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The valorization of discarded wool from dairy sheep breeding is a challenging issue. The most proposed strategies lie in the processing of keratin extracted from wool without reducing the molecular weight of the protein chains (the high molecular weight-HMW keratin). Here, the HMW keratin has been spun for the first time by solution blow spinning. A screening study of the process carried out with a 2-level full factorial design revealed that keratin filaments can be obtained by using the polyethylene oxide at 900 kDa, a 2 bar air pressure, and a 30 cm needle-collector distance. An annealing at 80 °C for 15 min, at pH 3.5 with citric acid contributes to increasing the viscosity of the keratin solutions thereby allowing the production of defect-free and water-stable filaments having diameters from 1 to 6 µm. A negligible toxic effect was observed after 24 and 48 h on HT29 epithelial cells and normal blood cells displayed behavior similar to the control demonstrating that the patches are hemocompatible. Therefore, the developed SBS process of keratin aqueous solutions could represent a valuable platform for developing patches that need to be blood-contacting and deposited in-situ.

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This report demonstrates a case study within the ASINA project, aimed at instantiating a roadmap with quantitative metrics for Safe(r) and (more) Sustainable by Design (SSbD) options. We begin with a description of ASINA's methodology across the product lifecycle, outlining the quantitative elements within: Physical-Chemical Features (PCFs), Key Decision Factors (KDFs), and Key Performance Indicators (KPIs). Subsequently, we delve in a proposed decision support tool for implementing the SSbD objectives across various dimensions-functionality, cost, environment, and human health safety-within a broader European context. We then provide an overview of the technical processes involved, including design rationales, experimental procedures, and tools/models developed within ASINA in delivering nano-silver-based antimicrobial textile coatings. The result is pragmatic, actionable metrics intended to be estimated and assessed in future SSbD applications and to be adopted in a common SSbD roadmap aligned with the EU's Green Deal objectives. The methodological approach is transparently and thoroughly described to inform similar projects through the integration of KPIs into SSbD and foster data-driven decision-making. Specific results and project data are beyond this work's scope, which is to demonstrate the ASINA roadmap and thus foster SSbD-oriented innovation in nanotechnology.

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The frontiers of antibacterial materials in the biomedical field are constantly evolving since infectious diseases are a continuous threat to human health. In this work, waste-wool-derived keratin electrospun nanofibers were blended with copper by an optimized impregnation procedure to fabricate antibacterial membranes with intrinsic biological activity, excellent degradability and good cytocompatibility. The keratin/copper complex electrospun nanofibers were multi-analytically characterized and the main differences in their physical-chemical features were related to the crosslinking effect caused by Cu2+. Indeed, copper ions modified the thermal profiles, improving the thermal stability (evaluated by differential scanning calorimetry and thermogravimetry), and changed the infrared vibrational features (determined by infrared spectroscopy) and the chemical composition (studied by an X-ray energy-dispersive spectroscopy probe and optical emission spectrometry). The copper impregnation process also affected the morphology, leading to partial nanofiber swelling, as evidenced by scanning electron microscopy analyses. Then, the membranes were successfully tested as antibacterial materials against gram-negative bacteria, Escherichia coli. Regarding cytocompatibility, in vitro assays performed with L929 cells showed good levels of cell adhesion and proliferation (XTT assay), and no significant cytotoxic effect, in comparison to bare keratin nanofibers. Given these results, the material described in this work can be suitable for use as antibiotic-free fibers for skin wound dressing or membranes for guided tissue regeneration.

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Cotton and polyamide 6,6 fabrics coated with chitosan, a natural biopolymer, have been tested against two different bacteria strains: Staphylococcus aureus as Gram-positive bacterium and Escherichia coli as Gram-negative bacterium. Using the ASTM standard method (Standard Test Method for Determining the Antimicrobial Activity of Antimicrobial Agents Under Dynamic Contact Conditions) for antibacterial testing, the treated fabrics is contacted for 1 h with the bacterial inoculum, the present study aims to investigate the possibility to reach interesting results considering shorter contact times. Moreover, the antibacterial activity of chitosan-treated fibers dyed with a natural dye, Carmine Red, was evaluated since chitosan has an interesting property that favors the attachment of the dye to the fiber (cross-linking ability). Finally, fabric samples were tested after washing cycles to verify the resistance of the dye and if the antibacterial property was maintained.

