Solvophobicity-Driven Mesoscale Structures: Stabilizer-Free Nanodispersions.

Solvophobicity-driven mesoscale structures that lack any stabilizers are perhaps the most common spontaneously formed stable colloidal objects (particles, droplets). In spite of this, they have been significantly overlooked for a long time and the knowledge on solvophobicity-driven mesoscale structures (SDMSs) is rather limited. Here, we follow up on our previous work on mesoscale solubility [Rak, D.; Sedlák, M. On the Mesoscale Solubility in Liquid Solutions and Mixtures. J. Phys. Chem. B 2019, 123, 1365-1374. 10.1021/acs.jpcb.8b10638] and aim at providing a fully consistent picture of the nature, formation, and stability of SDMSs. We investigate both aqueous and nonaqueous mixtures, showing that this phenomenon is universal and not limited to aqueous systems. An experimental regime diagram is constructed as a function of the concentration of the solvophobic component and the solvophobicity strength given by mixtures of various organic solvents. Aqueous mixtures are investigated using well-defined ternary systems comprising water, ethanol (or other organic solvents), and a series of linear alkanes serving as hydrophobes. This investigation covers unique long-time monitoring of SDMS stability (up to three years). Another parameter studied in detail is the temperature of the mixture. SDMSs are characterized in terms of their shape and size distributions obtained using orthogonal techniques. Last but not least, we bring some insights into the SDMS surface zeta potential─the key quantity behind the stability of SDMSs. We investigate zeta potential as a function of the mixture composition, pH, and temperature.

Small Extracellular Vesicles Derived from Human Chorionic MSCs as Modern Perspective towards Cell-Free Therapy.

Mesenchymal stem cells (MSCs) are of great interest to scientists due to their application in cell therapy of many diseases, as well as regenerative medicine and tissue engineering. Recently, there has been growing evidence surrounding the research based on extracellular vesicles (EVs), especially small EVs (sEVs)/exosomes derived from MSCs. EVs/exosomes can be secreted by almost all cell types and various types of EVs show multiple functions. In addition, MSCs-derived exosomes have similar characteristics and biological activities to MSCs and their therapeutic applications are considered as a safe strategy in cell-free therapy. The aim of this study was the characterization of MSCs isolated from the chorion (CHo-MSCs) of human full-term placenta, as well as the isolation and analysis of small EVs obtained from these cells. Accordingly, in this study, the ability of small EVs' uptake is indicated by synovial fibroblasts, osteoblasts and periosteum-derived MSCs. Improvement in the understanding of the structure, characteristics, mechanism of action and potential application of MSCs-derived small EVs can provide new insight into improved therapeutic strategies.

Comment on "Bulk Nanobubbles or Not Nanobubbles: That is the Question".

In a recent article, Jadhav and Barigou ( Langmuir 2020, 36 (7), 1699-1708) investigated the question of the existence of stable bulk nanobubbles in water generated by hydrodynamic cavitation, ultrasound cavitation, and the addition of an organic compound (namely, ethanol) to water. They firmly conclude that these procedures result in stable bulk nanobubbles. However, a number of previous works documented that the nanoentities observed in water upon such procedures are not nanobubbles. Here, we analyze work of Jadhav and Barigou and show that conclusions regarding the nanobubble nature of the nanoentities are incorrect and are due to the choice of experimental techniques with weak sensitivity, methodical issues in the use of otherwise proper experimental techniques, and ambiguous outcomes of the rest of experiments.

γ-Butyrolactone Copolymerization with the Well-Documented Polymer Drug Carrier Poly(ethylene oxide)-block-poly(ε-caprolactone) to Fine-Tune Its Biorelevant Properties.

