Silk Industry Waste Protein-Derived Sericin Hybrid Nanoflowers for Antibiotics Remediation via Circular Economy.

Hybrid protein-copper nanoflowers have emerged as promising materials with diverse applications in biocatalysis, biosensing, and bioremediation. Sericin, a waste biopolymer from the textile industry, has shown potential for fabricating such nanoflowers. However, the influence of the molecular weight of sericin on nanoflower morphology and peroxidase-like activity remains unexplored. This work focused on the self-assembly of nanoflowers using high- and low-molecular-weight (HMW and LMW) silk sericin combined with copper(II) as an inorganic moiety. The peroxidase-like activity of the resulting nanoflowers was evaluated using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and hydrogen peroxide (H2O2). The findings revealed that high-molecular-weight sericin hybrid nanoflowers (HMW-ShNFs) exhibited significantly higher peroxidase-like activity than low-molecular-weight sericin hybrid nanoflowers (LMW-ShNFs). Furthermore, HMW-ShNFs demonstrated superior reusability and storage stability, thereby enhancing their potential for practical use. This study also explored the application of HMW-ShNF for ciprofloxacin degradation to address the environmental and health hazards posed by this antibiotic in water. The results indicated that HMW-ShNFs facilitated the degradation of ciprofloxacin, achieving a maximum degradation of 33.2 ± 1% at pH 8 and 35 °C after 72 h. Overall, the enhanced peroxidase-like activity and successful application in ciprofloxacin degradation underscore the potential of HMW-ShNFs for a sustainable and ecofriendly remediation process. These findings open avenues for the further exploration and utilization of hybrid nanoflowers in various environmental applications.

Unveiling the potential of cellulose nanofibre based nitrogen fertilizer and its transformative effect on Vigna radiata (Mung Bean): nanofibre for sustainable agriculture.

Introduction: Fertilizer management is crucial to maintaining a balance between environmental health, plant health, and total crop yield. Farmers are overutilizing fertilizers with a mind set to enhance the productive capacity of the field, which adversely impacts soil fertility and causes serious environmental hazards. To mitigate the issues of over-utilization of fertilizers, controlled-release fertilizers were developed using nitrogen fertilizer (ammonium chloride) loaded on cellulose nanofibres (named CNF*N). Methodology: In this study, the effects of CNF*N were compared with commercial nitrogen fertilizer (ammonium chloride) on Vigna radiata (Mung) under greenhouse conditions. The pot experiment was conducted using six treatments: first treatment was control, where the plant was cultivated (T1); second treatment was T2, where the plant was cultivated with CNF to determine the impact of CNF on the plant; third was T3 where commercial ammonium chloride (24 mg/ 2 kg soil) was added to the plant; fourth was T4, where the plant was loaded with CNF, viz. CNF*N contains 4.8 mg of nitrogen; fifth was T5 CNF*N pellet contains 12 mg of nitrogen, and the last sixth treatment (T6) where CNF*N pellet containing 24 mg of nitrogen. Results: It indicated that the growth parameters were best achieved in T6 treatment. Plant height was at its maximum in the T6 treatment (44.4 ±0.1cm) after the second harvest, whereas the minimum plant height was observed in T1, which was 39.1 ±0.1 cm. Root-to-shoot weight ratio was also maximum in T6 (0.183± 0.02) and minimum in T1 (0.07± 0.01) after second harvesting. The significant difference among the treatments was determined with Tukey's honestly significant difference (HSD). The nitrogen content (available and total) was significantly higher in the T4, T5, and T6 treatments (0.22, 0.25, and 0.28%) as compared to the control treatments (T1 (0.12%), T2 (0.13%), and T3 (0.14%) during the second harvesting stage (90 days), as nitrogen plays a crucial role in the development of vegetative growth in Vigna radiata. The rate of controlled-release nitrogen-fertilizer was found to be optimal in terms of plant growth and soil nutrients; hence, it could potentially play a crucial role in improving soil health and the yield of the crop.

