Enzyme-Instructed Self-Assembly Enabled Monomer-Excimer Transition to Construct Higher Ordered Luminescent Supramolecular Assembly for Activity-based Bioimaging.

It is challenging to construct high-performing excimer-based luminescent analytic tools at low molecular concentrations. We report that enzyme-instructed self-assembly (EISA) enables the monomer-excimer transition of a coumarin dye (Cou) at low molecular concentrations, and the resulting higher ordered luminescent supramolecular assemblies (i.e., nanofibers) efficiently record the spatiotemporal details of alkaline phosphatase (ALP) activity in vitro and in vivo. Cou was conjugated to short self-assembly peptides with a hydrophilic ALP-responsive group. By ALP triggering, EISA actuated a nanoparticles-nanofibers transition at low peptide concentrations followed by monomer-excimer transition of Cou. Analysis of structure-property relationships revealed that the self-assembly motif was a prerequisite for peptides to induce the monomer-excimer transition of Cou. Luminescent supramolecular nanofibers of pYD (LSN-pYD) illuminated the intercellular bridge of cancer cells and distinguished cancer cells (tissues) from normal cells (tissues) efficiently and rapidly, promising potential use for the early diagnosis of cancer. This work extends the functions of EISA and provides a new application of supramolecular chemistry.

Self-assembling nitrilotriacetic acid nanofibers for tracking and enriching His-tagged proteins in living cells.

Specific and expeditious identification and enrichment of target proteins in living cells is often a challenging task. The hexahistidine (6His) tag is frequently used to label artificially engineered proteins produced in prokaryotic or eukaryotic cells. Utilizing the interaction between 6His-tag and nitrilotriacetic acid (NTA) mediated by divalent metal ions (Ni2+, Cu2+, Zn2+ or Co2+), we designed and synthesized a series of Nap-G/Biotin/ANA-FFpYGK-NTA probes that, assisted by alkaline phosphatase (ALP), self-assemble into nanofibers. The probe consists of an NTA group that specifically binds to 6His-tag, an FFpY group that promotes self-assembly facilitated by ALP, and a hydrophobic (Nap-G/ANA/Biotin) capping group for various applications. We demonstrate that the ANA-FFpYGK-NTA(Ni2+) nanofibers are fit for real-time tracking of His-tagged protein in living cells, and the Biotin-FFpYGK-NTA(Ni2+) nanofibers are for isolating His-tagged proteins and other proteins that they interact with.

Effect of gamma irradiation on physico-mechanical and structural properties of active Farsi gum-CMC films containing Ziziphora clinopodioides essential oil and lignocellulose nanofibers for meat packaging.

The objective of this study was to examine the effect of gamma irradiation (0, 2.5, and 5 kGy) on physico-mechanical and structural characteristics of films based on Farsi gum-carboxymethyl cellulose supplemented with Ziziphora clinopodioides essential oil (ZEO; 0%, 1%, and 2%) and lignocellulose nanofibers (LCNF; 0%, 1%, and 2%), and their application on fresh minced beef meat's shelf-life during refrigerated temperature (4 ± 1 °C) for 16 days. Gamma irradiation under the 60 Co source at 2.5 and 5 kGy doses did not have a significant effect on thickness, tensile strength, swelling index, oxygen permeability, and water vapor transmission rate of prepared films (P > 0.05). The best microbiological (total viable count, psychrotrophic bacterial count, Pseudomonas spp., Brochothrix thermosphacta, lactic acid bacteria, and Enterobacteriaceae) and chemical (thiobarbituric acid reactive substances, total volatile base nitrogen content, and peroxide value) properties were recorded for samples packaged with ZEO 2% + LCNF 2%, followed by ZEO 2% + LCNF 1%, ZEO 1% + LCNF 2%, and ZEO 1% + LCNF 1%. These results indicate acceptable extensions of hurdle technology for prolonged refrigeration of minced beef meat. PRACTICAL APPLICATION: The application of active packaging films has received considerable interest in extending the shelf-life of perishable foods during prolonged chilled storage. The effects of active Farsi gum-carboxymethyl cellulose films supplemented with Ziziphora clinopodioides essential oil 2% + lignocellulose nanofibers 2% resulted in delaying lipid oxidation and microbial spoilage growth of refrigerated minced beef meat and consequently extending the shelf-life during storage for at least 16 days.

