Natural Cross-linker Research Articles

Genipap Oil as a Natural Cross-Linker for Biodegradable and Low-Ecotoxicity Porous Absorbents via Reactive Extrusion.

Proteins derived from agroindustrial coproducts and a natural cross-linking agent (genipap oil containing genipin) were used to develop porous materials by reactive extrusion for replacing fossil-based absorbents. Incorporating genipap oil allowed the production of lightweight structures with high saline uptake (above 1000%) and competing retention capacity despite their porous nature. The mechanical response of the genipap-cross-linked materials was superior to that of the noncross-linked ones and comparable to those cross-linked using commercial genipin. The extruded products were hemocompatible and soil-biodegradable in less than 6 weeks. The compounds generated by the degradation process were not found to be toxic to the soil, showing a high bioassimilation capacity by promoting grass growth. The results demonstrate the potential of biopolymers and new green cross-linkers to produce fully renewable-based superabsorbents in hygiene products with low ecotoxicity. The study further promotes the production of these absorbents using low-cost proteins and continuous processing such as reactive extrusion.

Effect of Proteins on the Vulcanized Natural Rubber Crosslinking Network Structure and Mechanical Properties

Proteins are important factors affecting the properties of natural rubber. Therefore, investigating the effect of free and bonded proteins on the structure and mechanical properties of the vulcanized crosslinking network of natural rubber would provide a theoretical basis for the production of high mechanical resistance natural rubber. Herein, natural rubbers with different protein contents and types were prepared by high-speed centrifugation. And, the effects on their network structure, vulcanization, tensile strength, tearing strength and dynamic mechanical properties were investigated. The results showed that the reduction in protein content led to the decrement in the entanglement networks, crosslinking density and tensile and tear strengths of the vulcanized natural rubber. Moreover, the bonded proteins had an obvious influence on the vulcanization process, while free proteins played an important role in the crosslink densities. These results reveal that both bonded and free proteins are involved in the vulcanization process and the construction of the vulcanized crosslinking network structure of natural rubber, which enhances the mechanical properties such as the modulus and tensile strength of vulcanized natural rubber.

Ethanolic duea ching extract in combination with microbial transglutaminase: Strengthening effect on mince gel and cross-linking of myofibrillar proteins from Indian mackerel

Surimi or fish mince gel are popular seafood products worldwide. Lean fish are commonly used as raw material, while dark fleshed fish, especially Indian mackerel, has high fat content and dark color associated with poor gel quality. To improve gel strength, plant extract, particularly from duea ching (Ficus botryocarpa Miq.), rich in polyphenols and calcium could be used as natural protein cross-linker to strengthen gel network. Effect of ethanolic duea ching extract (EDCE) at varying levels in the absence and presence of microbial transglutaminase (MTGase) on gel property of Indian mackerel mince gel prepared from fillets pre-washed for different times (1 and 3 times) was studied. Heat-induced aggregation of myofibrillar protein (MP) from Indian mackerel fillets under different conditions was also investigated. Mince gel prepared from fillets pre-washed for 3 times added with 0.10 % EDCE and 0.5 unit/g of MTGase (3-PWM-MT-0.10) showed the highest breaking force, deformation and textural properties (P < 0.05). Lower autolysis of mince gel was obtained when EDCE, MTGase or in combination were added. However, addition of ECHE decreased whiteness of mince gel, particularly at higher concentrations (P < 0.05). Electrophoretic images revealed the disappearance of myosin heavy chain when MTGase or EDCE in conjunction with MTGase was incorporated. Addition of 0.10 % EDCE, MTGase or both could induce aggregation and enhance setting phenomenon of MP solution as indicated by higher absorbance at 660 nm and storage modulus (G′), respectively. Based on microstructure study, 3-PWM-MT-0.10 had more compact, finer and interconnected structure than other gels. Therefore, the use of EDCE and MTGase at the appropriate levels could be used for improving quality of mince gel from pre-washed Indian mackerel fillets.

