Keratinous Materials Research Articles

Bioinspired microstructure design simultaneously enhances strain-rate stiffening and toughening of composites

Keratinous biological materials, such as baleen, pangolin scales, and human hair, have similar constituent materials. However, their strain-rate dependent mechanical properties vary due to differences in their microstructures. Inspired by the microstructures observed in baleen, here we present novel microstructural designs of three types of lamellar-tubular fibers, namely the lamellar-tubular fiber (LTF), mineralized lamellar-tubular fiber (MLTF) and hollow mineralized-lamellar-tubular fiber (HMLTF). We systematically investigated their strain-rate dependent mechanical behaviors with comparison to the conventional cylinder-fiber (CF) reinforced composites. Through the finite element analysis (FEA), we found that the baleen-inspired composites have superior strain-rate stiffening and toughening effects than the conventional fiber reinforced composite. Rate-dependent constitutive models decoupling elastic and inelastic regimes were constructed for these bioinspired composites. Based on the FEA results, three constitutive parameters were obtained to quantitatively characterize the rate-dependent mechanical behaviors of these composites, especially the microstructure-induced difference. Furthermore, our study found that the baleen-inspired tubular microstructure improves stiffness and strain-rate stiffening through raising the stress levels of all phases and improves toughness and strain-rate toughening through enlarging the deformation in the inelastic region. This novel bioinspired design is hopeful to pave ways for the development of advanced composites with simultaneously enhanced strain-rate stiffening and toughening.

Optimal utilization of waste wool: Preparation of bionic moisture-absorbing keratin membrane with hierarchical porous structure to harvest atmospheric moisture

In addressing the challenge of water scarcity, solar-assisted atmospheric water harvesting (SAWH) has emerged as a promising strategy. Here, we have designed a hierarchical regenerated wool keratin membrane (LiCl@CNMC) that mimics the natural Murray network. Carbon nano tube, serving as solar absorbers, are uniformly deposited on the hierarchical keratin membrane. Hygroscopic salt LiCl is impregnated into the keratin membrane to achieve exceptional water uptake capacity. Under simulated sunlight exposure, the surface temperature can reach 83.5 °C. Furthermore, the LiCl@CNMC membrane exhibits outstanding water absorption (0.79–2.67 g g−1) and desorption performance at 30–80 % RH. The bionic water-collecting membrane can carry out 12 adsorption–desorption cycles every day under outdoor conditions, and yielding 11.92 g g−1 day−1 of clean water. The design of regenerated keratin membrane not only provides a direct way for the regeneration and utilization of keratin materials, but also provides valuable guidance and inspiration for the preparation and application of natural polymers found.

Engineered keratin/bFGF hydrogel to promote diabetic wound healing in rats

Keratin materials are promising in wound healing acceleration, however, it is a challenge for the keratin to efficiently therapy the impaired wound healing, such as diabetic foot ulcers. Here, we report a keratin/bFGF hydrogel for skin repair of chronic wounds in diabetic rats based on their characteristics of extracellular matrix and growth factor degradation in diabetic ulcer. Recombinant keratin 31 (K31), the most abundant keratin in human hair, exhibited the highly efficient performances in cell adhesion, proliferation and migration. More importantly, the introduction of bFGF into K31 hydrogel significantly enhances the properties of cell proliferation, wound closure acceleration, angiogenesis and skin appendages regeneration. Furthermore, the combination of K31 and bFGF can promote epithelial-mesenchymal transition by inhibiting the expression of E-cadherin and promoting the expression of vimentin and fibronectin. These findings demonstrate the engineered K31/bFGF hydrogel as a promising therapeutic agent for diabetic wound healing.

