Temperature- and pH-responsive poly(N-isopropylacrylamide-co-methacrylic acid) microgels as a carrier for controlled protein adsorption and release.

Herein, we report controlled protein adsorption and delivery of thermo- and pH-responsive poly(N-isopropylacrylamide-co-methacrylic acid) (PNIPAM-co-MAA) microgels at different temperatures, pH values and ionic strengths by employing bovine serum albumin (BSA) as a model protein. For these dual-responsive microgels, we found that the BSA adsorption was driven by several of six competing contributions, viz., physical diffusion (PD), hydrophobic interactions (HI), electrostatic attraction (EA), hydrogen bonding (HB) and temperature or pH-induced seizing action (SAT or SApH), depending on the temperature and pH of the solution. Compared to the pure PNIPAM microgels, the higher swelling degree of the PNIPAM-co-MAA microgels allowed a large amount of BSA loading under any experimental conditions. A largest BSA adsorption of 45.1 µg mg-1 was achieved at 40 °C and pH 4 due to the presence of all six contributions. The BSA adsorption and delivery could be further tuned by changing the crosslinking density within the microgels. The BSA binding onto the microgels was found to be ionic strength dependent, which could be attributed to the charge shielding of Na+ ions, salting out of BSA and aggregate formation of the microgels. The adsorbed BSA could be controllably released by adjusting the temperature and pH of the experiment, and with the help of sodium dodecyl sulphate (SDS) addition so as to eliminate each interaction between BSA and the microgels. Thus, this study can be useful to design a stimuli-responsive microgel-based carrier for controlled release of proteins.

Adsorption dynamics of thermoresponsive microgels with incorporated short oligo(ethylene glycol) chains at the oil-water interface.

Herein, we report a systematic study of the adsorption behaviour of short oligo(ethylene glycol) (OEG) chains incorporated into poly(N-isopropylaccrylamide) (PNIPAM) microgels at the dodecane-water interface as a function of the microgel concentration at two different temperatures: 298 and 313 K. The dynamic interfacial tension of the interface for the adsorption of these functional microgels is measured by means of a pendent drop method. We find that similar to pure PNIPAM microgels, the functionalized microgels initially get transported from the bulk to the interface, where they undergo the deformability dependent spreading process, and thus leading to a reduction of interfacial tension. However, the OEG chains significantly influence the dynamic processes of the microgels at the interface, enabling precise control over the interfacial activity. A tuneability of adsorption behaviour that is interpreted in terms of the diversity of structural and morphological features of the microgels, can be achieved by changing the temperature and/or the OEG chain length of the comonomer. While the temperature induced phase transition generally slows down the adsorption kinetics of the microgels, increasing the temperature from 298 to 313 K allows faster reduction of interfacial tension for the adsorption of the microgels with long OEG chains among the studied comonomers, making them a unique interfacially active functional material. Overall, incorporation of OEG chains allows tailoring the interfacial activity of microgels, thereby paving the way for the use of these microgels to act as effective Pickering emulsion stabilizers in a range of applications.

Protein Microgel-Stabilized Pickering Liquid Crystal Emulsions Undergo Analyte-Triggered Configurational Transition.

Herein, we report a novel approach that involves Pickering stabilization of micometer-sized liquid crystal (LC) droplets with biocompatible soft materials such as a whey protein microgel (WPM) to facilitate the analysis of analyte-induced configurational transition of the LC droplets. The WPM particles were able to irreversibly adsorb at the LC-water interface, and the resulting WPM-stabilized LC droplets possessed a remarkable stability against coalescence over time. Although the LC droplets were successfully protected by a continuous network of the WPM layer, the LC-water interface was still accessible for small molecules such as sodium dodecyl sulfate (SDS) that could diffuse through the meshes of the adsorbed WPM network or through the interfacial pores and induce an LC response. This approach was exploited to investigate the dynamic range of the WPM-stabilized LC droplet response to SDS. Nevertheless, the presence of the unadsorbed WPM in the aqueous medium reduced the access of SDS molecules to the LC droplets, thus suppressing the configuration transition. An improved LC response to SDS with a lower detection limit was achieved after washing off the unadsorbed WPM. Interestingly, the LC exhibited a detection limit as low as ∼0.85 mM for SDS within the initial WPM concentration ranging from 0.005 to 0.1 wt %. Furthermore, we demonstrate that the dose-response behavior was strongly influenced by the number of droplets exposed to the aqueous analytes and the type of surfactants such as anionic SDS, cationic dodecyltrimethylammonium bromide (DTAB), and nonionic tetra(ethylene glycol)monododecyl ether (C12E4). Thus, our results address key issues associated with the quantification of aqueous analytes and provide a promising colloidal platform toward the development of new classes of biocompatible LC droplet-based optical sensors.

