Optical glucose detection across the visible spectrum using anionic fluorescent dyes and a viologen quencher in a two-component saccharide sensing system

A very general system is described in which anionic fluorescent dyes possessing a wide range of absorbance and emission wavelengths are used in combination with a boronic acid-modified viologen quencher to sense glucose at pH 7.4 in buffered aqueous solution. The present study demonstrates this capability with the use of eleven anionic fluorescent dyes of various structural types. Signal modulation occurs as the monosaccharide binds to the viologen quencher and alters its efficiency in quenching the fluorescence of the anionic dyes. The degree of quenching and the magnitude of the glucose signal were found to correlate roughly with the number of anionic groups on the dye. Optimal quencher : dye ratios were determined for each dye to provide a fairly linear signal in response to changes in glucose concentration across the physiological range.     

Preparation and characterization of insulin-loaded acrylic hydrogels containing absorption enhancers

The objectives of this study were to prepare insulin-loaded acrylic hydrogel formulations containing various absorption enhancers, to perform in vitro and in vivo characterization of these formulations, and to evaluate the factors which affecting insulin availability on rectal delivery of insulin using this hydrogel system. The acrylic block copolymer of methacrylic acid and methacrylate, Eudispert, was used to make the hydrogel formulations. As absorption enhancers, 2,6-di-O-methyl-beta-cyclodextrin (DM-beta-CyD), lauric acid (C12), or the sodium salt of C12 (C12Na), were incorporated into the hydrogels. In an in vitro release test, the release rate of insulin from the hydrogels decreased as the polymer concentration of the hydrogel increased. The addition of C12Na to the hydrogel further increased the insulin release rate, which was greater at higher concentrations of the enhancer. A portion of the C12Na was found to remain bound to the acrylic polymer in dissolution medium. Serum insulin levels were determined at various time points after the administration of insulin solution or insulin-loaded (50 units/kg body weight) Eudispert hydrogels containing 5% (w/w) of C12, C12Na, or DM-beta-CyD to in situ loops in various regions of the rat intestine. The most effective enhancement of insulin release was observed with formulations containing C12Na. The bioavailability of insulin from the hydrogels was lower than that from the insulin solutions. Hydrogel formulations containing 7% or 10% Eudispert remained in the rectum for 5 h after rectal administration. However, the 5% (w/w) C12Na solution stained with Evan's-blue had diffused out and the dye had reached the upper intestinal tract within 2 h. Finally, the rectal administration of insulin-loaded hydrogels, containing 4%, 7%, or 10% (w/w) Eudispert and 5% (w/w) of enhancer (C12, C12Na, or DM-beta-CyD) to normal rats was shown to decrease serum glucose concentrations. The greatest effect was found with insulin-loaded 7% (Eudispert) hydrogel containing C12Na which having cosiderable large insulin release rate and bioadhesive characteristics.     

The effect of boronic acid acidity on performance of viologen-based boronic acids in a two-component optical glucose-sensing system

A two-component saccharide sensing system using the fluorescent dye, hydroxypyrene trisulfonic acid, combined with a boronic acid functional viologen as a receptor/quencher in pH 7.4 buffer solution has been further investigated. The effect of substituents on the acidity of the boronic acid was measured. The boronic acid pK_a changed in the expected manner when electron donating or withdrawing groups were present. The glucose binding constants were dependent on pK_a, but no simple correlation was observed for the Stern-Volmer quenching constants and the fluorescence signal modulation.     

Organic Dye Adsorption by Amphiphilic Tris‐Urea Supramolecular Hydrogel

The development of an effective adsorbent for cleansing polluted water is required for environmental purification. In this respect, a supramolecular hydrogel constructed by the self‐assembly of small molecules could be a strong candidate. Adsorption experiments of organic dyes were performed using supramolecular hydrogels of amphiphilic tris‐urea 1 . Cationic organic dyes were adsorbed efficiently; indeed, the adsorption of methylene blue was as high as 4.19 mol equivalents relative to 1 . Two luminescence peaks were observed in the rhodamine 6G‐adsorbed supramolecular hydrogels, and their ratios varied with the amount of dye adsorbed. Fluorescence microscopy images of the supramolecular hydrogel at lower dye levels exhibited fibrous fluorescence consistent with the fibrous aggregates of 1 . According to these results, adsorption may proceed gradually, that is, occurring initially on the fibers and later in the aqueous spaces of the supramolecular hydrogel.     