Biomedical Applications of Electro-Initiated Polymerisation on Ti6Al4†V Titanium Alloy using Silk Fibroin Coatings for Antibiotic Delivery and Improved Cell Metabolism.

Silk fibroin interactions with metallic surfaces can provide utility for medical materials and devices. Toward this goal, titanium alloy (Ti6Al4 V) was covalently grafted with polyacrylamide via electrochemically reducing 4-nitrobenzene diazonium salt in the presence of acrylamide. Analysis of the modified surfaces with FT-IR spectra, SEM and AFM were consistent with surface grafting. Functionalised titanium samples with a silk fibroin membrane, with and without impregnated therapeutics, were used to assess cytocompatibility and drug delivery. Initial cytocompatibility experiments using fibroblasts showed that the functionalised samples, both with and without silk fibroin coatings, supported significant increases between 72-136 % in cell metabolism, compared to the controls after 7 days. A 7-days release profiling showed consistent bacterial inhibition through gentamicin release with average inhibition zones of 239 mm2 . Over a 5-week period, silk fibroin coated samples, both with and without growth factors, supported better human mesenchymal stem cell metabolism with increases reaching 1031 % and 388 %, respectively, compared to samples without the silk fibroin coating with.

Rice straw-derived cellulose: a comparative study of various pre-treatment technologies and its conversion to nanofibres.

Rice straw is a waste product generated after the harvesting of rice crops and is commonly disposed of by burning it off in open fields. This study explored the potential for the extraction and conversion of cellulose to cellulose nanofibres (CNFs) to be used as smart delivery systems for fertilizers applications. In this study, alkali, steam explosion, and organosolv treatments were investigated for cellulose extraction efficiency. The morphological characterization of cellulose showed smooth fibrillar structures. Fourier transform infrared spectroscopy represented significant removal of non-cellulosic components in treatments. The crystallinity increased from 52.2 to 65% in CNFs after fibrillation. Cellulose nanofibres (CNFs) had an average diameter of 37.4 nm and - 25.2 mV surface charges as determined by SEM and zeta potential, respectively, which have desired properties for holding fertilizers. Therefore, this study paves the way for value-added uses of rice straw as alternatives to current environmentally harmful practices.

Controlled release fertilizer delivery system derived from rice straw cellulose nanofibres: a circular economy based solution for sustainable development.

Recently, the development of sustainable and environmentally friendly biomaterials has gained the attention of researchers as potential alternatives to petroleum-based materials. Biomaterials are a promising candidate to mitigate sustainability issues due to their renewability, biodegradability, and cost-effectiveness. Thus, the purpose of this study is to explore a cost-effective biomaterial-based delivery system for delivering fertilizers to plants. To achieve this, rice straw (agro-waste) was selected as a raw material for the extraction of cellulose. The cellulose was extracted through alkali treatment (12% NaOH), followed by TEMPO-based oxidation. The cellulose nanofibers were characterized using Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy, and transmission electron microscopy. In scanning electron microscopy, a loosening of the fibrillar structure in cellulose nanofibers (CNFs) was observed with a diameter of 17 ± 4 nm. The CNFs were loaded with nitrogen-based fertilizer (ammonium chloride) in 1:1, 1:2, and 2:1 (w/w) proportions. The loading was estimated through surface charge variation; in the case of the 1:1 sample, maximum reductions in surface charge were seen from -42.0 mV to -12.8 mV due to the binding of positive ammonium ions. In the release kinetics study, a controlled release pattern was observed at 1:1, which showed a 58% cumulative release of ammonium ions within 8 days. Thus, the study paves the way for value-added uses of rice straw as an alternative to the current environmentally harmful practices.

Bio-based cellulose nanofibers (CNFs) from rice straw via circular economy approach.Controlled release fertilizers for sustainable agriculture.Nanotechnology for precision agriculture and decarbonization via agricultural waste management.

Methods for Silk Property Analyses across Structural Hierarchies and Scales.

