Search Results (125)

Scaffolds designed for mechanically demanding soft tissue engineering applications should integrate mechanical support, efficient mass transfer, and good cellular compatibility. This work presents a one-pot method based on “radical-free click chemistry + carbodiimide coupling” to produce a double-network (DN) PEG–alginate cryogel. The PEG network is formed by a Michael addition reaction between thiol-based crosslinker and 8-arm PEG-acrylate. The second network is covalently crosslinked through EDC/NHS-mediated coupling of carboxyl groups in alginate and adipic acid dihydrazide (AAD). The subsequent freezing and gelation of the gel precursor at sub-zero temperatures results in an ice templated cryogel with an interconnected macroporous network. These cryogels demonstrate high elasticity, compressive modulus and rapid swelling equilibrium in aqueous environments, as well as controlled degradation under physiological conditions. Compared to the classical Ca²⁺ ion crosslinking systems, the covalent linking of the alginate in the double-network cryogel shows advantages in mechanical and structural stability. In addition, it is cell-compatible and allows culture of mesenchymal stem cells (MSCs) with homogeneous infiltration. Furthermore, the double-network cryogels supports chondrogenic differentiation of MSCs upon treatment with chondrogenic media or macrophage-conditioned media for a short period of time. These results indicate that crosslinking chemistry and polymer composition can be used to modulate the balance between mechanical performance and degradation behavior, while maintaining cytocompatibility and an interconnected macroporous network, thereby providing a scaffold design strategy for applications that require coordinated mechanical support and mass transfer, such as cartilage-related tissue engineering. Full article

Biosensors based on electrolyte-gated organic field-effect transistors (EGOFETs) have attracted considerable attention due to their advantages, including low cost, inherent signal amplification, and low-voltage operation. A critical step influencing sensing performance is the integration of specific receptors onto the device surface. Among various strategies, the covalent immobilization of biorecognition elements onto gold surfaces via thiol chemistry is one of the most widely used approaches. In this study, we report the optimization of a mixed self-assembled monolayer (SAM) composed of 11-mercaptoundecanoic acid (11-MUA) and 3-mercaptopropionic acid (3-MPA) for label-free detection of human IgG using EGOFETs. The quality of the SAM was systematically modulated by varying the total concentration from 10 to 400 mM and characterized using X-ray Photoelectron Spectroscopy (XPS), Electrochemical Impedance Spectroscopy (EIS), Cyclic Voltammetry (CV), and Atomic Force Microscopy (AFM). The results revealed that a concentration of 50 mM yielded a densely packed and well-ordered monolayer. After covalent immobilization of anti-IgG antibodies via 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) chemistry and subsequent blocking with ethanolamine and bovine serum albumin (BSA), the functionalized gate electrodes were integrated into poly(3-hexylthiophene) (P3HT)-based EGOFETs. Electrical measurements demonstrated that EGOFET biosensors functionalized with the 50 mM SAM achieved optimal sensing performance. The devices exhibited a highly linear response (R² = 0.998) over a wide concentration range from 1 fM to 10 nM, with a LOD of 2.82 fM, and showed excellent selectivity against non-target immunoglobulins A and M (IgA and IgM). This SAM concentration optimization strategy provides a versatile approach for engineering high-performance EGOFET biosensors, with potential applicability to a broad range of disease biomarkers. Full article

The increasing demand for continuous physiological monitoring has accelerated the development of high-sensitivity wearable electrochemical platforms. This study reports the fabrication of a multi-analyte electrochemical sensor based on single-walled carbon nanotubes (SWCNTs) for the detection of sweat-associated metabolites. To facilitate efficient heterogeneous electron transfer, glucose oxidase (Gox), lactate oxidase (Lox), and urease (Ure) were immobilized onto the SWCNT network through π–π interaction using 1-pyrenebutanoic acid succinimidyl ester (PBSE), followed by additional stabilization via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) coupling. The developed platform exhibited concentration-dependent resistance responses within the ranges of 0.02–0.20 mM for glucose, 20–100 mM for lactate, and 50–400 mM for urea under controlled experimental conditions. The resistance-based configuration enabled stable and reproducible signal modulation across these concentration intervals. Although direct testing with human sweat was not performed, the electrochemical behavior of key sweat-related metabolites was systematically evaluated as a preparatory step toward future wearable integration. Full article