Preparation and characterization of naftifine-loaded poly(vinyl alcohol)/sodium alginate electrospun nanofibers

In this study, naftifine (a topical antifungal drug) loaded poly(vinyl) alcohol (PVA)/sodium alginate (SA) nanofibrous mats were prepared using the single-needle electrospinning technique. The produced nanofibers were crosslinked with glutaraldehyde (GTA) vapor. The morphology and diameter of the electrospun nanofibers were studied by scanning electron microscopy (SEM). SEM images showed the smoothness of the nanofibers and indicated that the fiber diameter increased with crosslinking and drug loading. Atomic force microscopy (AFM) images confirmed the uniform production of the scaffolds, and elemental mapping via energy dispersive X-ray spectroscopy (EDS) showed the uniform distribution of the drug within the nanofibers. An attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy study demonstrated that naftifine has sufficient secondary interactions with the polymer blend. The crosslinking treatment decreased the burst drug release effectively and the release mechanism followed Korsmeyer-Peppas Super Case-II transport. Overall, these findings suggest the potential use of naftifine-loaded PVA/SA nanofibers as a topical antifungal drug delivery system.

Synthetic Genetic Circuits for Self-Actuated Cellular Nanomaterial Fabrication Devices.

Genetically controlled synthetic biosystems are being developed to create nanoscale materials. These biosystems are modeled on the natural ability of living cells to synthesize materials: many organisms have dedicated proteins that synthesize a wide range of hard tissues and solid materials, such as nanomagnets and biosilica. We designed an autonomous living material synthesizing system consisting of engineered cells with genetic circuits that synthesize nanomaterials. The circuits encode a nanomaterial precursor-sensing module (sensor) coupled with a materials synthesis module. The sensor detects the presence of cadmium, gold, or iron ions, and this detection triggers the synthesis of the related nanomaterial-nucleating extracellular matrix. We demonstrate that when engineered cells sense the availability of a precursor ion, they express the corresponding extracellular matrix to form the nanomaterials. This proof-of-concept study shows that endowing cells with synthetic genetic circuits enables nanomaterial synthesis and has the potential to be extended to the synthesis of a variety of nanomaterials and biomaterials using a green approach.

Amphotericin-B and vancomycin-loaded chitosan nanofiber for antifungal and antibacterial application

In the present study, a mucoadhesive non-woven fiber mat (d= 116 nm) was fabricated by the electrospinning method using chitosan (80% Wt), polyethylene oxide (10% Wt), cysteine (4% Wt) and drugs (6% Wt), respectively. In addition, a comparative study was conducted to define effect of drugs and mucoadhesive agent on the nanofiber formation. FTIR, SEM, DSC and DMA were used to investigate the chemical and physical properties of the nanofibers. In vitro release of the drugs was assessed over a 48-hour period by the total immersion method. Release data were fitted to kinetic models, including the zero-order, first-order, Higuchi matrix, and Hixson-Crowell. Zone inhibition investigations were used to describe the inhibition content of vancomycin and amphotericin B loaded in the mats. The SEM images displayed a slight decrease in the fiber diameter with adding drugs and mucoadhesive agents. FTIR spectra confirmed that any undesirable reaction between VAN-AMB and CS-PEO was not observed. DSC test recognized the uniform distribution of drugs in the polymeric bead of the fiber without any crystal form. The elasticity modulus of the nanofiber was in an acceptable range for oral mucosa (approximately 5 Mpa). The results indicated that biodegradable mucoadhesive nanofibrous membranes released high concentrations of VAN in the first 24 hours, but the AMB release was affected in more controlled phenomena

Hand-Held Volatilome Analyzer Based on Elastically Deformable Nanofibers.

