scholarly journals Development of Novel EE/Alginate Polyelectrolyte Complex Nanoparticles for Lysozyme Delivery: Physicochemical Properties and In Vitro Safety

Pharmaceutics ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 103 ◽  
Author(s):  
Sabrina Sepúlveda-Rivas ◽  
Hans Fritz ◽  
Camila Valenzuela ◽  
Carlos Santiviago ◽  
Javier Morales
Keyword(s):  
Physicochemical Properties ◽  
Hela Cells ◽  
Electrostatic Interactions ◽  
Polyelectrolyte Complex ◽  
Total Charge ◽  
Biologic Drugs ◽  
High Association ◽  
High Concentrations ◽  
The Impact

The number of biologic drugs has increased in the pharmaceutical industry due to their high therapeutic efficacy and selectivity. As such, safe and biocompatible delivery systems to improve their stability and efficacy are needed. Here, we developed novel cationic polymethacrylate-alginate (EE-alginate) pNPs for the biologic drug model lysozyme (Lys). The impact of variables such as total charge and charge ratios over nanoparticle physicochemical properties as well as their influence over in vitro safety (viability/proliferation and cell morphology) on HeLa cells was investigated. Our results showed that electrostatic interactions between the EE-alginate and lysozyme led to the formation of EE/alginate Lys pNPs with reproducible size, high stability due to their controllable zeta potential, a high association efficiency, and an in vitro sustained Lys release. Selected formulations remained stable for up to one month and Fourier transform-Infrared (FT-IR) showed that the functional groups of different polymers remain identifiable in combined systems, suggesting that Lys secondary structure is retained after pNP synthesis. EE-alginate Lys pNPs at low concentrations are biocompatible, while at high concentrations, they show cytotoxic for HeLa cells, and this effect was found to be dose-dependent. This study highlights the potential of the EE-alginate, a novel polyelectrolyte complex nanoparticle, as an effective and viable nanocarrier for future drug delivery applications.

The Role of Gold Nanoparticles in Sonosensitization of Human Cervical Carcinoma Cell Line under Ultrasound Irradiation: An In Vitro Study

2019 ◽  
Vol 59 ◽  
pp. 1-14 ◽  
Author(s):  
Ahmad Shanei ◽  
Hadi Akbari-Zadeh ◽  
Hamid Fakhimikabir ◽  
Neda Attaran
Keyword(s):  
Gold Nanoparticles ◽  
Hela Cells ◽  
Ultrasound Irradiation ◽  
Ultrasound Wave ◽  
Efficient Treatment ◽  
Ultrasound Waves ◽  
Cervical Carcinoma Cell Line ◽  
Ultrasound Intensity ◽  
High Concentrations

Abstract:Purpose: The objective of this investigation was to evaluate the combined effects of ultrasound irradiation as a non-invasive and non-ionizing radiation with gold nanoparticles as ultrasound sensitizers on the HeLa cells.Materials and Methods: First, Gold nanoparticles (GNPs) were prepared, and the characterizations of nanoparticles were analyzed using TEM and UV-vis. Different concentrations of nanoparticles (0.2, 1, 5, 25 and 50 μg/ml) were used. Then, cytotoxicity of the GNPs was studied on HeLa cells, and finally concentrations of 0.2, 1 and 5 µg/mL were chosen for supplementary studies. The effects of nanoparticles and ultrasound irradiation with different intensities (0.5, 1 and 1.5 W/cm2) as well as the combination of ultrasound radiation with various concentrations of nanoparticles on 24, 48 and 72 h post-experiment cells’ viability were estimated by MTT and trypan blue assay.Results: Results showed that the sonosensitizing effect of nanoparticles mainly depended on the intensity of ultrasound waves and the concentration of GNPs. By increasing the concentration of GNPs and ultrasound intensity, the nanoparticle's effect of sensitizer was also increased. Moreover, as expected, the highest effect was observed at the highest intensities (1.5 W/cm2) of ultrasound wave and high concentrations (5 μg/ml) of GNPs 72 h after exposure to ultrasound irradiation.Conclusions: It is revealed that GNPs can be used as sonosensitizers of HeLa cells to ultrasound irradiation in order to produce an efficient treatment.