Background/Objectives: Poor aqueous solubility and limited nasal permeability remain key challenges in the intranasal delivery of carbamazepine. In this study, biocompatible bovine serum albumin nanoparticles functionalized with sulfobutyl-β-cyclodextrin (SβCD-BSA NPs), comprising individually cytocompatible components with confirmed physical interactions), were formulated for intranasal delivery of carbamazepine (CBZ). Methods: The ethanolic desolvation method was utilised for the preparation of the nanoparticles, with the functional moiety incorporated during nanoparticle preparation. The effects of different molar ratios of SβCD-BSA and different ethanol volume ratios were studied. For crosslinking, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), a non-toxic crosslinker, was utilised. To determine the role of the SβCD, two preparation samples were formulated, with and without SβCD. Results: The formulation without SβCD incorporation had a mean particle size of 125 ± 0.64 nm, polydispersity index (PDI) of 0.34, encapsulation efficiency (EE%) of 61.5 ± 1.40%, and drug-loading ratio (DL%) of 31.9 ± 1.50%. Conversely, the SβCD-functionalized formulation showed a mean particle size of 128 ± 2.12 nm, PDI of 0.21 ± 0.03, EE of 64.6 ± 0.35%, and DL of 34.28 ± 1.60%. Statistical analysis revealed that the incorporation of SβCD resulted in a statistically significant increase in both DL% and EE% (p < 0.05). Conversely, the observed differences in particle size and PDI were not statistically significant (p > 0.05). This addition provides precise context regarding the comparability of the formulations while highlighting SβCD’s functional benefits in solubility and permeation. The interaction between CBZ and SβCD-BSA was confirmed using Fourier-transform infrared spectroscopy. Lastly, the prepared formulations were characterised by their physicochemical attributes and in vitro biopharmaceutical studies. It was discovered that SβCD plays a dual role, enhancing the solubility of CBZ in one scenario while promoting its nasal permeation, suggesting its potential use in epilepsy treatment. Conclusions: These findings highlight the potential of SβCD-BSA NPs as a versatile pharmaceutics platform for the intranasal delivery of poorly soluble CNS drugs. Full article

End-stage liver disease caused by advanced fibrosis and cirrhosis remains a major global burden, yet its treatment is limited by donor organ shortages. Bioengineered liver scaffolds offer a promising alternative, but their efficacy is often limited by thrombosis, insufficient vascularization, and poor graft integration due to inadequate endothelialization. To overcome these challenges, we employed LXW7 αvβ3 integrin targeting peptide with high endothelial cell specificity and low platelet affinity to enhance re-endothelialization and hemocompatibility of decellularized liver scaffold (DLS) and thereby improve hepatic integration and function. LXW7 was covalently conjugated to the decellularized rat liver scaffold via EDC/NHS-mediated carbodiimide coupling and subsequently reseeded with human umbilical vein endothelial cells (HUVECs) and cultured in a perfusion bioreactor to promote endothelialization. LXW7 immobilization significantly improved HUVECs attachment and proliferation, achieving approximately 81% vascular coverage, while sustaining the endothelial function. Ex vivo blood perfusion showed minimal thrombus formation and markedly reduced platelet adhesion, demonstrating enhanced hemocompatibility. Following confirmation of endothelialization, scaffolds were recellularized with hepatocellular carcinoma (HepG2) cells and HUVECs. LXW7 modified scaffolds promote organized hepatocyte distribution, sustained albumin expression, and increased urea secretion. In vivo implantation of LXW7-DLS into the omentum of mice promoted robust host endothelial recruitment and enhanced neovascularization, highlighting the scaffold’s excellent biocompatibility and good integration with surrounding tissues. Moreover, in vivo implantation of LXW7 recellularized scaffolds into a thioacetamide-induced fibrotic mouse liver resulted in reduced collagen deposition and lowered serum ALT/AST levels, demonstrating hepatic regeneration and extracellular matrix remodeling. Overall, our results showed that LXW7-modified DLS promotes stable endothelialization, improves hemocompatibility, and enhances hepatic function, underscoring its translational potential for the development of vascularized transplantable liver grafts. Full article