Silk from silkworms and spiders is an exceptionally important natural material, inspiring a range of new products and applications due to its high strength, elasticity, and toughness at low density, as well as its unique conductive and optical properties. Transgenic and recombinant technologies offer great promise for the scaled-up production of new silkworm- and spider-silk-inspired fibres. However, despite considerable effort, producing an artificial silk that recaptures the physico-chemical properties of naturally spun silk has thus far proven elusive. The mechanical, biochemical, and other properties of pre-and post-development fibres accordingly should be determined across scales and structural hierarchies whenever feasible. We have herein reviewed and made recommendations on some of those practices for measuring the bulk fibre properties; skin-core structures; and the primary, secondary, and tertiary structures of silk proteins and the properties of dopes and their proteins. We thereupon examine emerging methodologies and make assessments on how they might be utilized to realize the goal of developing high quality bio-inspired fibres.

Novel hybrid biocomposites for tendon grafts: The addition of silk to polydioxanone and poly(lactide-co-caprolactone) enhances material properties, in vitro and in vivo biocompatibility.

Biopolymers play a critical role as scaffolds used in tendon and ligament (TL) regeneration. Although advanced biopolymer materials have been proposed with optimised mechanical properties, biocompatibility, degradation, and processability, it is still challenging to find the right balance between these properties. Here, we aim to develop novel hybrid biocomposites based on poly(p-dioxanone) (PDO), poly(lactide-co-caprolactone) (LCL) and silk to produce high-performance grafts suitable for TL tissue repair. Biocomposites containing 1-15% of silk were studied through a range of characterisation techniques. We then explored biocompatibility through in vitro and in vivo studies using a mouse model. We found that adding up to 5% silk increases the tensile properties, degradation rate and miscibility between PDO and LCL phases without agglomeration of silk inside the composites. Furthermore, addition of silk increases surface roughness and hydrophilicity. In vitro experiments show that the silk improved attachment of tendon-derived stem cells and proliferation over 72 h, while in vivo studies indicate that the silk can reduce the expression of pro-inflammatory cytokines after six weeks of implantation. Finally, we selected a promising biocomposite and created a prototype TL graft based on extruded fibres. We found that the tensile properties of both individual fibres and braided grafts could be suitable for anterior cruciate ligament (ACL) repair applications.

Promoting Silk Fibroin Adhesion to Stainless Steel Surfaces by Interface Tailoring.

Bonding dissimilar materials has been a persistent challenge for decades. This paper presents a method to modify a stainless steel surface (316 L), routinely used in medical applications to enable the significant adhesion of a biopolymer (silk fibroin). The metallic surface was first covalently grafting with polyacrylamide, to enable a hydrogen bonding compatible surface. The polymerisation was initiated via the irreversible electrochemical reduction of a 4-nitrobenzene diazonium salt (20 mM), in the presence of an acrylamide monomer (1 M) at progressively faster scan rates (0.01 V/s to 1 V/s). Examination of the modified samples by FT-IR was consistent with successful surface modification, via observations of the acrylamide carbonyl (1600-1650 cm-1 ) was observed, with more intense peaks correlating to slower scan rates. Similar observations were made with respect to increasing surface polarity, assessed by water contact angle. Reductions of >60° were observed for the grafted surfaces, relative to the unmodified control materials, indicating a surface able to undergo significant hydrogen bonding. The adhesion of silk to the metallic surface was quantified using a lap shear test, effectively using silk fibroin as an adhesive. Adhesion improvements of 5-7-fold, from 4.1 MPa to 29.3 MPa per gram of silk fibroin, were observed for the treated samples, highlighting the beneficial effect of this surface treatment. The methods developed in this work can be transferred to any metallic (or conductive) surface and can be tailored to complement any desired interface.

Tuning the microstructure and mechanical properties of lyophilized silk scaffolds by pre-freezing treatment of silk hydrogel and silk solution.