scholarly journals Biphasic Effect of Pirfenidone on Angiogenesis

2022 ◽  
Vol 12 ◽  
Author(s):  
Donghao Gan ◽  
Wenxiang Cheng ◽  
Liqing Ke ◽  
Antonia RuJia Sun ◽  
Qingyun Jia ◽  
...  
Keyword(s):  
Vascular Endothelial Cells ◽  
Tube Formation ◽  
Endothelial Cell Migration ◽  
High Concentration ◽  
Low Concentrations ◽  
Biphasic Effect ◽  
High Concentrations ◽  
And Migration ◽  
The Impact

Pirfenidone (PFD), a synthetic arsenic compound, has been found to inhibit angiogenesis at high concentrations. However, the biphasic effects of different PFD concentrations on angiogenesis have not yet been elucidated, and the present study used an in vitro model to explore the mechanisms underlying this biphasic response. The effect of PFD on the initial angiogenesis of vascular endothelial cells was investigated through a Matrigel tube formation assay, and the impact of PFD on endothelial cell migration was evaluated through scratch and transwell migration experiments. Moreover, the expression of key migration cytokines, matrix metalloproteinase (MMP)-2 and MMP-9, was examined. Finally, the biphasic mechanism of PFD on angiogenesis was explored through cell signaling and apoptosis analyses. The results showed that 10–100 μM PFD has a significant and dose-dependent inhibitory effect on tube formation and migration, while 10 nM–1 μM PFD significantly promoted tube formation and migration, with 100 nM PFD having the strongest effect. Additionally, we found that a high concentration of PFD could significantly inhibit MMP-2 and MMP-9 expression, while low concentrations of PFD significantly promoted their expression. Finally, we found that high concentrations of PFD inhibited EA.hy926 cell tube formation by promoting apoptosis, while low concentrations of PFD promoted tube formation by increasing MMP-2 and MMP-9 protein expression predominantly via the EGFR/p-p38 pathway. Overall, PFD elicits a biphasic effect on angiogenesis through different mechanisms, could be used as a new potential drug for the treatment of vascular diseases.

scholarly journals The Impact of Nylon-3 Copolymer Composition on the Efficiency of siRNA Delivery to Glioblastoma Cells

Nanomaterials ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 986 ◽  
Author(s):  
Natascha Hartl ◽  
Friederike Adams ◽  
Gabriella Costabile ◽  
Lorenz Isert ◽  
Markus Döblinger ◽  
...  
Keyword(s):  
Particle Size ◽  
Electrostatic Interactions ◽  
Laser Doppler Anemometry ◽  
Sirna Delivery ◽  
Copolymer Composition ◽  
Distribution Analysis ◽  
Glioblastoma Cells ◽  
Promising Therapeutic Approach ◽  
The Impact

Glioblastoma multiforme is a devastating disease that has attracted enormous attention due to poor prognosis and high recurrence. Small interfering RNA (siRNA) in principle offers a promising therapeutic approach by the downregulation of disease-related genes via RNA interference. For efficient siRNA delivery to target sites, cationic polymers are often used in preclinical studies for the protection of siRNA and complex formation based on electrostatic interactions. In an effort to develop biocompatible and efficient nanocarriers with a translational outlook for optimal gene silencing at reduced toxicity, we synthesized two sets of nylon-3 copolymers with variable cationic content (DM or NM monomer) and hydrophobic subunits (CP monomer) and evaluated their suitability for in vitro siRNA delivery into glioblastoma cells. DM0.4/CP0.6 and NM0.4/CP0.6 polymers with similar subunit ratios were synthesized to compare the effect of different cationic subunits. Additionally, we utilized NM0.2/CP0.8 polymers to evaluate the impact of the different hydrophobic content in the polymer chain. The siRNA condensation ability and polymer–siRNA complex stability was evaluated by unmodified and modified SYBR gold assays, respectively. Further physicochemical characteristics, e.g., particle size and surface charge, were evaluated by dynamic light scattering and laser Doppler anemometry, whereas a relatively new method for polyplex size distribution analysis—tunable resistive pulse sensing—was additionally developed and compared to DLS measurements. Transfection efficiencies, the route of cell internalization, and protein knockdown abilities in glioblastoma cells were investigated by flow cytometry. Furthermore, cellular tolerability was evaluated by MTT and LDH assays. All the polymers efficiently condensed siRNA at N/P ratios of three, whereas polymers with NM cationic subunits demonstrated smaller particle size and lower polyplex stability. Furthermore, NM0.2/CP0.8 polyplexes with the highest hydrophobic content displayed significantly higher cellular internalization in comparison to more cationic formulations and successful knockdown capabilities. Detailed investigations of the cellular uptake route demonstrated that these polyplexes mainly follow clathrin-mediated endocytotic uptake mechanisms, implying high interaction capacity with cellular membranes. Taken together with conducive toxicity profiles, highly hydrophobic nylon-3 polymers provide an appropriate siRNA delivery agent for the potential treatment of glioblastoma.