Polymeric drug carriers exhibit excellent properties that advance drug delivery systems. In particular, carriers based on poly(ethylene oxide)-block-poly(ε-caprolactone) are very useful in pharmacokinetics. In addition to their proven biocompatibility, there are several requirements for the efficacy of the polymeric drug carriers after internalization, e.g., nanoparticle behavior, cellular uptake, the rate of degradation, and cellular localization. The introduction of γ-butyrolactone units into the hydrophobic block enables the tuning of the abovementioned properties over a wide range. In this study, a relatively high content of γ-butyrolactone units with a reasonable yield of ≈60% is achieved by anionic ring-opening copolymerization using 1,5,7-triazabicyclo[4.4.0]dec-5-ene as a very efficient catalyst in the nonpolar environment of toluene with an incorporated γ-butyrolactone content of ≈30%. The content of γ-butyrolactone units can be easily modulated according to the feed ratio of the monomers. This method enables control over the rate of degradation so that when the content of γ-butyrolactone increases, the rate of degradation increases. These findings broaden the application possibilities of polyester-polyether-based nanoparticles for biomedical applications, such as drug delivery systems.

(Non)Existence of Bulk Nanobubbles: The Role of Ultrasonic Cavitation and Organic Solutes in Water.

A drawback of studies on bulk nanobubbles is the absence of direct proof that the nano-objects reported are really nanobubbles. The aim of our work was to provide such a proof or disproof. We focused on two effects (processes) commonly considered in research on nanobubbles: ultrasonic cavitation and addition of organic compounds to water, which could create in principle a barrier at the gas/water interface contributing to the stability of nanobubbles. We found that both of these processes lead to the generation of nano-objects, which are, however, not bulk nanobubbles. Ultrasonication leads to the formation of fine metal nanoparticles originating from the disintegration of the surface of the metal ultrasonic probe. Addition of organic solutes to water leads to the formation of a population of nanoparticles/nanodroplets originating from the so-called mesoscale solubilization of hydrophobic compounds present in the added solute as molecularly dissolved impurities. Subsequent ultrasonication of such mixtures adds metal nanoparticles and only slightly modifies the size distribution of mesoscale particles. While our results do not dismiss existence of nanobubbles in general, described effects must be seriously taken into account, especially in the case of biomedical applications where they can result in serious side effects.

Self-assembly and nanostructure of poly(vinyl alcohol)-graft-poly(methyl methacrylate) amphiphilic nanoparticles.

HYPOTHESIS: Poly(vinyl alcohol)-g-poly(methyl methacrylate) (PVA-g-PMMA) amphiphilic copolymers self-assemble to form multi-micellar colloidal systems. EXPERIMENTS: A PVA-g-PMMA copolymer containing 16-17% w/w of PMMA was synthesized by the free radical graft polymerization of methyl methacrylate on a PVA backbone by utilizing cerium(IV) ammonium nitrate as initiator and tetramethylethylenediamine (TEMED) as initiation activator. The aggregation behavior of the copolymer in water was comprehensively characterized by dynamic light scattering (DLS) and static light scattering (SLS), small angle neutron scattering (SANS), asymmetrical flow field-flow fractionation (A4F) and transmission electron microscopy (TEM). The colloidal stability before and after non-covalent crosslinking of PVA domains with boric acid was assessed by DLS. Finally, nanoparticles were nano spray-dried. FINDINGS: This copolymer self-assembles in water to form a complex nanostructure of low aggregation number particles (ca. 12-15 nm in diameter) that aggregate into larger ones with size ca. 60-80 nm, as determined by SANS and TEM. In addition, boric acid-crosslinking preserves the nanoparticle size, while conferring full physical stability under extreme dilution conditions. Nano spray-drying consolidates the crosslnking and enables the production of a dry flowing powder that upon re-dispersion in water regenerates the nanoparticles without major size changes.

Chitosan-graft-poly(methyl methacrylate) amphiphilic nanoparticles: Self-association and physicochemical characterization.

In this work, we synthesized and characterized the self-assembly behavior of a chitosan-poly(methyl methacrylate) graft copolymer and the properties of the formed nanoparticles by static and dynamic light scattering, small-angle neutron scattering, and transmission electron microscopy. Overall, our results indicate that the hydrophobization of the chitosan side-chain with PMMA leads to a complex array of small unimolecular and/or small-aggregation number "building blocks" that further self-assemble into larger amphiphilic nanoparticles.

Rifampicin Nanoformulation Enhances Treatment of Tuberculosis in Zebrafish.