Impact of a Bio-Cross-Linking Agent Obtained from Spent Coffee Grounds on the Physicochemical and Thermal Properties of Gelatin/Κ-Carrageenan Hydrogels.

Gelatine hydrogels can be prepared using different cross-linking methods, such as enzymatic, physical or chemical. Unfortunately, in the case of chemical cross-linking, the typically utilized synthetic cross-linkers are harmful to human health and the environment. Therefore, in accordance with the principles of green chemistry and sustainable development, we have obtained compounds for the chemical cross-linking of hydrogel polymers from the processing of spent coffee grounds. In this study, gelatin/κ-carrageenan hydrogels are cross-linked using a bio-cross-linking agent from spent coffee grounds. Their physicochemical and thermal properties are compared with those of standard physical gels. The chemical cross-linking was confirmed based on FT-IR spectra, which demonstrated the formation of new covalent bonds between the oxidized polyphenols included in the extract from the spent coffee grounds and the amide groups present in the gelatine structure. Significant differences were also observed in morphology (SEM images) and other physico-chemical characteristics (gel fraction, swelling ability, hardness). The chemically cross-linked hydrogels in comparison to physically ones are characterized by a better developed porous network, a slightly higher gel fraction (64.03 ± 4.52% as compared to 68.15 ± 0.77%), and a lower swelling ratio (3820 ± 45% as compared to 1773 ± 35%), while TGA results show that they have better thermal stability. The research confirmed the possibility of using the developed natural cross-linking agent in the process of obtaining hydrogel materials based on bio-polymers.

Modulating the Properties of Brown Alga Alginate-Based Fibers Using Natural Cross-Linkers for Sustainable Textile and Fashion Applications.

Seaweed-derived alginate shows promise in the textile industry as a sustainable alternative to synthetic and natural materials. However, challenges arise due to its low mechanical strength. We addressed this limitation by sustainably extracting alginates from European brown algae and employing novel manufacturing methods. Using natural cross-linkers, such as chitosan, ferulic acid, and citric acid, we have successfully modulated the mechanical properties of alginate fibers. Mechanical properties of ferulic acid and citric acid-cross-linked alginate solutions were spinnable, producing fibers with a diameter of 73-75 μm. Ferulic acid cross-linked alginate fibers exhibited stiffness, with a tensile strength of 52.97 MPa and a strain percentage of 20.77, mechanical properties comparable to those of wool, polyester, and rayon. In contrast, citric acid-cross-linked fibers showed partial elasticity, with a tensile strength of 14.35 MPa and a strain percentage of 45.53, comparable to those of nylon. This ability to control the mechanical properties of seaweed-derived fibers represents a significant advancement for their application in sustainable textiles and the fashion industry.

Functionalised Sodium-Carboxymethylcellulose-Collagen Bioactive Bilayer as an Acellular Skin Substitute for Future Use in Diabetic Wound Management: The Evaluation of Physicochemical, Cell Viability, and Antibacterial Effects.

The wound healing mechanism is dynamic and well-orchestrated; yet, it is a complicated process. The hallmark of wound healing is to promote wound regeneration in less time without invading skin pathogens at the injury site. This study developed a sodium-carboxymethylcellulose (Na-CMC) bilayer scaffold that was later integrated with silver nanoparticles/graphene quantum dot nanoparticles (AgNPs/GQDs) as an acellular skin substitute for future use in diabetic wounds. The bilayer scaffold was prepared by layering the Na-CMC gauze onto the ovine tendon collagen type 1 (OTC-1). The bilayer scaffold was post-crosslinked with 0.1% (w/v) genipin (GNP) as a natural crosslinking agent. The physical and chemical characteristics of the bilayer scaffold were evaluated. The results demonstrate that crosslinked (CL) groups exhibited a high-water absorption capacity (>1000%) and an ideal water vapour evaporation rate (2000 g/m2 h) with a lower biodegradation rate and good hydrophilicity, compression, resilience, and porosity than the non-crosslinked (NC) groups. The minimum inhibitory concentration (MIC) of AgNPs/GQDs presented some bactericidal effects against Gram-positive and Gram-negative bacteria. The cytotoxicity tests on bilayer scaffolds demonstrated good cell viability for human epidermal keratinocytes (HEKs) and human dermal fibroblasts (HDFs). Therefore, the Na-CMC bilayer scaffold could be a potential candidate for future diabetic wound care.