This study reports on a hand-held volatilome analyzer for selective determination of clinically relevant biomarkers in exhaled breath. The sensing platform is based on electrospun polymer nanofiber-multiwalled carbon nanotube (MWCNT) sensing microchannels. Polymer nanofibers of poly(vinylidene fluoride) (PVDF), polystyrene (PS), and poly(methyl methacrylate) (PMMA) incorporated with MWCNT exhibits a stable response to interferences of humidity and CO2 and provides selective deformations upon exposure of exhaled breath target volatilomes acetone and toluene, exhibiting correlation to diabetes and lung cancer, respectively. The sensing microchannels "P1" (PVDF-MWCNT), "P2" (PS-MWCNT), and "P3" (PMMA-MWCNT) are integrated with a microfluidic cartridge (µ-card) that facilitates collection and concentration of exhaled breath. The volatilome analyzer consists of a conductivity monitoring unit, signal conditioning circuitries and a low energy display module. A combinatorial operation algorithm was developed for analyzing normalized resistivity changes of the sensing microchannels upon exposure to breath in the concentration ranges between 35 ppb and 3.0 ppm for acetone and 1 ppb and 10 ppm for toluene. Subsequently, responses of volatilomes from individuals in the different risk groups of diabetes were evaluated for validation of the proposed methodology. We foresee that proposed methodology provides an avenue for rapid detection of volatilomes thereby enabling point of care diagnosis in high-risk group individuals.

Evaluation of Mechanical Properties of Various Nanofibre Reinforced Bis-GMA/TEGDMA Based Dental Composite Resins / Evaluación de las propiedades mecánicas de varias resinas compuestas dentales basadas en Bis-GMA/TEGDMA reforzadas con nanofibras

ABSTRACT Objective: To investigate the mechanical properties of various mass fractions of Nylon 6 (N6), polymethyl-metacrylate (PMMA) and polyvinylidene-difluoride (PVDF) nanofibres reinforced bisphenol A-glycidyl methacrylate (Bis-GMA) and tri-ethylene glycol dimethacrylate (TEGDMA) based dental composite resins and to evaluate the penetration characteristics of the nanofibres into the resin. Methods: Nylon 6, PMMA and PVDF nanofibres were produced using the electrospinning method. The morphologies of the fabricated nanofibres were evaluated with a scanning electron microscope (SEM). The nanofibres were placed into the resin matrix at different mass fractions (3%, 5% and 7%). The three-point bending test was applied to nanofibre-reinforced dental composite resins and neat resin specimens. The flexural strength (Fs), flexural modulus (EY) and work of fracture (WOF) of the groups were found. The analysis of variance was used for the statistical analysis of the acquired data. Tukey 's multiple test was performed to compare the Fs, EY and WOF means. Fractured surfaces of the samples were observed by SEM, and fracture morphologies were evaluated. Results: Polymethyl-metacrylate nanofibres dissolved in the matrix, and a polymer alloy took place in the matrix. Fibre pull-out and fibre bridging mechanisms were observed by SEM images of the N6 and PVDF nanofibre-reinforced dental composites. The produced nanofibres enhanced the mechanical properties of the dental composite resins. Conclusion: Fibre pull-out and fibre bridging mechanisms on the fractured surfaces of samples may play a key role in the reinforcement of dental composite resins. However, polymer alloy of PMMA nanofibres increased the mechanical properties of the resin matrix.

RESUMEN Objetivo: Investigar las propiedades mecánicas de resinas compuestas dentales basadas en bisfenol A-diglicidildimetacrilato (Bis-GMA) y dimetacrilato trietilen-glicol (TEGDMA) reforzadas con nanofibras de fracciones de masa de Nylon 6 (N6), polimetilmetacrilato (PMMA) y fluoruro de polivinilideno (PVDF), y evaluar las características de la penetración de las nanofibras en la resina. Métodos: Se produjeron nanofibras de Nylon 6, PMMA y PVDF utilizando el método de electrohilado (electrospinning). Las morfologías de las nanofibras fabricadas fueron evaluadas con un microscopio electrónico de barrido (MEB). Las nanofibras fueron introducidas en la matriz de resina en diferentes fracciones de masa (3%, 5% y 7%). La prueba de flexión de tres puntos fue aplicada a las resinas compuestas dentales reforzadas por nanofibras y a las muestras de resina pura. La resistencia a la flexión (Rf), el módulo de flexión (EY) y el trabajo de fractura (WOF) de los grupos fueron halladas. El análisis de varianza se usó para el análisis estadístico de los datos adquiridos. Se realizó la prueba de comparaciones múltiples de Tukey con el propósito de comparar las medidas de Rf, EY y WOF. Las superficies fracturadas de las muestras fueron observadas mediante un MEB, y se evaluaron las morfologías de fractura. Resultados: Las nanofibras de polimetilmetacrilato se disolvieron en la matriz, y tuvo lugar una aleación de polímeros en la matriz. Los mecanismos de desprendimiento de fibras y puenteo de fibras fueron observados mediante imágenes de MEB de los compuestos dentales reforzados con nanofibras de N6 y PVDF. Las nanofibras producidas realzaron las propiedades mecánicas de las resinas compuestas dentales. Conclusión: Los mecanismos de desprendimiento de fibras y puenteo de fibras en las superficies fracturadas de las muestras pueden desempeñar un papel clave en el reforzamiento de las resinas de los compuestos dentales. Sin embargo, la aleación polimérica de las nanofibras de PMMA aumentó las propiedades mecánicas de la matriz de resina.