Bioconversão de resíduos de aves para produção de proteases por Aspergillus sp. isolado do solo da floresta Amazônica

ABSTRACT: Feathers are by-products that are generated in significant quantities by the poultry industry. Microbial bioconversion has been investigated as a promising strategy for the processing of feathers, since, along with the degradation of these keratinous materials, bioprocessing can result in value-added products. Thus, from the perspective of industrial microbiology, chicken feathers can be considered a raw material for obtaining microbial proteases. Within this context, this research investigated and characterized the production of extracellular proteases by Aspergillus sp., isolated from soil of the Amazon Rainforest. The enzymatic production was evaluated using several growth substrates (whole feathers, feather meal, human hair, casein, gelatin, peptone and chicken beaks). With highest enzyme production was obtained the feather meal (FM) and peptone. After 48 h of fermentation, FM degradation was 15.82%. The crude protease showed optimal activity at pH 5.0 and 37 °C and enzymatic activity was enhanced with the addition of 1 and 5 mM of CaCl2, MnSO4, KCl, MgSO4 and CuSO4. The detergents Tween 20 and Triton x-100, at concentrations 0.5 and 1% (v/v), tended to stimulate activity. The presence of 0.5 and 1% (v/v) of organic solvents (methanol, acetone, butanol, acetonitrile, isopropanol and DMSO), maintained the enzymatic activity. β-mercaptoethanolstimulated proteolytic activity in the enzymatic assays. This study suggested new direction for waste management with industrial applications giving rise to green technology for sustainable development.

Structure prediction, docking studies and molecular cloning of novel Pichia kudriavzevii YK46 metalloprotease (MetPr) for improvement of feather waste biodegradation

This study addresses the environmental risks associated with the accumulation of keratin waste from poultry, which is resistant to conventional protein degradation methods. To tackle this issue, microbial keratinases have emerged as promising tools for transforming resilient keratin materials into valuable products. We focus on the Metalloprotease (MetPr) gene isolated from novel Pichia kudriavzevii YK46, sequenced, and deposited in the NCBI GenBank database with the accession number OQ511281. The MetPr gene encodes a protein consisting of 557 amino acids and demonstrates a keratinase activity of 164.04 U/ml. The 3D structure of the protein was validated using Ramachandran's plot, revealing that 93% and 97.26% of the 557 residues were situated within the most favoured region for the MetPr proteins of template Pichia kudriavzevii strain 129 and Pichia kudriavzevii YK46, respectively. Computational analyses were employed to determine the binding affinities between the deduced protein and beta keratin. Molecular docking studies elucidated the optimal binding affinities between the metalloprotease (MetPr) and beta-keratin, yielding values of − 260.75 kcal/mol and − 257.02 kcal/mol for the template strains Pichia kudriavzevii strain 129 and Pichia kudriavzevii YK46, respectively. Subsequent molecular cloning and expression of the MetPr gene in E. coli DH5α led to a significantly higher keratinase activity of 281 ± 12.34 U/ml. These findings provide valuable insights into the potential of the MetPr gene and its encoded protein for keratin waste biotransformation, with implications for addressing environmental concerns related to keratinous waste accumulation.

Effective biodegradation on chicken feather by the recombinant KerJY-23 Bacillus subtilis WB600: A synergistic process coupled by disulfide reductase and keratinase

Keratin wastes are abundantly available but rich in hard-degrading fibrous proteins, and the keratinase-producing microorganisms have gained significant attention due to their biodegradation ability against keratinous materials. In order to improve the degradation efficiency of feather keratins, the keratinase gene (kerJY-23) from our previously isolated feather-degrading Ectobacillus sp. JY-23 was overexpressed in Bacillus subtilis WB600 strain. The recombinant KerJY-23 strain degraded chicken feathers rapidly within 48 h, during which the activities of disulfide reductase and keratinase KerJY-23 were sharply increased, and the free amino acids especially the essential phenylalanine and tyrosine were significantly accumulated in feather hydrolysate. The results of structural characterizations including scanning electron microscopy, Fourier transform infrared spectrum, X-ray diffraction, and X-ray photoelectron spectroscopy, demonstrated that the feather microstructure together with the polypeptide bonds and SS bonds in feather keratins were attacked and destroyed by the recombinant KerJY-23 strain. Therefore, the recombinant KerJY-23 strain contributed to feather degradation through the synergistic action of the secreted disulfide reductase to break the SS bonds and keratinase (KerJY-23) to hydrolyze the polypeptide bonds in keratins. This study offers a new insight into the underlying mechanism of keratin degradation, and provides a potential recombinant strain for the valorization of keratin wastes.

