Formation Of Nanofibrils Research Articles

Nanofilament organization in highly tough fibers based on lamin proteins

The escalating plastic pollution crisis necessitates sustainable alternatives, and one promising solution involves replacing petroleum-based polymers with fibrous proteins. This study focused on the recombinant production of intracellular fibrous proteins, specifically Caenorhabditis elegans lamin (Ce-lamin). Ce-lamins spontaneously organize within the cell nucleus, forming a network of nanofilaments. This intricate structure serves as an active layer that responds dynamically to mechanical strain and stress. Herein, we investigated the arrangement of nanofilaments into nanofibrils within wet-spun Ce-lamin fibers using alcoholic solutions as coagulants. Our goal was to understand their structural and mechanical properties, particularly in comparison with those produced with solutions containing Ca+2 ions, which typically result in the formation of nanofibrils with a collagen-like pattern. The introduction of ethanol solutions significantly altered this pattern, likely through rearrangement of the nanofilaments. Nevertheless, the resulting fibers exhibited superior toughness and strain, outperforming various synthetic fibers. The significance of the nanofilament structure in enhancing fiber toughness was emphasized through both the secondary structure transition during stretching and the influence of the Q159K point mutation. This study improves our understanding of the structural and mechanical aspects of Ce-lamin fibers, paving the way for the development of eco-friendly and high-quality fibers suitable for various applications, including medical implants and composite materials.

Self-Blinking Thioflavin T for Super-resolution Imaging.

Thioflavin T (ThT) is a typical dye used to visualize the aggregation and formation of fibrillar structures, e.g., amyloid fibrils and peptide nanofibrils. ThT has been considered to produce stable fluorescence when interacting with aggregated proteins. For single-molecule localization microscopy (SMLM)-based optical super-resolution imaging, a photoswitching/blinking fluorescence property is required. Here we demonstrate that, in contrast to previous reports, ThT exhibits intrinsic stochastic blinking properties, without the need for blinking imaging buffer, in stable binding conditions. The blinking properties (photon number, blinking time, and on-off duty cycle) of ThT at the single-molecule level (for ultralow concentrations) were investigated under different conditions. As a proof of concept, we performed SMLM imaging of ThT-labeled α-synuclein fibrils measured in air and PBS buffer.

Effects of molecular weight of hydrolysate on the formation of soy protein isolate hydrolysate nanofibrils: Kinetics, structures, and interactions

Enzymatic hydrolysis prior to protein fibrillation was an effective way to facilitate the formation of nanofibrils. This study aimed to investigate the effects of molecular weights of hydrolysate on the kinetics, structures, and interactions of soy protein isolate (SPI) hydrolysate nanofibrils. The results showed that hydrolysate with molecular weight > 10 kDa showed a distinct fibrillation kinetics curve and a higher apparent rate constant (27.72) during fibrillation, indicating their vital role in determining the fibrillation. Hydrolysate with molecular weight > 10 kDa could form nanofibrils with higher radius gyration (17.11 ± 0.77 Å) due to stronger hydrophobic interaction, showing a stronger fibrillation ability. Hydrolysate with molecular weight within 5–10 kDa exhibited enhanced π-π stacking interactions during fibrillation, thereby promoting the extension of nanofibrils, and contributing to the formation of more nanofibrils. Hydrolysate with molecular weight < 5 kDa tended to randomly aggregate during fibrillation, resulting in a significant loss of cross-β structures in nanofibrils. Therefore, hydrolysate with different molecular weights exhibited synergistic effects during fibrillation.

