Biopolymer Networks Research Articles

Capturing Dynamic Assembly of Nanoscale Proteins During Network Formation

AbstractThe structural evolution of hierarchical structures of nanoscale biomolecules is crucial for the construction of functional networks in vivo and in vitro. Despite the ubiquity of these networks, the physical mechanisms behind their formation and self‐assembly remains poorly understood. Here, this study uses photochemically cross‐linked folded protein hydrogels as a model biopolymer network system, with a combined time‐resolved rheology and small‐angle x‐ray scattering (SAXS) approach to probe both the load‐bearing structures and network architectures respectively thereby providing a cross‐length scale understanding of the network formation. Combining SAXS, rheology, and kinetic modeling, a dual formation mechanism consisting of a primary formation phase is proposed, where monomeric folded proteins create the preliminary protein network scaffold; and a subsequent secondary formation phase, where both additional intra‐network cross‐links form and larger oligomers diffuse to join the preliminary network, leading to a denser more mechanically robust structure. Identifying this as the origin of the structural and mechanical properties of protein networks creates future opportunities to understand hierarchical biomechanics in vivo and develop functional, designed‐for‐purpose, biomaterials.

Building rigid networks with prestress and selective pruning

Biopolymer networks from the intracellular to tissue scale display high rigidity and tensile stress while having coordinations well below the normal threshold for mechanical rigidity. The elastic filaments in these networks are often severed by enzymes in a tension-inhibited manner. The effects of such pruning on the mechanics of prestressed networks have not been studied. We show that networks pruned by a tension-inhibited method remain rigid at much lower coordinations than randomly pruned ones. These findings suggest a possible reason for the repeated evolution of tension-inhibited filament-severing proteins. Published by the American Physical Society 2024

Adsorption behavior of solids incorporated in alginate hydrogel beads using herbicides 2,4-D and paraquat as test molecules

This study investigates the behavior of adsorbent solids within three-dimensional networks of biopolymers. Two chemically different herbicides, 2,4-D and paraquat (PQ) were employed as test molecules. Alginate (ALG) hydrogels in bead form were synthesized, incorporating activated carbon (PSCA) and montmorillonite clay (MMT) simultaneously and in varying proportions, resulting in multicomponent beads. The Langmuir isotherm parameters, KL (L mmol−1) and qmax (mmol g−1), in the adsorption of herbicides on beads with an increasing amount of montmorillonite (MMT) were as follows: For PQ, KL values were 10.709, 61.951, 162.33, 223.27, and qmax values were 0.0711, 0.1484, 0.2135, 0.2497. For 2,4-D, KL values were 13.360, 8.3234, 7.4126, 5.4308, and qmax values were 0.9529, 0.7096, 0.6534, 0.5881. The values clearly demonstrates that, even when solids are mixed in the hydrogel network, their reactive sites remain exposed, interacting with molecules to which they have greater affinity. Adsorption was also investigated on identical materials using binary mixtures of herbicides, revealing trends similar to those observed for individual solutions. Moreover, it was demonstrated that the adsorption capacity of multicomponent beads is equivalent to that achieved by proportionally mixing beads synthesized exclusively with clay or activated carbon. This suggests the tangible possibility of creating on-demand adsorbents by synthesizing distinct hydrogels, each incorporating a specific solid. Subsequently, drying, storing, and mixing them based on the target contaminant for retention are feasible. Finally, a column was packed with a proportional mixture of beads and successfully retained PQ and 2,4-D using a flow-through system.

Injectable Carrageenan/Green Graphene Oxide Hydrogel: A Comprehensive Analysis of Mechanical, Rheological, and Biocompatibility Properties.

