31 P-magnetic resonance (MR) is an important diagnostic technique currently used for tissue metabolites assessing, but it also has great potential for visualizing the internal body structures. However, due to the low physiological level of phosphorus-containing biomolecules, precise imaging requires the administration of an exogenous probe. Herein, this work describes the synthesis and MR characterization of a pioneering metal-free 31 P-MR probe based on phosphorus-containing polymeric zwitterion. The developed probe (pTMPC) is a well-defined water-soluble macromolecule characterized by a high content of naturally rare phosphorothioate groups providing a high-intensity 31 P-MR signal clearly distinguishable from biological background both in vitro and in vitro. In addition, pTMPC can serve as a sensitive 31 P-MR sensor of pathological conditions in vivo because it undergoes oxidation-induced structural changes in the presence of reactive oxygen species (ROS). Add to this the favorable 1 H and 31 P T1 /T2 relaxation times and biocompatibility, pTMPC represents a conceptually new diagnostic, whose discovery opens up new possibilities in the field of 31 P-MR spectroscopy and imaging.
The formation of biomolecular coronas around nanoparticles as soon as they come in contact with biological media is nowadays well accepted. The self-developed biological outer surfaces can affect the targeting capability of the colloidal carriers as well as their cytotoxicity and cellular uptake behavior. In this framework, we explored the structural features and biological consequences of protein coronas around block copolymer assemblies consisting of a common pH-responsive core made by poly[2-(diisopropylamino) ethyl methacrylate] (PDPA) and hydrophilic shells of different chemical natures: zwitterionic poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) or highly hydrophilic poly(ethylene oxide) (PEO) and poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA). We demonstrated the presence of ∼50 nm protein coronas around the nanoparticles regardless of the chemical nature of the polymeric shells. The thickness is understood as the sum of the soft and hard layers and it is the actual interface seen by the cells. Although the soft corona composition is difficult to determine because the proteins are loosely bound to the outer surface of the assemblies, the tightly bound proteins (hard corona) could be identified and quantified. The compositional analysis of the hard corona demonstrated that human serum albumin (HSA), immunoglobulin G (IgG) and fibrinogen are the main components of the protein coronas, and serotransferrin is present particularly in the protein corona of the zwitterionic-stabilized assemblies. The protein coronas substantially reduce the cellular uptake of the colloidal particles due to their increased size and the presence of HSA which is known to reduce nanoparticle-cell adhesion. On the other hand, their existence also reduces the levels of cytotoxicity of the polymeric assemblies, highlighting that protein coronas should not be always understood as artifacts that need to be eliminated due to their positive outputs.
- MeSH
- Cell Adhesion MeSH
- Hydrogen-Ion Concentration MeSH
- Humans MeSH
- Mechanical Phenomena * MeSH
- Nanoparticles chemistry MeSH
- Polymers chemistry MeSH
- Surface Properties MeSH
- Protein Corona chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Ultra-low fouling and functionalizable coatings represent emerging surface platforms for various analytical and biomedical applications such as those involving examination of cellular interactions in their native environments. Ultra-low fouling surface platforms as advanced interfaces enabling modulation of behavior of living cells via tuning surface physicochemical properties are presented and studied. The state-of-art ultra-low fouling surface-grafted polymer brushes of zwitterionic poly(carboxybetaine acrylamide), nonionic poly(N-(2-hydroxypropyl)methacrylamide), and random copolymers of carboxybetaine methacrylamide (CBMAA) and HPMAA [p(CBMAA-co-HPMAA)] with tunable molar contents of CBMAA and HPMAA are employed. Using a model Huh7 cell line, a systematic study of surface wettability, swelling, and charge effects on the cell growth, shape, and cytoskeleton distribution is performed. This study reveals that ultra-low fouling interfaces with a high content of zwitterionic moieties (>65 mol%) modulate cell behavior in a distinctly different way compared to coatings with a high content of nonionic HPMAA. These differences are attributed mostly to the surface hydration capabilities. The results demonstrate a high potential of carboxybetaine-rich ultra-low fouling surfaces with high hydration capabilities and minimum background signal interferences to create next-generation bioresponsive interfaces for advanced studies of living objects.