Short oligo(ethylene glycol) chain incorporated thermoresponsive microgels: from structural analysis to modulation of solution properties.

Herein, we report synthesis of thermoresponsive poly(N-isopropylaccrylamide) (PNIPAM) microgels with short oligo(ethylene glycol) (OEG) chain comonomers (1 to 4/5 repeating unit) by surfactant-free precipitation copolymerization. The efficient incorporation of the comonomers was confirmed by a complete set of characterization methods viz., FTIR, 1H NMR, TEM, DLS, and viscometry. The structural heterogeneity and the distribution of the comonomers within the microgels were determined by means of 1H high-resolution transverse relaxation magnetization measurements. Interestingly, the incorporation of these short OEG chain comonomers led to the formation of a core-corona structure, in which the comonomers were mainly located in the core of the polymeric network with PNIPAM dangling chains at the microgel periphery. The experimental investigations of deswelling behaviours revealed that the OEG chains allowed precise control over the colloidal properties, including phase transition, particles size, swelling degree and polydispersity of the microgels. The tuneability of these properties that was interpreted in terms of polymeric hydrophobic/hydrophilic balance as well as structural diversity, could be achieved by changing the OEG chain length, comonomer feed and crosslinking density. Further, we found that the microgels with more hydrophilic OEG chains were able to show a higher relative swelling, and the same solid content thus led to a higher viscosity at all temperatures. The OEG chains remarkably improved the colloidal stability of the microgels in electrolyte solutions even at higher temperatures, thereby paving the way for the use of these microgels in a range of applications.

Adsorption and dilational rheology of mixed ß-casein/DoTAB layers formed by sequential and simultaneous adsorption at the water/hexane interface.

The interfacial behavior of ß-casein (ßCS) has been investigated in presence of the cationic surfactant dodecyl trimethyl ammonium bromide (DoTAB) at the water/hexane interface and compared to that obtained for the water/air interface. The used experimental technique is a drop profile analysis tensiometer specially equipped with a coaxial double capillary, which allows investigation of sequential adsorption of individual components besides the traditional simultaneous adsorption of two species. This method also provides the dilational rheological measurements based on low frequency harmonic drop oscillations. The tensiometric results show that the equilibrium states of the mixed ßCS/DoTAB layers built up on the two different routes do not differ significantly, that is, the general compositions of the mixed layers are similar. However, the results of dilational rheology for the two adsorption strategies are remarkably different indicating different dynamic characteristics of the adsorbed layers. These findings suggest that the respective mixed layers are more proteinlike if they are formed via sequential adsorption and more surfactant-like after simultaneous adsorption. In contrast to the W/A interface, at the W/H interface proteins remain at the interface once adsorbed and cannot be displaced just by competitive adsorption of surfactants.

Ultra-stable liquid crystal droplets coated by sustainable plant-based materials for optical sensing of chemical and biological analytes.