In this work, physically crosslinked injectable hydrogels based on carrageenan, locust bean gum, and gelatin, and mechanically nano-reinforced with green graphene oxide (GO), were developed to address the challenge of finding materials with a good balance between injectability and mechanical properties. The effect of GO content on the rheological and mechanical properties, injectability, swelling behavior, and biocompatibility of the nanocomposite hydrogels was studied. The hydrogels' morphology, assessed by FE-SEM, showed a homogeneous porous architecture separated by thin walls for all the GO loadings investigated. The rheology measurements evidence that G' > G″ over the whole frequency range, indicating the dominant elastic nature of the hydrogels and the difference between G' over G″ depends on the GO content. The GO incorporation into the biopolymer network enhanced the mechanical properties (ca. 20%) without appreciable change in the injectability of the nanocomposite hydrogels, demonstrating the success of the approach described in this work. In addition, the injectable hydrogels with GO loadings ≤0.05% w/v exhibit negligible toxicity for 3T3-L1 fibroblasts. However, it is noted that loadings over 0.25% w/v may affect the cell proliferation rate. Therefore, the nano-reinforced injectable hybrid hydrogels reported here, developed with a fully sustainable approach, have a promising future as potential materials for use in tissue repair.

Electric-field assisted cascade reactions to create alginate/carboxymethyl chitosan composite hydrogels with gradient architecture and reconfigurable mechanical properties

Rational designs of polysaccharide-based hydrogels with organ-like three-dimensional architecture provide a great possibility for addressing the shortages of allograft tissues and organs. However, spatial-temporal control over structure in bulk hydrogel and acquire satisfied mechanical properties remain an intrinsic challenge to achieve. Here, we show how electric-field assisted molecular self-assembly can be coupled to a directional reaction-diffusion (RD) process to produce macroscopic hydrogel in a controllable manner. The electrical energy input was not only to generate complex molecule gradients and initiate the molecular self-assembly, but also to guide/facilitate the RD processes for the gel rapid growth via a cascade construction interaction. The hydrogel mechanical properties can be tuned and enhanced by using an interpenetrating biopolymer network and multiple ionic crosslinkers, leading to a wide-range of mechanical modulus to match with biological organs or tissues. We demonstrate diverse 3D macroscopic hydrogels can be easily prepared via field-assisted directional reaction-diffusion and specific joint interactions. The humility-triggered dissipation of functional gradients and antibacterial performance confirm that the hydrogels can serve as an optically variable soft device for wound management. Therefore, this work provides a general approach toward the rational fabrication of soft hydrogels with controlled architectures and functionality for advanced biomedical systems.

Gamma irradiation-induced crosslinking and enhanced functionality of fish gelatin-agar edible films

Fish gelatin-agar edible films have potential for sustainable food packaging, but their limitations particularly water sensitivity require improvement. In this research, the effects of radiation from gamma rays at rates ranging from 0 to 40 kGy on the films' mechanical characteristics, microstructure, color, opacity, and water resistance were examined. Overall, gamma irradiation at doses of up to 30 kGy substantially improved the water resistance of the gelatin-agar film, as demonstrated by decreased water solubility, water absorption, and film vapor permeability. This could be due to crosslinking within the biopolymer network. However, at doses of 40 kGy, potential chain scission in the film's polymeric structure might occur, which counteracts further improvement and consequently starts degrading its water resistance. Similarly, the improvement of film's tensile strength and modulus of elasticity peaked at an optimal dose of 30 kGy, and then decreased at higher doses, while the film's elongation at break remained unaffected. FTIR and SEM analysis supported changes in functional group and structural morphology in these observations. Although irradiation caused some yellowing, an optimal dose range (at around 20–30 kGy) was identified to achieve improved functionality without degrading film integrity. These findings highlight the potential of gamma irradiation as a viable technique for modifying fish gelatin-agar films for enhanced performance in food packaging applications.

