Glutaraldehyde Molecules Research Articles

Improvement in the Sustained-Release Performance of Electrospun Zein Nanofibers via Crosslinking Using Glutaraldehyde Vapors.

Volatile active ingredients in biopolymer nanofibers are prone to burst and uncontrolled release. In this study, we used electrospinning and crosslinking to design a new sustained-release active packaging containing zein and eugenol (EU). Vapor-phase glutaraldehyde (GTA) was used as the crosslinker. Characterization of the crosslinked zein nanofibers was conducted via scanning electron microscopy (SEM), mechanical properties, water resistance, and Fourier transform infrared (FT-IR) spectroscopy. It was observed that crosslinked zein nanofibers did not lose their fiber shape, but the diameter of the fibers increased. By increasing the crosslink time, the mechanical properties and water resistance of the crosslinked zein nanofibers were greatly improved. The FT-IR results demonstrated the formation of chemical bonds between free amino groups in zein molecules and aldehyde groups in GTA molecules. EU was added to the zein nanofibers, and the corresponding release behavior in PBS was investigated using the dialysis membrane method. With an increase in crosslink time, the release rate of EU from crosslinked zein nanofibers decreased. This study demonstrates the potential of crosslinking by GTA vapors on the controlled release of the zein encapsulation structure containing EU. Such sustainable-release nanofibers have promising potential for the design of fortified foods or as active and smart food packaging.

Cross-linked lipase particles with improved activity; application of a non-toxic linker for cross-linking

The preparation of cross-linked enzymes is often performed through enzyme aggregation or crystallization prior to cross-linking by glutaraldehyde as the most common linker. Isocyanide-based multi-component reactions (ICMRs) provide a one-step process to prepare cross-linked preparations of enzymes directly from their soluble forms. ICMRs accept a variety of functional groups to be involved in the coupling reaction. We here introduce cysteamine as a novel non-toxic alternative for glutaraldehyde to be used as a cross-linker in cross-linking reactions implemented by ICMRs. The cross-linked enzymes (CLEs) of Rhizomucor miehei lipase (RML) and Thermomyces lanuginosa lipase (TLL) were prepared with high immobilization yields (up to 97%). The results showed the maximum activities of 43.7 U/mg and 213.35 U/mg for the immobilized derivatives of RML and TLL, respectively, presenting an 11-fold and 10-fold improvement compared to the activity of the corresponding soluble lipases. The immobilized derivatives showed improved thermal and co-solvent stability. Additionally, the CLEs of TLL showed a selectivity of 19.9 in the selective hydrolysis of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil. The used linker provides an alternative strategy for CLEs production, particularly where the traditional cross-linking method has the possible risk of reaction contamination by weakly bonded glutaraldehyde molecules.

In Situ "Humanization" of Porcine Bioprostheses: Demonstration of Tendon Bioprostheses Conversion into Human ACL and Possible Implications for Heart Valve Bioprostheses.

This review describes the first studies on successful conversion of porcine soft-tissue bioprostheses into viable permanently functional tissue in humans. This process includes gradual degradation of the porcine tissue, with concomitant neo-vascularization and reconstruction of the implanted bioprosthesis with human cells and extracellular matrix. Such a reconstruction process is referred to in this review as “humanization”. Humanization was achieved with porcine bone-patellar-tendon-bone (BTB), replacing torn anterior-cruciate-ligament (ACL) in patients. In addition to its possible use in orthopedic surgery, it is suggested that this humanization method should be studied as a possible mechanism for converting implanted porcine bioprosthetic heart-valves (BHV) into viable tissue valves in young patients. Presently, these patients are only implanted with mechanical heart-valves, which require constant anticoagulation therapy. The processing of porcine bioprostheses, which enables humanization, includes elimination of α-gal epitopes and partial (incomplete) crosslinking with glutaraldehyde. Studies on implantation of porcine BTB bioprostheses indicated that enzymatic elimination of α-gal epitopes prevents subsequent accelerated destruction of implanted tissues by the natural anti-Gal antibody, whereas the partial crosslinking by glutaraldehyde molecules results in their function as “speed bumps” that slow the infiltration of macrophages. Anti-non gal antibodies produced against porcine antigens in implanted bioprostheses recruit macrophages, which infiltrate at a pace that enables slow degradation of the porcine tissue, neo-vascularization, and infiltration of fibroblasts. These fibroblasts align with the porcine collagen-fibers scaffold, secrete their collagen-fibers and other extracellular-matrix (ECM) components, and gradually replace porcine tissues degraded by macrophages with autologous functional viable tissue. Porcine BTB implanted in patients completes humanization into autologous ACL within ~2 years. The similarities in cells and ECM comprising heart-valves and tendons, raises the possibility that porcine BHV undergoing a similar processing, may also undergo humanization, resulting in formation of an autologous, viable, permanently functional, non-calcifying heart-valves.

