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In this work, PVA nanocomposite films containing cellulose nanocrystals (CNC) and different amounts of lignin nanoparticles (LNP), prepared via a facile solvent cast method, were crosslinked by adding glutaraldehyde (GD). The primary objective was to investigate the effects of crosslinker and bio-based nanofillers loading on thermal, mechanical, antioxidant and water barrier behaviour of PVA nanocomposite films for active food packaging. Thermogravimetric analysis showed improved thermal stability, due to the strong interactions between LNP, CNC and PVA in the presence of GD, while Wide-angle X-ray diffraction results confirmed a negative effect on crystallinity, due to enhanced crosslinking interactions between the nanofillers and PVA matrix. Meanwhile, the tensile strength of PVA-2CNC-1LNP increased from 26 for neat PVA to 35.4 MPa, without sacrificing the ductility, which could be explained by a sacrificial hydrogen bond reinforcing mechanism induced by spherical-like LNP. UV irradiation shielding effect was detected for LNP containing PVA films, also migrating ingredients from PVA nanocomposite films induced radical scavenging activity (RSA) in the produced films in presence of LNP. Furthermore, PVA-CNC-LNP films crosslinked by GD showed marked barrier ability to water vapour.

Ficin extract has been immobilized on different 4% aminated-agarose beads. Using just ion exchange, immobilization yield was poor and expressed activity did not surpass 10% of the offered enzyme, with no significant effects on enzyme stability. The treatment with glutaraldehyde of this ionically exchanged enzyme produced an almost full enzyme inactivation. Using aminated supports activated with glutaraldehyde, immobilization was optimal at pH 7 (at pH 5 immobilization yield was 80%, while at pH 9, the immobilized enzyme became inactivated). At pH 7, full immobilization was accomplished maintaining 40% activity versus a small synthetic substrate and 30% versus casein. Ficin stabilization upon immobilization could be observed but it depended on the inactivation pH and the substrate employed, suggesting the complexity of the mechanism of inactivation of the immobilized enzyme. The maximum enzyme loading on the support was determined to be around 70 mg/g. The loading has no significant effect on the enzyme stability or enzyme activity using the synthetic substrate but it had a significant effect on the activity using casein; the biocatalysts activity greatly decreased using more than 30 mg/g, suggesting that the near presence of other immobilized enzyme molecules may generate some steric hindrances for the casein hydrolysis.

A continuous adsorption study in a fixed-bed column was carried out using a chitosan–glutaraldehyde biosorbent for the removal of the textile dye Direct Blue 71 from an aqueous solution. The biosorbent was prepared from shrimp shells and characterized by scanning electron microscopy, X-ray diffraction, and nuclear magnetic resonance spectroscopy. The effects of chitosan–glutaraldehyde bed height (3–12 cm), inlet Direct Blue 71 concentration (15–50 mg l−1), and feed flow rate (1–3 ml min−1) on the column performance were analyzed. The highest bed capacity of 343.59 mg Direct Blue 71 per gram of chitosan–glutaraldehyde adsorbent was obtained using 1 ml min−1 flow rate, 50 mg l−1 inlet Direct Blue 71 concentration, and 3 cm bed height. The breakthrough curve was analyzed using the Adams–Bohart, Thomas, and bed depth service time mathematical models. The behaviors of the breakthrough curves were defined by the Thomas model at different conditions. The bed depth service time model showed good agreement with the experimental data, and the high values of correlation coefficients (R2 ≥ 0.9646) obtained indicate the validity of the bed depth service time model for the present column system.

