PRODAN Photophysics as a Tool to Determine the Bilayer Properties of Different Unilamellar Vesicles Composed of Phospholipids.

Vesicles formed by phospholipids are promising candidates for drug delivery. It is known that the lipid composition affects properties such as the rigidity-fluidity of the membrane and that it influences the bilayer permeability, but sometimes sophisticated techniques are selected to monitor them. In this work, we study the bilayer of different unilamellar vesicles composed of different lipids (1,2-dioleoyl-sn-glycero-3-phosphocholine, DOPC, and lecithin) and diverse techniques such as extruder and electrospun templates and using 6-propionyl-2-(N,N-dimethyl) aminonaphthalene (PRODAN) and its photophysics. Moreover, we were able to monitor the influence of cholesterol on the bilayers. We demonstrate that the bilayer properties can be evaluated using the emission feature of the molecular probe PRODAN. This fluorescent probe gives relevant information on the polarity and fluidity of the microenvironment for unilamellar vesicles formed by two different methods. The PRODAN emission at 434 nm suggests that the bilayer properties significantly change if DOPC or lecithin is used in the vesicle preparation especially in their fluidity. Moreover, cholesterol induces alterations in the bilayer's structural and microenvironmental properties to a greater or lesser degree in both vesicles. Thus, we propose an easy and elegant way to evaluate physicochemical properties, which is fundamental for manufacturing vesicles as a drug delivery system, simply by monitoring the molecular probe emission band centered at 434 nm, which corresponds to the PRODAN species deep inside the bilayer.

Amphiphilic Ionic Liquids Capable to Formulate Organized Systems in an Aqueous Solution, Designed by a Combination of Traditional Surfactants and Commercial Drugs.

The present review describes the state of the art in the conversion of pharmaceutically active ingredients (API) in amphiphilic Ionic Liquids (ILs) as alternative drug delivery systems. In particular, we focus our attention on the compounds generated by ionic exchange and without original counterions which generate different systems in comparison with the simple mixtures. In water, these new amphiphiles show similar or even better properties as surfactants in comparison with their precursors. Cations such as 1-alkyl-3-methyl-imidazolium and anions such as dioctyl sulfosuccinate or sodium dodecyl sulfate appear as the amphiphilic components most studied. In conclusion, this work shows interesting information on several promissory compounds and they appear as an interesting challenge to extend the application of ILs in the medical field.

Production of Pd nanoparticles in microemulsions. Effect of reaction rates on the particle size.

In the synthesis of metallic nanoparticles in microemulsions, we hypothesized that the particle size is controlled by the reaction rate and not by the microemulsion size. Thus, the changes observed in the particle sizes as reaction conditions, such as concentrations, temperatures, the type of surfactant used, etc., are varied which should not be correlated directly to the modification of these conditions but indirectly to the changes they produce in the reaction rates. In this work, the microemulsions were formulated with benzene and water as continuous and dispersed phases, respectively, using n-dodecyltrimethylammonium bromide (DTAB) and n-octanol as the surfactant and cosurfactant. Using time-resolved UV-vis spectroscopy, we measured the reaction rates in the production of palladium (Pd) nanoparticles inside the microemulsions at different reactant concentrations and temperatures, keeping all the other parameters constant. The measured reaction rates were then correlated with the particle sizes measured by transmission electron microscopy (TEM). We found that the nanoparticle size increases linearly as the reaction rate increases, independently of the actual reactant concentration or temperature. We proposed a simple model for the observed kinetics where the reaction rate is controlled mainly by the diffusion of the reducing agent. With this model, we predicted that the particle size should depend indirectly, via the reaction kinetics, on the micelle radius, the water volume and the total microemulsion volume. Some of these predictions were indeed observed and reported in the literature.

Is it Necessary for the Use of Fluorinated Compounds to Formulate Reverse Micelles in a Supercritical Fluid? Searching the Best Cosurfactant to Create "Green" AOT Reverse Micelle Media.

Herein, we report the effect of employing two different alcohols, such as n-pentanol and 2,2,3,3,4,4,5,5-octafluoro pentanol (from now on F-pentanol), into 1,4-bis-2-ethylhexylsulfosuccinate (AOT) reverse micelles (RMs), to determine the interfacial activity and establish the best candidate to act as a cosurfactant in supercritical RMs. Dynamic light scattering (DLS), Fourier transform infrared (FT-IR), and fluorescence emission spectroscopy allowed us to determine and understand the behavior of alkanols in RMs. As a result, we found interesting displacements of alkanol molecules within the RMs, suggesting that the electrostatic interaction between SO3- and Na+ weakens because of new interactions of n-pentanol with SO3- through H-bonds, changing the curvature of the micellar interface. According to FT-IR and DLS studies, F-pentanol forms a RM polar core interacting through intermolecular H-bonds, suggesting no perturbations of the AOT RM interface. Hence, n-pentanol was selected as a cosurfactant to form supercritical RMs, which is confirmed by red edge excitation shift studies, using C343 as a molecular probe. Herein, we were able to create RMs under supercritical conditions without the presence of modified surfactants, fluorinated or multitailed compounds, which, to the best of our knowledge, was not shown before.