Biopolymer networks packed with microgels combine strain stiffening and shape programmability

Biomaterials that can be reversibly stiffened and shaped could be useful in broad biomedical applications where form-fitting scaffolds are needed. Here we investigate the combination of strong non-linear elasticity in biopolymer networks with the reconfigurability of packed hydrogel particles within a composite biomaterial. By packing microgels into collagen-1 networks and characterizing their linear and non-linear material properties, we empirically determine a scaling relationship that describes the synergistic dependence of the material's linear elastic shear modulus on the concentration of both components. We perform high-strain rheological tests and find that the materials strain stiffen and also exhibit a form of programmability, where no applied stress is required to maintain stiffened states of deformation after large strains are applied. We demonstrate that this non-linear rheological behavior can be used to shape samples that do not spontaneously relax large-scale bends, holding their deformed shapes for days. Detailed analysis of the frequency-dependent rheology reveals an unexpected connection to the rheology of living cells, where models of soft glasses capture their low-frequency behaviors and polymer elasticity models capture their high-frequency behaviors.

Vimentin is a key regulator of cell mechanosensing through opposite actions on actomyosin and microtubule networks

The cytoskeleton is a complex network of interconnected biopolymers consisting of actin filaments, microtubules, and intermediate filaments. These biopolymers work in concert to transmit cell-generated forces to the extracellular matrix required for cell motility, wound healing, and tissue maintenance. While we know cell-generated forces are driven by actomyosin contractility and balanced by microtubule network resistance, the effect of intermediate filaments on cellular forces is unclear. Using a combination of theoretical modeling and experiments, we show that vimentin intermediate filaments tune cell stress by assisting in both actomyosin-based force transmission and reinforcement of microtubule networks under compression. We show that the competition between these two opposing effects of vimentin is regulated by the microenvironment stiffness. These results reconcile seemingly contradictory results in the literature and provide a unified description of vimentin’s effects on the transmission of cell contractile forces to the extracellular matrix.

Eco-friendly disposable porous absorbents from gluten proteins through diverse plastic processing techniques

The production of biodegradable gluten-based protein foams showing complete natural degradation in soil after 26 days is reported, as an alternative to commercial foams in disposable sanitary articles that rely on non-biodegradable materials. The foams were developed from an extensive evaluation of different foaming methodologies (oven expansion, compression moulding, and extrusion), resulting in low-density foams (ca. 400 kg/m3) with homogenous pore size distributions. The products showed the ability to absorb 3–4 times their weight, reaching ranges for their use as absorbents in single-use disposable sanitary articles. An additional innovative contribution is that these gluten foams were made from natural and non-toxic wheat protein, glycerol, sodium and ammonium bicarbonate, making them useful as fossil-plastic-free replacements for commercial products without the risk of having micro-plastic and chemical pollution. The impact of different processing conditions on forming the porous biopolymer network is explained, i.e., temperature, pressure, and extensive shear forces, which were also investigated for different pH/chemical conditions. The development of micro-plastic-free foams mitigating environmental pollution and waste while using industrial co-products is fundamental for developing large-scale production of single-use items. A sanitary pad prototype is demonstrated as an eco-friendly material alternative that paves the way for sustainable practices in manufacturing, and contributes to the global effort in combating plastic pollution and waste management challenges, Sustainable Development Goals: 12, 13, 14, and 15.

Effects of local incompressibility on the rheology of composite biopolymer networks.

Fibrous networks such as collagen are common in biological systems. Recent theoretical and experimental efforts have shed light on the mechanics of single component networks. Most real biopolymer networks, however, are composites made of elements with different rigidity. For instance, the extracellular matrix in mammalian tissues consists of stiff collagen fibers in a background matrix of flexible polymers such as hyaluronic acid (HA). The interplay between different biopolymer components in such composite networks remains unclear. In this work, we use 2D coarse-grained models to study the nonlinear strain-stiffening behavior of composites. We introduce a local volume constraint to model the incompressibility of HA. We also perform rheology experiments on composites of collagen with HA. Theoretically and experimentally, we demonstrate that the linear shear modulus of composite networks can be increased by approximately an order of magnitude above the corresponding moduli of the pure components. Our model shows that this synergistic effect can be understood in terms of the local incompressibility of HA, which acts to suppress density fluctuations of the collagen matrix with which it is entangled.