Human exposure to carbon-based fibrous nanomaterials: A review.

In an emerging field of nanotechnologies, assessment of exposure to carbon nanotubes (CNT) and carbon nanofibers (CNF) is an integral component of occupational and environmental epidemiology, risk assessment and management, as well as regulatory actions. The current state of knowledge on exposure to carbon-based fibrous nanomaterials among workers, consumers and general population was studied in frame of the International Agency for Research on Cancer (IARC) Monographs-Volume 111 "Some Nanomaterials and Some Fibres". Completeness and reliability of available exposure data for use in epidemiology and risk assessment were assessed. Occupational exposure to CNT/CNF may be of concern at all stages of the material life-cycle from research through manufacture to use and disposal. Consumer and environmental exposures are only estimated by modeled data. The available information of the final steps of the life-cycle of these materials remains incomplete so far regarding amounts of handled materials and levels of exposure. The quality and amount of information available on the uses and applications of CNT/CNF should be improved to enable quantitative assessment of human exposure to these materials. For that, coordinated effort in producing surveys and exposure inventories based on harmonized strategy of material test, exposure measurement and reporting results is strongly encouraged.

Characterization of Potential Exposures to Nanoparticles and Fibers during Manufacturing and Recycling of Carbon Nanotube Reinforced Polypropylene Composites.

Carbon nanotube (CNT) polymer composites are widely used as raw materials in multiple industries because of their excellent properties. This expansion, however, is accompanied by realistic concerns over potential release of CNTs and associated nanoparticles during the manufacturing, recycling, use, and disposal of CNT composite products. Such data continue to be limited, especially with regards to post-processing of CNT-enabled products, recycling and handling of nanowaste, and end-of-life disposal. This study investigated for the first time airborne nanoparticle and fibers exposures during injection molding and recycling of CNT polypropylene composites (CNT-PP) relative to that of PP. Exposure characterization focused on source emissions during loading, melting, molding, grinding, and recycling of scrap material over 20 cycles and included real-time characterization of total particle number concentration and size distribution, nanoparticle and fiber morphology, and fiber concentrations near the operator. Total airborne nanoparticle concentration emitted during loading, melting, molding, and grinding of CNT-PP had geometric mean ranging from 1.2 × 10(3) to 4.3 × 10(5) particles cm(-3), with the highest exposures being up to 2.9 and 300.7 times above the background for injection molding and grinding, respectively. Most of these emissions were similar to PP synthesis. Melting and molding of CNT-PP and PP produced exclusively nanoparticles. Grinding of CNT-PP but not PP generated larger particles with encapsulated CNTs, particles with CNT extrusions, and respirable fiber (up to 0.2 fibers cm(-3)). No free CNTs were found in any of the processes. The number of recycling runs had no significant impact on exposures. Further research into the chemical composition of the emitted nanoparticles is warranted. In the meanwhile, exposure controls should be instituted during processing and recycling of CNT-PP.

Stabilizing oil-in-water emulsions with regenerated chitin nanofibers.