Enhancing Gelatine Hydrogel Robustness with Sacran-Aldehyde: A Natural Cross-Linker Approach

Tunable hydrogels have gained significant attention in the bioengineering field due to their designer preparation approach. Towards this end, gelatine stands out as a promising candidate owing to its desirable attributes, such as biocompatibility, ability to support cell adhesion and proliferation, biodegradability, and cost-effectiveness. This study presents the preparation of a robust gelatine hydrogel employing sacran aldehyde (SDA) as a natural cross-linker. The resulting SDA-cross-linked gelatine hydrogels (GSDA) display an optimal compressive stress of 0.15 MPa at 50% strain, five times higher than pure gelatine hydrogel. As SDA cross-linking concentration is increased, the swelling capacity of GSDA declines. This decline in swelling capacity, from 80% to 40%, is a result of strong crosslinking of gelatin with SDA. Probing further with FT-IR spectroscopy and SEM at the micron scale unveiled a dual-cross-linking mechanism within the hydrogels. This mechanism encompasses both short- and long-range covalent cross-linking, along with thermo-induced physical cross-linking, resulting in a significant enhancement of the load-bearing capacity of the fabricated hydrogels.

Proanthocyanidin surface preconditioning of dental pulp stem cell spheroids enhances dimensional stability and biomineralization invitro.

Lack of adequate mechanical strength and progressive shrinkage over time remain challenges in scaffold-free microtissue-based dental pulp regeneration. Surface collagen cross-linking holds the promise to enhance the mechanical stability of microtissue constructs and trigger biological regulations. In this study, we proposed a novel strategy for surface preconditioning microtissues using a natural collagen cross-linker, proanthocyanidin (PA). We evaluated its effects on cell viability, tissue integrity, and biomineralization of dental pulp stem cell (DPSCs)-derived 3D cell spheroids. Microtissue and macrotissue spheroids were fabricated from DPSCs and incubated with PA solution for surface collagen cross-linking. Microtissue viability was examined by live/dead staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, with transverse dimension change monitored. Microtissue surface stiffness was measured by an atomic force microscope (AFM). PA-preconditioned microtissues and macrotissues were cultured under basal or osteogenic conditions. Immunofluorescence staining of PA-preconditioned microtissues was performed to detect dentin sialophosphoprotein (DSPP) and F-actin expressions. PA-preconditioned macrotissues were subjected to histological analysis, including haematoxylin-eosin (HE), alizarin red, and Masson trichrome staining. Immunohistochemistry staining was used to detect alkaline phosphatase (ALP) and dentin matrix acidic phosphoprotein 1 (DMP-1) expressions. PA preconditioning had no adverse effects on microtissue spheroid viability and increased surface stiffness. It reduced dimensional shrinkage for over 7 days in microtissues and induced a larger transverse-section area in the macrotissue. PA preconditioning enhanced collagen formation, mineralized nodule formation, and elevated ALP and DMP-1 expressions in macrotissues. Additionally, PA preconditioning induced higher F-actin and DSPP expression in microtissues, while inhibition of F-actin activity by cytochalasin B attenuated PA-induced dimensional change and DSPP upregulation. PA surface preconditioning of DPSCs spheroids demonstrates excellent biocompatibility while effectively enhancing tissue structure stability and promoting biomineralization. This strategy strengthens tissue integrity in DPSC-derived spheroids and amplifies osteogenic differentiation potential, advancing scaffold-free tissue engineering applications in regenerative dentistry.