Collagen sponges are widely used for oral tissue regeneration, due to their extracellular matrix-mimetic architecture and excellent biocompatibility. However, in practical biomedical applications, collagen sponges may exhibit hydration-induced structural instability, and there can be associated inflammatory responses under physiological conditions, potentially compromising their regenerative performance. In this study, we investigated how two cross-linking strategies—transglutaminase (TG) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide (EDC/NHS)—modulate the structural stability and inflammatory profiles of collagen sponges. TG-cross-linked sponges exhibited microstructural collapse, associated with macrophage activation and engagement of the Itgαvβ3/5–Src–RhoC–NF-κB signaling axis. In contrast, EDC/NHS-cross-linked sponges preserved a stable porous architecture, effectively suppressing this signaling cascade and establishing a low-inflammatory microenvironment. These findings elucidate a key mechanism by which cross-linking regulates the microstructural integrity of collagen scaffolds and provides in vitro-derived preliminary design principles for developing next-generation collagen biomaterials with low-inflammatory properties. Full article

Alginate is a natural polysaccharide extracted from brown algae and is commonly used as a biomaterial scaffold in tissue engineering. In this study, we performed phenol functionalization of sodium alginate based on chemical modification methods using 1-ethyl-(3-dimethylaminopropyl)carbodiimide/N-hydroxybutanediimide/2-(N-morpholino) ethanesulfonic acid (EDC/NHS/MES) and tyramine. The presence of phenol groups was confirmed by spectrophotometry and Fourier Transform Infrared. We successfully prepared hydrogels using a horseradish peroxidase/hydrogen peroxide (HRP/H₂O₂) enzymatic system as well as an sodium persulfate (SPS)/ruthenium light-crosslinking system. Optimization identified 1 mM ruthenium and 4 mM SPS as the most effective photo crosslinking conditions. At the same time, 1 mM H₂O₂ and 10 U/mL HRP are considered optimal conditions for the enzyme-linked reaction. Rheological measurements monitored the gelation process, revealing that the viscosity, storage modulus, and loss modulus of the material increased by at least one hundredfold after crosslinking. Thixotropy results demonstrated excellent recovery of the material. Texture analysis indicated that the crosslinked material possessed notable strength and toughness, highlighting its potential applications in tissue engineering after 3D bioprinting. Full article

This study investigates the valorization of two agri-food residues, specifically olive pomace (alperujo, A) and banana peel (B), into efficient N-doped carbon-based catalysts for polluted wastewater treatment. The residues were converted into hydrochar (HA and HB), which were subsequently N-doped using polyethylenimine (PEI) in combination with cross-linkers (glutaraldehyde (GTA) or 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC)) to optimize their catalytic properties. The enhanced hydrochars were utilized as catalysts for the removal of organic pollutants from water by activation of peroxymonosulfate (PMS). Characterization techniques, including CHNS, FTIR, XPS, SEM and electrochemical analysis, were employed to understand the physicochemical properties of the materials. The catalytic activity was evaluated using Reactive Black 5 (RB5) as a model pollutant, with the N-doped alperujo-derived hydrochar cross-linked with EDC (N-HA-EDC) showing the best performance, achieving 80% removal in 60 min and an adsorption capacity of 97 mg/g. The versatility of this functionalization approach was assessed through tests with three pharmaceuticals, corroborating the adaptability and efficacy of the catalyst and demonstrating its potential for wastewater treatment applications. This study provides insights into the development of sustainable, cost-effective carbocatalysts, aligning with circular economy and zero waste principles. Full article

