Dyeing Improvement and Stability of Antibacterial Properties in Chitosan-Modified Cotton and Polyamide 6,6 Fabrics.

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.

Wool Keratin Nanofibers for Bioinspired and Sustainable Use in Biomedical Field.

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).

Wool Keratin-Based Nanofibres-In Vitro Validation.

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.

Design of Asymmetric Nanofibers-Membranes Based on Polyvinyl Alcohol and Wool-Keratin for Wound Healing Applications.

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.

Polyvinyl alcohol/silver electrospun nanofibers: Biocidal filter media capturing virus-size particles.

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.

Safety Assessment of Polypyrrole Nanoparticles and Spray-Coated Textiles.

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.

Topographical and Biomechanical Guidance of Electrospun Fibers for Biomedical Applications.

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.

Comparative Study on Protein-Rich Electrospun Fibers for in Vitro Applications.

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.

Antibacterial properties of polypyrrole-treated fabrics by ultrasound deposition.

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.

Highly polydisperse keratin rich nanofibers: Scaffold design and in vitro characterization.

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.

Biomineralization of a titanium-modified hydroxyapatite semiconductor on conductive wool fibers.

Metal ions are frequently incorporated into crystalline materials to improve their electrochemical properties and to confer new physicochemical properties. Naturally-occurring phosphate apatite, which is formed geologically and in biomineralization processes, has extensive potential applications and is therefore an attractive functional material. In this study, we generate a novel building block for flexible optoelectronics using bio-inspired methods to deposit a layer of photoactive titanium-modified hydroxyapatite (TiHA) nanoparticles (NPs) on conductive polypyrrole(PPy)-coated wool yarns. The titanium concentration in the reaction solution was varied between 8-50 mol% with respect to the phosphorous, which led to titanate ions replacing phosphate in the hydroxyapatite lattice at levels up to 17 mol%. PPy was separately deposited on wool yarns by oxidative polymerization, using two dopants: (i) anthraquinone-2,6-disulfonic acid to increase the conductivity of the PPy layer and (ii) pyroglutamic acid, to reduce the resistivity of the wool yarns and to promote the heterogeneous nucleation of the TiHA NPs. A specific titanium concentration (25 mol% wrt P) was used to endow the TiHA NPs on the PPy-coated fibers with a desirable band gap value of 3.68 eV, and a specific surface area of 146 m2 g-1. This is the first time that a thin film of a wide-band gap semiconductor has been deposited on natural fibers to create a fiber-based building block that can be used to manufacture flexible electronic devices.

Silver-doped keratin nanofibers preserve a titanium surface from biofilm contamination and favor soft-tissue healing.

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.

Electrospun Lipid Binding Proteins Composite Nanofibers with Antibacterial Properties.

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.

Chitosan coated cotton gauze for antibacterial water filtration.

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.

Study on the adsorption of chromium (VI) by hydrolyzed keratin/polyamide 6 blend nanofibres.

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.

Study on the conversion of wool keratin by steam explosion.

A wool fiber sample was submitted to chemical-free steam explosion in view of potential exploitation of keratin-based industrial and farm wastes. Fiber keratin was converted into a dark-yellow sludge that was submitted to phase separation by filtration, centrifugation, and precipitation of the soluble materials from the supernatant liquid. The resulting products, when compared with the original wool, showed the extent of disruption of the histology structure, reduction of the molecular weight to water-soluble peptides and free amino acids, and change of the structure of the remainder of the protein associated with breaking of disulfide bonds and decomposition of the high-sulfur-content protein fraction.