Modification of pea starch using ternary mixtures of natural crosslinking agents: Vanillin-chitosan-betaine and vanillin-gelatin-betaine

Different methods of starch modification have been proposed to broaden its application. In this study, the effects of ternary mixtures of natural crosslinking agents: chitosan-betaine-vanillin and gelatin-betaine-vanillin on the properties of pea starch were explored. These combinations of substances were selected because they have complementary crosslinking mechanisms. The effects of the ternary crosslinker mixtures on the gelatinization, mechanical properties, thermal stability, and microstructure of pea starch were compared. Both combinations of crosslinkers enhanced the gelatinization viscosity, viscoelasticity, gel hardness, and thermal stability of the pea starch, by an amount that depended on the ratio of the different components in the ternary mixtures. In all cases, the crystal structure of the starch granules disappeared after gelatinization. The modified starch had a more compact and uniform microstructure than the non-modified version, especially when it was crosslinked by vanillin, gelatin, and betaine. The improvement in the gelation properties of the starch were primarily attributed to hydrogen bonding, electrostatic attraction, and Schiff base crosslinking of the various components present. Gelatin enhanced the gel strength more than chitosan, which was probably because of greater hydrogen bonding. Our findings suggest that the properties of starch can be enhanced by adding ternary mixtures of natural crosslinkers.

Rare-Earth-Metal-Free Solid-State Fluorescent Carbonized-Polymer Microspheres for Unclonable Anti-Counterfeit Whispering-Gallery Emissions from Red to Near-Infrared Wavelengths.

Colloidal carbon dots (CDs) have garnered much attention as metal-free photoluminescent nanomaterials, yet creation of solid-state fluorescent (SSF) materials emitting in the deep red (DR) to near-infrared (NIR) range poses a significant challenge with practical implications. To address this challenge and to engineer photonic functionalities, a micro-resonator architecture is developed using carbonized polymer microspheres (CPMs), evolved from conventional colloidal nanodots. Gram-scale production of CPMs utilizes controlled microscopic phase separation facilitated by natural peptide cross-linking during hydrothermal processing. The resulting microstructure effectively suppresses aggregation-induced quenching (AIQ), enabling strong solid-state light emission. Both experimental and theoretical analysis support a role for extended π-conjugated polycyclic aromatic hydrocarbons (PAHs) trapped within these microstructures, which exhibit a progressive red shift in light absorption/emission toward the NIR range. Moreover, the highly spherical shape of CPMs endows them with innate photonic functionalities in combination with their intrinsic CD-based attributes. Harnessing their excitation wavelength-dependent photoluminescent (PL) property, a single CPM exhibits whispering-gallery modes (WGMs) that are emission-tunable from the DR to the NIR. This type of newly developed microresonator can serve as, for example, unclonable anti-counterfeiting labels. This innovative cross-cutting approach, combining photonics and chemistry, offers robust, bottom-up, built-in photonic functionality with diverse NIR applications.

Influence of Crosslinking Concentration on the Properties of Biodegradable Modified Cassava Starch-Based Films for Packaging Applications.

Chitosan/modified cassava starch/curcumin (CS/S/Cur) films with a crosslinker were developed via the solvent casting technique for the application of food packaging. The effects of citric acid (CA) as a natural crosslinker were assessed at different concentrations (0-10.0%, w/w, on a dry base on CS and S content). To measure the most favorable film, chemical structure and physical, mechanical, and thermal properties were investigated. Successful crosslinking between CS and S was seen clearly in the Fourier Transform Infrared (FTIR) spectra. The properties of the water resistance of the CS/S/Cur films crosslinked with CA were enhanced when compared to those without CA. Furthermore, it was found that the addition of CA crosslinking would improve the mechanical properties of composite films to some extent. It had been reported that the CA crosslinking level of 7.5 wt% of CS/S/Cur film demonstrated high performance in terms of physical properties. The tensile strength of the crosslinked film increased from 8 ± 1 MPa to 12 ± 1 MPa with the increasing content of CA, while water vapor permeability (WVP), swelling degree (SD), and water solubility (WS) decreased. An effective antioxidant scavenging activity of the CS/S/Cur film decreased with an increase in CA concentrations. This study provides an effective pathway for the development of active films based on polysaccharide-based film for food packaging applications.