scholarly journals Comparative Toxicity of Fly Ash: An In Vitro Study

Molecules ◽  
2021 ◽  
Vol 26 (7) ◽  
pp. 1926
Author(s):  
Elvira Rozhina ◽  
Ilnur Ishmukhametov ◽  
Läysän Nigamatzyanova ◽  
Farida Akhatova ◽  
Svetlana Batasheva ◽  
...  
Keyword(s):  
Fly Ash ◽  
Hela Cells ◽  
In Vitro Study ◽  
The United States ◽  
Dark Field ◽  
Jurkat Cells ◽  
Silicate Mineral ◽  
Micrometer Size ◽  
The Impact

Fly ash produced during coal combustion is one of the major sources of air and water pollution, but the data on the impact of micrometer-size fly ash particles on human cells is still incomplete. Fly ash samples were collected from several electric power stations in the United States (Rockdale, TX; Dolet Hill, Mansfield, LA; Rockport, IN; Muskogee, OK) and from a metallurgic plant located in the Russian Federation (Chelyabinsk Electro-Metallurgical Works OJSC). The particles were characterized using dynamic light scattering, atomic force, and hyperspectral microscopy. According to chemical composition, the fly ash studied was ferro-alumino-silicate mineral containing substantial quantities of Ca, Mg, and a negligible concentration of K, Na, Mn, and Sr. The toxicity of the fly ash microparticles was assessed in vitro using HeLa cells (human cervical cancer cells) and Jurkat cells (immortalized human T lymphocytes). Incubation of cells with different concentrations of fly ash resulted in a dose-dependent decrease in cell viability for all fly ash variants. The most prominent cytotoxic effect in HeLa cells was produced by the ash particles from Rockdale, while the least was produced by the fly ash from Chelyabinsk. In Jurkat cells, the lowest toxicity was observed for fly ash collected from Rockport, Dolet Hill and Muscogee plants. The fly ash from Rockdale and Chelyabinsk induced DNA damage in HeLa cells, as revealed by the single cell electrophoresis, and disrupted the normal nuclear morphology. The interaction of fly ash microparticles of different origins with cells was visualized using dark-field microscopy and hyperspectral imaging. The size of ash particles appeared to be an important determinant of their toxicity, and the smallest fly ash particles from Chelyabinsk turned out to be the most cytotoxic to Jukart cells and the most genotoxic to HeLa cells.

scholarly journals IMPACT OF VARIOUS 2, 4-D CONCENTRATIONS AND DIFFERENT VITAMIN MIXTURES ON in vitro CULTURE OF COCONUT (Cocos nucifera L.) BY UTILIZING SEEDLINGS SHOOT TIP

2020 ◽  
Vol 19 (2) ◽  
pp. 135-145
Author(s):  
Sherif Fathy ElGioushy ◽  
Liu Rui ◽  
Haikuo Fan
Keyword(s):  
In Vitro Culture ◽  
Cocos Nucifera ◽  
Shoot Tips ◽  
Shoot Tip ◽  
Survival Percentage ◽  
High Concentrations ◽  
Cocos Nucifera L ◽  
The Impact ◽  
Vitamin Mixture