Natural chitin is a highly crystalline biopolymer with poor aqueous solubility. Thus direct application of chitin is rather limited unless chemical modifications are made to improve its solubility in aqueous media. Through a simple dissolution and regeneration process, we have successfully prepared chitin nanofibers with diameters around 50nm, which form a stable suspension at concentrations higher than 0.50% and a self-supporting gel at concentrations higher than 1.00%. Additionally, these nanofibers can stabilize oil-in-water emulsions with oil fraction more than 0.50 at chitin usage level of 0.01g/g oil. The droplet sizes of the resulting emulsions decrease with increasing chitin concentrations and decreasing oil fraction. Confocal laser scanning micrographs demonstrate the adsorption of chitin nanofibers on the emulsion droplet surface, which indicates the emulsion stabilization is through a Pickering mechanism. Our findings allow the direct application of chitin in the food industry without chemical modifications.

Airborne fiber size characterization in exposure estimation: Evaluation of a modified transmission electron microcopy protocol for asbestos and potential use for carbon nanotubes and nanofibers.

BACKGROUND: Airborne fiber size has been shown to be an important factor relative to adverse lung effects of asbestos and suggested in animal studies of carbon nanotubes and nanofibers (CNT/CNF). MATERIALS AND METHODS: The International Standards Organization (ISO) transmission electron microscopy (TEM) method for asbestos was modified to increase the statistical precision of fiber size determinations, improve efficiency, and reduce analysis costs. Comparisons of the fiber size distributions and exposure indices by laboratory and counting method were performed. RESULTS: No significant differences in size distributions by the ISO and modified ISO methods were observed. Small but statistically-significant inter-lab differences in the proportion of fibers in some size bins were found, but these differences had little impact on the summary exposure indices. The modified ISO method produced slightly more precise estimates of the long fiber fraction (>15 µm). CONCLUSIONS: The modified ISO method may be useful for estimating size-specific structure exposures, including CNT/CNF, for risk assessment research.

Occupational exposure assessment in carbon nanotube and nanofiber primary and secondary manufacturers: mobile direct-reading sampling.

UNLABELLED: RESEARCH SIGNIFICANCE: Toxicological evidence suggests the potential for a wide range of health effects from exposure to carbon nanotubes (CNTs) and carbon nanofibers (CNFs). To date, there has been much focus on the use of direct-reading instruments (DRIs) to assess multiple airborne exposure metrics for potential exposures to CNTs and CNFs due to their ease of use and ability to provide instantaneous results. Still, uncertainty exists in the usefulness and interpretation of the data. To address this gap, air-monitoring was conducted at six sites identified as CNT and CNF manufacturers or users and results were compared with filter-based metrics. METHODS: Particle number, respirable mass, and active surface area concentrations were monitored with a condensation particle counter, a photometer, and a diffusion charger, respectively. The instruments were placed on a mobile cart and used as area monitors in parallel with filter-based elemental carbon (EC) and electron microscopy samples. Repeat samples were collected on consecutive days, when possible, during the same processes. All instruments in this study are portable and routinely used for industrial hygiene sampling. RESULTS: Differences were not observed among the various sampled processes compared with concurrent indoor or outdoor background samples while examining the different DRI exposure metrics. Such data were also inconsistent with results for filter-based samples collected concurrently at the same sites [Dahm MM, Evans DE, Schubauer-Berigan MK et al. (2012) Occupational exposure assessment in CNT and nanofiber primary and secondary manufacturers. Ann Occup Hyg; 56: 542-56]. Significant variability was seen between these processes as well as the indoor and outdoor backgrounds. However, no clear pattern emerged linking the DRI results to the EC or the microscopy data (CNT and CNF structure counts). CONCLUSIONS: Overall, no consistent trends were seen among similar processes at the various sites. The DRI instruments employed were limited in their usefulness in assessing and quantifying potential exposures at the sampled sites but were helpful for hypothesis generation, control technology evaluations, and other air quality issues. The DRIs employed are nonspecific, aerosol monitors, and, therefore, subject to interferences. As such, it is necessary to collect samples for analysis by more selective, time-integrated, laboratory-based methods to confirm and quantify exposures.

Occupational exposure assessment in carbon nanotube and nanofiber primary and secondary manufacturers.