The Pathophysiology of Keratoconus.

Keratoconus is a progressive disease characterized by changes in corneal shape, resulting in loss of visual function. There remains a lack of comprehensive understanding regarding its underlying pathophysiology. This review aims to bridge this gap by exploring structural failures and inflammatory processes involved in the etiology and progression of keratoconus. A literature review was conducted using PubMed and Google Scholar databases, screening for articles published in English using the keyword combinations of "keratoconus" with "pathophysiology," "pathology," "metabolism," "inflammatory," "oxidative stress," "cytokines," "enzymes," "collagen," and "cornea." Articles published between January 1, 1970, and June 1, 2023, were queried and reviewed, with greater emphasis placed on more recent data. Fifty-six relevant studies were examined to develop a thorough review of the pathophysiological mechanisms at play in keratoconus. Biomechanical structural failures in the cornea seem to be the primary militating factors in keratoconus etiology and progression. These include disruptions in the arrangement in the collagen lamellae, a decrease in collagen levels, a decrease in natural collagen crosslinking, and changes in lysosomal enzyme activity. Immunologic changes have also been identified in keratoconus, challenging the traditional view of the condition as noninflammatory. Elevated levels of proinflammatory cytokines like IL-1b, IL-6, IL-17, and TNF-α have been observed, along with increased apoptosis of keratocytes. Increased oxidative stress leads to the activation of collagenase and gelatinase enzymes. Keratoconus is a complex condition influenced by both structural defects and inflammatory processes. Understanding these mechanisms can inform clinical management and potentially lead to more effective treatments.

Role of crosslinkers on the properties of bio‐based wood adhesives

AbstractAs people became more conscious of the risks associated with pollution in the early 21st century, eco‐friendly adhesives made from plant oil began to be used. Bio‐based adhesives are widely used due to their low price, easy accessibility, and high biodegradability; however, their low adhesive strength is an issue. This work incorporates three natural crosslinkers: tannic acid (TA), sorbitol (SR), and cellulose (CL) to address the drawbacks of bio‐based adhesives, such as low adhesive strength, inferior output in humidity and longer curing time that result from excess water. Methylene diphenyl diisocyanate (MDI) was used to react with soybean oil‐based polyol (SOP) plus the crosslinkers to make bio‐based polyurethane adhesives. The maximum bonding strength of the soybean‐based adhesive containing 10 wt.% of SR (SR‐10%) was measured at 6.19 MPa which is about 44% higher than the sample without SR. The higher strength is due to the crosslinking ability of SR. The effect of crosslinking is also observed in the study of polar protic and polar aprotic solvents used to dissolve the adhesive. As compared with the polar aprotic solvent tetrahydrofuran, TA‐containing adhesive samples showed a maximum tensile strength of 3.69 MPa in methanol; a polar protic solvent that can participate in further hydrogen bonding to form a higher crosslinked complex. The glass transition temperature (Tg) increases with the increase in crosslinker amount. The observed increase in Tg may be attributable to an increase in crosslinking density, which reduces polymer chain flexibility and causes a stronger material.Highlights Bio‐adhesives based on soybean oil and diisocyanate were synthesized. Natural crosslinking agents such as tannic acid, sorbitol, and cellulose were used. The effect of crosslinking using polar protic and aprotic solvents was studied. The glass transition temperature increases with the increase in crosslinker amount.