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In recent years, multifunctional inorganic-organic hybrid materials have been widely investigated in order to determine their potential synergetic, antagonist, or independent effects in terms of reactivity. The aim of this study was to design and characterize a new hybrid material by coupling well-known photocatalytic TiO2 nanoparticles with sodium surfactin (SS), a biosurfactant showing high binding affinity for metal cations as well as the ability to interact with and disrupt microorganisms' cell membranes. We used both chemical and colloidal synthesis methodologies and investigated how different TiO2:SS weight ratios affected colloidal, physicochemical, and functional properties. We discovered a clear breaking point between TiO2 and SS single-component trends and identified different ranges of applicability by considering different functional properties such as photocatalytic, heavy metal sorption capacity, and antibacterial properties. At low SS contents, the photocatalytic properties of TiO2 are preserved (conversion of organic dye = 99% after 40 min), and the hybrid system can be used in advanced oxidation processes, taking advantage of the additional antimicrobial SS properties. At high SS contents, the TiO2 photoactivity is inhibited, and the hybrid can be usefully exploited as a UV blocker in cosmetics, avoiding undesired oxidative effects (UV adsorption in the range between 300-400 nm). Around the breaking point (TiO2:SS 1:1), the hybrid material preserves the high surface area of TiO2 (specific surface area around 180 m2/g) and demonstrates NOx depletion of up to 100% in 80 min, together with improved adhesion of hybrid antibacterial coating. The last design demonstrated the best results for the concurrent removal of inorganic, organic, and biological pollutants in water/soil remediation applications.

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Keratin is a biocompatible and biodegradable protein as the main component of wool and animal hair fibers. Keratin-based materials support fibroblasts and osteoblasts growth. Keratin has been extracted by sulphitolysis, a green method (no harmful chemicals) with a yield of 38-45%. Keratin has been processed into nanofibers from its solutions by electrospinning. Electrospinning is a versatile and easy-to-use technique to generate nanofibers. It is an eco-friendly and economical method for the production of randomly and uniaxially oriented polymeric nanofibers. Thanks to their high specific surface area, nanofibers have great potential in the biomedical field. Keratin nanofibers have received significant attention in biomedical applications, such as tissue engineering and cell growth scaffolds, for their biocompatibility and bio-functionality. Accordingly, we propose an extensive overview of recent studies focused on the optimization of keratinbased nanofibers, emphasizing their peculiar functions for cell interactions and the role of additive phases in blends or composite systems to particularize them as a function of specific applications (i.e., antibacterial).

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Protein-based nanofibres are commonly used in the biomedical field to support cell growth. For this study, the cell viability of wool keratin-based nanofibres was tested. Membranes were obtained by electrospinning using formic acid, hexafluoroisopropanol, and water as solvents. For aqueous solutions, polyethylene oxide blended with keratin was employed, and their use to support in vitro cell interactions was also validated. Morphological characterization and secondary structure quantification were carried out by SEM and FTIR analyses. Although formic acid produced the best nanofibres from a morphological point of view, the results showed a better response to cell proliferation after 14 days in the case of fibres from hexafluoroisopropanol solution. Polyethylene oxide in keratin nanofibres was demonstrated, over time, to influence in vitro cell interactions, modifying membranes-wettability and reducing the contact between keratin chains and water molecules, respectively.

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The development of asymmetric membranes-i.e., matching two fibrous layers with selected composition and morphological properties to mimic both the epidermis and dermis-currently represents one of the most promising strategies to support skin regeneration during the wound healing process. Herein, a new asymmetric platform fabricated by a sequential electrospinning process was investigated. The top layer comprises cross-linked polyvinylalcohol (PVA) nanofibers (NFs)-from water solution-to replicate the epidermis's chemical stability and wettability features. Otherwise, the bottom layer is fabricated by integrating PVA with wool-keratin extracted via sulfitolysis. This protein is a biocompatibility polymer with excellent properties for dermis-like structures. Morphological characterization via SEM supported by image analysis showed that the asymmetric membrane exhibited average fiber size-max frequency diameter 450 nm, range 1.40 µm-and porosity suitable for the healing process. FTIR-spectrums confirmed the presence of keratin in the bottom layer and variations of keratin-secondary structures. Compared with pure PVA-NFs, keratin/PVA-NFs showed a significant improvement in cell adhesion in in vitro tests. In perspective, these asymmetric membranes could be promisingly used to confine active species (i.e., antioxidants, antimicrobials) to the bottom layer to support specific cell activities (i.e., proliferation, differentiation) in wound healing applications.