PH-equilibration of human hair: Kinetics and pH-dependence of the partition ratios for H+ − and OH− -ions based on a Freundlich isotherm

Hair is an insoluble, fibrous, α-keratinous, protein composite material, providing outer coverage, e.g., for mammals. In the context of a wider study on the effects of pH on human hair properties, we investigated the time-dependence of pH-equilibration study across the acid and the basic pH-range, using appropriate pure solutions of hydrochloric acid and sodium hydroxide. The results show that pH-equilibration follows essentially equal 1st-order kinetics across the pH-range. The characteristic process time does not change significantly and is in the range of 2.5–5 h. The analysis enables to determine the equilibrium uptakes of H+- and OH− -ions. These follow the expected U-shaped path across the pH-range. For both acidic and alkaline conditions, data are well described by two very similar sorption isotherms of the Freundlich-type. In consequence, partition ratios for both ions are highest near neutrality (pH 7: >6000) and drop off strongly towards low and high pHs (<50). Hair is thus a very strong ‘sink’ for H+ and OH−. This observation fundamentally challenges traditional views of limited ion uptake, namely, in the mid-pH-range due to hindered diffusion. It also does not support considerations on special roles of certain pHs, specific groups of amino acids, or morphological components. Our analysis thus suggests that established views of the interaction of hair and pH need to be reconsidered, The Freundlich isotherm approach appears to provide a versatile tool to refine our understanding of the interactions of hair and possibly other keratinous materials (horn, nail, feathers) with acids and bases.

The weathering and scavenging of keratin

Keratin is scarcely found in the archaeological record and is typically only preserved if conditions are conducive to the preservation of other organic materials. However, the rate of decomposition of keratinous materials, such as horn sheaths and hooves, is unknown. This study measures the rates of keratin decay and scavenging by animals of bison hooves over a two-year period in two environments: one at low elevation with warm temperatures and high humidity levels and the other at high elevation with generally cooler temperatures and low humidity levels. We find that keratin decays at a faster rate in humid environments, and a keratinous ecofact such as a hoof should be expected to decay, if exposed to the elements, within 5 years in warm, humid, low elevation environments. In cool, low humidity high elevation environments, we find that keratinous ecofacts should be expected to decay within 20 years. These decomposition rates allow us to put constraints on conditions of keratin preservation in the archaeological record.

Characterization of the keratin/polyamide 6 composite fiber's structure and performance prepared by the optimized spinning process based on the rheological analysis

The complex chemical structure of polypeptide and the imperfection of processing technology cause the mechanical properties of regenerated keratin to be hard and brittle. This defect seriously affects the application prospects of keratin materials. To solve the above problems, α-lipoic acid modified keratin (KER) was blended with Polyamide 6 (PA6) and prepared into composite fibers via the wet-spinning method in this work. The spinnability and spinning conditions of the KER/PA6 blend solution were analyzed by rheological theory. The results illustrated that keratin solution will easily form a gel state under certain temperatures and concentrations, which was not conducive to the preparation of regenerated fiber. When the temperature was 45 °C and the mass fraction was 10 %, the viscosity and rheology of the solution were appropriate. The rheological properties of the blend solution showed that too much keratin would make the solution easy to gel, which was not conducive to the preparation of regenerated fibers and may affect the fiber properties. On this basis, the prepared composite fibers were characterized to explore the macromolecular aggregation state of keratin and PA6 in fibers. FT-IR and XRD results proved that there was no chemical reaction between keratin and PA6 in the composite fibers, which belonged to physical blending. At the same time, the two polymers had good compatibility and can be blended at the molecular level. SEM, DSC, and tensile strength test results indicated that when the proportion of keratin was too high, the structure and properties of the composite fibers will have obvious defects, which was consistent with the rheological analysis. Therefore, the blend ratio of keratin/PA6 was determined to be 3:7. Under this condition, the fibers exhibited a homogeneous structure and good thermal properties, especially its mechanical properties were close to wool fibers. The KER/PA6 composite fibers show important research value and can also provide technical reference for the development of regenerated biomass materials.

Keratin as a sustainable biopolymer for waste water treatment

Keratin is one of the most abundant natural polymers with potential application in various fields but is usually seen discarded as waste generated from poultry farms along roadsides and landfills. These are indeed the cheapest source of keratin protein which could be used for various applications. Owing to the structural properties, keratinous materials are now being exploited in wastewater treatment systems as adsorbents. The rich amino acid content having hydroxyl, carboxyl and amino groups has been found to be beneficial in removing contaminants from waste waters like heavy metals and dyes. Research based on this idea has received peak attention to a point where formulations of different adsorbent materials like nanofibre, biofilms and biocomposite from keratinous raw materials are now available for commercial use. This review summarises the application of keratin as an efficient adsorbent for waste water treatment providing an insight into its structure, forms of keratin used for treatments and mechanism of adsorption of different components in waste water.