Swelling equilibria and dye adsorption studies of chemically crosslinked superabsorbent acrylamide/maleic acid hydrogels

Superswelling acrylamide (AAm)/maleic acid (MA) hydrogels were prepared by free radical polymerization in aqueous solution of AAm with MA as comonomer with some multifunctional crosslinkers such as trimethylolpropane triacrylate and 1,4-butanediol dimethacrylate. AAm/MA hydrogels were used in experiments on swelling and adsorption of a water-soluble monovalent cationic dye such as Basic Blue 17 (Toluidin Blue). As a result of dynamic swelling tests, the influence of relative content of MA on the swelling properties of the hydrogel systems was examined. AAm/MA hydrogels were swollen in the range 1660-6050% in water, while AAm hydrogels swelled in the range 780-1360%. Equilibrium water content of AAm/MA hydrogels were calculated in the range 0.8873-0.9837. Water intake of hydrogels followed a non-Fickian type diffusion. The uptake of the cationic dye, BB-17 to AAm/MA hydrogels is studied by batch adsorption technique at 25 °C. In the experiments of the adsorption equilibrium, S-type adsorption in Giles's classification system was found. The binding ratio of hydrogel/dye systems was gradually increased with the increase of MA content in the AAm/MA hydrogels.     

The interaction of boronic acid-substituted viologens with pyranine: the effects of quencher charge on fluorescence quenching and glucose response

The fluorescence sensing of several monosaccharides using boronic acid-substituted viologen quenchers in combination with the fluorescent dye pyranine (HPTS) is reported. In this two-component sensing system, fluorescence quenching by the viologen is modulated by monosaccharides to provide a fluorescence signal. A series of viologen quenchers with different charges were prepared and tested for their ability both to quench the fluorescence of HPTS and to sense changes in glucose concentration in aqueous solution at pH 7.4. Both quenching efficiency and sugar sensing were found to be strongly dependent upon viologen charge. The molar ratio between HPTS and each of the viologen quenchers was varied in order to obtain an optimal ratio that provided a fairly linear fluorescence signal across a physiological glucose concentration range. Both the quenching and sugar sensing results are explained by electrostatic interaction between dye and quencher.     

Fabrication of poly(ethylene glycol)‐based hydrogels entrapping enzyme‐immobilized silica nanoparticles

In this study, we immobilized enzymes by combining covalent surface immobilization and hydrogel entrapment. A model enzyme, glucose oxidase (GOX), was first covalently immobilized on the surface of silica nanoparticles (SNPs) via 3‐aminopropyltriethoxysilane (APTES), and the resultant SNP‐immobilized enzyme was physically entrapped within photopolymerized hydrogels prepared from two different molecular weights (MWs) (575 and 8000 Da) of poly(ethylene glycol)(PEG). The hydrogel entrapment resulted in a decrease in reaction rate and an increase in apparent K_m of SNP‐immobilized GOX, but these negative effects could be minimized by using hydrogel with a higher MW PEG, which provides higher water content and larger mesh size. The catalytic rate of the PEG 8000 hydrogel was about ten times faster than that of the PEG 575 hydrogel because of enhanced mass transfer. Long‐term stability test demonstrated that SNP‐immobilized GOX entrapped within hydrogel maintained more than 60% of its initial activity after a week, whereas non‐entrapped SNP‐immobilized GOX and entrapped GOX without SNP immobilization maintained less than 20% of their initial activity. Incorporation of SNPs into hydrogel enhanced the mechanical strength of the hydrogel six‐fold relative to bare hydrogels. Finally, a hydrogel microarray entrapping SNP‐immobilized GOX was fabricated using photolithography and successfully used for quantitative glucose detection. Copyright © 2009 John Wiley & Sons, Ltd.     

Preparation of interpenetrating polymer network composed of poly(ethylene glycol) and poly(acrylamide) hydrogels as a support of enzyme immobilization

Poly(ethylene glycol)(PEG)‐based interpenetrating polymeric network (IPN) hydrogels were prepared for the application of enzyme immobilization. Poly(acrylamide)(PAAm) was chosen as the other network of IPN hydrogel and different concentration of PAAm networks were incorporated inside the PEG hydrogel to improve the mechanical strength and provide functional groups that covalently bind the enzyme. Formation of IPN hydrogels was confirmed by observing the weight per cent gain of hydrogel after incorporation of PAAm network and by attenuated total reflectance/Fourier transform infrared (ATR/FTIR) analysis. Synthesis of IPN hydrogels with higher PAAm content produced more crosslinked hydrogels with lower water content (WC), smaller M_c and mesh size, which resulted in enhanced mechanical properties compared to the PEG hydrogel. The IPN hydrogels exhibited tensile strength between 0.2 and 1.2 MPa while retaining high levels of hydration (70–81% water). For enzyme immobilization, glucose oxidase (GOX) was immobilized to PEG and IPN hydrogel beads. Enzyme activity studies revealed that although all the hydrogels initially had similar enzymatic activity, enzyme‐immobilizing PEG hydrogels lost most of the enzymatic activity within 2 days due to enzyme leaching while IPN hydrogels maintained a maximum 80% of the initial enzymatic activity over a week due to the covalent linkage between the enzyme and amine groups of PAAm. Copyright © 2008 John Wiley & Sons, Ltd.     