- MeSH
- Coated Materials, Biocompatible * chemistry pharmacology MeSH
- Cytoskeleton metabolism MeSH
- Humans MeSH
- Cell Line, Tumor MeSH
- Polymers * chemistry pharmacology MeSH
- Wettability MeSH
- Materials Testing * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Seven retention models have been selected to describe a dual-retention behavior of ten dopamine-related compounds on polymer-based monolithic stationary phase with zwitterion sulfobetaine functionality. Regression quality, as well as a statistical significance of individual regression parameters, have been evaluated. Better regression performance showed two four-parameter models when compared to three-parameter models. On the other hand, limited number of experimental points disqualified statistical robustness of four-parameter models. Among three-parameter models, retention description introduced by Horváth and Liang provided comparable quality of regression at significantly improved robustness. Multivariate analysis of the best three-parameter models provided the description of physicochemical properties of dopamine precursors and metabolites. Principal component analysis and logistic regression allowed structural characterization of dopamine-related compounds based solely on regression parameters extracted from an isocratic elution data. Both polarity and type of functional groups has been correctly assigned for 3-methoxytyramine that has not been part of an evaluation study. Among applied dual-retention models, Horváth´s model, initially developed to describe a retention of ionic compounds on nonpolar stationary phases, provided robust regression of experimental data and allowed an extraction of structural characteristics of dopamine-related compounds.
Nonthrombogenic modifications of membranes for extracorporeal membrane oxygenators (ECMOs) are of key interest. The absence of hemocompatibility of these membranes and the need of anticoagulation of patients result in severe and potentially life-threatening complications during ECMO treatment. To address the lack of hemocompatibility of the membrane, surface modifications are developed, which act as barriers to protein adsorption on the membrane and, in this way, prevent activation of the coagulation cascade. The modifications are based on nonionic and zwitterionic polymer brushes grafted directly from poly(4-methyl-1-pentene) (TPX) membranes via single electron transfer-living radical polymerization. Notably, this work introduces the first example of well-controlled surface-initiated radical polymerization of zwitterionic brushes. The antifouling layers markedly increase the recalcification time (a proxy of initiation of coagulation) compared to bare TPX membranes. Furthermore, platelet and leukocyte adhesion is drastically decreased, rendering the ECMO membranes hemocompatible.
Polymer vesicles formed by a pair of oppositely charged diblock copolyelectrolytes (PICsomes) are considered as a good alternative to polymersomes formed by amphiphilic copolymers. Here, we report on inherent stability and in vitro biocompatibility of PICsomes prepared from a pair of oppositely charged zwitterionic-ionic copolymers, in which the ionic block is a strong polyelectrolyte. Our results demonstrated that the PICsomes are highly stable over a wide range of pH and temperatures. Direct microscopic observations revealed that the PICsomes retain their morphology in the presence of human serum. In vitro studies using human skin fibroblasts (HSFs) showed that the polymer vesicles are not cytotoxic and do not affect cell proliferation and adhesion. A model hydrophilic dye was effectively incorporated into the PICsomes by simple mixing. Using confocal microscopy observations, we demonstrated that the dye-loaded PICsomes are efficiently internalized by the cells and are located predominantly in endo/lysosomal compartments. Thus, the PICsomes have promising potential for use as nanocontainers for substances of biomedical interest.
In this paper, the concentration of N-isopropylacrylamide in the polymerization mixture has been varied to prepare several polymethacrylate monolithic capillary columns. Polymer monoliths combining N-isopropylacrylamide with zwitterion monomer, as well as various dimethacrylate crosslinking monomers have been prepared and characterized. Uracil, thiourea, phenol, toluene, ethylbenzene, propylbenzene, and butylbenzene have been used to characterize retention of prepared capillary columns in the mobile phases with 40-95% of acetonitrile and at working temperatures ranging from 25 to 60°C. By an optimization of six-parameter polynomial models we have found that the retention of small molecules is affected mainly by the concentration of the acetonitrile in the mobile phase with very low contribution of working temperature and combined effect of acetonitrile concentration and temperature. Concentration of the mobile phase controlled also enthalpy of the retention. On the other hand, entropic contribution was almost insensitive to the change of the mobile phase composition, especially for mobile phases containing more than 60% of acetonitrile.