Equine hoof wall: Structure, properties, and bioinspired designs

The horse hoof wall exhibits exceptional impact resistance and fracture control due to its unique hierarchical structure which contains tubular, lamellar, and gradient configurations. In this study, structural characterization of the hoof wall was performed revealing features previously unknown. Prominent among them are tubule bridges, which are imaged and quantified. The hydration-dependent viscoelasticity of the hoof wall is described by a simplified Maxwell-Weichert model with two characteristic relaxation times corresponding to nanoscale and mesoscale features. Creep and relaxation tests reveal that the specific hydration gradient in the hoof keratin likely leads to reduced internal stresses that arise from spatial stiffness variations. To better understand realistic impact modes for the hoof wall in-vivo, drop tower tests were executed on hoof wall samples. Fractography revealed that the hoof wall's reinforced tubular structure dominates at lower impact energies, while the intertubular lamellae are dominant at higher impact energies. Broken fibers were observed on the surface of the tubules after failure, suggesting that the physically intertwined nature of the tubule reinforcement and intertubular matrix improves the toughness of this natural fiber reinforced composite. The augmented understanding of the structure-mechanical property relationship in dynamic loading led to the design of additively manufactured bioinspired structures, which were evaluated in quasistatic and dynamic loadings. The inclusion of gradient structures and lamellae significantly reduced the damage sustained in drop tower tests, while tubules increased the energy absorption of samples tested in compact tension. The samples most similar to the hoof wall displayed remarkably consistent fracture control properties. Statement of significanceThe horse hoof wall, capable of withstanding large, repeated, dynamic loads, has been touted as a candidate for impact-resistant bioinspiration. However, our understanding of this biological material and its translation into engineered designs is incomplete. In this work, new features of the horse hoof wall are quantified and the hierarchical failure mechanisms of this remarkable material under near-natural loading conditions are uncovered. A model of the hoof wall's viscoelastic response, based on studies of other keratinous materials, was developed. The role of hydration, strain rate, and impact energy on the material's response were elucidated. Finally, multi-material 3D printed designs based on the hoof's meso/microstructure were fabricated and exhibited advantageous energy absorption and fracture control relative to control samples.

Renal Epidermoid Cyst: A Case Report

In contrast to epidermoid cysts of the dermis, epidermoid cysts of the kidneys are exceptional. This report explains an uncommon patient with renal epidermoid cyst who presented with flank pain and previous history of renal stones. Our patient underwent excisional resection. The histopathological evaluation discovered cyst wall covering by squamous epithelium and containing keratinous materials compatible with the epidermoid cyst. Based on pathology findings, the patient didn’t receive any more treatment. Because pathology evaluation is necessary for the diagnosis or rules out the possibility of malignancy, biopsy before surgery is advised.

An environment-friendly chemical modification method for thiol groups on polypeptide macromolecules to improve the performance of regenerated keratin materials

The R&D and application of natural biomass resources is one of the important ways to solve the world's energy shortage and environmental pollution. However, the complex processing technology and poor mechanical properties seriously hinder the development of the biomass polymers. A kind of natural antioxidant, alpha-lipoic acid, was used to modify wool keratin by grafting thiols in this study, so as to ameliorated the physical properties of regenerated materials. The optimal process parameters and modification mechanism were explored. Research results indicated that grafting ratio of lipoic acid can reach more than 13% and the polypeptide chains were more stretched in solution, when 30% lipoic acid was added into keratin solution at 40 °C with a reaction time of 2 h. In addition, lipoic acid grafted on thiol groups can effectively weaken the intra- and intermolecular forces of keratin, this change made the arrangement of macromolecular chains more orderly. The physical structure of modified keratin became uniform and compact, and the materials had obtained better thermal and mechanical properties. This research provided a novel and environment-friendly method for the modification technology of keratin, and it was beneficial to expand the application fields of biomass materials.