Understanding the influence of the arabinoxylan-rich psyllium (Plantago ovata) husk on dough elasticity and bread staling: Interplay between biopolymer and water dynamics

Bread staling is a complex phenomenon in which multiple interconnected mechanisms involving water operate, including the formation of amylopectin crystallites sequestering water, crumb-to-crust moisture migration, and the escalating immobilization of water reducing gluten plasticization. Considering the pivotal role of water-gluten-starch interactions, the integration of the highly-soluble, water-avid, arabinoxylan-rich psyllium husk (PSY) into a wheat dough/bread system was explored, to understand the impact of PSY on the water and biopolymer dynamics that could lead to lower staling. With the addition of PSY, optimal dough development required more water (from 68.0 to 119.5 % in control and 10 % wheat flour replacement with PSY, respectively). LF-NMR showed a parallel increase in the proton population attributed to free water entrapped by the continuous network of biopolymers surrounding granular starch (A23), and its relaxation time (T23). PSY resulted in freshly baked bread with similar specific volume, porosity and starch gelatinization than the control, but with a much higher proportion of intergranular free water (A23). Importantly, PSY did not affect amylopectin retrogradation nor shortened the relaxation time of water protons in exchange with gluten, suggesting that PSY did not withdraw plasticizing water from starch or gluten, respectively. During storage, PSY resulted in a dose-dependent delay of both crumb hardening (correlated to T23, r = −0.76, p < 0.05) and loss of cohesiveness (correlated to T23, r = 0.71, p < 0.05 and A23, r = 0.74, p < 0.05). PSY also resulted in greater crumb resilience, which was related to the self-assembly of PSY molecules and its impact on dough elasticity (with up to two-fold increase in recovery strain).

Exploring Dielectric and Magnetic Properties of Ni and Co Ferrites through Biopolymer Composite Films

This study explores the synthesis and characterization of chitosan/gelatine films incorporating nickel ferrite (NiFe2O4) and cobalt ferrite (CoFe2O4) nanoparticles. The magnetic nanoparticles exhibit superparamagnetic behaviour, making them attractive for various applications, including biomedical uses. The X-ray diffraction analysis confirmed the successful synthesis of NiFe2O4 and CoFe2O4 nanoparticles, and the scanning electron micrographs illustrated well-dispersed ferrite nanoparticles within the biopolymer network, despite the formation of some aggregates attributed to magnetic interactions. Magnetization loops revealed lower saturation magnetization values for the composites, attributed to the chitosan/gelatine coating and the dielectric studies, indicating increased dielectric losses in the presence of ferrites, particularly pronounced in the case of NiFe2O4, suggesting interactions at the interface region between the polymer and ferrite particles. The AC conductivity shows almost linear frequency dependence, associated with proton polarization and conduction processes, more significant at higher temperatures for samples with ferrite particles.

Synthesis of Gum Arabic-Based Biopolymer Network and Determination of Its Toxicity Properties in In Vitro - In Vivo Model Systems

In this study, gum arabic based network polymers were prepared using epoxy functional PEG structures. The basic physicochemical properties of these structures, their structural characterization, thermal properties and morphological properties were investigated. Toxicity properties of constructs synthesized on zebrafish (Danio rerio (Hamilton)) offspring were determined in vivo. In addition, in vitro toxicity tests were performed on L929 fibroblast cells. When the general properties of these structures were examined. Structural and thermal properties were better with increasing cross-linker rates ratios (1%, 3%, 5%). According to the toxicity test performed on zebrafish juveniles; GA-PEG-Epox (1%) constructs are non-toxic to zebrafish juveniles. The mortality rate of GA-PEG-Epox (3%) and GA-PEG-Epox (5%) structures was observed as 12.5% and 20.8%, respectively. It was observed that the structures were not toxic to zebrafish juveniles. MTT test performed on L929 fibroblast cells, high cell viability (&amp;gt;90%) was observed in all synthesized structures. These results are evaluated as Grade 1 according to ISO standards.