Herein, we demonstrate for the first time the synthesis of ultra-stable, spherical, nematic liquid crystal (LC) droplets of narrow size polydispersity coated by sustainable, biodegradable, plant-based materials that trigger a typical bipolar-to-radial configurational transition in dynamic response to chemical and biological analytes. Specifically, a highly soluble polymer, potato protein (PoP) and a physically-crosslinked potato protein microgel (PoPM) of ∼100 nm in diameter, prepared from the PoP, a byproduct of the starch industry, were compared for their ability to coat LC droplets. Although both PoP and PoPM were capable of reducing the interfacial tension between water and n-tetradecane <30 mN m-1, PoPM-coated LC droplets showed better stability than the PoP-coated droplets via a Pickering-like mechanism. Strikingly, the Pickering LC droplets outperformed PoP-stabilized droplets in terms of dynamic response with 5× lower detection limit to model chemical analytes (sodium dodecyl sulphate, SDS) due to the difference in SDS-binding features between the protein and the microgel. To eliminate the effect of size polydispersity on the response, monodisperse Pickering LC droplets of diameter ∼16 µm were additionally obtained using microfluidics, which mirrored the response to chemical as well as biological analytes, i.e., primary bile acid, an important biomarker of liver diseases. We demonstrate that these eco-friendly microgels used for creating monodisperse, ultra-stable, LC complex colloids are powerful templates for fabricating next generation, sustainable optical sensors for early diagnosis in clinical settings and other sensing applications.

Chitosan Hydrogels with Embedded Thermo- and pH-Responsive Microgels as a Potential Carrier for Controlled Release of Drugs.

We report a promising strategy based on chitosan (CS) hydrogels and dual temperature- and pH-responsive poly(N-isopropylacrylamide-co-methacrylic acid) (PNIPAM-co-MAA) microgels to facilitate release of a model drug, moxifloxacin (MFX). In this protocol, first, the microgels were prepared using a free radical copolymerization method, and subsequently, these carboxyl-group-rich soft particles were incorporated inside the hydrogel matrix using an EDC-NHS amidation method. Interestingly, the resulting microgel-embedded hydrogel composites (MG-HG) acting as a double barrier system largely reduced the drug release rate and prolonged the delivery time for up to 68 h, which was significantly longer than that obtained using microgels or hydrogels alone (20 h). On account of the dual-responsive features of the embedded microgels and the variation of water-solubility of drug molecules as a function of pH, MFX could be released in a controllable manner by regulating the temperature and pH of the delivery medium. The release kinetics followed a Korsmeyer-Peppas model, and the drug delivery mechanism was described by Fickian diffusion. Both the gel precursors and the hydrogel composites exhibited low cytotoxicity against mammalian cell lines (HeLa and HEK-293) and no deleterious hemolytic activity up to a certain higher concentration, indicating excellent biocompatibility of the materials. Thus, the unprecedented combination of modularity of physical properties caused by soft particle entrapment, unique macromolecular architecture, biocompatibility, and the general utility of the stimuli-responsive polymers offers a great promise to use these composite materials in drug delivery applications.

Physicochemistry of interaction between the cationic polymer poly(diallyldimethylammonium chloride) and the anionic surfactants sodium dodecyl sulfate, sodium dodecylbenzenesulfonate, and sodium N-dodecanoylsarcosinate in water and isopropyl alcohol-water media.

The physicochemistry of interaction of the cationic polymer poly(diallyldimethylammonium chloride) (PDADMAC) with the anionic surfactants sodium dodecyl sulfate, sodium dodecylbenzenesulfonate, and sodium N-dodecanoylsarcosinate was studied in detail using tensiometry, turbidimetry, calorimetry, viscometry, dynamic light scattering (DLS), and scanning electron microscopy (SEM). Fair interaction initially formed induced small micelles of the surfactants and later on produced free normal micelles in solution. The interaction process yielded coacervates that initially grew by aggregation in the aqueous medium and disintegrated into smaller species at higher surfactant concentration. The phenomena observed were affected by the presence of isopropyl alcohol (IP) in the medium. The hydrodynamic sizes of the dispersed polymer and its surfactant-interacted species were determined by DLS measurements. The surface morphologies of the solvent-removed PDADMAC and its surfactant-interacted complexes from water and IP-water media were examined by the SEM technique. The morphologies witnessed different patterns depending on the composition and the solvent environment. The head groups of the dodecyl chain containing surfactants made differences in the interaction process.