This work aims to understand how pre-freezing treatments (-20 °C, -80 °C or -196 °C (liquid nitrogen)) affect the microstructure, mechanical properties and secondary structure of silk scaffolds prepared from lyophilization of silk hydrogels and silk solutions. It is found that in comparison with silk solutions, silk hydrogels at the same silk fibroin concentrations produce scaffolds with more nanofibrous structures when they are pre-frozen at the different temperatures. Although pre-freezing with liquid nitrogen can produce nanofibrous scaffolds from either a silk solution (low concentration of 2%) or silk hydrogel (produced from 2 to 6% silk fibroin solutions), aligned macro-channels can be produced only from silk hydrogels. In addition, scaffolds obtained from silk hydrogels are dominated by ß-sheets due to the crystallization process for gel network formation, while scaffolds prepared from silk solutions are largely amorphous. The findings of this work are important to tune the microstructure and mechanical properties of silk scaffolds.

Toughening Wet-Spun Silk Fibers by Silk Nanofiber Templating.

Regenerated silk fibers typically fall short of silkworm cocoon fibers in mechanical properties due to reduced fiber crystal structure and alignment. One approach to address this has been to employ inorganic materials as reinforcing agents. The present study avoids the need for synthetic additives, demonstrating the first use of exfoliated silk nanofibers to control silk solution crystallization, resulting in all-silk pseudocomposite fibers with remarkable mechanical properties. Incorporating only 0.06 wt% silk nanofibers led to a ≈44% increase in tensile strength (over 600 MPa) and ≈33% increase in toughness (over 200 kJ kg-1 ) compared with fibers without silk nanofibers. These remarkable properties can be attributed to nanofiber crystal seeding in conjunction with fiber draw. The crystallinity nearly doubled from ≈17% for fiber spun from pure silk solution to ≈30% for the silk nanofiber reinforced sample. The latter fiber also shows a high degree of crystal orientation with a Herman's orientation factor of 0.93, a value which approaches that of natural degummed B. mori silk cocoon fiber (0.96). This study provides a strong foundation to guide the development of simple, eco-friendly methods to spin regenerated silk with excellent properties and a hierarchical structure that mimics natural silk.

Mechanical, structural and biodegradation characteristics of fibrillated silk fibres and papers.

We characterised fibres and papers of microfibrillated silk from Bombyx mori produced by mechanical and enzymatic process. Milling increased the specific surface area of fibres from 1.5 to 8.5 m2/g and that enzymatic pre-treatment increased it further to 16.5 m2/g. These fibrils produced a uniform, significantly strong (tenacity 55 Nm/g) and stiff (Young's modulus > 2 GPa) papers. Enzymatic pre-treatment did not reduce molecular weight and tensile strength of papers but significantly improved fibrillation. Silk remained highly crystalline throughout the fibrillation process. Protease biodegradation was more rapid after fibrillation. Biodegradation was impacted by structural change due to enzymatic pre-treatment during the fibrillation. Biodegraded silk had much higher thermal degradation temperature. The unique combination of high strength, slow yet predicable degradation and controllable wicking properties make the materials ideally suited to biomedical and healthcare applications.

Spinning Regenerated Silk Fibers with Improved Toughness by Plasticizing with Low Molecular Weight Silk.

Low-molecular weight (LMW) silk was utilized as a LMW silk plasticizer for regenerated silk, generating weak physical crosslinks between high-molecular weight (HMW) silk chains in the amorphous regions of a mixed solution of HMW/LMW silk. The plasticization effect of LMW silk was investigated using mechanical testing, Raman spectroscopy, and wide-angle X-ray scattering (WAXS). Small amounts (10%) of LMW silk resulted in a 19.4% enhancement in fiber extensibility and 37.8% increase in toughness. The addition of the LMW silk facilitated the movement of HMW silk chains during drawing, resulting in an increase in molecular chain orientation when compared with silk spun from 100% HMW silk solution. The best regenerated silk fibers produced in this work had an orientation factor of 0.94 and crystallinity of 47.82%, close to the values of natural degummedBombyx mori silk fiber. The approach and mechanism elucidated here can facilitate artificial silk systems with enhanced properties.

Silk Protein Paper with In Situ Synthesized Silver Nanoparticles.