Algae-Based Biopolymers for Batteries and Biofuel Applications in Comparison with Bacterial Biopolymers-A Review.

Algae-based biopolymers can be used in diverse energy-related applications, such as separators and polymer electrolytes in batteries and fuel cells and also as microalgal biofuel, which is regarded as a highly renewable energy source. For these purposes, different physical, thermochemical, and biochemical properties are necessary, which are discussed within this review, such as porosity, high temperature resistance, or good mechanical properties for batteries and high energy density and abundance of the base materials in case of biofuel, along with the environmental aspects of using algae-based biopolymers in these applications. On the other hand, bacterial biopolymers are also often used in batteries as bacterial cellulose separators or as biopolymer network binders, besides their potential use as polymer electrolytes. In addition, they are also regarded as potential sustainable biofuel producers and converters. This review aims at comparing biopolymers from both aforementioned sources for energy conversion and storage. Challenges regarding the production of algal biopolymers include low scalability and low cost-effectiveness, and for bacterial polymers, slow growth rates and non-optimal fermentation processes often cause challenges. On the other hand, environmental benefits in comparison with conventional polymers and the better biodegradability are large advantages of these biopolymers, which suggest further research to make their production more economical.

Biobased Interpenetrating Polymer Network Membranes for Sustainable Molecular Sieving.

There is an urgent need for sustainable alternatives to fossil-based polymer materials. Through nanodomain engineering, we developed, without using toxic cross-linking agents, interpenetrating biopolymer network membranes from natural compounds that have opposing polarity in water. Agarose and natural rubber latex were consecutively self-assembled and self-cross-linked to form patchlike nanodomains. Both nano-Fourier transform infrared (nano-FTIR) spectroscopy and computational methods revealed the biopolymers' molecular-level entanglement. The membranes exhibited excellent solvent resistance and offered tunable molecular sieving. We demonstrated control over separation performance in the range of 227-623 g mol-1 via two methodologies: adjusting the molecular composition of the membranes and activating them in water. A carcinogenic impurity at a concentration of 5 ppm, which corresponds to the threshold of toxicological concern, was successfully purged at a negligible 0.56% pharmaceutical loss. The biodegradable nature of the membranes enables an environmentally friendly end-of-life phase; therefore, the membranes have a sustainable lifecycle from cradle to grave.

Ionic Liquids as Designed, Multi-Functional Plasticizers for Biodegradable Polymeric Materials: A Mini-Review.

Measures to endorse the adoption of eco-friendly biodegradable plastics as a response to the scale of plastic pollution has created a demand for innovative products from materials from Nature. Ionic liquids (ILs) have the ability to disrupt the hydrogen bonding network of biopolymers, increase the mobility of biopolymer chains, reduce friction, and produce materials with various morphologies and mechanical properties. Due to these qualities, ILs are considered ideal for plasticizing biopolymers, enabling them to meet a wide range of specifications for biopolymeric materials. This mini-review discusses the effect of different IL-plasticizers on the processing, tensile strength, and elasticity of materials made from various biopolymers (e.g., starch, chitosan, alginate, cellulose), and specifically covers IL-plasticized packaging materials and materials for biomedical and electrochemical applications. Furthermore, challenges (cost, scale, and eco-friendliness) and future research directions in IL-based plasticizers for biopolymers are discussed.

De novo design of modular protein hydrogels with programmable intra- and extracellular viscoelasticity