Efficient Portable Urea Biosensor Based on Urease Immobilized Membrane for Monitoring of Physiological Fluids.

Numerous studies have addressed the utilization of glutaraldehyde (GA) as a homobifunctional cross-linker. However, its applicability has been impeded due to several issues, including the tendency of GA molecules to undergo polymerization. Herein, a portable urea biosensor was developed for the real-time monitoring of the flow of physiological fluids; this was achieved by using disuccinimidyl cross-linker-based urease immobilization. Urease was immobilized on a porous polytetrafluoroethylene (PTFE) solid support using different disuccinimidyl cross-linkers, namely disuccinimidyl glutarate (DSG), disuccinimidyl suberate (DSS) and bis-N-succinimidyl-(pentaethylene glycol) ester (BS(PEG)5). A urease activity test revealed that DSS exhibited the highest urease immobilizing efficiency, whereas FT-IR analysis confirmed that urease was immobilized on the PTFE membrane via DSS cross-linking. The membrane was inserted in a polydimethylsiloxane (PDMS) fluidic chamber that generated an electrochemical signal in the presence of a flowing fluid containing urea. Urea samples were allowed to flow into the urea biosensor (1.0 mL/min) and the signal was measured using chronoamperometry. The sensitivity of the DSS urea biosensor was the highest of all the trialed biosensors and was found to be superior to the more commonly used GA cross-linker. To simulate real-time monitoring in a human patient, flowing urea-spiked human serum was measured and the effective urease immobilization of the DSS urea biosensor was confirmed. The repeatability and interference of the urea biosensor were suitable for monitoring urea concentrations typically found in human patients.

Synthesis of a novel arginine-modified starch resin and its adsorption of dye wastewater.

In this work, corn starch (St) was firstly grafted with polyacrylamide (PAM) to obtain StAM, which was subsequently immobilized with arginine to obtain a guanidine-containing starch-based resin, StAM–Arg. The synthesized products were characterized via Fourier transform infrared spectroscopy (FT-IR), 13C-NMR nuclear magnetic resonance (13C-NMR), scanning electron microscopy (SEM), X-ray diffraction (XRD), gel permeation chromatography (GPC), X-ray photoelectron spectroscopy (XPS) and thermogravimetric analysis (TGA). StAM–Arg exhibited a significantly enhanced adsorption capacity for acid fuchsin (AF), acid orange G (AOG), and acid blue 80 (AB80) compared with zeolite, diatomite, St and StAM, and it also exhibited broad-spectrum adsorption for different dyes. Weak acidic conditions were favorable for the resin to adsorb acid dyes. The decolorization rate (DR) by StAM–Arg for mixed wastewater reached 82.49%, which was higher than that of activated carbon (DR = 58.09%). StAM–Arg showed high resistance to microbial degradation, resulting in significantly improved structural stability for the resin. Its antibacterial rate (AR) for E. coli was up to 99.73%. After 7 days in simulated natural water, the weight loss ratio (WR) of StAM–Arg was 14.5%, which was much lower than that of St (WR = 66.53%). The introduced guanidine groups were considered to be the major reason for the observed improvements. Furthermore, the cationic guanidine could trap the acid dyes via ion-exchange reactions, while effectively inhibiting or eliminating the growth of bacteria on the adsorbent surface. The above advantages, including good dyestuff adsorption properties, high structural stability and prolonged service life, make StAM–Arg overcome the inherent drawbacks of the existing natural polymer adsorbents and have good application prospect in the treatment of textile wastewater.