A comparative study of antimicrobial activity of differently-synthesized chitosan nanoparticles against bovine mastitis pathogens.

The greatest concern in dairy farming nowadays is bovine mastitis (BM), which results mainly from bacterial colonization of the mammary gland. Antibiotics are the most widely used strategy for its prevention and treatment, but overuse has led to growing antimicrobial resistance. Pathogens have also developed other mechanisms to persist in the udder, such as biofilm formation and internalization into bovine epithelial cells. New therapies are therefore needed to reduce or replace antibiotic therapies. In a previous study, we found that chitosan nanoparticles (Ch-NPs) have considerable potential for the treatment of BM. The aim of the present study was to evaluate the antimicrobial activity of differently-synthesized Ch-NPs against BM pathogens and their toxicity in bovine cells in vitro, to further explore the attributes of Ch-NPs for the prevention and treatment of intramammary infections. We also looked into their ability to inhibit biofilm formation and prevent the internalization of S. aureus into mammary epithelial cells. Finally, since an interesting approach for BM prevention is to enhance the host's immune response, we studied whether Ch-NPs could promote the release of pro-inflammatory cytokines in mammary epithelial cells. The results reveal that the bactericidal effect of Ch-NPs on BM pathogens and their ability to inhibit biofilm formation are size-dependent, with smaller particles being more efficient. In contrast, their effect on the viability of the cell lines is not size-dependent and all samples tested were non-toxic. The smallest Ch-NPs successfully prevented the internalization of S. aureus into the cells, but did not promote the production of pro-inflammatory cytokines. These findings make it possible to conclude that Ch-NPs are a great bactericidal agent which can prevent the main mechanisms developed by BM pathogens to persist in the udder.

How the external solvent in biocompatible reverse micelles can improve the alkaline phosphatase behavior.

In the last decade, the nature of the nonpolar solvents that can be part of reverse micelles (RMs) has been the topic of several investigations to improve their applications. In this sense, the hydrolysis of 1-naphthyl phosphate catalyzed by the enzyme alkaline phosphatase (AP) was used as a probe to investigate the effect of the change of the external solvent on RMs formulated with the anionic surfactant sodium diethylhexyl sulfosuccinate (AOT). As external nonpolar solvents, two biocompatible lipophilic esters, isopropyl myristate and methyl laurate, and the traditional nonpolar solvents, n-heptane and benzene, were used. The results were compared among the RMs investigated and with the reaction in homogeneous media. Thus, the effect of the nanoconfinement as well as the impact of the replacement of a conventional external nonpolar solvent by biocompatible solvents were analyzed. The results indicate that the catalytic efficiency in the AOT RMs is larger than that in homogeneous media, denoting a different hydration level over the AP enzyme, which is directly related to the different degrees of nonpolar solvent penetration to the RM interface. Our findings demonstrated that toxic solvents such as n-heptane and benzene can be replaced by nontoxic ones (isopropyl myristate or methyl laurate) in AOT RMs without affecting the performance of micellar systems as nanoreactors, making them a green and promising alternative toward efficient and sustainable chemistry.

Influence of the AOT Counterion Chemical Structure on the Generation of Organized Systems.

The impact of the imidazolium counterion structure on the organized systems formed by the surfactant 1,4-bis-2-ethylhexylsulfosuccinate, AOT, both in aqueous solutions and in nonpolar solvents is investigated. With this in mind, we investigated if the ionic liquid-like (IL-like) surfactant 1-ethyl-3-methylimidazolium 1,4-bis-2-ethylhexylsulfosuccinate, emim-AOT, forms direct micelles or vesicles in water. Dynamic light scattering, zeta potential, conductivity, fluorescence spectroscopy, and UV-visible spectroscopy measurements were performed to characterize the organized systems in aqueous solutions. We also studied the self-aggregation of emim-AOT, 1-butyl-3-methylimidazolium 1,4-bis-2-ethylhexylsulfosuccinate, bmim-AOT, and of 1-hexyl-3-methylimidazolium 1,4-bis-2-ethylhexylsulfosuccinate, hmim-AOT, in nonpolar solvents. The results obtained showed that the IL-like surfactant emim-AOT forms direct micelles in water, as sodium 1,4-bis-2-ethylhexylsulfosuccinate (Na-AOT) does. However, emim-AOT aggregates are larger, have a lower surface charge, are more stable, and have a more polar and less fluid micellar interface than Na-AOT micelles. It was also observed that emim-AOT and hmim-AOT form reverse micelles in nonpolar solvents. The size of the imidazolium cations dramatically influences the size of the reverse micelles and their ability to solubilize water.

Hydrolysis Reactions of Two Benzoyl Chlorides as a Probe to Investigate Reverse Micelles Formed by the Ionic Liquid-Surfactant bmim-AOT.