Supramolecular Hydrolase Mimics in Equilibrium and Kinetically Trapped States

AbstractThe folding of proteins into intricate three‐dimensional structures to achieve biological functions, such as catalysis, is governed by both kinetic and thermodynamic controls. The quest to design artificial enzymes using minimalist peptides seeks to emulate supramolecular structures existing in a catalytically active state. Drawing inspiration from the nuanced process of protein folding, our study explores the enzyme‐like activity of amphiphilic peptide nanosystems in both equilibrium and non‐equilibrium states, featuring the formation of supramolecular nanofibrils and nanosheets. In contrast to thermodynamically stable nanosheets, the kinetically trapped nanofibrils exhibit dynamic characteristics (e.g., rapid molecular exchange and relatively weak intermolecular packing), resulting in a higher hydrolase‐mimicking activity. We emphasize that a supramolecular microenvironment characterized by an optimal local polarity, microviscosity, and β‐sheet hydrogen bonding is conducive to both substrate binding and ester bond hydrolysis. Our work underscores the pivotal role of both thermodynamic and kinetic control in impacting biomimetic catalysis and sheds a light on the development of artificial enzymes.

Supramolecular Hydrolase Mimics in Equilibrium and Kinetically Trapped States.

The folding of proteins into intricate three-dimensional structures to achieve biological functions, such as catalysis, is governed by both kinetic and thermodynamic controls. The quest to design artificial enzymes using minimalist peptides seeks to emulate supramolecular structures existing in a catalytically active state. Drawing inspiration from the nuanced process of protein folding, our study explores the enzyme-like activity of amphiphilic peptide nanosystems in both equilibrium and non-equilibrium states, featuring the formation of supramolecular nanofibrils and nanosheets. In contrast to thermodynamically stable nanosheets, the kinetically trapped nanofibrils exhibit dynamic characteristics (e.g., rapid molecular exchange and relatively weak intermolecular packing), resulting in a higher hydrolase-mimicking activity. We emphasize that a supramolecular microenvironment characterized by an optimal local polarity, microviscosity, and β-sheet hydrogen bonding is conducive to both substrate binding and ester bond hydrolysis. Our work underscores the pivotal role of both thermodynamic and kinetic control in impacting biomimetic catalysis and sheds a light on the development of artificial enzymes.

Suicide gene delivery by morphology-adaptable enantiomeric peptide assemblies for combined ovarian cancer therapy

Suicide gene therapy is a promising therapeutic model for ovarian cancer (OC), while suffering from poor gene delivery and limited therapeutic efficacy. To address this concern, here we reported the GSH-responsive morphology-transformable enantiomeric peptide assemblies as delivering vehicles for suicide genes and co-delivery of paclitaxel (PTX). Connecting a lipid-like amphiphile and a hydrophilic arginine segment through disulfide bonds led to the enantiomeric peptides. The enantiomeric peptide assemblies are able to simultaneously uptake plasmid DNA (pDNA) and PTX based on electrostatic and hydrophobic interactions. The resulting co-assemblies underwent GSH-responsive disulfide cleavage and thereby promoting their assembly from nanoparticles to nanofibers, leading to the co-release of pDNA and PTX. Cellular and animal studies confirmed the co-delivery of pDNA and PTX into OC cells and the cell apoptosis by the enantiomeric peptides. In addition, in vitro and in vivo experiments supported the advanced uptake and cytotoxicity for L-type peptide vehicles by OC cells, and their great potential for OC-imaging, growth-inhibition and apoptosis-induction compared to D-counterpart. Our results demonstrate that the GSH-responsive morphology-transformable chiral peptide assemblies accurately and simultaneously release suicide genes and chemodrugs at tumor sites, thus providing a new strategy for the development of delivering vehicles for suicide gene and establishment of new therapeutic models for ovarian cancer. Statement of significanceAppropriate delivery carriers are essential for the clinical translation of cancer gene therapy, including the emerging suicide gene therapy. By combining the advantages of morphological transformable vehicles with the chirality peptides towards their bioactivity, we developed the GSH-responsive morphology-transformable enantiomeric peptide assemblies as delivering vehicles for suicide genes and co-delivery of paclitaxel. The GSH-responsive assembly of the enantiomeric peptides allows for precise release of plasmid DNA and paclitaxel in cancer cells, and promotes the formation of nanofibrils that facilitate gene entering nuclei for transfection. The enantiomeric peptide-based vehicles show the chirality-dependent capability for inducing cell apoptosis and inhibiting tumor growth. Our findings demonstrate a new strategy for developing therapeutic models for ovarian cancer.