POTENTIAL TEST OF GREEN BETEL (PIPER BETLE L.) LEAF EXTRACT CREAM IN COMBINATION WITH BASIL LEAF EXTRACT (OCIMUMSANCTUM L.) AS ANTI-ACNE

Acne problem is the most common problem that many people fight for. Even though is not a dangerous thing, but that problem will make the sufferer feel unconfident. Acne vulgaris is an inflammatory disorder with blockage and accumulation of the Keratin materials that caused by Propionibacterium acnes bacteria. Now on, Herbal plant is the most selected one as an acne treatment because of the lack effect for the sufferer than a medical treatment. The plants are Basil (Ocimum sanctum L.) and green Lemongrass (Piper Betle L.). Both of them have a Saponin and Flavonoid as an antimicroba. Therefore, in this research will combine that two of the plants which is then formulated in to the cream form. The cream formulation made with 3 different concentration variances were 15%, 30% dan 45%. The type of the research is an in vitro of experimental research method. Where the cream formula making process done by manually and to measure that inhibition to Propionibacterium acnes bacteria. That in the concentration of 15% has an inhibition 20.02 mm, the concentration of 30% was 21.24 mm and the concentration of 45% was 25.05 mm. Inhibition zone &gt; 20 mm categorized as very strong inhibition. So that, could be conclude the cream Basil Leaf Extract (Ocimum sanctum L.) in combination with Green Betle Leaf Extract (Piper Betle L.) had a potential and most effective concentration as an anti-acne was the third formulation (45% concentration) with an inhibition was 25.5 mm and categorized as a very strong inhibition to Propionibacterium acnes.

Molecular interaction and partitioning in α-keratin using 1H NMR spin-lattice (T1) relaxation times.

The interactions between small molecules and keratins are poorly understood. In this paper, a nuclear magnetic resonance method is presented to measure changes in the 1H T1 relaxation times of small molecules in human hair keratin to quantify their interaction with the fibre. Two populations of small-molecule compounds were identified with distinct relaxation times, demonstrating the partitioning of the compounds into different keratin environments. The changes in relaxation time for solvent in hair compared with bulk solvent were shown to be related to the molecular weight (MW) and the partition coefficient, LogP, of the solvent investigated. Compounds with low MWs and high hydrophilicities had greater reductions in their T1 relaxation times and therefore experienced increased interactions with the hair fibre. The relative population sizes were also calculated. This is a significant step towards modelling the behaviour of small molecules in keratinous materials and other large insoluble fibrous proteins.

Potential Applications of Textile Wastes and By-products in Preparation of Textile Auxiliaries

Ever since the Stone Age, humans have preferred natural products and materials for better health and comfortability. Wool and natural silk have been used in the textile clothing thousands of years ago. The performance and appearance attributes of wool and silk make them superior for many applications. On the other hand, the textile industries are technically, economically, socially, and environmentally related to some of the sustainable development goals (SDGs) of the United Nations Agenda 2030. For instance, the item number 12 in the SDGs aims to ensure sustainable consumption and production pattern. This necessitates proper utilization of all natural products as well as recycling and utilization of any waste thereof. This article review deals with retrieval of wastes or by-products during wet processing of proteinic fibres; namely wool and silk. Different ways of proper utilization of the retrieved materials will be proposed. Specifically, we will focus mainly on three main by-products; namely keratinous materials, wool wax, and sericin. State of the art and future prospects for the utilization of these materials in the production of textile auxiliaries; Viz. softeners, binders, and flame-retardants, were discussed.

Comparative Genomics Analysis of Keratin-Degrading Chryseobacterium Species Reveals Their Keratinolytic Potential for Secondary Metabolite Production.

A promising keratin-degrading strain from the genus Chryseobacterium (Chryseobacterium sp. KMC2) was investigated using comparative genomic tools against three publicly available reference genomes to reveal the keratinolytic potential for biosynthesis of valuable secondary metabolites. Genomic features and metabolic potential of four species were compared, showing genomic differences but similar functional categories. Eleven different secondary metabolite gene clusters of interest were mined from the four genomes successfully, including five common ones shared across all genomes. Among the common metabolites, we identified gene clusters involved in biosynthesis of flexirubin-type pigment, microviridin, and siderophore, showing remarkable conservation across the four genomes. Unique secondary metabolite gene clusters were also discovered, for example, ladderane from Chryseobacterium sp. KMC2. Additionally, this study provides a more comprehensive understanding of the potential metabolic pathways of keratin utilization in Chryseobacterium sp. KMC2, with the involvement of amino acid metabolism, TCA cycle, glycolysis/gluconeogenesis, propanoate metabolism, and sulfate reduction. This work uncovers the biosynthesis of secondary metabolite gene clusters from four keratinolytic Chryseobacterium species and shades lights on the keratinolytic potential of Chryseobacterium sp. KMC2 from a genome-mining perspective, can provide alternatives to valorize keratinous materials into high-value bioactive natural products.