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In response to the nowadays battle against SARS-CoV-2, we designed a new class of high performant filter media suitable to advance the facemask technology and provide new efficient widespread solutions against virus propagation. By means of the electrospinning technology we developed filter media based on polyvinyl alcohol (PVA) nanofibers doped with AgNPs combining three main performance requirements: high air filtration efficiency to capture nanometer-size particles, low airflow resistance essential to ensure breathability and antimicrobial activity to inactivate aerosolized microorganisms. PVA/AgNPs electrospun nanofibers were produced by electrospinning the dispersion of colloidal silver into the PVA water solution. A widespread physicochemical characterization was addressed to the Ag colloidal suspension. The key functional performances of the electrospun nanofibers were proven by water stability, antibacterial activity, and filtration efficiency and pressure drop measurements performed under conditions representative of facemasks. We assessed a total bacterial depletion associated to a filtering efficiency towards nano-aerosolized particles of 97.7% higher than required by the EN149 standard and a pressure drop in line with FFP1 and FFP2 masks, even at the highest filtration velocity. Such results pave the way to the application of PVA/AgNPs electrospun nanofibers in facemasks as advanced filtering media for protecting against airborne microorganisms.

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Polypyrrole (PPy) nanoparticles (NPs) are used for the coating of materials, such as textiles, with biomedical applications, including wound care and tissue engineering, but they are also promising antibacterial agents. In this work, PPy NPs were used for the spray-coating of textiles with antimicrobial properties. The functional properties of the materials were verified, and their safety was evaluated. Two main exposure scenarios for humans were identified: inhalation of PPy NPs during spray (manufacturing) and direct skin contact with NPs-coated fabrics (use). Thus, the toxicity properties of PPy NPs and PPy-coated textiles were assessed by using in vitro models representative of the lung and the skin. The results from the materials' characterization showed the stability of both the PPy NP suspension and the textile coating, even after washing cycles and extraction in artificial sweat. Data from an in vitro model of the air-blood barrier showed the low toxicity of these NPs, with no alteration of cell viability and functionality observed. The skin toxicity of PPy NPs and the coated textiles was assessed on a reconstructed human epidermis model following OECD 431 and 439 guidelines. PPy NPs proved to be non-corrosive at the tested conditions, as well as non-irritant after extraction in artificial sweat at two different pH conditions. The obtained data suggest that PPy NPs are safe NMs in applications for textile coating.

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Electrospinning is gaining increasing interest in the biomedical field as an eco-friendly and economic technique for production of random and oriented polymeric fibers. The aim of this review was to give an overview of electrospinning potentialities in the production of fibers for biomedical applications with a focus on the possibility to combine biomechanical and topographical stimuli. In fact, selection of the polymer and the eventual surface modification of the fibers allow selection of the proper chemical/biological signal to be administered to the cells. Moreover, a proper design of fiber orientation, dimension, and topography can give the opportunity to drive cell growth also from a spatial standpoint. At this purpose, the review contains a first introduction on potentialities of electrospinning for the obtainment of random and oriented fibers both with synthetic and natural polymers. The biological phenomena which can be guided and promoted by fibers composition and topography are in depth investigated and discussed in the second section of the paper. Finally, the recent strategies developed in the scientific community for the realization of electrospun fibers and for their surface modification for biomedical application are presented and discussed in the last section.

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Electrospinning is the leading technology to fabricate fibrous scaffolds that mimic the architecture of the extracellular matrix of natural tissues. In order to improve the biological response, a consolidated trend involves the blending of synthetic polymers with natural proteins to form protein-rich fibers that include selected biochemical cues able to more actively support in vitro cell interaction. In this study, we compared protein-rich fibers fabricated via electrospinning by the blending of poly ε-caprolactone (PCL) with three different proteins, i.e., gelatin, zein, and keratin, respectively. We demonstrated that the peculiar features of the proteins used significantly influence the morphological properties, in terms of fiber size and distribution. Moreover, keratin drastically enhances the fiber hydrophilicity (water contact angle equal to 44.3° ± 3.9°) with positive effects on cell interaction, as confirmed by the higher proliferation of human mesenchymal stem cells (hMSC) until 7 days. By contrast, gelatin and zein not equally contribute to the fiber wettability (water contact angles equal to 95.2° ± 1.2° and 76.3° ± 4.0°, respectively) due to morphological constraints, i.e., broader fiber diameter distribution ascribable to the non-homogeneous presence of the protein along the fibers, or chemical constrains, i.e., large amount of non-polar amino acids. According to in vitro experimental studies, which included SEM and confocal microscopy analyses and vitality assay, we concluded that keratin is the most promising protein to be combined with PCL for the fabrication of biologically instructive fibers for in vitro applications.