Relating the macroscopic properties of protein-based materials to their underlying component microstructure is an outstanding challenge. Here, we exploit computational design to specify the size, flexibility, and valency of de novo protein building blocks, as well as the interaction dynamics between them, to investigate how molecular parameters govern the macroscopic viscoelasticity of the resultant protein hydrogels. We construct gel systems from pairs of symmetric protein homo-oligomers, each comprising 2, 5, 24, or 120 individual protein components, that are crosslinked either physically or covalently into idealized step-growth biopolymer networks. Through rheological assessment, we find that the covalent linkage of multifunctional precursors yields hydrogels whose viscoelasticity depends on the crosslink length between the constituent building blocks. In contrast, reversibly crosslinking the homo-oligomeric components with a computationally designed heterodimer results in viscoelastic biomaterials exhibiting fluid-like properties under rest and low shear, but solid-like behavior at higher frequencies. Exploiting the unique genetic encodability of these materials, we demonstrate the assembly of protein networks within living mammalian cells and show via fluorescence recovery after photobleaching (FRAP) that mechanical properties can be tuned intracellularly in a manner similar to formulations formed extracellularly. We anticipate that the ability to modularly construct and systematically program the viscoelastic properties of designer protein-based materials could have broad utility in biomedicine, with applications in tissue engineering, therapeutic delivery, and synthetic biology.

Clots reveal anomalous elastic behavior of fiber networks.

The adaptive mechanical properties of soft and fibrous biological materials are relevant to their functionality. The emergence of the macroscopic response of these materials to external stress and intrinsic cell traction from local deformations of their structural components is not well understood. Here, we investigate the nonlinear elastic behavior of blood clots by combining microscopy, rheology, and an elastic network model that incorporates the stretching, bending, and buckling of constituent fibrin fibers. By inhibiting fibrin cross-linking in blood clots, we observe an anomalous softening regime in the macroscopic shear response as well as a reduction in platelet-induced clot contractility. Our model explains these observations from two independent macroscopic measurements in a unified manner, through a single mechanical parameter, the bending stiffness of individual fibers. Supported by experimental evidence, our mechanics-based model provides a framework for predicting and comprehending the nonlinear elastic behavior of blood clots and other active biopolymer networks in general.

Sprayed water-based lignin colloidal nanoparticle-cellulose nanofibril hybrid films with UV-blocking ability.

In the context of global climate change, the demand for new functional materials that are sustainable and environmentally friendly is rapidly increasing. Cellulose and lignin are the two most abundant raw materials in nature, and are ideal components for functional materials. The hydrophilic interface and easy film-forming properties of cellulose nanofibrils make them excellent candidates for natural biopolymer templates and network structures. Lignin is a natural UV-shielding material, as it contains a large number of phenolic groups. In this work, we have applied two routes for spray deposition of hybrid films with different laminar structures using surface-charged cellulose nanofibrils and water-based colloidal lignin particles. As the first route, we prepare stacked colloidal lignin particles and cellulose nanofibrils hybrid film through a layer-by-layer deposition. As the second route, we spray-deposite premixed colloidal lignin particles and cellulose nanofibrils dispersion to prepare a mixed hybrid film. We find that cellulose nanofibrils act as a directing agent to dominate the arrangement of the colloidal lignin particles in a mixed system. Additionally, cellulose nanofibrils eliminate the agglomerations and thus increase the visible light transparency while retaining the UV shielding ability. Our research on these colloidal lignin and cellulose nanofibril hybrid films provides a fundamental understanding of using colloidal lignin nanoparticles as functional material on porous cellulose-based materials, for example on fabrics.

Rigidity percolation in a random tensegrity via analytic graph theory.

Functional structures from across the engineered and biological world combine rigid elements such as bones and columns with flexible ones such as cables, fibers, and membranes. These structures are known loosely as tensegrities, since these cable-like elements have the highly nonlinear property of supporting only extensile tension. Marginally rigid systems are of particular interest because the number of structural constraints permits both flexible deformation and the support of external loads. We present a model system in which tensegrity elements are added at random to a regular backbone. This system can be solved analytically via a directed graph theory, revealing a mechanical critical point generalizing that of Maxwell. We show that even the addition of a few cable-like elements fundamentally modifies the nature of this transition point, as well as the later transition to a fully rigid structure. Moreover, the tensegrity network displays a collective avalanche behavior, in which the addition of a single cable leads to the elimination of multiple floppy modes, a phenomenon that becomes dominant at the transition point. These phenomena have implications for systems with nonlinear mechanical constraints, from biopolymer networks to soft robots to jammed packings to origami sheets.