In this experiment, seedlings as sources for shoot tips was assembled from the coconut research institute (CRI) of Chinese Academy of tropical agricultural sciences germplasm, Wenchang, Hainan, China. The shoot tips were cultured on Y3 medium to study the impact of ten concentrations of 2, 4-D and three vitamin mixtures on survival, death and development % on in vitro culture establishment stage of coconut (Cocos nucifera L.). The obtained results revealed that all 2,4-D concentrations 4,7, 75, 100, 120 and 140 mg/L + any vitamin mixture did not give any development and high concentrations of 2, 4-D increased the death %. In any case, it can be concluded that the rate of differences in survival, death and development percentages exhibited by 2, 4-D concentrations was more pronounced than the analogous ones resulted by vitamin mixtures. At any rate, utilizing of (2, 4-D at concentration 25 mg/L+ vitamin mixture 1 or 2) exhibited the greatest values of survival percentage and reduced death percentage. Just as, increased the percentages of development in in vitro culture establishment stage of coconut (Cocos nucifera L.) through seedlings shoot tip.

scholarly journals Antimicrobial Activity of Encapsulated Mexican Oregano (Lippia berlandieri Schauer) Essential Oil Applied on Bagels

2020 ◽  
Vol 4 ◽  
Author(s):  
Aída Araceli Salgado-Nava ◽  
Ruth Hernández-Nava ◽  
Aurelio López-Malo ◽  
María Teresa Jiménez-Munguía
Keyword(s):  
Antimicrobial Activity ◽  
Essential Oil ◽  
Physicochemical Properties ◽  
Wheat Flour ◽  
Penicillium Expansum ◽  
Arabic Gum ◽  
Oregano Essential Oil ◽  
Significant Difference ◽  
High Concentrations

Several in vitro studies of Mexican oregano essential oil (MOEO) have demonstrated its effectiveness as an antimicrobial. On the other hand, the encapsulation technique of emulsification could be used to mask the intense flavor of MOEO, protect it from physicochemical degradation, and release gradually its antimicrobial components. In the present study, emulsions were elaborated with two encapsulating agents, pectin or Arabic gum, with five concentrations of MOEO (0%, 5%, 7.5%, 10%, or 15% w/w). Emulsions were characterized by density, viscosity, particle size distribution, creaming index, and peroxide index. Emulsions were applied, in a 3:1 ratio, on the surface of two formulations of bagel, one with wheat flour (BW) and another with wheat flour and oats (BWO). Bagels were characterized by water activity, moisture content, and hardness. The antimicrobial activity of MOEO encapsulated by emulsification on Penicillium expansum on the surface of the bagels was determined. In addition, a sensory analysis of the bagels was carried out using a hedonic scale. All physicochemical properties of the emulsions had a significant difference (p < 0.05) because of the encapsulating agent used (pectin or Arabic gum). Emulsions stabilized with Arabic gum were more stable over time than the ones with pectin. The application of the emulsions on the surface of the bagels did not show a statistical difference (p > 0.05) in the physicochemical properties of the breads. The growth of P. expansum was delayed for up to 5 or 6 days in bagels with wheat flour and Arabic gum as encapsulating agent. Both systems, BW and BWO, atomized with MOEO emulsions at high concentrations (MOEO 15% w/w) were well accepted by the panel of judges when these were evaluated on day 0 (freshly baked bread) but not after 6 days of their elaboration.

scholarly journals Influence of Different Peritoneal Dialysis Fluids on theIn VitroActivity of Cefepime, Ciprofloxacin, Ertapenem, Meropenem and Tobramycin againstEscherichia Coli