Poly-l-lysine-Functionalized Green-Light-Emitting Carbon Dots as a Fluorescence Turn-on Sensor for Ultrasensitive Detection of Endotoxin.

Herein, we report a facile, ultrasensitive, and selective fluorescence turn-on sensing strategy based on green-light-emitting functional nanodots for the detection of bacterial lipopolysaccharide (LPS) endotoxin. In this protocol, first, the pure carbon dots (CDs) with a fairly high quantum yield were prepared by microwave-assisted pyrolysis of citric acid in the presence of urea. Subsequently, the carboxyl-group-rich surfaces of the CDs were allowed to conjugate with the poly-l-lysine (PLL) using an EDC-NHS amidization method to obtain the PLL-modified CDs (PLL-CDs). The LPS could specifically bind to the PLL at the PLL-CD surfaces, and this binding enabled an electron transfer from the phosphate groups of LPS to the carbon core through the PLL bridge, thus resulting in a fluorescence enhancement. Interestingly, this fluorescent turn-on sensor provided a detection limit of 68.3 fM in PBS (pH 7.4), which is the lowest ever reported among all of the synthetic assays for LPS detection. Furthermore, our fluorescent probe was able to show a remarkable selectivity toward LPS over a range of commonly known interfering substances. Thus, this study demonstrated the feasibility of using specific LPS binding to PLL to drive molecular recognition in aqueous medium and offered an effective fluorescence turn-on sensing strategy to detect bacterial endotoxin in diverse clinical and biological applications.

Physicochemistry of the interaction between inulin and alkyltrimethylammonium bromides in aqueous medium and the formed coacervates.

Inulin, a polydisperse reserve polysaccharide, has prospective uses in food, pharmacy, and industry. Its uses and applications often encounter interactions with lipids and amphiphiles. Reports on such interactions are scarcely found in literature. In the present study, we have examined the nature of interactions between inulin and cationic amphiphiles, alkyltrimethylammonium bromides (CnTAB: n=12, 14, 16, 18), over a fair range of concentrations for both the polymer and the amphiphile. At low concentration, small induced amphiphile aggregates form complexes with inulin; at moderate concentration, the complexed inulin self-aggregates leading to coacervate formation, and at higher concentration, the amphiphile forms free micelles in solution. Tensiometric, conductometric, viscometric, and turbidimetric methods have been employed to study the above phenomena. The isolated coacervates of inulin with C18TAB were examined by scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force microscopy (AFM), differential scanning calorimetry (DSC), thermogravimetry (TG), and differential thermal analysis (DTA) to ascertain their morphology, structure, and thermal stability.

Microgel-stabilized liquid crystal emulsions enable an analyte-induced ordering transition.

We develop a new strategy that involves the formation of microgel (MG) decorated liquid crystal (LC) droplets, which show remarkable stability. This system facilitates the analysis of the LC droplets that undergo an analyte-triggered conformational transition, thus optimizing the quantitation of aqueous analytes.

The solution behavior of poly(vinylpyrrolidone): its clouding in salt solution, solvation by water and isopropanol, and interaction with sodium dodecyl sulfate.

This article deals with the solution properties of poly(vinylpyrrolidone) (PVP) in salt and surfactant environment. The cloud point (CP) of PVP has been found to be induced by the salts NaCl, KCl, KBr, Na2SO4, MgSO4, and Na3PO4. On the basis of CP values for a salt at different [PVP], the energetics of the clouding process have been estimated. The effect of the surfactant, sodium dodecyl sulfate (SDS), on the salt-induced CP has also been studied, and reduction in CP at low [SDS] and increase in CP at high [SDS] have been observed. The water vapor adsorption of PVP has been determined by isopiestic method. The results display a BET Type III isotherm whose analysis has helped to obtain the monolayer capacity of PVP and formation of multilayer on it. The solvation of PVP in a solution of water and a water-isopropanol mixture has been determined by conductometry from which contribution of the individual components were estimated. The interaction of PVP with SDS in solution led to formation of a complex entity, which has been studied also by conductometry adopting a binding-equilibrium scheme. SDS has been found to undergo two types of binding as monomers in the pre- critical aggregation concentration (CAC) range and as small clusters in the post CAC region. The stoichiometries of binding and binding constant were evaluated.