Novel Cationic Poly[AAm/NVP/DAPB] Hydrogels for Removal of Some Textile Anionic Dyes from Aqueous Solution

Novel cationic poly [AAm/NVP/DAPB] hydrogels (YH₁ – YH₅) were synthesized by free radical solution polymerization of AAm, NVP and DAPB by increasing the concentration of DAPB in the feed. According to swelling experiments, hydrogel YH₅ with higher DAPB content gave relative higher swelling percentage compared to other hydrogels. The hydrogel YH₅ was characterized by FTIR, TGA and SEM analysis. The anionic dye solution such as Orange-II, Reactive Golden Yellow and Acid Yellow were used as an adsorption medium to investigate the usability of the hydrogel for the removal of anionic dyes. The effect of pH of the adsorption medium, initial concentration of the dye, adsorbent dose, %swelling and contact time was also investigated. Furthermore, the Langmuir and Freundlich adsorption isotherms were applied and they showed a good fit to the experimental data. From the results, the uptake of dyes within the hydrogel increased in the following order: Reactive Golden Yellow>Orange-II>Acid Yellow.     

SWELLING BEHAVIOR OF ACRYLAMIDE HYDROGEL IN DIFFERENT SOLVENTS AND pHs

Swelling property of acrylamide hydrogels, prepared from aqueous solutions of acrylamide monomer having concentrations in the range of 10-60 wt% by γ-ray irradiation method using a Co-60 gamma radiation source at doses ranging 1-30.0 kGy, has been investigated under various swelling media. These swelling media were basically solvents (solutions), produced by dissolving methanol, ethanol, glucose, sucrose, sodium chloride and sodium persulfate individually with distilled water, and solutions prepared with pHs = 3, 7 and 10. The investigation was performed in order to observe the effect of these solvents and pHs as well as the influence of monomer concentrations, radiation doses and times on swelling behavior of hydrogels. Swelling values were found higher for hydrogels prepared with lower monomer concentrations (ca.20 wt%) and radiation doses (ca. 5 kGy) and showed a leveling off tendency within 24 h. The glucose solvent and the buffer solution of pH = 10 revealed significant increase of swelling of hydrogels as compared to other solutions. Results are explained based on crosslinking density in hydrogel, polymer-solvent/polymer-polymer interactions in solutions,permeability of molecules in solutions and ionization capacity of hydrogel in pH.     

Silica hydrogel formation and aging monitored by pyrene-based fluorescence probes

Fluorescent probes pyrene (Py), di(1-pyrenylmethyl)ether (DiPyM) and newly synthesized 2,3-bis-[4-(1-pyrenemethoxy)methylphenyl]butane (DiPyS) were used to monitor the formation and aging of silica hydrogel prepared from poly(glyceryl silicate) (PGS) sol. The fluorescence emission spectra of these probes are sensitive to their environment and this feature is utilized for monitoring the evolution of silica hydrogels prepared in this work. The polarity of hydrogel matrix during sol–gel transition assessed by all three pyrene probes decreases in the first stage of hydrogel formation, for about 2 h, followed by a gradual increase in polarity and reaching the constant level after 24 h for at least 2 weeks. The process of crosslinking was assessed by DiPyM and DiPyS. These fluorescent probes possess two pyrene structures, which ability to form a dynamic intramolecular excimer can be used to monitor the degree of hydrogel crosslinking with time. These data support the polarity measurements that the hydrogel network is predominatly formed within the first 2 h, stabilized within the 24 h, and that there is a minor increase in the network density for about 10 days until reaching the constant level. In addition, utilizing the second-order diffraction of scattered excitation light may also be used to obtain an adequate information about the silica hydrogel evolution. In summary, this paper demonstrates that pyrene-type fluorescent probes represent simple and precise tool for characterization of formation and aging of the silica hydrogels.     