Synthesis and Characterization of pH Triggered Imine Functional Low Molecular Weight Hydrogel

AbstractA C‐3 symmetric imine functional nonpolymeric low molecular weight gelator is prepared using microwave assisted Schiff base reaction. The synthesized molecule forms a hydrogel at 5 wt% under slightly acidic pH. The material was characterized using an inversion tube test, thermogravimetric analysis (TGA), powder X‐ray diffraction (PXRD) and FT‐IR spectroscopy. Rheological studies show the material forms a strong hydrogel with formation of nanofibrils supported by field‐emission scanning electron microscopy (FE‐SEM). To the best of our knowledge this is the first example of imine based trigonal low molecular weight hydrogelator reported.

Silk Assembly against Hydrophobic Surfaces─Modeling and Imaging of Formation of Nanofibrils.

A detailed insight about the molecular organization behind spider silk assembly is valuable for the decoding of the unique properties of silk. The recombinant partial spider silk protein 4RepCT contains four poly-alanine/glycine-rich repeats followed by an amphiphilic C-terminal domain and has shown the capacity to self-assemble into fibrils on hydrophobic surfaces. We herein use molecular dynamic simulations to address the structure of 4RepCT and its different parts on hydrophobic versus hydrophilic surfaces. When 4RepCT is placed in a wing arrangement model and periodically repeated on a hydrophobic surface, β-sheet structures of the poly-alanine repeats are preserved, while the CT part is settled on top, presenting a fibril with a height of ∼7 nm and a width of ∼11 nm. Both atomic force microscopy and cryo-electron microscopy imaging support this model as a possible fibril formation on hydrophobic surfaces. These results contribute to the understanding of silk assembly and alignment mechanism onto hydrophobic surfaces.

Phototherapeutic effect of transformable peptides containing pheophorbide a on colorectal cancer

Photodynamic therapy (PDT) and photothermal therapy (PTT) have attracted research interest for their noninvasive nature and selective treatment of tumor tissues. They are effective through the generation of reactive oxygen species (ROS) or heat. Nevertheless, several problems, including low bioavailability and long-lasting cutaneous photosensitivity, have limited their clinical application. In this study, we reported an in situ self-assembly strategy that could improve various biological properties of the photosensitizer in vivo. A photosensitizer connected to a receptor-mediated smart peptide can self-assemble into nanoparticles (NPs) under the force of hydrophobic interaction and then transform into a nanofibrillar network after attaching to the tumor cell surface with the help of the β-sheet-forming peptide KLVFF. The supramolecular structural changes deeply affected the PDT and PTT properties of the photosensitizer on tumors. After being aggregated into the nanostructure, the water solubility and targeting ability of the photosensitizer was ameliorated. Moreover, the improvement of the photothermal conversion efficiency, ROS generation, and tumor retention followed the formation of nanofibrils (NFs). This self-assembly strategy showed the ability of supramolecular nanofibrils to improve the bioavailability of photosensitizers, which provides a new potential treatment avenue for various cancer therapies.

Self‐Assembled Nanostructures of Quantum Dot/Conjugated Polymer Hybrids for Photonic Synaptic Transistors with Ultralow Energy Consumption and Zero‐Gate Bias

AbstractHerein, it is reported the influence of solution processing and treatments, such as adding marginal solvent, ultrasonication, and UV treatment, on the resulting perovskite (CsPbBr3) quantum dot (QD)/poly(3‐hexylthiophene) (P3HT) composite nanofibril films (CNFs) to improve the charge dissociation and photonic synaptic performance. A photonic synaptic transistor with CNFs can perform fundamental functions, including short‐term plasticity, long‐term plasticity, spike‐number‐dependent, and spike‐time‐dependent plasticity, to mimic sensing, computing, and memory functions. Notably, a synaptic device with CNFs presents an ultralow energy consumption of 0.18 fJ and zero‐gate operation. The superior performance of synaptic devices with CNFs can be attributed to two factors: (i) homogeneous axial distribution of the QDs and (ii) the formation of P3HT nanofibrils and co‐aggregates. Therefore, enhanced interfacial charge transfer between QDs and P3HT, ensuring decent carrier transport capability, is achieved. Collectively, the composite artificial synapse successfully provides an effective guide that offers a new perspective for the fabrication of one‐dimensional self‐assembled nanostructure‐based artificial synapses emulating human‐like memory, neuromorphic computing, and artificial intelligent systems.