Industrial sustainability of microbial keratinases: production and potential applications.

Keratinases are proteolytic enzymes with a particular ability to cleave peptide bonds in keratin, and in other proteins. Due to their broad-spectrum of activity, keratinases are considered viable substitutes for chemical and thermal treatments of protein-rich industrial by-products. Among these protein residues, special attention has been given to keratinous materials (feathers, hair, horns, etc.), which disposal through harsh conditions methods, such as acid/alkaline hydrolysis or incineration, is not considered ecologically safe. Microbial keratinolytic enzymes allow for keratin degradation under mild conditions, resulting in keratin hydrolysates containing undamaged amino acids and peptides. In this review article, we offer perspectives on the relevance of these unique biocatalysts and their revolutionary ascent in industries that generate keratin-rich wastes. Additionally, we share insights for applications of keratinases and protein hydrolysates in agriculture, animal feed, cosmetics, phamaceuticals, detergent additives, leather processing, and others. Due to the scientific importance of keratinases and their potential use in green technologies, searching for bacterial and fungal species that efficiently produce these enzymes may contribute to the sustainability of industries.

Keratin Biomaterials Improve Functional Recovery in a Rat Spinal Cord Injury Model.

Laboratory study using a rat T9 contusion model of spinal cord injury (SCI). The purpose of this study was to evaluate which method of delivery of soluble keratin biomaterials would best support functional restoration through the macrophage polarization paradigm. SCI is a devastating neurologic event with complex pathophysiological mechanisms that currently has no cure. After injury, macrophages and resident microglia are key regulators of inflammation and tissue repair exhibiting phenotypic and functional plasticity. Keratin biomaterials have been demonstrated to influence macrophage polarization and promote the M2 anti-inflammatory phenotype that attenuates inflammatory responses. Anesthetized female Lewis rats were subjected to moderate T9 contusion SCI and randomly divided into: no therapy (control group), an intrathecally injected keratin group, and a keratin-soaked sponge group (n = 11 in all groups). Functional recovery assessments were obtained at 3- and 6-weeks post-injury (WPI) using gait analysis performed with the DigiGait Imaging System treadmill and at 1, 3, 7, 14, 21, 28, 35, and 42 days post-injury by the Basso, Beattie, Bresnahan (BBB) locomotor rating scale. Histology and immunohistochemistry of serial spinal cord sections were performed to assess injury severity and treatment efficacy. Compared to control rats, applying keratin materials after injury improved functional recovery in certain gait parameters and overall trended toward significance in BBB scores; however, no significant differences were observed with tissue analysis between groups at 6 WPI. Results suggest that keratin biomaterials support some locomotor functional recovery and may alter the acute inflammatory response by inducing macrophage polarization following SCI. This therapy warrants further investigation into treatment of SCI.Level of Evidence: N/A.

Structure of Keratin.

Keratins, as a group of insoluble and filament-forming proteins, mainly exist in certain epithelial cells of vertebrates. Keratinous materials are made up of cells filled with keratins, while they are the toughest biological materials such as the human hair, wool and horns of mammals and feathers, claws, and beaks of birds and reptiles which usually used for protection, defense, hunting and as armor. They generally exhibit a sophisticated hierarchical structure ranging from nanoscale to centimeter-scale: polypeptide chain structures, intermediated filaments/matrix structures, and lamellar structures. Therefore, more and more attention has been paid to the investigation of the relationship between structure and properties of keratins, and a series of biomimetic materials based on keratin came into being. In this chapter, we mainly introduce the hierarchical structure, the secondary structure, and the molecular structure of keratins, including α- and β-keratin, to promote the development of novel keratin-based biomimetic materials designs.