Synergistic impact of natural gums and crosslinkers on the properties of oilseed meals based biopolymeric films

The waste material utilization from available agricultural resources can be beneficial in the field of economic, social, and environmental well-being. One of the main industrial crops used to manufacture oil from oilseeds worldwide is agricultural waste, such as the cake made from oilseeds. In this study, de-oiled cakes are used to create biopolymeric films. Three widely accessible oilseed meals viz. flaxseed, soybean, and mustard were gathered, ground, and sieved. A film forming suspension of defatted meals along with natural gums (acacia and xanthan gum) and crosslinkers (citric acid and glutaraldehyde) were formed. The suspension was cast into petri dishes and dried to produce smooth and even films. The physical, functional, color, thermal and morphological properties of the oilseed meals-gums crosslinked biopolymeric film were evaluated and statistical analysis was performed. The solubility was found to be decreased and tensile strength was increased with the addition of citric acid and increase in tensile strength. There was significant difference observed in the values of elongation at break after addition of citric acid as crosslinker. The research shows how oilseed meals enriched with natural gum and crosslinkers may be converted into biopolymeric films, which can then be used in food packaging to lessen reliance on petroleum-based, non-biodegradable plastics.

Long-term function of a novel autologous transcatheter pulmonary heart valve implant in an adult animal model.

Current heart valve implants entail major disadvantages in the treatment for younger patients or those with congenital heart defects. Evaluation of novel transcatheter pulmonary valve implant made from autologous pericardium with natural crosslinking agent in an in vitro setup and in vivo animal model METHODS: Valves were tested in a pulse duplicator according to ISO-standard 5840. For in vivo studies computer tomography was performed to measure sheep's native pulmonary valve dimensions. Pericardium was harvested by thoracotomy, personalized implants were manufactured and deployed in pulmonary valve position of the same sheep. Every 3 months implant functionality was evaluated by intracardiac echocardiography, intracardiac pressure measurements and cardiac magnetic resonance imaging (cMRI). Implants were explanted for macroscopic and histological examination. In vitro experiments showed compliance with regulatory requirements in terms of valve opening and insufficiency. Five sheep successfully received an autologous valve implant. Two animals had to be euthanized due to trauma sustained in the stable. Long-term valve function was excellent in three out of four animals with median implant cMRI regurgitation fraction of 9% (n = 4) at 3 months, 8% (n = 3) at 6, 8% (n = 3) at 9, 12% (n = 3) at 13, 8% (n = 2) at 17% and 8% (n = 2) at 20.5 months after implantation. Despite good adherence to neighboring tissue and endothelization, histological assessment revealed some signs of degeneration. Transcatheter pulmonary valve implants showed promising function for up to 20.5 months encouraging research to further improve this approach.

Enhancing the angiogenesis and osteogenesis of low elastic modulus Ti-Nb-Zr-Sn alloy surface through grafting growth factors-containing gelatin nanoparticles

The study aimed to apply a multiple surface modification for low elastic modulus Ti-24 % Nb-4 % Zr-8 % Sn (Ti2448) alloy, to enhance its biological properties for bone implant applications. We employed a combined sandblasting/acid etching/alkaline etching process to modify the Ti2448 alloy surfaces with mixed nano/submicron/micron-scaled pits and super wettability. We prepared gelatin nanoparticles (GNPs) loaded with growth factors: vascular endothelial growth factor/platelet-derived growth factor (VEGF/PDGF) and/or bone morphogenetic protein-2 (BMP-2). The growth factors-containing GNPs were then sequentially grafted onto the surface-modified Ti2448 alloy in a layered (single layer: BMP-2 or VEGF/PDGF; double layer: inner BMP-2 and outer VEGF/PDGF) or mixed BMP-2/VEGF/PDGF manner. Surface characterizations, including morphology, roughness, surface energy, and cytotoxicity, of Ti2448 alloy were analyzed. For biological response, we investigated the tube formation ability of human umbilical vein endothelial cells (HUVECs) and the cell responses (adhesion, proliferation, and mineralization) of human bone marrow mesenchymal stem cells (hMSCs). The results demonstrated the successful preparation of GNPs with 200∼300 nm particle size and their successful loading with the growth factors using natural crosslinker genipin. The growth factor-containing GNPs were effectively grafted onto the surface-modified hydrophilic Ti2448 alloy without significantly reducing its wettability (water contact angle remained below 40°). The results of the study showed that the GNPs-grafted Ti2448 alloy surfaces, with the GNPs loaded with VEGF/PDGF and/or BMP-2, exhibited enhanced biological properties. The GNPs-grafted surfaces were found to be non-toxic and capable of promoting tube formation of HUVECs, particularly when single VEGF/PDGF layer or double (inner BMP-2/ outer VEGF/PDGF) layer was loaded on GNPs (enhancing the cumulative tube length approximately 1.4 times that of no GNPs-grafted surfaces). Moreover, the GNPs-grafted surfaces enhanced the adhesion-related proteins and extracellular matrix mineralization of hMSCs, particularly with the loading of single VEGF/PDGF or mixed VEGF/PDGF/BMP-2 on GNPs (enhancing αvβ3 integrin patch number > 3.5 times, vinculin patch number > 2 times, and mineralization approximately 1.4 times). Based on these findings, we concluded that the proposed Ti2448 alloy surfaces, when grafted with VEGF/PDGF- and/or BMP-2-containing GNPs, have the potential for bone implant applications. The surface modifications on the Ti2448 alloy may promote angiogenesis in the early stage of bone repair (through VEGF/PDGF) and osteogenesis in the late stage (through BMP-2), thereby accelerating the overall bone healing process.