Temperature and Heating Time of Forming Process of Nanofibrils of Whey Protein Isolate

Nanofibrils are nano-sized fibers. One of the materials that can be used to make nanofibrils is whey protein isolate. Nanofibrils can be formed by heating whey protein isolate at a certain temperature and for a certain duration. This study examines the effect of heating temperature and heating time on the formation of nanofibrils. WPI was formed into nanofibrils by heating the WPI solutions at 70 °C, 80 °C and 90 °C for 0, 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 hours. WPI nanofibrils in the solutions is observed by placing the solutions in between a cross-polariser. Based on the results of the study, it shows that heating at 70 °C was observed for 2 to 20 hours and did not form nanofibrils whey protein isolate. The nanofibrils were formed for heating at 80 °C for 4 hours, but the population was still small; along with the length of heating the population of nanofibrils is increasing. Heating at 90 °C for 2 hours the nanofibrils began to form; the population is increasing along with the length of heating. Based on the image, the nanofibrils heating at 80 °C is similar to 90 °C heating. Heating 80 °C is sufficient for the process of forming whey protein nanofibrils.

Effect mechanism of ultrasound pretreatment on fibrillation Kinetics, physicochemical properties and structure characteristics of soy protein isolate nanofibrils

Self-assembly of soy proteins into nanofibrils is gradually considered as an effective method to improve their technical and functional properties. Ultrasound is a non-thermal, non-toxic and environmentally friendly technology that can modulate the formation of protein nanofibrils through controlled structural modification. In this research, the effect of ultrasound pretreatment on soy protein isolate nanofibrils (SPIN) was evaluated by fibrillation kinetics, physicochemical properties and structure characteristics. The results showed that the optimum ultrasound condition (20% amplitude, 15 min, 5 s on-time and 5 s off-time) could increase the formation rate of SPIN by 38.66%. Ultrasound reduced the average particle size of SPIN from 191.90 ± 5.40 nm to 151.83 ± 3.27 nm. Ultrasound could increase the surface hydrophobicity to 1547.67 in the initial stage of nanofibrils formation, and extend the duration of surface hydrophobicity increased, indicating ultrasound could expose more binding sites, creating more beneficial conditions for nanofibrils formation. Ultrasound could change the secondary and tertiary structure of SPIN. The reduction of α-helix content of ultrasound-pretreated soy protein isolate nanofibrils (USPIN) was 12.1% (versus 5.3% for SPIN) and the increase of β-sheet content was 5.9% (versus 3.5% for SPIN) during fibrillation. Ultrasound could accelerate the formation of SPIN by promoting the unfolding of SPI, exposure of hydrophobic groups and formation of β-sheets. Microscopic images revealed that USPIN generated a curlier and looser shape. And ultrasound reduced the zeta potential, free sulfhydryl groups content and viscosity of SPIN. SDS-PAGE results showed that ultrasound could promote the conversion of SPI into low molecular weight peptides, providing building blocks for the nanofibrils formation. The results indicated that ultrasound pretreatment could be a promising technology to accelerate SPIN formation and promote its application in food industry, but further research is needed for the improvement of the functional properties of SPIN.