Silver nanoparticles (AgNPs) are in situ synthesized for the first time on microfibrillated silk (MFS) exfoliated from domesticated Philosamia cynthia ricini (eri) and Bombyx mori (mulberry) silkworm silk fibers. The process is rapid (hours time), does not rely on harmful chemicals, and produces robust and flexible AgNPs coated MFS (MFS-AgNPs) protein papers with excellent handling properties. None of these can be achieved by approaches used in the past to fabricate AgNPs silk systems. MFS bonds the AgNPs strongly, providing good support and stabilization for the NPs, leading to strong wash fastness. The mechanical properties of the MFS-AgNPs papers largely do not change compared to the MFS papers without nanoparticles, except for some higher concentration of AgNPs in the case of mulberry silk. The improved tensile properties of eri silk papers with or without AgNPs compared to mulberry silk papers can be attributed to the higher degree of fibrillation achieved in eri silk and its inherent higher ductility. MFS-AgNPs from eri silk also exhibit strong antibacterial activity. This study provides the basis for the development of smart protein papers based on silk fiber and functional nanomaterials.

3D printing of silk microparticle reinforced polycaprolactone scaffolds for tissue engineering applications.

Polycaprolactone (PCL) scaffolds have been widely investigated for tissue engineering applications, however, they exhibit poor cell adhesion and mechanical properties. Subsequently, PCL composites have been produced to improve the material properties. This study utilises a natural material, Bombyx mori silk microparticles (SMP) prepared by milling silk fibre, to produce a composite to enhance the scaffolds properties. Silk is biocompatible and biodegradable with excellent mechanical properties. However, there are no studies using SMPs as a reinforcing agent in a 3D printed thermoplastic polymer scaffold. PCL/SMP (10, 20, 30 wt%) composites were prepared by melt blending. Rheological analysis showed that SMP loading increased the shear thinning and storage modulus of the material. Scaffolds were fabricated using a screw-assisted extrusion-based additive manufacturing system. Scanning electron microscopy and X-ray microtomography was used to determine scaffold morphology. The scaffolds had high interconnectivity with regular printed fibres and pore morphologies within the designed parameters. Compressive mechanical testing showed that the addition of SMP significantly improved the compressive Young's modulus of the scaffolds. The scaffolds were more hydrophobic with the inclusion of SMP which was linked to a decrease in total protein adsorption. Cell behaviour was assessed using human adipose derived mesenchymal stem cells. A cytotoxic effect was observed at higher particle loading (30 wt%) after 7 days of culture. By day 21, 10 wt% loading showed significantly higher cell metabolic activity and proliferation, high cell viability, and cell migration throughout the scaffold. Calcium mineral deposition was observed on the scaffolds during cell culture. Large calcium mineral deposits were observed at 30 wt% and smaller calcium deposits were observed at 10 wt%. This study demonstrates that SMPs incorporated into a PCL scaffold provided effective mechanical reinforcement, improved the rate of degradation, and increased cell proliferation, demonstrating potential suitability for bone tissue engineering applications.

Exfoliating B. mori silk into high aspect ratio nanofibrils facilitated by response surface methodology.

Silk fiber is formed by an assembly of fibrils. The fibrils can be isolated by a top-down mechanical process called microfibrillation and the fibrils are known as microfibrillated silk (MFS). The process involves chopping, milling, enzyme treatment and high-pressure homogenization. The milling is an important manufacturing step and to optimize the milling step, a response surface methodology was used in this work where the influence of fiber content in milled suspension, milling time and alkaline concentration were investigated. Output responses for the optimization were diameter distribution of fibrils, size and percentage of different diameter fractions, and the aspect ratio. The main and interaction effects of the milling parameters on these responses were statistically analysed. Milling time was the most significant factor for producing finer fibrils while the fiber content in milling had the maximum impact in reducing the number of large fibrils. Milling time had a positive correlation with the aspect ratio. The optimized milling resulted in MFS with an average diameter of 55.35 nm and 90% of MFS less than 100 nm based on high-magnification SEM image analysis. The aspect ratio of the MFS was 137. The MFS suspension was stable over the pH range 3-11.

Using In Situ Polymerization to Increase Puncture Resistance and Induce Reversible Formability in Silk Membranes.