Control of functionalization of supports for subsequent assembly of titania nanoparticle films

For self‐assembling of TiO2 nanoparticles in multiple layers by layer‐by‐layer deposition to be applied to TiO2 thin films with defined and homogeneous thickness for large‐scale applications, the proper functionalization of substrate surface is a prerequisite to guarantee sufficient adhesion. The substrates selected and tested in the present paper were conductive, fluorine‐doped tin oxide (FTO) glass, nonconductive silica glass, and titanium alloy. The current study focusses on the analytical control of the stepwise functionalization of the substrates with 3‐aminopropyltriethoxysilane and glutaraldehyde (GA) for both the FTO glass and silica glass and with 3‐aminepropyl phosphonic acid and GA for Ti alloy. The analyses have been conducted by means of surface sensitive methods, X‐ray photoelectron spectroscopy, Auger electron spectroscopy, and time‐of‐flight secondary ions mass spectrometry. Chemical composition of surface of functionalized substrates shows differences in the degree and type of modification in dependence on substrate. It could be demonstrated that the best functionalized substrates were the conductive FTO glasses. The analysis of the functionalized Ti substrates has revealed that the surface coverage with 3‐aminepropyl phosphonic acid and GA molecules is an inhomogeneous one, and further optimization of the two‐step functionalization on the Ti alloy substrate is necessary.

Construction of an Acetylcholinesterase Sensor Based on Synthesized Paramagnetic Nanoparticles, a Simple Tool for Neurotoxic Compounds Assay.

Magnetic particles (MPs) have been widely used in biological applications in recent years as a carrier for various molecules. Their big advantage is in repeated use of immobilized molecules including enzymes. Acetylcholinesterase (AChE) is an enzyme playing crucial role in neurotransmission and the enzyme is targeted by various molecules like Alzheimer’s drugs, pesticides and warfare agents. In this work, an electrochemical biosensor having AChE immobilized onto MPs and stabilized through glutaraldehyde (GA) molecule was proposed for assay of the neurotoxic compounds. The prepared nanoparticles were modified by pure AChE and they were used for the measurement anti-Alzheimer’s drug galantamine and carbamate pesticide carbofuran with limit of detection 1.5 µM and 20 nM, respectively. All measurements were carried out using screen-printed sensor with carbon working, silver reference, and carbon auxiliary electrode. Standard Ellman’s assay was used for validation measurement of both inhibitors. Part of this work was the elimination of reversible inhibitors represented by galantamine from the active site of AChE. For this purpose, we used a lower pH to get the original activity of AChE after inhibition by galantamine. We also observed decarbamylation of the AChE-carbofuran adduct. Influence of organic solvents to AChE as well as repeatability of measurement with MPs with AChE was also established.

Developing and physicochemical evaluation of cross-linked electrospun gelatin–glycerol nanofibrous membranes for medical applications

This study aims to develop optimal cross-linked electrospun gelatin–glycerol (GEL-GLY) nano-fibrous mats suitable for tissue engineering and wound dressing applications. The optimized procedure involves heating the gelatin and gelatin–glycerol solutions up to 90 °C. The electrospinning process was performed, followed by further cross-linking of electrospun films in a container containing glutaraldehyde (GTA) vapor. The results of X-ray diffraction (XRD), Fourier transformed infrared (FTIR), and Differential thermal analysis (DTA) confirmed that heating gelatin solution up to 90 °C in the presence of glycerol affected the cross-linking efficiency and interactions between GTA molecules and gelatin chains. Scanning Electron Microscope (SEM) analysis showed that GEL-GLY nano-fibrous mats with weight ratios less than or equal (12:3 w/w) exhibited a regular morphology with defect free in addition to increasing the degradation time, cross-linking efficiency, and swelling degree of electrospun gelatin/glycerol.

ChemInform Abstract: Glutaraldehyde in Bio‐Catalysts Design: A Useful Crosslinker and a Versatile Tool in Enzyme Immobilization

AbstractReview: 173 refs.