Nanocellulose-mediated transparent high strength conductive hydrogel based on in-situ formed polypyrrole nanofibrils as a multimodal sensor

A simple method was provided to prepare a transparent, highly conductive, mechanically reinforced, stretchable, and compressible hydrogel. In this system, pyrrole (Py) monomers were gently polymerized and uniformly deposited on the surface of cellulose nanofiber (CNF) via the improved in-situ polymerization. In the opaque PPy@CNF suspension, acrylamide monomers (AM) were dissolved and radical-polymerized to construct the PPy@CNF-PAM hydrogel with the in-situ formation of PPy nanofibrils in the presence of excess ammonium persulfate (APS). The in-situ formed PPy nanofibrils were well intertwined with the CNF and PAM chains, and a highly conductive path was established and permitted visible light to pass through. The amphipathic CNF took along and dispersed PPy aggregates well, and reinforced the hydrogel after formation of PPy nanofibrils. In view of the improved mechanical compressive, stretchable properties and excellent electrical conductivity (4.5 S/m), the resulting hydrogels could serve as a potential electrical device in a range of applications.

Water-in-Water Emulsions, Ultralow Interfacial Tension, and Biolubrication

Water-in-Water Emulsions, Ultralow Interfacial Tension, and Biolubrication

Hierarchical approach for the rational construction of helix-containing nanofibrils using α,β-peptides.

The rational design of novel self-assembled nanomaterials based on peptides remains a great challenge in modern chemistry. A hierarchical approach for the construction of nanofibrils based on α,β-peptide foldamers is proposed. The incorporation of a helix-promoting trans-(1S,2S)-2-aminocyclopentanecarboxylic acid residue in the outer positions of the model coiled-coil peptide led to its increased conformational stability, which was established consistently by the results of CD, NMR and FT-IR spectroscopy. The designed oligomerization state in the solution of the studied peptides was confirmed using analytical ultracentrifugation. Moreover, the cyclopentane side chain allowed additional interactions between coiled-coil-like structures to direct the self-assembly process towards the formation of well-defined nanofibrils, as observed using AFM and TEM techniques.

Characterization of cellulose nano/microfibril reinforced polypropylene composites processed via solid‐state shear pulverization

AbstractPolypropylene (PP) composites with cellulose nanofibril (CNF), lignocellulose nanofibril (LCNF) or bleached kraft wood pulp (BWKP) cellulose microfibril compatibilized by maleic anhydride grafted polypropylene were produced by different approaches. Samples containing 10–30 wt% filler were prepared by mixing cellulose aqueous suspensions and PP through solid‐state shear pulverization. The flakes obtained were compounded by twin‐screw melt extrusion (MSE) followed by injection molding (IM) into standard specimens. Reference samples with 30 wt% CNF or LCNF were prepared through a standard procedure, which involved freeze‐drying of nanofibrils aqueous suspensions before MSE and IM, whereas a sample with 30 wt% BWKP was prepared directly by MSE and IM. Injection‐molded specimens were characterized by scanning electron microscopy, differential scanning calorimetry, thermogravimetry, and tensile and notched impact tests. The thermal behavior of the PP matrix in the composites was not significantly affected by the filler type and content and processing route as well. CNF and LCNF reinforced PP composites showed inferior mechanical properties, due to the formation of nanofibrils agglomerates in the PP matrix, regardless of the processing route adopted. Highly filled (30 wt%) BWKP composites presented superior mechanical properties with significant increase in the modulus, tensile strength, and impact strength due to the formation of well‐dispersed microfibrils bundles in the PP matrix. A comparative analysis showed that the composites developed in this study present similar or even superior mechanical performance for given filler content to other natural fiber reinforced PP composites reported in the literature.

Formation, Structure, and Mechanical Performance of Silk Nanofibrils Produced by Heat-Induced Self-Assembly.

The heat-induced self-assembly of silk fibroin (SF) is studied by combing fluorescence assessment, infrared nanospectroscopy, wide-angle X-ray scattering, and Derjaguin-Muller-Toporov coupled with atomic force microscopy. Several fundamental issues regarding the formation, structure, and mechanical performance of silk nanofibrils (SNFs) under heat-induced self-assembly are discussed. Accordingly, SF in aqueous solution is rod-like in shape and not micellar. The formation of SNFs occurs through nucleation-dependent aggregation, but the assembly period is variable and irregular. SF shows inherent fractal growth, and this trend is critical for the short-term assembly. The long-term assembly of SF, however, mainly involves an elongation growth process. SNFs produced by different methods, such as ethanol treatment and heat incubation, have similar secondary structure and mechanical properties. These investigations improve the in-depth understanding of fundamental issues related to self-assembly of SNFs, and thus provide inspiration and guidance in designing of silk nanomaterials.