UNLABELLED: RESEARCH SIGNIFICANCE: Toxicological evidence suggests the potential for a wide range of health effects, which could result from exposure to carbon nanotubes (CNTs) and carbon nanofibers (CNFs). The National Institute for Occupational Safety and Health (NIOSH) has proposed a recommended exposure limit (REL) for CNTs/CNFs at the respirable size fraction. The current literature is lacking exposure information, with few studies reporting results for personal breathing zone (PBZ) samples in occupational settings. To address this gap, exposure assessments were conducted at six representative sites identified as CNT/CNF primary or secondary manufacturers. METHODS: Personal and area filter-based samples were collected for both the inhalable mass concentration and the respirable mass concentration of elemental carbon (EC) as well as CNT structure count analysis by transmission electron microscopy to assess exposures. When possible, full-shift PBZ samples were collected; area samples were collected on a task-based approach. RESULTS: The vast majority of samples collected in this study were below the proposed REL (7 µg m(-3)). Two of the three secondary manufacturers' surveyed found concentrations above the proposed REL. None of the samples collected at primary manufacturers were found to be above the REL. Visual and microscopy-based evidence of CNTs/CNFs were found at all sites, with the highest CNT/CNF structure counts being found in samples collected at secondary manufacturing sites. The statistical correlations between the filter-based samples for the mass concentration of EC and CNT structure counts were examined. A general trend was found with a P-value of 0.01 and a corresponding Pearson correlation coefficient of 0.44. CONCLUSIONS: CNT/CNF concentrations were above the proposed NIOSH REL for PBZ samples in two secondary manufacturing facilities that use these materials for commercial applications. These samples were collected during dry powder handling processes, such as mixing and weighing, using fairly large quantities of CNTs/CNFs.

Tuning size scale and crystallinity of PCL electrospun fibres via solvent permittivity to address hMSC response.

The effect of solvent permittivity on the fibre morphology of PCL electrospun membranes for tissue engineering applications is studied. Morphological results indicate that polar solvents with higher permittivity are able to promote the formation of sub-micrometric fibres, while apolar solvents yield microfibres with an average fibre diameter of 2.86 ± 0.31 µm. Polymer/solvent interactions and electrospinning process parameters influence the mechanism of fibre and bead formation. It is shown that the dielectric properties of solvents influence the fibre size scale and crystallinity and directly contribute to the biological response of stem cells. Solvent permittivity is a key factor in controlling the morphological and physical properties of electrospun fibre meshes.

Exposure and emissions monitoring during carbon nanofiber production--Part I: elemental carbon and iron-soot aerosols.

Production of carbon nanofibers and nanotubes (CNFs/CNTs) and their composite products is increasing globally. High volume production may increase the exposure risks for workers who handle these materials. Though health effects data for CNFs/CNTs are limited, some studies raise serious health concerns. Given the uncertainty about their potential hazards, there is an immediate need for toxicity data and field studies to assess exposure to CNFs/CNTs. An extensive study was conducted at a facility that manufactures and processes CNFs. Filter, sorbent, cascade impactor, bulk, and microscopy samples, combined with direct-reading instruments, provided complementary information on air contaminants. Samples were analyzed for organic carbon (OC) and elemental carbon (EC), metals, and polycyclic aromatic hydrocarbons (PAHs), with EC as a measure of CNFs. Transmission electron microscopy with energy-dispersive X-ray spectroscopy also was applied. Fine/ultrafine iron-rich soot, PAHs, and carbon monoxide were production byproducts. Direct-reading instrument results were reported previously [Evans DE et al. (Aerosol monitoring during carbon nanofiber production: mobile direct-reading sampling. Ann Occup Hyg 2010;54:514-31.)] Results for time-integrated samples are reported as companion papers in this Issue. OC and EC, metals, and microscopy results are reported here, in Part I, while results for PAHs are reported in Part II [Birch ME. (Exposure and Emissions Monitoring during Carbon Nanofiber Production-Part II: Polycyclic Aromatic Hydrocarbons. Ann. Occup. Hyg 2011; 55: 1037-47.)]. Respirable EC area concentrations inside the facility were about 6-68 times higher than outdoors, while personal breathing zone samples were up to 170 times higher.