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Antimicrobial textiles can contribute to the fighting against antibiotic resistance pathogenic microorganisms. Polypyrrole is a conjugated polymer that exerts a biocidal action thanks to positive charges on its backbone chain produced during it synthesis. In this work, dispersions of stable polypyrrole nanoparticles were produced by chemical oxidative polymerization at room temperature in water. An ultrasound-assisted coating process was then used to effectively treat a polyester fabric with the nanoparticles to obtain an optimal antibacterial coating which efficiently eradicates the bacteria. The results showed that the treated fabric with about 4 g/m2 of polypyrrole had log bacteria reductions of 6.0 against Staphylococcus aureus and 7.5 against Escherichia coli. The combination of a polypyrrole synthesis in the form of water nanoparticles dispersions and a continuous coating of fabrics supported by ultrasound overcomes some issues of upscaling of the traditional in-situ chemical deposition used until now for the production of polypyrrole-coated textiles.

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The use of bioactive proteins such as keratin has been successfully explored to improve the biological interface of scaffolds with cells during the tissue regeneration. In this work, it is optimized the fabrication of nanofibers combining wool keratin extracted by sulfitolysis, with polycaprolactone (PCL) in order to design bicomponent fibrous matrices able to exert a self-adapting pattern of signals-morphological, chemical, or physical-confined at the single fiber level, to influence cell and bacteria interactions. It is demonstrated that the blending of highly polydisperse keratin with PCL (50:50) improves the stability of the electrospinning process, promoting the formation of nanofibers-144.1 ± 43.9 nm-without the formation of defects (i.e., beads, ribbons) typically recognized in the fabrication of keratin ones. Moreover, keratin drastically increases the fiber hydrophilicity-compared with PCL fiber alone-thus improving the hMSC adhesion and in vitro proliferation until 14 days. Moreover, the growth of bacterial strains (i.e., Escherichia coli and Staphylococcus aureus) seems to be not specifically inhibited by the contribution of keratin, so that the integration of further selected compounds (i.e., metal ions) is suggested to more efficiently fight against bacteria resistance, to make them suitable for the regeneration of different interfaces and soft tissues (i.e., skin and cornea). © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1803-1813, 2019.

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Peri-implantitis is a severe condition affecting the success of transmucosal dental implants: tissue healing is severely limited by the inflammatory processes that come about to control homeostasis in the surrounding tissues. The main cause of peri-implantitis is bacterial biofilm infection; gingival fibroblasts play a pivotal role in regulating the inflammatory cascades. A new technology aimed at preventing bacterial colonization of titanium (Ti) implants, and enhancing the spread of gingival fibroblasts, is presented. Using electro-spinning, mirror-polished Ti disks were uniformly coated with keratin fibers obtained from discarded wool via sulfitolysis. The keratin-coated surfaces were then doped with silver (Ag) to introduce antibacterial properties, using different concentrations of silver nitrate as a precursor (0.01, 0.05 and 0.1 M). The resulting specimens were characterized in terms of morphology and chemical composition by FESEM, FTIR and XPS, revealing silver concentrations between 1.7 and 1.9%. Silver release into the medium was evaluated in the presence of cells (α-MEM) or bacteria (LB) by ICP; release was 0.2-1.4 mg l-1 for α-MEM, and 10-40 mg l-1 for LB. The antibacterial properties of the Ag-doped specimens were tested against a multidrug-resistant Staphylococcus aureus biofilm through morphology (FESEM) and metabolic assay (XTT); reduction in viability was significant (p < 0.05; >80% reduction within 72 h). Lastly, the cytocompatibility of the specimens was confirmed using human primary gingival fibroblasts, whose viability, spread and matrix deposition were found to be comparable to those of untreated Ti polished controls (p > 0.05). Thus, Ag surface enrichment was effective in reducing viability and maturation of S. aureus biofilm, without compromising human cell viability. Moreover, cell spread was found to be very sensitive to keratin fiber stimulation. The strategy thus appears to be very promising to introduce surface features in line with the main requirements for transmucosal dental implants.