Shape-stabilized poly(ethylene glycol) phase change materials with self-assembled network scaffolds for thermal energy storage

We propose a facile method for preparing novel poly(ethylene glycol) (PEG) shape-stabilized phase change materials (PCMs) using self-assembled network scaffolds formed by gels for thermal energy storage. 1,3:2,4-dibenzylidene sorbitol (DBS) is capable of self-organizing into 3-D nanofibrillar networks in organic solvents to form gels at relatively low concentrations. In this study, we induced the formation of such nanofibrils in PEG. After PEG is heated to its melting temperature, the DBS gels (nanofibrillar structures) can capture most of the molten PEG, significantly stabilizing the shape and preventing leakage. The fabricated PCMs can sustain their shape up to at least until 96 °C, as determined using rheological measurement. The PEG with a molecular weight of 8000 g/mol containing DBS systems exhibited the best thermal properties, including latent heat, leakage percentage and thermal conductivity, which could be useful in developing novel phase change materials for thermal energy storage.

The Effect of Limited Proteolysis by Trypsin on the Formation of Soy Protein Isolate Nanofibrils

Nanofibril system constructed by protein self-assembly is widely used in the food industry because of purposive functional properties. Soy protein isolate nanofibrils (SPINs) were reported to form via heating at pH 2.0. In this research, the soy protein isolate (SPI) hydrolysate prepared by trypsin was used as a raw material for the formation of nanofibrils called soy protein isolate hydrolysate nanofibrils (SPIHNs). Microscopic images demonstrated the formation of two nanofibrils. Based on circular dichroism spectroscopy and Thioflavin T (ThT) fluorescence spectral, we concluded thatβ-sheet played an important role in SPIN and SPIHN’s structural composition. At the same time, theα-helix in SPI had not been destroyed, thereby favoring the formation of SPIHN. The surface hydrophobicity of SPIHN continued to increase during the heating process and reached the highest value when heating 8 h. SDS-PAGE analysis showed that peptides produced by enzyme-modified SPI affected the formation of SPIHN. These results proposed that enzymatic hydrolysis prior to acidic during fibrillation process affected the fibrillation of SPI, and the peptides formed by enzymatic hydrolysis were more efficient for the self-assembly process. This study will provide a theoretical basis for the future research of SPI nanofibril functionality.

Effect of heating temperature and time on the formation of 11S globulin nanofibril from Bogor nut (Vigna subterranean (L.) Verdc.) for food ingredients

Bogor beans (Bambara groundnuts) are thought to originate from Bambara, Timbuktu, Mali, West Africa. Introduced to Indonesia in the 20th century, and the name of the Bogor nut was given because it was widely sold in the city of Bogor. This research aims to study the formation of nanofibrils from 11S globulins to increase the benefits of Bogor nut as a food ingredient. Temperature treatment (65, 75, 85 °C) and heating time (6.12, 18, 24, 30 hours) at a concentration of 1% (w/w) protein solution, at pH 2, was applied to observe nanofibril formation. The formation of nanofibril was observed by measuring the viscosity of protein solutions before and after heating with a rheometer. A transmission electron microscope observed the morphology of nanofibrils formed at various heating times. The results showed that 11S globulin nanofibril formation was achieved at 85 °C heating temperatures. The morphology of 11S globulin nanofibrils from a 1% protein solution (w/w) with a heating temperature of 85 °C, is visible and significant in number after 24 hours of heating. The increase in the viscosity of the nanofibril solution from the protein solution indicates that the nanofibril from Bogor nut 11S globulin has potential as a food thickener.