Chronic lower-extremity wounds in patients undergoing exoskeleton-assisted rehabilitation require materials that can both protect tissue and enable real-time physiological monitoring. Conventional dressings lack dynamic sensing capability, while current conductive hydrogels often compromise either adhesion or electronic performance. Here, we present a biointegrated hydrogel (CPSD) composed of carboxymethyl chitosan (CMCS) and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) forming the conductive backbone, integrated with dopamine-functionalized sodium alginate (SD); the network is assembled via electrostatic complexation and carbodiimide (EDC/NHS)-mediated covalent crosslinking. The resulting hydrogel exhibits a dense, tissue-conformal porous network with tunable swelling, stable mechanical integrity, and high photothermal conversion efficiency. In vitro assays confirmed potent antioxidant activity, strong antibacterial performance (>90% under near-infrared), and excellent cytocompatibility and hemocompatibility. CPSD shows bulk conductivity ~1.6 S·m⁻¹, compressive modulus ~15 kPa, lap-shear adhesion on porcine skin ~9.5 kPa, and WVTR ~75 g·m⁻²·h⁻¹, supporting stable biointerfaces for motion/sEMG sensing. Integrated into a lower-limb exoskeleton, CPSD hydrogels adhered securely during motion and reliably captured electromyographic and strain signals, enabling movement-intent detection. These results highlight CPSD hydrogel as a multifunctional interface material for next-generation closed-loop rehabilitation systems and mobile health monitoring. Full article

Spiky Au-Ag nanostars offer intense plasmonic enhancement due to their sharp-tipped morphology, enabling powerful surface-enhanced Raman scattering (SERS). Here, we report a liquid-phase SERS platform that addresses current limitations in cancer biomarker detection, such as low sensitivity and dependence on Raman reporters. Nanostar concentration was tuned by simple centrifugation (10, 30, and 60 min), and their SERS performance was evaluated using methylene blue (MB) and mercaptopropionic acid (MPA) as probe molecules. Signal intensity scaled with nanostar content, enabling sensitive detection. Optimized nanostars were functionalized with MPA, 1-Ethyl-3-(3-dimethylamino1-Ethyl-3-(3dimethylaminopropyl1) carbodiimide (EDC), and N-Hydroxy succinimide (NHS) for covalent attachment of monoclonal anti-α-fetoprotein antibodies (AFP-Ab), facilitating the detection of AFP antigens across 167–38 ng/mL (antibody) and 500–0 ng/mL (antigen) ranges. The limit of detection (LOD) for the antigens was determined to be 16.73 ng/mL. Unlike conventional SERS systems, this aqueous, surfactant-free platform exploits the intrinsic vibrational modes of AFP, enabling sensitive and rapid biomarker detection with strong potential for early cancer diagnostics. Full article

Early diagnosis of Alzheimer’s disease (AD) is essential for effective treatment; however current diagnostic methods are often complex, costly, and unsuitable for point-of-care testing. Graphene-based biosensors offer an alternative due to their affordability, versatility, and high conductivity. However, graphene’s conductivity can be compromised when its carbon lattice is oxidized to introduce functional groups for biomolecule immobilization. This study addresses this challenge by developing an electrochemical immunosensor using carboxyl-modified commercial graphene foam (COOH-GF) electrodes. The conductivity of graphene is preserved by enabling efficient COOH modification through π–π non-covalent interactions, while antibody immobilization is optimized via EDC-NHS carbodiimide chemistry. The immunosensor detects tau-441, an AD biomarker, using differential pulse voltammetry (DPV), achieving a detection range of 1 fM–1 nM, with a limit of detection (LOD) of 0.14 fM both in PBS and human serum. It demonstrates high selectivity against other AD-related proteins, including tau-217, tau-181, amyloid beta (Aβ_1-40 and Aβ_1-42), and 1% BSA. These findings underscore its potential as a highly sensitive, cost-effective tool for early AD diagnosis. Full article

The enzymatic resolution of chiral sofosbuvir precursors is a critical step in producing stereoisomerically pure ProTide drugs, essential for their therapeutic efficacy. In this study, a mutated phosphotriesterase (W131M-PTE) was immobilized onto various polymeric macroporous beads, including commercial immobead 150P (IB), modified (IB-EDA and IB-MTD), and synthetic polyacrylamide (PAM) beads functionalized with glutaraldehyde (PAM-GA) or 1-Ethyl-3-(3-dimethyl aminopropyl) carbodiimide (PAM-EDC). The immobilization efficiency, stability, and reusability of the enzyme were systematically evaluated. Among the tested supports, PAM-EDC demonstrated superior performance, retaining high enzymatic activity across multiple cycles and achieving a 92% yield of the (Sp)-diastereomer. The study highlights the potential of immobilized W131M-PTE as a cost-effective and scalable solution for chiral separation in pharmaceutical manufacturing, with implications for broader applications in ProTide drug production. Full article

