on prostate cancer cells. These NPs undergo controlled release of both drugs over a period of 48-72 h. Targeted NPs were internalized by the PSMA-expressing LNCaP cells via endocytosis, and formation of cisplatin 1,2-d(GpG) intrastrand cross-links on nuclear DNA was verified. In vitro toxicities demonstrated superiority of the targeted dual-drug combination NPs over NPs with single drug or nontargeted NPs. This work reveals the potential of a single, programmable nanoparticle to blend and deliver a combination of drugs for cancer treatment.

In this work, a pH-driven method was used to prepare zein-soy protein isolate (SPI) composite nanoparticles (NPs). The mass ratio of SPI to zein influenced the Z-average size (Z-ave). Once the zeta potential stabilized, SPI was completely coated on the periphery of the zein NPs. The optimal mass ratio of zein:SPI was found to be 2:3. After determining the structure using TEM, curcumin (Cur) and/or diosmetin (Dio) were loaded into zein-SPI NPs for co-encapsulation or individual delivery. The co-encapsulation of Cur and Dio altered their protein conformations, and both Cur and Dio transformed from a crystalline structure to an amorphous form. The protein conformation change increased the number of binding sites between Dio and zein NPs. As a result, the encapsulation efficiency (EE%) of Dio improved from 43.07% to 73.41%, and thereby increased the loading efficiency (LE%) of zein-SPI NPs to 16.54%. Compared to Dio-loaded zein-SPI NPs, Cur/Dio-loaded zein-SPI NPs improved the storage stability of Dio from 61.96% to 82.41% within four weeks. The extended release of bioactive substances in the intestine during simulated gastrointestinal digestion improved the bioavailability. When exposed to a concentration of 0-800 µg/mL blank-loaded zein-SPI NPs, the viability of HepG2 and LO-2 cells was more than 90%, as shown in MTT assay tests. The zein-SPI NPs are non-toxic, biocompatible, and have potential applications in the food industry.

The in silico characterization of bioactive substances which are constituents of functional foods or nutraceuticals is the methodology described in this communication. The aim of this work was to show the potential of the TOPSMODE approach as chemical-informatics method to study the structure/clastogenic activity (chromosomal aberrations) and identify structural alerts related to genotoxicity. The results of QSAR studies were analyzed for several classes of phenolic compounds (flavonoids, phenolic acids and coumarins) for which was required the use of software STATISTIC and MODESLAB and a mathematical model encoding topological information of a substructural level. It was observed that the criteria for maximum clastogenicity are the methoxy and hydroxyl polisubstitutions (methoxy > hydroxyl) and the polarity of the substituents. This was observed for all the analyzed subclasses. The confirmation of these results is based on the percentage of good classification for the used external databases, the recognition of physical chemical descriptors of polarity (μ1Pol y μ2Pol) and in the calculation of the fragments contribution. It can be conclude that QSAR methods, and in particular the used topographical approach, may constitute a predictive tool for the design and evaluation of bioactive components of functional foods or nutraceuticals.

Nutraceuticals and functional foods: Whole versus processed foods

In recently, there has been a great interest in natural antioxidants as bioactive components of food, nutraceuticals or potential drugs against several diseases. In our study, 88 extracts from various parts of plants from European Asteraceae and Cichoriaceae were assayed for radical scavenging activity by means of DPPH (1,1-diphenyl-2-picryl hydrazyl radical) test using the SIA (Sequential injection analysis) method developed for this purpose in our laboratory. DPPH radical scavenging activity of all tested plant extracts was evaluated according to the IC50 parameter. 29 extracts exhibited IC50 value lower than 0.1 mg/mL. The leaves of Leuzea carthamoides (IC50 = 0.046 mg/mL) were chosen as the most promising sample for a subsequent phytochemical study, which resulted in isolation of seven natural compounds, namely, 4′,5,7-trihydroxy-6-methoxyflavone (hispidulin) (1), 5, 7, 3′, 4′- tetrahydroxyflavanone (eriodictyol) (2), 3′,4′,5,7-pentahydroxy-6-methoxyflavonol (patuletin) (3), eriodictyol-7-β-glucopyranoside (4), 6-hydroxykaempferol-7-O-(6″-O-acetyl-β-D-glucopyranoside) (5), 4-hydroxybenzoic acid (6) and 3,4-dihydroxybenzoic acid (protocatechuic acid) (7). Antioxidant activity of the isolated compounds was evaluated by DPPH test and ferric reducing antioxidant power (FRAP) test and compared with trolox and quercetin. Both tests evaluated the flavonoid (5) as the most active antioxidant. This result was confirmed by comparison with known data concerning the structure/activity relationships of flavonoids.