Synthesis and functionalization of dendron‐polymer conjugate based hydrogels via sequential thiol‐ene “click” reactions

Fabrication and functionalization of hydrogels from well‐defined dendron‐polymer‐dendron conjugates is accomplished using sequential radical thiol‐ene “click” reactions. The dendron‐polymer conjugates were synthesized using an azide‐alkyne “click” reaction of alkene‐containing polyester dendrons bearing an alkyne group at their focal point with linear poly(ethylene glycol)‐bisazides. Thiol‐ene “click” reaction was used for crosslinking these alkene functionalized dendron‐polymer conjugates using a tetrathiol‐based crosslinker to provide clear and transparent hydrogels. Hydrogels with residual alkene groups at crosslinking sites were obtained by tuning the alkene‐thiol stoichiometry. The residual alkene groups allow efficient postfunctionalization of these hydrogel matrices with thiol‐containing molecules via a subsequent radical thiol‐ene reaction. The photochemical nature of radical thiol‐ene reaction was exploited to fabricate micropatterned hydrogels. Tunability of functionalization of these hydrogels, by varying dendron generation and polymer chain length was demonstrated by conjugation of a thiol‐containing fluorescent dye. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2016 , 54, 926–934     

Amphoteric surface hydrogels derived from hydrogen-bonded multilayers: reversible loading of dyes and macromolecules

We used hydrogen-bonded multilayers of poly(N-vinylpyrrolidone) (PVPON) and poly(methacrylic acid) (PMAA) as precursors for producing surface-bound hydrogels and studied their pH-dependent swelling and protein uptake behavior using in situ attenuated total reflection Fourier transform infrared spectroscopy and in situ ellipsometry. The hydrogels were produced by selective chemical cross-linking between PMAA units using carbodiimide chemistry and ethylenediamine (EDA) as a cross-linking reagent, followed by complete removal of PVPON from the film obtained by exposing the film to pH 7.5. As shown by in situ ellipsometry, hydrogels exhibit distinctive polyampholytic swelling as a function of pH, with minimum swelling at pH 4.2-5.7, and increased film thickness at both lower and higher pH values. Film swelling at lower pH values occurs as a result of the presence of amino groups within the hydrogels, which originate from the one-end attachment of the EDA cross-linker to PMAA chains. The pH-switching of hydrogel swelling was fast and reversible. The degree of hydrogel swelling could be also controlled by varying the time allowed for cross-linking. The produced hydrogels were able to absorb large amounts of dyes and proteins of opposite charge reversibly, in response to pH variations. Finally, we demonstrate that proteins included within the hydrogel can easily be replaced with linear polycations. These surface hydrogels hold promise for bioseparation and controlled delivery applications.     

Layered hydrogel of poly(γ-glutamic acid), sodium alginate, and chitosan: fluorescence observation of structure and cytocompatibility

In this study, a novel layered hydrogel composing of poly (γ-glutamic acid) (PGA), chitosan (CS), and alginate (AL) were prepared. Furthermore, PGA, CS, and AL were labeled with different fluorescent dyes. The bilayer structure of hydrogel was then revealed using these fluorescent labeled polymers. To mimic the stability of these hydrogels in physiological fluids, the dissolution of PGA and the release of Ca2+ from these hydrogels in normal saline were also monitored. The results showed that by adding CS to the hydrogel, the dissolution of PGA was decreased by 67%, and the release of Ca2+ was reduced by 40%. In addition, the hydrogel exhibited no cytotoxicity for L929 fibroblasts.     

Adsorption of methyl violet in aqueous solutions by poly(N-vinylpyrrolidone-co-methacrylic acid) hydrogels

N-vinylpyrrolidone(VP)/methacrylic acid (MAA) monomer mixtures containing different quantities of methacrylic acid were irradiated with gamma-radiation to form hydrogels. Mixtures which contain 5, 10, 15% and irradiated with 3.4 kGy were used for swelling and diffusion studies in water and solutions of methyl violet and for the adsorption of methyl violet from aqueous solutions. Diffusion of water and methyl violet within hydrogels were found to be of non-Fickian character. In the dye adsorption experiments, the effects of pH, concentration of aqueous solution of dye and the composition of hydrogels on the adsorption process were investigated. When the pH and concentration of aqueous solutions of dye and the MAA content in hydrogels increased, the adsorption increased.     

Adsorption of Dyes Using Multi-walled Carbon Nanotube Hydrogel

The hydrophilic multi-walled carbon nanotube(MWCNT)hydrogel was prepared using acrylic acid,acrylamide and hydrophilic MWCNT.The orthogonal experiment was applied to optimize the synthetic conditions.The MWCNT hydrogel was characterized by Fourier transform infrared spectrophotometer(FTIR)and scanning electron microscopy(SEM)analysis.The MWCNT hydrogel was used as the adsorbent to adsorb water-soluble cationic dye.This study evaluated the adsorption performance of hydrogels on four dyes of safranine Ts crystal violet,malachite green and methylene blue in water.The effects of the amount of hydrogel,the size of hydrogel,pH;and the temperature on the adsorption performance were investigated.The adsorption kinetic and adsorption isotherm curves were measured.The experimental results show that the MWCNT hydrogel can be easily separated from water and the adsorption capacity is much greater compared to the hydrogel without MWCNT.The MWCNT hydrogels can be used in wastewater treatment with a great potential.     