Tensiometry and dilational rheology of mixed ß-lactoglobulin/ionic surfactant adsorption layers at water/air and water/hexane interfaces.

Oscillating drop tensiometry was applied to study adsorbed interfacial layers at water/air and water/hexane interfaces formed from mixed solutions of ß-lactoglobulin (BLG, 1 µM in 10 mM buffer, pH 7 - negative net charge) and the anionic surfactant SDS or the cationic DoTAB. The interfacial pressure Π and the dilational viscoelasticity modulus |E| of the mixed layers were measured for mixtures of varying surfactant concentrations. The double capillary technique was employed which enables exchange of the protein solution in the drop bulk by surfactant solution (sequential adsorption) or by pure buffer (washing out). The first protocol allows probing the influence of the surfactant on a pre-adsorbed protein layer thus studying the protein/surfactant interactions at the interface. The second protocol gives access to the residual values of Π and |E| measured after the washing out procedure thus bringing information about the process of protein desorption. The DoTAB/BLG complexes exhibit higher surface activity and higher resistance to desorption in comparison with those for the SDS/BLG complexes due to hydrophobization via electrostatic binding of surfactant molecules. The neutral DoTAB/BLG complexes achieve maximum elastic response of the mixed layer. Mixed BLG/surfactant layers at the water/oil interface are found to reach higher surface pressure and lower maximum dilational elasticity than those at the water/air surface. The sequential adsorption mode experiments and the desorption study reveal that binding of DoTAB to pre-adsorbed BLG globules is somehow restricted at the water/air surface in comparison with the case of complex formation in the solution bulk and subsequently adsorbed at the water/air surface. Maximum elasticity is achieved with washed out layers obtained after simultaneous adsorption, i.e. isolation of the most surface active DoTAB/BLG complex. These specific effects are much less pronounced at the W/H interface.

A new water-soluble copper(II) complex as a selective fluorescent sensor for azide ion.

{[Cu(2)(H(2)L)(OH)(H(2)O)].(ClO(4))(2)(H(2)O)} (1) serves as a selective azide ion fluorescent sensor in aqueous medium. 1 binds with N(3)(-) to give [Cu(6)(HL)(2)(mu(1,1)-N(3))(6)] (2) which imposes rigidity and decreases the non-radiative decay of the excited state to give rise to fluorescent enhancement. 1 is highly selective over other various anions in aqueous medium.

Physicochemical studies on the biopolymer inulin: a critical evaluation of its self-aggregation, aggregate-morphology, interaction with water, and thermal stability.

Physicochemical properties viz., aggregation, molar mass, shape, and size of chicory inulin in solution were determined by fluorimetry, DLS, SLS, TEM, and viscometry methods. The thermal stability of the biopolymer was examined by TGA, DTA, and DSC measurements. The water vapor adsorption of desiccated inulin was also studied by the isopiestic method, and the data were analyzed in the light of the BET equation. On the basis of the obstruction to ion conductance by the inulin aggregates in solution and analysis of the data, the extent of hydration of inulin in solution was estimated. The result was coupled with the intrinsic viscosity, [eta], of inulin to ascertain the shape of the biopolymer aggregates in aqueous solution. The critical aggregation concentration (cac) of inulin in aqueous as well as in salt solution was assessed by fluorimetry. The weight average molar mass, Mw , of inulin monomer and its aggregate was found to be 4468 and 1.03 x 10(6) g/mol, respectively, in aqueous solution. This aggregated mass was 2.4 x 10(6) g/mol in 0.5M NH(4)SCN solution. The [eta] values of the soft supramolecular aggregates in solution (without and with salt) were small and comparable with globular proteins evidencing spherical geometry of the biopolymer aggregates as supported by the TEM results. In DMSO, rod-like aggregates of inulin was found by the TEM study. The [eta] of the biopolymer in the DMSO medium was therefore, higher than that in the aqueous medium. Unlike aqueous medium, the aggregation in DMSO was not associated with a cac.