Reinforcement of nanostructured polyacrylamide hydrogels through the generation of secondary physical network using the nanoparticles’ functional groups

The main drawback regarding hydrogels utilization in biomedical engineering is represented by their weak mechanical properties and it is often addressed either by using nanospecies as fillers or by generating secondary networks that strengthen the material. Our study validates a new synthesis route of obtaining double-network nanostructured materials making use of the functional groups of an adsorbed glyco-protein on carbon-based nanospecies. Following a simple procedure, the decorated nanospecies were embedded in a synthetic polymer and subsequently formed a secondary hydrogen-bond based network through incubation in natural polyphenol solutions of various concentrations. The natural macromolecule's adsorption was evaluated through FTIR and DLS measurements. When compared to the single-network nanostructured materials, the double-network hydrogels exhibited lower affinity to aqueous media and improved ability to dissipate energy upon mechanical stress, depending on the amount of the employed natural crosslinker. As a result of generating the second network through incubation in polyphenol aqueous solution, the surface storage and loss moduli exceed the values of the bulk ones, as indicated by rheology and nanoindentation tests. The properties of the double-network hydrogels can also be tailored towards more hydrophilic surfaces with increased roughness through the control of the supplementary crosslinker amount. Considering the studied physical-chemical properties, and especially their mechanical characteristics, the obtained materials hold great promise for biomedical applications, particularly for the fabrication of scaffolds for wound dressing, soft or low load bearing tissue regeneration and repair.

A microscale 3D organ on a chip for recapitulating reciprocal neuroendocrine crosstalk between the hypothalamus and the pituitary gland

Conventional 2D or even recently developed 3D in vitro culture models for hypothalamus and pituitary gland cannot successfully recapitulate reciprocal neuroendocrine communications between these two pivotal neuroendocrine tissues known to play an essential role in controlling the body’s endocrine system, survival, and reproduction. In addition, most current vitro culture models for neuroendocrine tissues fail to properly reflect their complex multicellular structure. In this context, we developed a novel microscale chip platform, termed the ‘hypothalamic–pituitary (HP) axis-on-a-chip,’ which integrates various cellular components of the hypothalamus and pituitary gland with biomaterials such as collagen and hyaluronic acid. We used non-toxic blood coagulation factors (fibrinogen and thrombin) as natural cross-linking agents to increase the mechanical strength of biomaterials without showing residual toxicity to overcome drawbacks of conventional chemical cross-linking agents. Furthermore, we identified and verified SERPINB2 as a reliable neuroendocrine toxic marker, with its expression significantly increased in both hypothalamus and pituitary gland cells following exposure to various types of toxins. Next, we introduced SERPINB2-fluorescence reporter system into loaded hypothalamic cells and pituitary gland cells within each chamber of the HP axis on a chip, respectively. By incorporating this SERPINB2 detection system into the loaded hypothalamic and pituitary gland cells within our chip platform, Our HP axis-on-chip platform can better mimic reciprocal neuroendocrine crosstalk between the hypothalamus and the pituitary gland in the brain microenvironments with improved efficiency in evaluating neuroendocrine toxicities of certain drug candidates.