Biocatalytic reversible control of the stiffness of DNA-modified responsive hydrogels: applications in shape-memory,self-healing and autonomous controlled release of insulin

The enzymes glucose oxidase (GOx), acetylcholine esterase (AchE) and urease that drive biocatalytic transformations to alter pH, are integrated into pH-responsive DNA-based hydrogels. A two-enzyme-loaded hydrogel composed of GOx/urease or AchE/urease and a three-enzyme-loaded hydrogel composed of GOx/AchE/urease are presented. The biocatalytic transformations within the hydrogels lead to the dictated reconfiguration of nucleic acid bridges and the switchable control over the stiffness of the respective hydrogels. The switchable stiffness features are used to develop biocatalytically guided shape-memory and self-healing matrices. In addition, loading of GOx/insulin in a pH-responsive DNA-based hydrogel yields a glucose-triggered matrix for the controlled release of insulin, acting as an artificial pancreas. The release of insulin is controlled by the concentrations of glucose, hence, the biocatalytic insulin-loaded hydrogel provides an interesting sense-and-treat carrier for controlling diabetes.

Biocatalytic control over the stiffness of pH-responsive hydrogels is applied to develop shape-memory, self-healing and controlled release matrices.     

丝素蛋白-聚氨酯复合水凝胶的制备及性能研究

以聚乙二醇(PEG)、聚氧化丙烯二醇(PPG)、异弗尔酮二异氰酸酯(IPDI)为主要原料制备聚氨酯预聚体(PU),与丝素蛋白水溶液(SF)交联制得丝素蛋白-聚氨酯(SF-PU)复合水凝胶.分别利用ATR、SEM对水凝胶组成、结构及微观形貌进行表征;DSC、吸水溶胀测试探讨了丝素蛋白与聚氨酯的质量比(SF/PU)以及聚氨酯中不同软段质量比(PEG/PPG)对SF-PU水凝胶热性能、溶胀性能的影响.结果表明,SF-PU水凝胶具有多孔结构;样品中不同的SF/PU、PEG/PPG均对材料的玻璃化转变温度、结晶度及溶胀性能产生影响,且当水凝胶组分为SF/PU=1/25、PEG/PPG=2/1时,平衡溶胀比(ESR)可达到440%;水凝胶在溶胀初始阶段符合菲克扩散模型,整个溶胀过程遵循溶胀动力学2级方程.     

Aam/Aac水凝胶对碱性藏花红染料的吸附特性研究

以丙烯酰胺(AAm)、丙烯酸(AAc)合成了单体配比分别为1，2、1／1、2，1的AAm/AAcc水凝胶，采用分光光度计法研究了此水凝胶对水溶性单价阳离子染料碱性藏花红的吸附特性，测定了它们的吸附动力学曲线和吸附等温线：探讨了水凝胶单体组成对该染料吸附性能的影响：并且用静电场理论解释了解吸后水凝胶更优的再吸附特性，研究表明，AAm，AAc水凝胶可作为染料污水处理中一种良好的吸附剂。     

A novel single-step fabrication technique to create heterogeneous poly(ethylene glycol) hydrogel microstructures containing multiple phenotypes of mammalian cells

In this study, a novel method for the one-step fabrication of stacked hydrogel microstructures using a microfluidic mold is presented. The fabrication of these structures takes advantage of the laminar flow regime in microfluidic devices, limiting the mixing of polymer precursor solutions. To create multilayered hydrogel structures, microfluidic devices were rotated 90 degrees from the traditional xy axes and sealed with a cover slip. Two discreet fluidic regions form in the channels, resulting in the multilayered hydrogel upon UV polymerization. Multilayered patterned poly(ethylene glycol) hydrogel arrays (60 mum tall, 250 mum wide) containing fluorescent dyes, fluorescein isothiocyanate, and tetramethylrhodamine isothiocyanate were created for imaging purposes. Additionally, this method was used to generate hydrogel layers containing murine fibroblasts and macrophages. The cell adhesion promoter, RGD, was added to hydrogel precursor solution to enhance fibroblast cell spreading within the hydrogel matrix in one layer, but not the other. We were able to successfully generate patterns of hydrogels containing multiple phenotypes by using this technique.