Interfacial and bulk behavior of sodium dodecyl sulfate in isopropanol-water and in isopropanol-poly(vinylpyrrolidone)-water media.

The surface activity of isopropanol (IP) and poly(vinylpyrrolidone) (PVP) at the air/water interface has been studied. The self-aggregation of sodium dodecyl sulfate (SDS) in IP-water as well as in IP-PVP-water media has been investigated using physical methods, viz., tensiometry, conductometry, calorimetry, and viscometry. The interaction of SDS with PVP in IP-water medium as well as its self-aggregation (or micellization) in the presence of PVP has been assessed. The results reveal a fair degree of surface activity of IP in aqueous medium, which is only moderate for PVP. The critical micellar concentration (CMC) of SDS passes through a minimum at (v/v) % IP = 6.62. SDS interacts with PVP, yielding a critical aggregation concentration (CAC) at a low [SDS], independent of IP content in the medium. At a higher [SDS], free micelle formation takes place in solution, which is lower in mixed solvent than in water and is independent of solvent composition by tensiometry, but not by conductometry and calorimetry. The viscosity of micelle-interacted PVP in solution takes a long time to stabilize, whereas, for non-interacting additives, such as NaCl and cetyltrimethylammonium bromide (CTAB), it is time independent.

Transition metal carbene chemistry. 4. Nucleophilic attachment of DABCO to fischer carbene complexes in MeCN.

Rate constants for the attachment of DABCO (1,4-diazabicyclo[2.2.2]octane) to Fischer carbene complexes of the type (CO)(5)Cr=C(XR)C(6)H(4)Z (X = O and S) in dry MeCN at 25 degrees C are reported. Hammett rho values are 2.18 +/- 0.13 and 0.89 +/- 0.07 for DABCO reactions with (CO)(5)Cr=C(OMe)C(6)H(4)Z (10-Cr-Z) and (CO)(5)Cr=C(SMe)C(6)H(4)Z (11-Cr-Z), respectively. The rho values for the reaction of 10-Cr-Z and 11-Cr-Z with CH(CN)(2)(-) in 50% MeCN-50% H(2)O (v/v) are comparable to the present reactions. The reaction of DABCO with 10-Cr-Z is more closely related to the reaction of (n-Bu)(3)P with (CO)(5)W=C(OMe)C(6)H(5)-Z (23) which also provided a rho value 2.22. The much higher rho values and hence much higher reactivity of methoxy carbene complexes over the corresponding thiomethyl derivatives fit a pattern observed previously for alkoxide ion, OH(-), amine, and thiolate ion nucleophiles, and a rational explanation comes from the consideration of the substituent effects not only on the transition state but also on the reactant. A major difference between 10-Cr-Z and 11-Cr-Z is that the pi-donor effect of the methoxy group is much stronger than that of thiomethyl group. This leads to a substantial contribution of the zwitterionic form to the structure of 10-Cr-Z with much more localized positive charge on the methoxy group than the negative charge on the (CO)(5)Cr moiety. This leads to overall destabilization by an electron-withdrawing phenyl substituents resulting an increase in reactivity. The ethoxycarbene complexes are somewhat less reactive than their methoxy counterparts due to the somewhat more ground state stabilization through its stronger pi donor effect and partly due to steric crowding exerted by the slightly larger ethoxy group in the transition state. Higher k(1)(W)/k(1)(Cr) ratios for (thiomethyl)carbene complexes than methoxy or ethoxycarbene complexes are related to the intrinsic rate constant which is higher for ((thiomethyl)carbene)tungsten complexes than the corresponding Cr ones resulting in an enhanced ratio.