A novel electrochemical detection method utilizing a cost-effective hybrid-modified electrode has been established. A glassy carbon (GC) modified electrode was tested for its ability to measure electrochemical tTG antibody levels, which are essential for diagnosing and monitoring Celiac disease (CD). Tissue transglutaminase protein biomolecules are immobilized on a quantum dots-polypyrrole nanocomposite in the improved electrode. Initial, quantum dots (QDs) were obtained from Bombyx mori silk fibroin and embedded in polypyrrole film. Using carbodiimide coupling, a polyamidoamine (PAMAM) dendrimer was linked with GQDs-polypyrrole film to improve sensor sensitivity. The tissue transglutaminase (tTG) antigen was cross-linked onto PAMAM using N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC)-N-hydroxy succinimide (NHS) chemistry to develop a nanoprobe that can detect human serum anti-tTG antibodies. The physicochemical characteristics of the synthesized nanocomposite were examined by FTIR, UV-visible, FE-SEM, EDX, and electrochemical studies. The novel electrode measures anti-tissue antibody levels in real time using human blood serum samples. The modified electrode has great repeatability and an 8.7 U/mL detection limit. Serum samples from healthy people and CD patients were compared to standard ELISA kit assays. SPSS and Excel were used for statistical analysis. The improved electrode and detection system can identify anti-tissue antibodies up to 80 U/mL. Full article

Methylene blue is a cationic organic dye commonly found in wastewater, groundwater, and surface water due to industrial discharge into the environment. This emerging pollutant is notably persistent and can pose risks to both human health and the environment. In this study, we developed a Surface Plasmon Resonance Biosensor employing a BK7 prism coated with 3 nm chromium and 50 nm of gold in the Kretschmann configuration, specifically for the detection of methylene blue. For the first time, laccases immobilized on a gold surface were utilized as bio-receptors for this organic dye. The enzyme was immobilized using carbodiimide bonds with EDC/NHS crosslinkers, allowing for the analysis of samples with minimal preparation. The method demonstrated validation with a limit of detection (LOD) of 4.61 mg L⁻¹ and a limit of quantification (LOQ) of 15.37 mg L⁻¹, a working range of 0–100 mg L⁻¹, and an R² value of 0.9614 during real-time analysis. A rainwater sample spiked with methylene blue yielded a recovery rate of 122.46 ± 4.41%. The biosensor maintained a stable signal over 17 cycles and remained effective for 30 days at room temperature. Full article

Microgel particles can play a key role, e.g., in drug delivery systems, tissue engineering, advanced (bio)sensors or (bio)catalysis. Amine-functionalized microgels are particularly interesting in many applications since they can provide pH responsiveness, chemical functionalities for, e.g., bioconjugation, unique binding characteristics for pollutants and interactions with cell surfaces. Since the incorporation of amine functionalities in controlled amounts with predefined architectures is still a challenge, here, we present a novel method for the synthesis of responsive core–shell nanogels (d_h < 100 nm) with a poly(N-isopropylacrylamide) (pNIPAm) core and a polyamine shell. To achieve this goal, a surface-functionalized pNIPAm nanogel was first prepared in a semi-batch precipitation polymerization reaction. Surface functionalization was achieved by adding acrylic acid to the reaction mixture in the final stage of the precipitation polymerization. Under these conditions, the carboxyl functionalities were confined to the outer shell of the nanogel particles, preserving the core’s temperature-responsive behavior and providing reactive functionalities on the nanogel surface. The polyamine shell was prepared by the chemical coupling of polyethyleneimine to the nanogel’s carboxyl functionalities using a water-soluble carbodiimide (EDC) to facilitate the coupling reaction. The efficiency of the coupling was assessed by varying the EDC concentration and reaction temperature. The molecular weight of PEI was also varied in a wide range (M_w = 0.6 to 750 kDa), and we found that it had a profound effect on how many polyamine repeat units could be immobilized in the nanogel shell. The swelling and the electrophoretic mobility of the prepared core–shell nanogels were also studied as a function of pH and temperature, demonstrating the successful formation of the polyamine shell on the nanogel core and its effect on the nanogel characteristics. This study provides a general framework for the controlled synthesis of core–shell nanogels with tunable surface properties, which can be applied in many potential applications. Full article