2016 ◽  
Vol 36 (6) ◽  
pp. 662-668 ◽  
Author(s):  
Manuel Kussmann ◽  
Linda Schuster ◽  
Sarah Wrenger ◽  
Petra Pichler ◽  
Gottfried Reznicek ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Peritoneal Dialysis ◽  
Antimicrobial Agents ◽  
Highly Active ◽  
Microbiological Outcome ◽  
Mueller Hinton ◽  
Baxter Healthcare ◽  
High Concentrations ◽  
The Impact

BackgroundPeritonitis is a major problem among patients on peritoneal dialysis (PD). The influence of diverse PD fluids on the activity of frequently used antibiotics has been insufficiently investigated. Thus, the present study set out to investigate the impact of different PD fluids on the activity of cefepime, ciprofloxacin, ertapenem, meropenem, and tobramycin against Escherichia coli.MethodsTime-kill curves in 4 different PD fluids (Dianeal PDG4, Extraneal, Nutrineal PD4 and Physioneal 40, all Baxter Healthcare Corp., Deerfield, IL, USA) were performed over 24 hours with 4 different concentrations (1 x minimum inhibitory concentration [MIC], 4 x MIC, 8 x MIC, 30 x MIC) of each antibiotic evaluated and without antibiotics as control. Cation-adjusted Mueller Hinton broth (CA-MHB) was used as comparator solution.ResultsIn all PD fluids investigated, bacterial growth and antimicrobial activity of all antibiotics tested was significantly reduced compared with the CA-MHB comparator solution. Except at high concentrations of 30 x MIC, cefepime, ertapenem and meropenem demonstrated a strongly reduced activity in all PD fluids investigated. Ciprofloxacin and tobramycin were highly active and bactericidal in all PD fluids and demonstrated dose-dependent activity.ConclusionThe antimicrobial activity of cefepime, ertapenem and meropenem is limited or even nullified in certain PD fluids in vitro, whereas ciprofloxacin and tobramycin show excellent activity. The choice of PD fluids can impact the activity of antimicrobial agents and might influence microbiological outcome. Further studies are required to verify the clinical relevance of our findings.

scholarly journals Enhanced Cellular Uptake in an Electrostatically Interacting Fucoidan–L-Arginine Fiber Complex

Polymers ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1795
Author(s):  
Vinothini Arunagiri ◽  
Hsieh-Chih Tsai ◽  
Haile Fentahun Darge ◽  
Endiries Yibru Hanurry ◽  
Chang Yi Lee ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Hela Cells ◽  
Electrostatic Interactions ◽  
In Vitro Study ◽  
Sulfated Polysaccharide ◽  
Industrial Applications ◽  
Resonance Spectroscopy ◽  
Brown Macroalgae ◽  
Negative Surface

Fucoidan is an abundant marine sulfated polysaccharide extracted from the cell wall of brown macroalgae (seaweed). Recently, fucoidan has been highly involved in various industrial applications, such as pharmaceuticals, biomedicals, cosmetics, and food. However, the presence of a sulfate group (negative surface charge) in the fucoidan structure limits its potential and biological activity for use in biomedical applications during cellular uptake. Thus, we aimed to improve the uptake of fucoidan by using an L-arginine uptake enhancer within an in vitro study. A Fucoidan–L-Arginine (Fuc-L-Arg) fiber complex was prepared via α-helical electrostatic interactions using a freeze-drying technique and confirmed using field-emission scanning electron microscopy, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectroscopy. In addition, fucoidan was conjugated with cyanine 3 (Cy3) dye to track its cellular uptake. Furthermore, the results of Fuc-L-Arg (1:1, 1:2.5) complexes revealed biocompatibility >80% at various concentrations (5, 10, 25, 50, 100 µg/mL). Owing to the higher internalization of the Fuc-L-Arg (1:5) complex, it exhibited <80% biocompatibility at higher concentrations (25, 50, 100 µg/mL) of the complex. In addition, improved cellular internalization of Fuc-L-Arg complexes (1:5) in HeLa cells have been proved via flow cytometry quantitative analysis. Hence, we highlight that the Fuc-L-Arg (1:5) fiber complex can act as an excellent biocomplex to exhibit potential bioactivities, such as targeting cancers, as fucoidan shows higher permeability in HeLa cells.

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