Magneto-fluorescent hybrid of dye and SPION with ordered and radially distributed porous structures

We have reported the development of a silica based magneto-fluorescent hybrid of a newly synthesized dye and superparamagnetic iron oxide nanoparticles with ordered and radially distributed porous structure. The dye is synthesized by a novel yet simple synthetic approach based on Michael addition between dimer of glutaraldehyde and oleylamine molecule. The surfactant used for phase transformation of the dye from organic to aqueous phase, also acts as a structure directing agent for the porous structure evolution of the hybrid with radial distribution. The evolution of the radially distributed pores in the hybrids can be attributed to the formation of rod-like micelles containing nanoparticles, for concentration of micelles greater than critical micelle concentration. A novel water extraction method is applied to remove the surfactants resulting in the characteristic porous structure of the hybrid. Adsorption isotherm analysis confirms the porous nature of the hybrids with pore diameter ∼2.4nm. A distinct modification in optical and magnetic property is observed due to interaction of the dye and SPION within the silica matrix. The integration of multiple structural components in the so developed hybrid nanosystem results into a potential agent for multifunctional biomedical application.

Glutaraldehyde in bio-catalysts design: a useful crosslinker and a versatile tool in enzyme immobilization

Glutaraldehyde is one of the most widely used reagents in the design of biocatalysts. It is a powerful crosslinker, able to react with itself, with the advantages that this may bring forth. In this review, we intend to give a general vision of its potential and the precautions that must be taken when using this effective reagent. First, the chemistry of the glutaraldehyde/amino reaction will be commented upon. This reaction is still not fully clarified, but it seems to be based on the formation of 6-membered heterocycles formed by 5 C and one O. Then, we will discuss the production of intra- and inter-molecular enzyme crosslinks (increasing enzyme rigidity or preventing subunit dissociation in multimeric enzymes). Special emphasis will be placed on the preparation of cross-linked enzyme aggregates (CLEAs), mainly in enzymes that have low density of surface reactive groups and, therefore, may be problematic to obtain a final solid catalyst. Next, we will comment on the uses of glutaraldehyde in enzymes previously immobilized on supports. First, the treatment of enzymes immobilized on supports that cannot react with glutaraldehyde (only inter and intramolecular cross-linkings will be possible) to prevent enzyme leakage and obtain some enzyme stabilization via cross-linking. Second, the cross-linking of enzymes adsorbed on aminated supports, where together with other reactions enzyme/support crosslinking is also possible; the enzyme is incorporated into the support. Finally, we will present the use of aminated supports preactivated with glutaraldehyde. Optimal glutaraldehyde modifications will be discussed in each specific case (one or two glutaraldehyde molecules for amino group in the support and/or the protein). Using preactivated supports, the heterofunctional nature of the supports will be highlighted, with the drawbacks and advantages that the heterofunctionality may have. Particular attention will be paid to the control of the first event that causes the immobilization depending on the experimental conditions to alter the enzyme orientation regarding the support surface. Thus, glutaraldehyde, an apparently old fashioned reactive, remains the most widely used and with broadest application possibilities among the compounds used for the design of biocatalyst.

Phospholipids covalently attached to silica particles as stationary phase in nano-liquid chromatography

Silica particles were covalently modified with phospholipids and used as packing material for nano-liquid chromatography (nano-LC). This modification involved aminopropylsilylation of the raw silica particles using 3-(aminopropyl)-triethoxysilane, covalent binding of glutaraldehyde molecules to the aminopropylsilylated particles, and finally covalent binding of different phospholipid vesicles containing primary amino groups to the iminoaldehyde silica particles. Capillaries with an inner diameter of 100μm were packed with phospholipid-coated silica particles using a slurry packing method. The packed capillaries were tested in nano-LC with UV-detection for the separation of acidic, neutral, and basic model analytes. The effect of the buffer ion on the retention factor of the analytes was evaluated using buffer solutions with constant ionic strength and pH. In addition, the effect of the volume of methanol in the mobile phase was studied. The calculated distribution coefficients (logKD) of the model compounds were in agreement with those reported in the literature. A good correlation between logKD values and octanol/water partitioning coefficients (Po/w) for neutral hydrophobic analytes was obtained proving the applicability of the method for predicting partitioning of the compounds with the biomembranes.