In this study, two different materials—alginate and glutaraldehyde-activated chitosan beads—were used for the co-immobilization of α-amylase, protease, and pectinase. Firstly, optimization of multienzyme immobilization with Na alginate beads was carried out. Optimum Na alginate and CaCl 2 concentration were found to be 2.5% and 0.1 M, respectively, and optimal enzyme loading ratio was determined as 2:1:0.02 for pectinase, protease, and α-amylase, respectively. Next, the immobilization of multiple enzymes on glutaraldehyde-activated chitosan beads was optimized (3% chitosan concentration, 0.25% glutaraldehyde with 3 h of activation and 3 h of coupling time). While co-immobilization was successfully performed with both materials, the specific activities of enzymes were found to be higher for the enzymes co-immobilized with glutaraldehyde-activated chitosan beads. In this process, glutaraldehyde was acting as a spacer arm. SEM and FTIR were used for the characterization of activated chitosan beads. Moreover, pectinase and α-amylase enzymes immobilized with chitosan beads were also found to have higher activity than their free forms. Three different enzymes were co-immobilized with these two materials for the first time in this study.

Biologically inspired self-healing structural color hydrogels were developed by adding a glucose oxidase (GOX)- and catalase (CAT)-filled glutaraldehyde cross-linked BSA hydrogel into methacrylated gelatin (GelMA) inverse opal scaffolds. The composite hydrogel materials with the polymerized GelMA scaffold could maintain the stability of an inverse opal structure and its resultant structural colors, whereas the protein hydrogel filler could impart self-healing capability through the reversible covalent attachment of glutaraldehyde to lysine residues of BSA and enzyme additives. A series of unprecedented structural color materials could be created by assembling and healing the elements of the composite hydrogel. In addition, as both the GelMA and the protein hydrogels were derived from organisms, the composite materials presented high biocompatibility and plasticity. These features of self-healing structural color hydrogels make them excellent functional materials for different applications.

This comparative study aims to identify a biocompatible and effective crosslinker for preparing gelatin sponges. Glutaraldehyde (GTA), genipin (GP), 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide (EDC), and microbial transglutaminase (mTG) were used as crosslinking agents. The physical properties of the prepared samples were characterized, and material degradation was studied in vitro with various proteases and in vivo through subcutaneous implantation of the sponges in rats. Adipose-derived stromal stem cells (ADSCs) were cultured and inoculated onto the scaffolds to compare the cellular biocompatibility of the sponges. Cellular seeding efficiency and digestion time of the sponges were also evaluated. Cellular viability and proliferation in scaffolds were analyzed by fluorescence staining and MTT assay. All the samples exhibited high porosity, good swelling ratio, and hydrolysis properties; however, material strength, hydrolysis, and enzymolytic properties varied among the samples. GTA–sponge and GP–sponge possessed high compressive moduli, and EDC–sponge exhibited fast degradation performance. GTA and GP sponge implants exerted strong in vivo rejections, and the former showed poor cell growth. mTG–sponge exhibited the optimal comprehensive performance, with good porosity, compressive modulus, anti-degradation ability, and good biocompatibility. Hence, mTG–sponge can be used as a scaffold material for tissue engineering applications.

•Biopolymeric materials are preferred for medical applications.•Biopolymeric materials lack the desired mechanical properties and aqueous stability.•Crosslinking is carried out to improve properties but is potentially cytotoxic.•Green chemicals and new technologies for crosslinking need to be developed. Biomaterials made from proteins, polysaccharides, and synthetic biopolymers are preferred but lack the mechanical properties and stability in aqueous environments necessary for medical applications. Crosslinking improves the properties of the biomaterials, but most crosslinkers either cause undesirable changes to the functionality of the biopolymers or result in cytotoxicity. Glutaraldehyde, the most widely used crosslinking agent, is difficult to handle and contradictory views have been presented on the cytotoxicity of glutaraldehyde-crosslinked materials. Recently, poly(carboxylic acids) that can crosslink in both dry and wet conditions have been shown to provide the desired improvements in tensile properties, increase in stability under aqueous conditions, and also promote cell attachment and proliferation. Green chemicals and newer crosslinking approaches are necessary to obtain biopolymeric materials with properties desired for medical applications.