In this work, two hydrolysis reactions were used as a probe to investigate the properties of reverse micelles (RMs) formed by the ionic liquid-surfactant 1-butyl-3-methylimidazolium 1,4-bis-2-ethylhexylsulfosuccinate (bmim-AOT). The results were compared with those found for RMs generated with sodium 1,4-bis-2-ethylhexylsulfosuccinate (Na-AOT). As external nonpolar solvents, n-heptane (n-Hp), isopropyl myristate (IPM), and methyl laurate (ML) were used. Thus, the effect of changing the Na+ cation by bmim+ was analyzed, as well as the impact of the replacement of a conventional external nonpolar solvent by biocompatible solvents. The kinetics of the hydrolysis reactions of 4-methoxybenzoyl chloride (OMe) and 4-(trifluoromethyl)benzoyl chloride (CF3) were studied. The results indicate that the replacement of the Na+ counterion by bmim+ in AOT RMs alters the rates of reactions carried out in them and produces changes in the reaction mechanism. In bmim-AOT RMs, the bmim+ cation is located between the surfactant molecules; this has an important influence on the reaction intermediates' stability and, therefore, in the reaction rates and mechanisms. Also, the results indicate that when IPM is used as an external solvent instead of ML or n-Hp, interfacial water molecules have larger nucleophilicity due to the higher interface penetration of IPM.

Characterization of Reverse Micelles Formulated with the Ionic-Liquid-like Surfactant Bmim-AOT and Comparison with the Traditional Na-AOT: Dynamic Light Scattering, 1H NMR Spectroscopy, and Hydrolysis Reaction of Carbonate as a Probe.

The present study investigated how the presence of butylmethylimidazolium cation (bmim+) alters the interfacial properties of reverse micelles (RMs) created with the ionic liquid-like surfactant 1-butyl-3-methylimidazolium 1,4-bis-2-ethylhexylsulfosuccinate (bmim-AOT), in comparison to sodium 1,4-bis-2-ethylhexylsulfosuccinate (Na-AOT) RMs, employing dynamic light scattering (DLS) and 1H NMR techniques. Moreover, through the hydrolysis reaction of bis(4-nitrophenyl)carbonate inside both RMs as reaction probe, interfacial properties changes were explored in more detail. The kinetic solvent isotope effect was also analyzed. Micellar systems were formed using n-heptane as external nonpolar solvent and water as the polar component. According to the DLS studies, water is encapsulated inside the organized media; however, a different tendency is observed depending on the cationic component of the surfactant. For Na-AOT system, the results suggest that the micellar shapes are probably spherical, while in the case of bmim-AOT, a transition from ellipsoidal to spherical micelles could be occurring when water is added. 1H NMR data show that water is structured differently when Na+ cation is replaced by bmim+; in bmim-AOT RMs, the interaction of water with the surfactant is weaker and the water hydrogen-bonding network is less disturbed than in Na-AOT RMs. Kinetic studies reveal that the hydrolysis reaction in bmim-AOT RMs was much more favorable in comparison to Na-AOT RMs. In addition, when water content decreases in bmim-AOT RMs, the hydrolysis reaction rate increases and the solvent isotope effect remains constant, while for Na-AOT solutions, both the reaction rate and the solvent isotope effect decrease. Our results indicate that bmim+ cation would be located in the surfactant layer in such a way the negative charge density in the interface is less than that in Na-AOT RMs, and the reaction is more favorable. Additionally, as 1H NMR studies reveal, the interfacial water molecules would be more available in bmim-AOT RMs to participate in the nucleophilic attack. Therefore, the present study evidences how the replacement of Na+ counterion by bmim+ alters the composition of the interface of AOT RMs.

Use of Ionic Liquids-like Surfactants for the Generation of Unilamellar Vesicles with Potential Applications in Biomedicine.

The goal of this work is to understand the influence of the counterion nature on the organized systems formed by 1,4-bis-2-ethylhexylsulfosuccinate surfactants in aqueous solutions and how these aggregates will influence the deoxyribonucleic acid (DNA)-surfactant interactions. With this in mind, two ionic liquid-like surfactants were investigated: 1-butyl-3-methylimidazolium 1,4-bis-2-ethylhexylsulfosuccinate (bmim-AOT) and 1-hexyl-3-methylimidazolium 1,4-bis-2-ethylhexylsulfosuccinate (hmim-AOT). Measurements of dynamic light scattering, ζ-potential, transmission electron microscopy, and fluorescence and UV-visible spectroscopy were performed to study the characteristics of the vesicles formed by bmim-AOT and hmim-AOT. Regarding the determination of the interaction of the surfactants with DNA, circular dichroism was used. The results obtained showed that bmim-AOT and hmim-AOT ionic liquid-like surfactants spontaneously form unilamellar vesicles in water at very low surfactant concentrations. The characteristics of these aggregates are dependent on the length of the tail of the counterions. The length of the hydrophobic chains of the counterions also influences the DNA-surfactant interactions through hydrophobic effects.