Spectroscopy studies of functionalized oxidized porous silicon surface for biosensing applications

In this paper we report detailed Fourier transform infra-red (FTIR) and Raman spectroscopic results obtained after the modification of a porous silica surface using different steps of functionalization and protein grafting. After the elaboration of porous silicon (PS) layers, with sufficient adjusted pore size and well defined porosity, we carried out complete thermal oxidation and we optimized the silanization step by varying the concentration of 3-aminopropyltriethoxysilane (APTES) molecules using different immersion and rinsing durations. Then we introduced a coupling agent, glutaraldehyde (GL) molecules, which has a great affinity for bovine serum albumin (BSA) molecule grafting. FTIR and Raman spectroscopic analyses present complementary spectra that allow us to get an idea about the chemical links and vibrational modes that appear during the functionalization process and after protein attachment. This proves that the biological molecules are covalently attached on the modified porous silica surface. Moreover, a modelling study of the reflectance spectra allows values of the volume fraction and refractive index variations to be estimated by assigning them with the number of APTES, glutaraldehyde and protein layers after each step. These results are well correlated to those obtained by the FTIR and Raman analyses. This work on porous silica single-layers is carried out in order to exploit it for the elaboration of functionalized optical waveguides to obtain a sensitive label-free optical biosensor.

The Effect of Interlayer Adhesion on the Mechanical Behaviors of Macroscopic Graphene Oxide Papers

High mechanical performances of macroscopic graphene oxide (GO) papers are attracting great interest owing to their merits of lightweight and multiple functionalities. However, the loading role of individual nanosheets and its effect on the mechanical properties of the macroscopic GO papers are not yet well understood. Herein, we effectively tailored the interlayer adhesions of the GO papers by introducing small molecules, that is, glutaraldehyde (GA) and water molecules, into the gallery regions. With the help of in situ Raman spectroscopy, we compared the varied load-reinforcing roles of nanosheets, and further predicted the Young's moduli of the GO papers. Systematic mechanical tests have proven that the enhancement of the tensile modulus and strength of the GA-treated GO paper arose from the improved load-bearing capability of the nanosheets. On the basis of Raman and macroscopic mechanical tests, the influences of interlayer adhesions on the fracture mechanisms of the strained GO papers were inferred.

Controllable synthesis, characterization and optical properties of colloidal PbS/gelatin core–shell nanocrystals

Native quantum dots (QDs) made up of semiconductor nanocrystals (NCs) are toxic in nature but due to their excellent optical properties, they have proven themselves to be an attractive choice in biological labeling and targeting. In order to improve the general biocompatibility of lead sulfide (PbS) NCs, we present a new and simple procedure for preparing PbS/gelatin core-shell nanoparticles cross-linked with glutaraldehyde (GA) molecules. The phase composition, morphology, luminescence and in vitro photostability of the samples were characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR), transmission electron microscope (TEM) and fluorescence spectroscopy, respectively. The XRD analysis showed that the PbS NCs were of the cubic structure, the mean crystallite size was calculated to be 13.5 nm and the calculated lattice constant using Bragg's equation was 0.5950 nm, which was very close to its value in the standard card (JCPDS No. 5-592). In vitro test revealed that compared with bare PbS NCs, the photostability of the core-shell nanostructure remarkably improved. In addition, possible formation mechanisms of the PbS/gelatin nanoparticles were discussed in detail. Consequently, the advantages of high stability as well as high fluorescent intensity and biocompatibility make the core-shell nanoparticles promising candidates for in vivo biological targeting applications.

Alternative Reaction Mechanism for the Cross-Linking of Gelatin with Glutaraldehyde

FT-IR, in combination with residual amino group determination using a fluorescence technique, has been used to investigate the chemical functional groups involved in the cross-linking reaction between glutaraldehyde and gelatin molecules. The results suggest that, at high pH values (i.e., close to the pK(a) of lysine), the cross-linking reaction is mainly governed by the well-known Schiff base formation, whereas at low pH (i.e., when the amino groups of lysine are protonated), the reaction may also involve the -OH groups of hydroxyproline and hydroxylysine, leading to the formation of hemiacetals.