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Electrospinning is here used for the first time to prepare nanofibers including a host/guest complex in a keratin/poly(ethylene oxide) matrix. The host is a lipid binding protein and the guest is an insoluble bactericidal molecule, irgasan, bound within the protein internal cavity. The obtained nanofibers, characterized by scanning electron microscopy, exhibit excellent antibacterial activity toward Gram positive and negative bacteria, even with a moderate protein/irgasan cargo. Solution NMR studies, employed to provide molecular information on the cargo system, points to a micromolar affinity, compatible with both the electrospinning process and slow guest release. The versatility of the carrier protein, capable of interacting with a variety of druggable hydrophobic molecules, is exploitable for the development of innovative biomedical devices, whose properties can be tuned by the selected guest.

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Communicable diseases can be transmitted by contaminated water. Water decontamination process is fundamental to eliminate microorganisms. In this work, cotton gauzes were coated with chitosan using an UV-curing process or cationized by introduction of quaternary ammonium groups and tested, in static and dynamic conditions, as water filter for biological disinfection against both Gram-negative and Gram-positive bacteria. Both materials showed good antibacterial activity, in static assessment, instead in dynamic conditions, chitosan treated gauze showed a high antimicrobial efficiency in few seconds of contact time. This composite could be a good candidate for application as biological filter.

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The aim of this study was the surface modification of wool fibers to confer a multifunctional finishing to the fabrics, improving the textile value and its applications without damage of comfort properties. The attention was focused on an economical and environmental friendly process to obtain an effective treatment with good durability to washing. Chitosan in acetic acid solution was applied by padding, and grafted by ultraviolet radiation, through radical reactions promoted by a photoinitiator. 2% chitosan grafted was enough to confer satisfactory antimicrobial activity (67% reduction of Escherichia coli) after an oxidative wool pre-treatment and 1h impregnation at 50 °C. Moreover treated wool fabrics showed a strong dyeability increase toward acid dye. However the evaluation of the treatment durability to laundering showed different behavior depending on the nature of the surfactants. Finally, anti-felting properties with respect to untreated fabrics were revealed, while no effect was shown toward anti-pilling properties.

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In the present paper, a proteomic method for species determination in fibres has been developed. Keratin was extracted from yak, wool and cashmere fibres and digested by trypsin, providing peptide mixtures that were analyzed by liquid chromatography coupled with electrospray mass spectrometry (LC/ESI-MS) in order to identify peptidic species-specific markers able to differentiate the fibres. Several suitable peptide markers were identified and validated in different fibres of different origin and having undergone different technological treatments, showing 100% specificity and 100% selectivity. Most of the peptide markers were also identified by means of high-resolution mass spectrometry, confirming the origin from species-specific keratin sequences. Some peptides were also used for the quantification of the different species in mixed fibres by LC/ESI-MS. Validation experiments and blind tests confirmed their ability to act as very specific quantitative and qualitative markers. The method here developed is a valid complement to the standard benchmark methods for fibre identification and quantification and will be very useful for assessing the authenticity of textile products.

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In this study, nanofibre mats for chemical adsorption of heavy metals were prepared by electrospinning blends of hydrolyzed keratin (HK) and polyamide 6 (PA6) in formic acid. Viscosity measurements of the spinning solutions and morphological analyses of the fracture sections of the same polymer blends cast into films, suggested intermolecular interactions and good compatibility between HK and PA6. The mats made of continuous randomly oriented blend nanofilaments of HK/PA6 50/50 wt, with a mean diameter of about 200 nm, were tested as chromium (VI) ion adsorbents. The parameters investigated included initial chromium ion concentration, pH, contact time and adsorbent dosage. The maximum adsorption capacity occurred at acidic pH. The pseudo-first order, the pseudo-second order and the intraparticle diffusion models were used to describe the kinetics of adsorption process. It was found that kinetic data fit the pseudo-second order model and follow the intraparticle diffusion model, although diffusion is not the only rate control step. Adsorption data fit well the Freundlich isotherm model and the maximum adsorption capacity was found 55.9 mg/g. Moreover, the mean free energy (E) of adsorption ranges between 8 and 16 kJ/mol, so that the adsorption mechanism for HK-based nanofibres was explained as an ion-exchange process.