COTINUS COGGYGRIA ETHANOL EXTRACT AS CROSSLINKING AGENT IN FORMULATION OF THE PROTEIN FILMS

The study explores the potential application of smoketree (Cotinus coggygria Scop) ethanol extract (STE) as a natural cross-linking and active component in the formulation of films based on collagen (CL), gelatin (GL) and soy protein isolate (SPI). After casting and drying of the respective film-forming solutions, elastic and homogeneous films were obtained. A significant color difference was found for all protein films with added STE compared to the control series (mainly due to an increase in yellowness). The degree of cross-linking was highest in CLSTE (60 %), followed by GLSTE (27 %) and SPISTE (18 %). The antioxidant activity of the films, determined by the DPPH radical scavenging assay, increased significantly after the addition of STE. The observed increase in tensile strength (TS) for the films with STE ranged from double (CLSTE and SPISTE) to more than 4 times (GLSTE). In addition, there was a significant improvement in the light barrier properties of the extract-incorporated films, but also a reduction in their transparency up to 1.5 times compared to their respective controls. No significant influence was observed on physical characteristics such as thickness, moisture absorption and total soluble matter. Research results show that STE can be successfully used as a natural cross-linking agent in protein films, resulting in improved light barrier properties as well as increased strength and antioxidant activity.

DOTAGEL: a hydrogen and amide bonded, gelatin based, tunable, antibacterial, and high strength adhesive synthesized in an unoxidized environment.

The development of bioadhesives that concurrently exhibit high adhesion strength, biocompatibility, and tunable properties and involve simple fabrication processes continues to be a significant challenge. In this study, a novel bioadhesive named DOTAGEL is synthesized by crosslinking gelatin (GA), dopamine (DA), and tannic acid (TA) in an unoxidized environment due to the advantage of controlling the degree of protonation in GA and TA, as well as controlling the degree of intermolecular amide and hydrogen bonding in the acidic medium. DOTAGEL (DA + TA + GA) shows superior adhesion strengths of 104.6 ± 46 kPa on dry skin and 35.6 ± 4.5 kPa on wet skin, up to 13 attachment-detachment cycles, retains adhesion strength under water for up to 10 days and is capable of joining two cut parts of internal organs of mice. Moreover, DOTAGEL shows strong antibacterial properties, self-healing, and biocompatibility since it contains TA, a natural and antibacterial cross-linker with abundant hydroxyl groups and the capability of forming non-covalent bonds in an unoxidized environment, and dopamine hydrochloride, a mussel inspired biomaterial containing both the amine and catechol groups for amide bonding and hydrogen bonding with TA and GA. The cross-linking among 20% (w/v) GA, 0.2% (w/v) DA, and 20% (w/v) TA is done by the centrifugation process at room temperature. Two different acids, hydrochloric acid and acetic acid, were used for tuning the pH of the medium, which led to two different samples named DOTAGEL/AA and DOTAGEL/HCL. The degree of cross-linking and mechanical and biochemical properties, like adhesion strength, degradation rate, antibacterial properties, stickiness, etc., are tuned by adjusting the pH of the medium. DOTAGEL/HCL showed 6.5 times faster degradation in 10 days, a faster release rate in the antibacterial study, 2 times adhesion strength in a dry medium, and more stickiness. The novelty lies not only in increased adhesion strength but also in the single-step fabrication process of the adhesive in the acidic medium. This research proposes the formation of a tunable antibacterial adhesive that is capable of working on wet surfaces within the body and that has the potential to become a successful tissue adhesive with a wide range of possibilities in controlled drug delivery at wound sites and other biomedical applications.