Real-time liquid crystal-based glutaraldehyde sensor

Abstract We develop a liquid crystal (LC)-based optical sensor for detecting glutaraldehyde vapor by using real-time orientational responses of 4-cyano-4′-pentylbiphenyl (5CB) to glutaraldehyde. The sensor comprises a thin layer of 5CB (∼20 μm) supported on an amine-decorated surface. When a glutaraldehyde molecule diffuses through the layer of 5CB, it reacts with a surface amine group and forms an imine bond. Subsequently, the resulting free aldehyde group from the glutaraldehyde molecule triggers an orientational transition of 5CB, which manifests as an optical signal visible to the naked eye. Because the performance of the LC-based sensor critically depends on the diffusion of glutaraldehyde in 5CB, we also measured the solubility and diffusivity of glutaraldehyde in 5CB. Based on the diffusivity, an optimal LC sensor with a 20-μm-thick film of 5CB can respond to 207 ppmv glutaraldehyde within 20 s and shows a full response in 4 min. It also shows good specificity. Only a very weak response is observed when this sensor is exposed to acetic acid, and no response is recorded for methanol, ethanol or formaldehyde.

Influência da cola Bioglue® na deiscência de anastomose colônica: estudo experimental

A grande parte da morbimortalidade associada com a cirurgia colorretal, é associada com a deiscência anastomótica. Trabalhos experimentais sobre a utilidade de adesivos tissulares nas anastomoses colônicas são controversos, assim como estudos clínicos prospectivos randomizados são ausentes. O adesivo cirúrgico BioGlue®, formado por dois componentes - albumina sérica bovina purificada e glutaraldeído, forma uma ligação co-valente entre esses dois componentes e as proteínas teciduais no local de aplicação. O objetivo do estudo é avaliar a eficácia da BioGlue® na prevenção da deiscência anastomótica colônica em ratos. Foram utilizados 30 ratos machos da raça Wistar albino. A anastomose colocolônica foi confeccionada com sutura em pontos separados com polipropileno 5-0 (grupo 1) e aplicação da cola BioGlue® envolvendo a anastomose (grupo 2). Avaliaram-se a formação e extensão das aderências, a pressão de ruptura nas anastomoses e as alterações histológicas. Apenas um animal do grupo 1 (7%) faleceu, sendo constatada na necropsia, obstrução intestinal com grande distensão de alças. A mortalidade no grupo 2, por outro lado, foi de 10 animais (67%), sendo observado: distensão de alças intestinais, vazamento anastomótico e, em algumas situações, franca peritonite fecal por deiscência quase que total da anastomose. O presente trabalho nos permitiu concluir que, o uso da Bioglue® nas anastomoses colônicas realizadas em ratos, promoveu um aumento na morbimortalidade que foi estatisticamente significante comparado à sutura convencional.

Different mechanisms of protein immobilization on glutaraldehyde activated supports: Effect of support activation and immobilization conditions

The precise control of the conditions during support activation with glutaraldehyde has enabled the modification of the amino groups of the matrix with one or two glutaraldehyde molecules. Moreover, the use of aminated supports implies that below each glutaraldehyde molecule, there are amino groups and, therefore, these supports could be considered as heterofunctional matrices; an anion exchanger bearing glutaraldehyde groups for covalent immobilization. Thus, by using several enzymes as models, it has been found that at low ionic strength the protein immobilization on glutaraldehyde activated supports proceeds via a first ionic interchange of the protein on the amino groups of the support, followed by the covalent reaction. Moreover, the dimeric form of glutaraldehyde seems to be much more reactive than its monomeric counterpart, permitting the immobilization of proteins even at very high ionic strength. Although in all cases the immobilization of the enzymes on both monomer and dimeric matrices promoted a significant increment in the enzyme stability, it was found that the stabilization depends on the degree of activation (monomer or dimer), and it is necessary to analyze each individual enzyme before selecting any of the immobilization protocols.

Microcantilever-Based Biosensor for Detection of Various Biomolecules

We propose an optical and an electrical methods for detecting various biomolecules effectively with microcantilevers. The microcantilevers were fabricated employing surface micromachining technique that has attractive advantages in terms of cost efficiency, simplicity and ability of fabricating in array. The fluid cell system for injection of biomolecular solution is fabricated using poly(dimethylsiloxane) (PDMS) and a fused silica glass. The microcantilever is deflected with respect to the difference of the surface stress caused by the formation of self-assembled biomolecules on the gold coated side of the microcantilever. It detected cystamine dihydrochloride and glutaraldehyde molecules and analyzed individual concentrations of the cystamine dihydrochloride solution. We confirm that the deflections of bending-up or bending-down are occurred by the biomolecule adsorption and microcantilever can be widely used to a micro total analysis system (µ-TAS) and a lab-on-a-chip for a potential detection of various biomolecules.