Mycobacterium tuberculosis, the etiologic agent of tuberculosis, is an intracellular pathogen of alveolar macrophages. These cells avidly take up nanoparticles, even without the use of specific targeting ligands, making the use of nanotherapeutics ideal for the treatment of such infections. Methoxy poly(ethylene oxide)- block-poly(ε-caprolactone) nanoparticles of several different polymer blocks' molecular weights and sizes (20-110 nm) were developed and critically compared as carriers for rifampicin, a cornerstone in tuberculosis therapy. The polymeric nanoparticles' uptake, consequent organelle targeting and intracellular degradation were shown to be highly dependent on the nanoparticles' physicochemical properties (the cell uptake half-lives 2.4-21 min, the degradation half-lives 51.6 min-ca. 20 h after the internalization). We show that the nanoparticles are efficiently taken up by macrophages and are able to effectively neutralize the persisting bacilli. Finally, we demonstrate, using a zebrafish model of tuberculosis, that the nanoparticles are well tolerated, have a curative effect, and are significantly more efficient compared to a free form of rifampicin. Hence, these findings demonstrate that this system shows great promise, both in vitro and in vivo, for the treatment of tuberculosis.

On the Mesoscale Solubility in Liquid Solutions and Mixtures.

We report on a mesoscale solubility reflecting the fact that solubility is achieved not only by the well-known "like likes like" or "like dissolves like" based on molecular solvation but also on mesoscale solubilization of dislike compounds characterized in that the solubility (homogeneous distribution over the whole volume of the system) is achieved on a mesoscale level ranging from tens to hundreds of nanometers. It is shown that mesoscale solubility is a spontaneously occurring, literally everywhere present phenomenon, which was hidden and overlooked for a long time. This paper reveals the physical mechanism of mesoscale solubilization comprising nucleation and aggregation accompanied by the development of significant surface zeta potentials on nanoprecipitates, giving them a long-term stability. We show that mesoscale solubilization is common for aqueous as well as nonaqueous systems. Experiments with organic solvents not capable of self-ionization (self-dissociation) instead of water also shed light on the mechanism of the generation of surface zeta potentials at hydrophobic interfaces. We identified the key parameters enabling the mesoscale solubilization and mapped its occurrence as their function. Mesoscale structures including their formation kinetics, long-term stability, and different types of solubilization procedures were characterized by scattering and ultramicroscopic visualization comprising sizing and counting.

Self-assembly thermodynamics of pH-responsive amino-acid-based polymers with a nonionic surfactant.

The behavior of pH-responsive polymers poly(N-methacryloyl-l-valine) (P1), poly(N-methacryloyl-l-phenylalanine) (P2), and poly(N-methacryloylglycyne-l-leucine) (P3) has been studied in the presence of the nonionic surfactant Brij98. The pure polymers phase-separate in an acidic medium with critical pHtr values of 3.7, 5.5, and 3.4, respectively. The addition of the surfactant prevents phase separation and promotes reorganization of polymer molecules. The nature of the interaction between polymer and surfactant depends on the amino acid structure in the side chain of the polymer. This effect was investigated by dynamic light scattering, isothermal titration calorimetry, electrophoretic measurements, small-angle neutron scattering, and infrared spectroscopy. Thermodynamic analysis revealed an endothermic association reaction in P1/Brij98 mixture, whereas a strong exothermic effect was observed for P2/Brij98 and P3/Brij98. Application of regular solution theory for the analysis of experimental enthalpograms indicated dominant hydrophobic interactions between P1 and Brij98 and specific interactions for the P2/Brij98 system. Electrophoretic and dynamic light scattering measurements support the applicability of the theory to these cases. The specific interactions can be ascribed to hydrogen bonds formed between the carboxylic groups of the polymer and the oligo(ethylene oxide) head groups of the surfactant. Thus, differences in polymer-surfactant interactions between P1 and P2 polymers result in different structures of polymer-surfactant complexes. Specifically, small-angle neutron scattering revealed pearl-necklace complexes and "core-shell" structures for P1/Brij98 and P2/Brij98 systems, respectively. These results may help in the design of new pH-responsive site-specific micellar drug delivery systems or pH-responsive membrane-disrupting agents.

On the origin of mesoscale structures in aqueous solutions of tertiary butyl alcohol: the mystery resolved.