Transdermal hydrogel films were fabricated from gellan gum, chitosan, and agar–agar, employing glutaraldehyde as a covalent crosslinker. The obtained formulation exhibited structural stability, pH-sensitive swelling, and high biocompatibility without the participation of metal ions. FTIR spectra showed the emergence of a characteristic imine (C=N) vibration near 1630 cm−1, confirming covalent network formation through Schiff-base reactions. SEM imaging revealed a homogeneous porous architecture (45–120 μm) that enhances moisture absorption and molecular diffusion. The swelling ratio reached 410 ± 12% at pH 9.18 and 275 ± 9% at pH 4.01, evidencing pronounced pH responsiveness. Mechanical strength measured 0.82 ± 0.03 MPa with elongation of 42 ± 2%, ensuring flexibility for skin application. The temperature-controlled release of methylene blue achieved 78 ± 4% at 40 °C after 24 h, consistent with diffusion-limited transport. This gellan–chitosan–agar hydrogel network crosslinked with glutaraldehyde represents a stable, pH-responsive, and biocompatible platform suitable for wound care and transdermal drug delivery.

A magnetic Schiff’s base chitosan-glutaraldehyde/Fe3O4 composite (CHT-GLA/ZnO/Fe3O4) was developed by incorporating zinc oxide (ZnO) nanoparticles into its structure to prepare an efficient adsorbent for the removal of remazol brilliant blue R (RBBR) dye. The CHT-GLA/ZnO/Fe3O4 was characterized by the following methods: CHN, BET, FTIR, XRD, SEM–EDX, pHpzc, and potentiometric titrations. Box-Behnken design based on response surface methodology was used to optimize the effects of the A: ZnO nanoparticles loading (0–50%), B: dose (0.02–0.1 g), C: pH (4–10), D: temperature (30–60 °C), and time E: (10–60 min) on the synthesis of the magnetic adsorbent and the RBBR dye adsorption. The experimental data of kinetics followed the pseudo-second order model, while isotherms showed better fit to Freundlich and Temkin models. The maximum adsorption capacity of the target nanocomposite (CHT-GLA/Fe3O4 containing 25% ZnO or CHT-GLA/ZnO/Fe3O4-25) was reached of 176.6 mg/g at 60 °C. The adsorption mechanism of RBBR onto CHT-GLA/ZnO/Fe3O4 nanocomposite can be attributed to multi-interactions including electrostatic attractions, hydrogen bonding, Yoshida H-bonding, and n-π interactions. This study offers a promising hybrid nanobiomaterial adsorbent in environmental nanotechnology to separate and remove the contaminants such as organic dyes from wastewater.

Polymerized allergen extracts are widely used in allergen immunotherapy (AIT) to reduce allergenicity and enhance safety. However, the impact of cross-linking stoichiometry on the structural and allergenic properties of these polymers remains insufficiently characterized. This study systematically investigates how varying glutaraldehyde-to-protein ratios influence the resulting polymers and their potential relevance for AIT. Pollen extracts from and were polymerized using five different glutaraldehyde-to-protein ratios. The resulting polymers were structurally characterized using SDS-PAGE, nuclear magnetic resonance (NMR), transmission electron microscopy (TEM), and mass spectrometry (MS). Allergenicity was evaluated by measuring the immunoglobulin E (IgE) reactivity through enzyme-linked immunosorbent assay (ELISA), Western blotting, and mast cell activation test (MAT). Immunogenicity was assessed by analyzing the serum-specific IgG and lymphocyte responses in mice immunized with the least allergenic polymer formulation. All cross-linking conditions produced polymers with distinct differences in size, morphology, and yield. Despite retaining similar peptide profiles to native allergens, as confirmed by MS, the polymers exhibited increased stability, as shown by NMR, and significantly reduced allergenicity, according to ELISA and MAT. Notably, increased polymer size and density, as determined by NMR and TEM, correlated with lower allergenicity. The most extensively cross-linked high-density polymers, optimized for minimal allergenicity, elicited immunogenic responses comparable to those induced by native extracts when tested against unmodified allergens. Cross-linking stoichiometry critically shapes the structural and immunological properties of polymerized allergen extracts. Adjusting the glutaraldehyde-to-protein ratio to produce highly polymerized, dense polymers enables a well-balanced profile of safety and efficacy for use in AIT.