Porous polymer monoliths have been used to develop an online solid-phase extraction with liquid chromatography method for determination of dopamine in urine as well as for a continuous monitoring of dopamine in flowing system. A polymerization mixture containing 4-vinylphenylboronic acid monomer has been used to prepare a trapping column based on specific ring formation reaction with dopamine cis-diol functionality. Additionally, a monolithic stationary phase with zwitterion functionality has been used to prepare capillary column for the separation of dopamine. Experimental conditions including molarity, pH, and flow rate of the loading buffer together with a valve switching time have been optimized to provide the highest recovery for dopamine. Experimental setup has been used to determine dopamine in a urine. By using both calibration curve and standard addition method, the dopamine level was determined to be 1.19 and 1.28 mg/L, respectively. Further, we have used experimental design to optimize coupling of two extraction monolithic loops to separation capillary column with monolithic phase for a comprehensive monitoring of dopamine. After multivariate analysis, sample loading flow-rate and a flow-rate of flushing buffer were selected as the most significant variables. Optimized experimental setup was applied to continuously monitor dopamine degradation.
- MeSH
- Dopamine urine MeSH
- Solid Phase Extraction * MeSH
- Humans MeSH
- Polymerization MeSH
- Polymers MeSH
- Chromatography, High Pressure Liquid * MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
A monolithic sulfobetaine polymethacrylate micro-column BIGDMA-MEDSA designed in our laboratory, shows dual retention mechanism: In acetonitrile-rich mobile phase, hydrophilic interactions control the retention (HILIC system), whereas in more aqueous mobile phases the column shows essentially reversed-phase behavior with major role of hydrophobic interactions. The zwitterionic polymethacrylate micro-column can be used in the first dimension of two-dimensional LC in alternating reversed-phase (RP) and HILIC modes, coupled with an alkyl-bonded core-shell or silica-based monolithic column in the second dimension, for HILIC×RP and RP×RP comprehensive two-dimensional separations. During the HILIC×RP period, a gradient of decreasing acetonitrile gradient is used for separation in the first dimension, so that at the end of the gradient the polymeric monolithic micro-column is equilibrated with a highly aqueous mobile phase and is ready for repeated sample injection, this time for separation under reversed-phase gradient conditions with increasing concentration of acetonitrile in the first dimension. The fast repeating reversed-phase gradients on a short silica-monolithic or core-shell column in the second dimension can be optimized independently of the actual running first-dimension gradient program. As the alternating HILIC and RP separations on the first-dimension zwitterionic methacrylate column are based on complementary retention mechanisms, the instrumental setup essentially represents two coupled two-dimensional systems. It is first time that such an automated dual LCxLC approach is reported. The novel system allows obtaining three-dimensional data in a relatively short time and can be applied not only to multidimensional gradient separations of flavones and related polyphenolic compounds.
- MeSH
- Acetonitriles MeSH
- Betaine analogs & derivatives MeSH
- Chromatography, Liquid instrumentation methods MeSH
- Chromatography, Reverse-Phase instrumentation methods MeSH
- Flavones isolation & purification MeSH
- Hydrophobic and Hydrophilic Interactions MeSH
- Hydroxybenzoates isolation & purification MeSH
- Polymethacrylic Acids * MeSH
- Silicon Dioxide * MeSH
- Solvents MeSH
- Water MeSH
- Publication type
- Journal Article MeSH
Zwitterionic methacrylate based polymeric monolithic columns were prepared in two-step polymerizations, with reduced polymerization times. Characteristic properties such as hydrodynamic permeability, porosity, retention factors, and pore size distribution charts were used for column evaluation. A scaffold column was fabricated by polymerization of poly(lauryl methacrylate-co-tetraethyleneglycol dimethacrylate) and was used without further modification as a support for a poly(N,N-dimethyl-N-methacryloxyethyl-N-(3-sulfopropyl)ammonium betaine-co-bisphenol A glycerolate dimethacrylate) second monolith layer with zwitterionic functionality, for HILIC separations. An additional internal structure was formed by the second monolithic layer. The fabrication procedure was reproducible with RSD<5%. Field emission scanning electron microscopy has also been used to investigate column pore morphology, using a novel technique where the polymeric material is imaged directly, without coverage with a conducting film or particles. The new polar monolithic columns were used for HILIC separations of phenolic acids, flavones, nucleosides, and bases of nucleic acids, with similar efficiencies but different selectivities for zwitterionic methacrylate monolithic columns recently prepared by single step polymerization.
- MeSH
- Chemistry Techniques, Analytical instrumentation MeSH
- Chromatography, Liquid instrumentation MeSH
- Hydrophobic and Hydrophilic Interactions MeSH
- Polymethacrylic Acids chemistry MeSH
- Nucleosides analysis MeSH
- Permeability MeSH
- Polymerization MeSH
- Polymers chemistry MeSH
- Porosity MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