We have performed a detailed experimental study on aqueous solutions of tertiary butyl alcohol which were a subject of long-standing controversies regarding the puzzling presence of virtually infinitely stable large-scale structures in such solutions occurring at length scales exceeding appreciably dimensions of individual molecules, referred to also as mesoscale structures. A combination of static and dynamic light scattering yielding information on solution structure and dynamics and gas chromatography coupled with mass spectrometry yielding information on chemical composition was used. We show that tertiary butyl alcohol clearly exhibiting such structures upon mixing with water does not contain any propylene oxide, which was previously considered as a source of these structures (an impurity expected to be present in all commercial samples of TBA). More importantly, we show that no mesoscale structures are generated upon addition of propylene oxide to aqueous solutions of TBA. The ternary system TBA/water/propylene oxide exhibits homogeneous mixing of the components on mesoscales. We show that the source of the mesoscale structures is a mesophase separation of appreciably more hydrophobic compounds than propylene oxide. These substances are explicitly analytically identified as well as their disappearance upon filtering out the mesoscale structures by nanopore filtration. We clearly show which substances are disappearing upon filtration and which are not. This enables us to estimate with rather high probability the chemical composition of the mesoscale structures. Visualization of large-scale structures via nanoparticle tracking analysis is also presented. Video capturing the mesoscale particles as well as their Brownian motion can be found in the Supporting Information .

Large-scale inhomogeneities in solutions of low molar mass compounds and mixtures of liquids: supramolecular structures or nanobubbles?

In textbooks, undersaturated solutions of low molar mass compounds and mixtures of freely miscible liquids are considered as homogeneous at larger length scales exceeding appreciably dimensions of individual molecules. However, growing experimental evidence reveals that it is not the case. Large-scale structures with sizes on the order of 100 nm are present in solutions and mixtures used in everyday life and research practice, especially in aqueous systems. These mesoscale inhomogeneities are long-lived, and (relatively slow) kinetics of their formation can be monitored upon mixing the components. Nevertheless, the nature of these structures and mechanisms behind their formation are not clear yet. Since it was previously suggested that these can be nanobubbles stabilized by adsorbed solute at the gas/solvent interface, we devote the current study to addressing this question. Static and dynamic light scattering was used to investigate solutions and mixtures prepared at ordinary conditions (equilibrated with air at 1 atm), prepared with degassed solvent, and solutions and mixtures degassed after formation of large structures. The behavior of large structures in strong gravitational centrifugal fields was also investigated. Systems from various categories were chosen for this study: aqueous solutions of an inorganic ionic compound (MgSO4), organic ionic compound (citric acid), uncharged organic compound (urea), and a mixture of water with organic solvent freely miscible with water (tert-butyl alcohol). Obtained results show that these structures are not nanobubbles in all cases. Visualization of large-scale structures via nanoparticle tracking analysis is presented. NTA results confirm conclusions from our previous light scattering work.

Bromidobis(1,10-phenanthroline-κN,N')copper(II) dicyanamidate.

The title compound, [CuBr(C(12)H(8)N(2))(2)][N(CN)(2)], is formed of discrete [CuBr(phen)(2)](+) complex cations and uncoordinated [N(CN)(2)](-) anions (phen is 1,10-phenanthroline). The Cu atom is five-coordinated in a distorted trigonal-bipyramidal geometry by two phen mol-ecules and one bromide ligand, which coordinates in the equatorial plane at a distance of 2.5228 (4) Šand lying along with the Cu and the amide N atoms on a twofold rotation axis. The two axial Cu-N distances [1.9926 (15) Å] are slightly shorter than the two equatorial Cu-N bonds [2.0979 (15) Å]. The structure is stabilized by a weak C-H⋯N hydrogen bond, with a cyanide N atom of the dicyanamide ligand as an acceptor, and π-π inter-actions between nearly parallel phenyl and pyridine rings of two adjacent phen mol-ecules [centroid-centroid distance = 3.589 (1) Å], and between π electrons of the dicyanamide anion and the pyridine ring [N⋯Cg(pyridine) = 3.511 (3) Å; C-N⋯Cg(pyridine) = 80.2 (2)°].