Silk fibroin is an excellent biopolymer for application in a variety of areas, such as textiles, medicine, composites and as a novel material for additive manufacturing. In this work, silk membranes were surface modified by in situ polymerization of aqueous acrylic acid, initiated by the reduction of various aryldiazonium salts with vitamin C. Treatment times of 20 min gave membranes which possessed increased tensile strength, tensile modulus, and showed significant increased resistance to needle puncture (+131%), relative to 'untreated' standards. Most interestingly, the treated silk membranes were able to be reversibly formed into various shapes via the hydration and plasticizing of the surface bound poly(acrylic acid), by simply steaming the modified membranes. These membranes and their unique properties have potential applications in advanced textiles, and as medical materials.

Protein Paper from Exfoliated Eri Silk Nanofibers.

The exfoliation of silk fiber is an attractive method to produce silk micro- and nanofibers that retain the secondary structure of native silk. However, most fibrillation methods used to date require the use of toxic and/or expensive solvents and the use of high energy. This study describes a low cost, scalable method to produce microfibrillated silk nanofibers without the use of toxic chemicals by controlling the application of shear using commercially scalable milling and homogenization equipment. Manipulation of the degumming conditions (alkaline concentration and degumming temperature) and the shear in milling and/or homogenization enabled control over the degree of fibrillation. The microfibrillated silk was then characterized to determine structural change during processing and the stability of the resulting suspensions at different pH. Silk nanofibers obtained from milling degummed silk were characterized using atomic force microscopy. Nanofibers obtained both with and without high-pressure homogenization were then used to produce silk "protein paper" through casting. Silk degumming conditions played a critical role in determining the degree of microfibrillation and the properties of the cast silk papers. The silk papers produced from homogenized nanofibers showed excellent mechanical properties, high water absorption, and wicking properties. The silk papers were excellent for supporting the attachment and growth of human skin keratinocytes, demonstrating application possibilities in healthcare such as wound healing.

Tunable Biodegradable Silk-Based Memory Foams with Controlled Release of Antibiotics.

Sustained, local delivery of the antibiotic ciprofloxacin under different formats from porous silk protein-based memory foam systems was studied. Similarly, protease XIV was incorporated during processing to provide control of the degradation kinetics of the silk materials. In vitro antibiotic release studies combined with degradation assessments were utilized to assess the mechanisms and kinetics of release from the silk materials. The sequestered protease XIV affected the degradation profiles of the silk foams yet did not impact the release kinetics of the ciprofloxacin, which was controlled by solubility and diffusion of the drug. The ability to tune the release of ciprofloxacin between 1 and 200 days, combined with the option to modulate the degradation rate up to 80% in 2 weeks via incorporation of a protease, suggests utility for drug release devices. Further, we anticipate that this approach could also be extended to other medical implant needs and other drugs.

Improving the Tensile Properties of Wet Spun Silk Fibers Using Rapid Bayesian Algorithm.

Wet spinning of silkworm silk has the potential to overcome the limitations of the natural spinning process, producing fibers with exceptional mechanical properties. However, the complexity of the extraction and spinning processes have meant that this potential has so far not been realized. The choice of silk processing parameters, including fiber degumming, dissolving, and concentration, are critical in producing a sufficiently viscous dope, while avoiding silk's natural tendency to gel via self-assembly. This study utilized recently developed rapid Bayesian optimization to explore the impact of these variables on dope viscosity. By following the dope preparation conditions recommended by the algorithm, a 13% (w/v) silk dope was produced with a viscosity of 0.46 Pa·s, approximately five times higher than the dope obtained using traditional experimental design. The tensile strength, modulus, and toughness of fibers spun from this dope also improved by a factor of 2.20×, 2.16×, and 2.75×, respectively. These results represent the outcome of just five sets of experimental trials focusing on just dope preparation. Given the number of parameters in the spinning and post spinning processes, the use of Bayesian optimization represents an exciting opportunity to explore the multivariate wet spinning process to unlock the potential to produce wet spun fibers with truly exceptional mechanical properties.